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08 January 2010, Volume 14 Issue 2 Previous Issue    Next Issue

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In vitro culture of high-purity osteoclasts induced by macrophage colony stimulating factor/receptor activator of nuclear factor kappa B ligand: Optimal dosage investigation Bao Hong-wei, Sun Ji-fu, Wang Qing 2010, 14 (2):  191-195.  doi: 10.3969/j.issn.1673-8225.2010.02.001 Abstract ( 301 )   PDF (1258KB) ( 479 )   Save BACKGROUND: Macrophage colony-stimulating factor (M-CSF)/receptor activator of nuclear kappa B ligand (RANKL), two types of cytokines co-induce myeloid stem cells to form osteoclasts, is a kind of new method to harvest osteoclasts with high purity and quantity, but there is lack of uniform cultivation standard. OBJECTIVE: To construct an effective M-CSF/RANKL induced mice myeloid stem cells inducing osteoclast differentiation cultivation system. METHODS: Myeloid stem cells were obtained from ICR mice and then cultured for 24 hours in a-minimum essential medium containing M-CSF, at cell density of 107/L, 108/L, 109/L. Then 10 µg/L M-CSF and 20, 50, 100 µg/L RANKL were added into culture medium. Tartaric-resistant acid phosphatase stained was performed to observe the transition process from stem cell to osteoclast, as well as cell morphology and stain situation after culture, and positive stained osteoclasts were counted. We compared the influence of different induction conditions to the quantity of osteoclast. RESULTS AND CONCLUSION: A small quantity of osteoclasts contained many red positive beads in the intracytoplasm were observed at 3 days. There were positive beads with hypochromatic dikaryon in cells. A large amount of positively stained osteoclasts were seen after 6-day culturing, which maintained dikaryon. After 9-day culturing, positively stained colossal multinuclear cells occurred, became larger and maintained three nuclei. At certain cell density, 100 µg/L RANKL could induce to form more osteoclasts compared with other 2 concentrations (P < 0.05); at certain RANKL concentration, the osteoclasts formation at cells density of 108/L was dramatically greater than other 2 cell densities (P < 0.05); the number of osteoclasts was the most when the concentration of RANKL was 100 µg/L and cell density of 108/L (P < 0.05). When osteoclasts are induced by M-CSF/RANKL from murine myeloid stem cells, the best concentration of RANKL is 100 µg/L and cells density is 108/L. Related Articles | Metrics

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Effect of compound vibration on bone quality of ovariectomized rats Deng Xuan-geng, Chen Jian-ting, Feng Ying 2010, 14 (2):  196-200.  doi: 10.3969/j.issn.1673-8225.2010.02.002 Abstract ( 252 )   PDF (1377KB) ( 331 )   Save BACKGROUND: Whole body vibration which is used to prevent and treat osteoporosis recently is uncomfortable due to its strong vibration strength. The designed compound vibration with low vibration strength could effectively prevent bone quality decrease of ovariectomized rats. OBJECTIVE: To verify the effect of compound vibration with low vibration strength on the bone quality of ovariectomized rats. METHODS: A total of 32 4-month-old female SD rats were used in this study. All SD rats were randomly divided into 4 groups: normal control group, ovariectomized control group, vibration Ⅰ group (45 Hz-55 Hz, 0.05 g-0.1 g), and vibration Ⅱ group (45 Hz-55 Hz, 0.12 g-0.21 g), with 8 rats for each group. The intervention to each vibration group was one a day, 20 minutes/times, and 5 days per week. The resting interval was shorted than 2 days, and the whole vibration intervention lasted 13 weeks. Bone mineral density (BMD) before and after vibration, ex vivo bone microstructure parameters and biomechanics parameters were measured in this study. RESULTS AND CONCLUSION: Lumbar BMD of ovariectomized control group was decreased (P < 0.05), but the BMD in the normal control group and the two vibration groups was increased significantly. Femoral BMD of all groups was increased, but there was no significant difference among the four groups. Bone microstructure parameters of the three ovariectomized groups were decreased; however, parameters including number, thickness and distance of bone trabecula and bone volume fraction in the vibration II group were significantly improved compared to ovariectomized control group. Moreover, bone strength of lumbar vertebra in the two vibration groups was significantly increased (P=0.025, 0.006), but there was no significant difference compared to normal control group. Compound vibration with low strength could effectively prevent BMD decrease of ovariectomized rats, lighten the degree of bone microstructure damage, keep bone strength, improve bone quality of ovariectomized rats, and potentially prevent osteoporosis. References | Related Articles | Metrics

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Role of osteoblasts in the sense of glucose Zhao Hong-li, Zhao Hong-qiu, Fu Xiao-yi, Meng Rong-sen, Zhang Ying, Yan Li, Cheng Hua, Xu Ming-tong, Zhang Shao-ling, Fu Zu-zhi 2010, 14 (2):  201-204.  doi: 10.3969/j.issn.1673-8225.2010.02.003 Abstract ( 260 )   PDF (1111KB) ( 434 )   Save BACKGROUND: Recent study showed that osteocalcin may elevate Insulin secretion and sensitivity, prevent the fat accumulation, play a role in the metablism of glucose and lipid. Undercarboxylated osteocalcin works as the main role. OBJECTIVE: To investigate the effect of different concentrations of glucose on osteoblast undercarboxylated osteocalcin. METHODS: The rib trabeculae were resected and broken, trypsinizated and washed completely by PBS. Bone surface and non-adhesive floating cells in cleaning fluid were observed with inverted microscope. Rib trabeculae was washed by DMEM culture medium once, and cultured in culture bottle. The culture liquid was replaced by new one once a week. The osteoblast was moved from the sclerite a week later. The cells were fused monolayer and could be subcultured 4 to 6 weeks later. The active second or third generation cells were inoculated to 6-pore plate forming 5 groups. Osteoblast were stimulated by 5.6 mmol/L, 7.6 mmol/L, 9.6 mmol/L, 12.6 mmol/L, 20.6 mmol/L glucose medium respectively after the 80% cells were fused, the vitamin K2 was added into the culture liquid until the concentration of it to be 10-5 mol/L. Supernatant was collected after half hour culturing, the undercarboxylated osteocalcin level were detected with RIA test kit, and corrected it as the total the undercarboxylated osteocalcin, calculated the carboxylated incomplete osteocalcin rate. RESULTS AND CONCLUSION: The rate of osteoblast carboxylated incomplete osteocalcin was different under different concentration glucose. The rate of 7.6 mmol/L, 9.6 mmol/L, 20.6 mmol/L concentration glucose groups were higher than that of 5.6 mmol/L glucose group [(0.27±0.02)%, (0.29±0.04)%, (0.12±0.02)%, P < 0.05]. It is indicated that osteoblast could sense the change of glucose concentration by regulating the secretion of the undercarboxylated osteocalcin between the concentration of 5.6mmol/L to 9.6mmol/L, while the carboxylated incomplete osteocalcin decreased as the concentration of glucose increased.  Related Articles | Metrics

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Effects of gallium nitrate on collagen and bone calcium protein in rat models of estrogen deficiency-induced osteoporosis Guo Yu-xia, Yang Da-zhi, Wang Duo, Zhen Wan-xin, Liu Hong-tao, Gao Guo-yong 2010, 14 (2):  205-208.  doi: 10.3969/j.issn.1673-8225.2010.02.004 Abstract ( 309 )   PDF (1381KB) ( 448 )   Save BACKGROUND: Gallium is a non-essential trace element in the human body. In vivo experiments have confirmed that gallium can directly inhibit bone osteolysis, prevent bone calcium release, increase bone calcium content, serves as a new drug treatment of metabolic bone disease, its anti-bone transformation mechanism remains unclear. OBJECTIVE: To observe the effects of gallium nitrate on collagen and bone calcium protein in osteoporotic rat model. METHODS: Ninety female SD rats were divided into control group (n = 20) and osteoporosis group (n = 70) at random. Control group rats were sutured to close abdominal cavity after bilateral ovarian was exposed. Osteoporosis group rats received the bilateral ovariectomy to produce osteoporotic rat models, which then were assigned into 4 groups by random digits table: osteoporotic control group (n = 16) by intraperitoneal injection of saline, 3 times per week; Low-dose gallium salt group (n = 16) by intraperitoneal injection of 1 mg/kg of gallium nitrate, 3 times per week; High-dose gallium salt group (n = 15) by intraperitoneal injection of 2 mg/kg of gallium nitrate, 3 times per week; Estrogen group (n = 15) by intraperitoneal injection of estradiol, 3 times per week. After 12 weeks of the treatment, the bone collagen, osteocalcin protein and hydroxyproline levels in bone specimens were detected. RESULTS AND CONCLUSION: Compared with control group, the content of collagen in osteoporosis control group was reduced (P < 0.05), the contents of aminohexose and hydroxyproline increased (P < 0.05), no significant differences were observed in the content of sulfate-base for both groups. Following gallium and estradiol treatment, the collagen contents enhanced (P < 0.05), while the contents of aminohexose and hydroxyproline reduced (P < 0.05). High-dose gallium salt group had a remarkable curative effect compared with low-dose gallium salt group (P < 0.05), and was similar to estradiol group (P > 0.05). It is indicated that gallium nitrate can improve bone metabolism status with osteoporosis through increasing the content of collagen and decreasing the content of hydroxyproline, 2 mg/kg gallium nitrate are similar to estrogen treatment. Related Articles | Metrics

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Effect of platelet-rich plasma, activated by different concentrations of thrombin, on the repair of cranial defects Lin Min-kui, Chen Xiao-ling, Zhao Xin, Yan Fu-hua 2010, 14 (2):  209-213.  doi: 10.3969/j.issn.1673-8225.2010.02.005 Abstract ( 304 )   PDF (1494KB) ( 414 )   Save BACKGROUND: The biological functions of platelet-rich plasma (PRP) are affected by multiple factors, such as individual difference, PRP concentration, PRP carrier, PRP-activated methods and so on. OBJECTIVE: To evaluate the effect of PRP, activated by different concentrations of thrombin, on the repair of cranial defects. METHODS: Whole blood of the central artery of rabbit ears was extracted to prepare PRP, which was then diluted so that the final platelet count was about 5 times of the whole blood. Four whole-thickness layer of cranial defects at an 8-mm diameter were created in 16 New Zealand rabbits and randomly grafted with β-tricalcium phosphate (β-TCP) and PRP, activated by 60 U/mL thrombin; β-TCP and PRP, activated by 1 000 U/mL thrombin; β-TCP and PRP; β-TCP alone. At 1 and 3 months following implantation, X-ray analysis and microscopic observation were performed to onserve cranial repair, the area percent of new bone formation was calculated. RESULTS AND CONCLUSION: At one month post-surgery, the edge of defects was clear in each group, with varying degrees of new bone formation surrounding the defects, β-TCP particles partially degraded and the degradation lesion was replaced by new bone, only a small amount of bone lacunae was seen, fiber wrapped around the defect center β-TCP, only a small number of specimens showed new bone formation; X-ray showed a clear boundary and uniform defect density; the percentage of new bone formation in the PRP groups were higher than β-TCP groups (P < 0.05). However, there was no significant difference between PRP group groups (P > 0.05). At 3 months post-surgery, the defect boundary was unclear in each group, the new bone formation increased, the β-TCP particles surrounding defects partially or all degraded and were replaced by new bones, some regions appeared trabecular bone, bone lacuna in new bone was increased, the central defect of the majority of specimens exhibited new bone formation; X-ray showed defect boundary was unclear in each group, defect surrounding density was higher than the center defect, and bone mineral density was equivalent to other normal parts; the percentage of new bone formation in the PRP groups was significantly higher than that in the β-TCP groups (P < 0.05), PRP + β-TCP group was higher than the other 3 groups (P < 0.05), there was no significant difference between two thrombin groups (P > 0.05) . It is indicated that although PRP improves the repair of cranial defects, 60 and 1 000 U/mL of thrombin has no effects on PRP repairing cranial defects in New Zealand white rabbits, compared with PRP+β-TCP group, possible the absence of the optimal concentration of thrombin. Related Articles | Metrics

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Expression of articular chondrocytes in rabbits transfected by retroviral vector-mediated transforming growth factor beta 1 gene in vitro  Lin Shu-zhong, Liu Jun, Xiang Chuan, Wei Xiao-chun 2010, 14 (2):  214-217.  doi: 10.3969/j.issn.1673-8225.2010.02.006 Abstract ( 349 )   PDF (1405KB) ( 335 )   Save BACKGROUND: The functional gene fragments integrate into gene vector, which is then transfected into target cells or joint cavity, through the transgenic target cells continue to secrete a large number of functional gene product, local therapeutic concentrations could be maintained within a long period of time, thus repairing articular cartilage injury. OBJECTIVE: To transfect rabbit articular chondrocytes using recombinant retroviral vector-mediated transforming growth factor-β1 (TGFβ1) in vitro, and to observe its expression and its effect on biological characters of chondrocytes. METHODS: Rabbit chondrocytes were isolated by use of trypsin digestion method. Vector was PLNCX2 Hind Ⅲ/Not Ⅰ doubly digested and dephosphorylated, connected with some multiple cloning sites and RFP gene following pDsRed2 double digestion, to build PLNCX2-RFP. TGFβ1 gene was amplified from the PGEMT-TGF and connected with PLNCX2-RFP following double digestion, to build PLNCX2-TGFβ1-RFP. Subsequent to packaging retroviral vector, viral supernatant titer was detected. The cultured and transfected chondrocytes in rabbit knee joint were divided into 3 groups: control group (without any transfection), transfected PLNCX2 group and transfected PLNCX2-TGFβ1-RFP group, continued screening 2 weeks to observe the cellular changes. Cell supernatant transfected stably were collected for detecting the effect of gene transfection on the chondrocytes with NO detection kit, ELISA assay was applied to determine human TGFβ1 expression in cell culture supernatant. RESULTS AND CONCLUSION: The recombinant gene PLNCX2-TGFβ1-RFP was identified correct sequence by the enzyme digestion sequencing TGFβ1 and RFP, which showed that the eukaryotic expression vector PLNCX2-TGFβ1-RFP had been successfully built as expectation. They were then transfected into packaging cells and cultured, the virus titer was defined as 1×106 CFU. Following stable transfection of cartilage cells, red fluorescence can be observed, proving successful transfection. After continuous screening 2 weeks, the scattered adherent cells formed positive clones, and gradually diffusely integrated, cell clusters appeared with common dual cores, the cells proliferated actively. NO concentration in the transfected PLNCX2-TGFβ1-RFP group was higher than that of transfected PLNCX2 group (P < 0.05), no difference was significant between control group and transfected PLNCX2 group. The control group and the group transfected PLNCX2 showed no TGFβ1 expression, while TGFβ1 concentration was (28.08±3.73) ng/L in the transfected PLNCX2-TGFβ1-RFP group. PLNCX2 retroviral vector-mediated human TGFβ1 can be effectively transfected into rabbit knee joint cartilage cells and obtain stable expression, while the transfected cartilage cells proliferate actively. Related Articles | Metrics

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Differentiation of bone marrow mesenchymal stem cells into fibrochondrocyte phenotype Cai Gui-quan, Cui Yi-min, Chen Xiao-dong 2010, 14 (2):  218-222.  doi: 10.3969/j.issn.1673-8225.2010.02.007 Abstract ( 283 )   PDF (1479KB) ( 451 )   Save BACKGROUND: The meniscus has limited ability in repairing itself after being injured. However, tissue engineering provides a new way to meniscus repair after injury. Bone marrow mesenchymal stem cells (BMSCs), which possess the potential of multi-directional differentiations, can be ideal seed cells in meniscus tissue engineering. OBJECTIVE: To investigate the feasibility of differentiation of in vitro cultured porcine BMSCs into fibrochondrocyte phenotypes in inductive medium. METHODS: BMSCs were isolated with whole bone marrow culture method. Then, BMSCs of the third passage were digested and incubated in a medium containing transforming growth factor-β1, insulin-like growth factor-Ⅰ, dexamethasone and ascorbic acid in a 24-well plate at a density of 2.0×104/cm2 in the experimental group. While in the control group, the DMEM-LG complete culture medium containing no inductive factor were used instead. At day 7, 14 and 21 after induction respectively, Toluidine blue staining and immunocytochemical staining were performed to detect differentiation. MAIN OUTCOME MEASURES: ①Population double time (PDT) of BMSCs; ②Morphological changes of BMSCs under light microscope; ③Proteoglycan expression; ④Collagen type Ⅰand type Ⅱ expression. RESULTS AND CONCLUSION: ①The PDT of the second passage BMSCs was 2 days, which was the shortest. The PDT prolonged relatively after the fourth passage, which were 5 to 9 days. ②The BMSCs changed from a spindle-like appearance into a polygonal shape after induction. ③In the experimental group, toluidine blue staining resulted in hyacinthine-stained cytoplasm and the blue was even deeper in the area where cells were dense; The degree of staining increased with the increasing induction time. While in the control group, only nucleus of BMSCs were stained blue. ④Collagen type Ⅰ immunocytochemical staining was positive in both the experimental and the control group and there was no difference of significance between various induction time. No collagen type Ⅱwas seen expressed in the control group, while in the experimental group it was seen to be expressed steadily after 14 days of induction. It is indicated tlat BMSCs can be induced to synthesize fibrochondrocyte-characterized extracellular matrixes in vitro, which suggests that BMSCs are available as seed cells in meniscus tissue engineering.   Related Articles | Metrics

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Construction of immortalized human precartilaginous stem cell lines Yin De-long, Chen An-min, Guo Feng-jin, Wang Jun-fang, Cheng Hao 2010, 14 (2):  223-226.  doi: 10.3969/j.issn.1673-8225.2010.02.008 Abstract ( 343 )   PDF (1413KB) ( 379 )   Save BACKGROUND: The precartilaginous stem cells are limited regarding in vitro proliferative capacity, but the immortalized cell lines can provide a large number of stable immortalized cells, and simian virus 40 large T antigen gene (SV40Tag) is one of gene fragments which are commonly used and effective in vitro immortalized cells. OBJECTIVE: To construct human immortalized precartilaginous stem cells (IPSCs) using human precartilaginous stem cells induced by SV40LTAg gene. METHODS: The human immortalized precartilaginous stem cells were isolated from aborted fetus and purified with enzyme digestion and immunomagnetic beads screening method. By using liposome-mediated gene transfection technology, plasmid pCMVSV40T/PUR containing SV40Tag was transfected in primary embryonic precartilaginous stem cells, while non-transfected cells served as negative controls. Positive clones were cultured to observe the cell morphology and the passage recovery, to calculate cell survival rate and population doubling time, to draw cell growth curve. Immunofluorescence cytochemistry was used to detect the expression of IPSCs fibroblast growth factor receptor 3, the expressions of SV40Tag and fibroblast growth factor receptor 3 in the human precartilaginous stem cells were determined by RT-PCR. RESULTS AND CONCLUSION: Morphology of human IPSCs seemed coincidence with primary human precartilaginous stem cells. The survival rate of human IPSCs was not influenced by subculture, freezing and recovery, but the survival rate was descended in the human precartilaginous stem cells at the 6th and 10th passages (P < 0.01). Compared with cells at the 6th and 10th passages, the proliferation of human IPSCs was greater, with short population doubling time and high growth rate (P < 0.01). The immunofluorescence showed that fibroblast growth factor receptor 3 was positive in human IPSCs at the second passage, and the RT-PCR results of fibroblast growth factor receptor 3 revealed a specific amplification band at 400 bp, while that of SV40Tag revealed at 560 bp. No band was seen in the primary cells. It is indicated that SV40Tag human IPSCs can be constructed successfully using immunomagnetic bead screening technology and liposome transfection technique. Related Articles | Metrics

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Influence of growth factors and bone marrow mesenchymal stem cells on lung revascularization and restoration of smoking-induced emophysematous model in rats Li Bao-ping, Xing Wan-hong, Lu Peng-yan, Zhang Lei, Ma Jie 2010, 14 (2):  227-232.  doi: 10.3969/j.issn.1673-8225.2010.02.009 Abstract ( 293 )   PDF (2265KB) ( 379 )   Save BACKGROUND: Preliminary study has proved that the bone marrow-derived mesenchymal stem cells (MSCs) in a rat emphysema model produced by use of trypsin alone can "homing" to the lesioned lung tissues, and participate in the formation of pulmonary arteries to promote lung tissue repair. Basic fibroblast growth factor (bFGF) and vascular endothelial growth factor (VEGF) play equally a powerful role in promoting angiogenesis. OBJECTIVE: To observe the influence of bFGF, VEGF and MSCs in regeneration of pulmonary capillary and pathological repair of pulmonary emphysema rats. METHODS: Except normal control group, the remaining 5 groups of rats were exposed to tobacco smoke and received a single intratracheally instillation of porcine pancreatic elastase to induce emphysema models. Following successful modeling, rats of bFGF group were intratracheally injected with 400 U bFGF and rats of VEGF group with 2 μg VEGF, once a week for three times. MSCs group was injected 1 mL suspension of 4×109/L MSCs into tail vein. MSCs+VEGF group was injected MSCs into tail vein and intratracheally injected VEGF (2 ug, three times) at the same time. Model control and normal control groups were intratracheally injected with equal volume of sodium chloride. Four weeks after treatment, arterial blood gas analysis was performed to observe pathological and morphological changes of lung tissues. CD34+ expression in lung tissues was determined using immunohistochemistry method. RESULTS AND CONCLUSION: Compared with model control group, PaO2 values dramatically increased in VEGF group (P < 0.05), while other indices remained unchanged (P > 0.05); there were no obvious changes in each index in other groups (P > 0.05). Gross and microscopic observations showed that, lung was smooth, pale pink, and elastic in normal control group, with uniform size of pulmonary alveoli on cross-section; pathological changes of chronic obstructive pulmonary emphysema existed in model control group, but improved in other 4 groups. Compared with model control group, mean pulmonary alveoli number and CD34+ relative positive area dramatically increased in bFGF, VEGF, MSCs, MSCs+VEGF groups (P < 0.05), mean linear intercept and mean alveoli area were significantly reduced (P < 0.05). No significant difference was observed in each index among these 4 groups (P > 0.05). bFGF, VEGF and MSCs could improved the pathology of pulmonary emphysema models produced by tobacco smoking and intratracheally instillation of porcine pancreatic elastase. The possible mechanism of recovering the pulmonary emphysema is the proliferation of pulmonary capillary and enlargement of pulmonary artery, improved blood flow in the lung, improved ventilation/perfusion shunt, reduced pulmonary alveolus size and volume of the lung through self-compensation. Related Articles | Metrics

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Vascular endothelial growth factor expression and angiogenesis in human ovarian tissue after different cryopreservations Fu Jin-mei, Wang Yan, Xiao Zhun, Li Shang-wei 2010, 14 (2):  233-236.  doi: 10.3969/j.issn.1673-8225.2010.02.010 Abstract ( 379 )   PDF (1271KB) ( 298 )   Save BACKGROUND: The cryopreservation of human ovarian tissue has become an attractive method to preserve female fertility. Human ovarian tissue experiences neovascularization after transplantation to recover blood supply, cryopreservation and resuscitation technique is a key for the neovascularization of human ovarian tissue following transplantation. OBJECTIVE: To investigate vascular endothelial growth factor (VEGF) expression and microvessel density in human ovarian tissue following novel needle immersed vitrification (NIV) and slow-freezing, to explore the influence of two cryopreservation methods play in the neovascularization of human ovarian tissue after transplantation. METHODS: Eight normal human ovarian tissues from patients with carcinoma of endometrium were cut into 12 fragments in the size of 1.5 mm × 1.5 mm × 1.0 mm, then randomly assigned to 3 groups: fresh control group, NIV group and slow-freezing group. In the NIV group, pieces of ovarian tissue strips were dehydrated in an equilibration solution consisting of 7.5% ethylene glycol and 7.5% dimethyl sulfoxide in TCM-199 supplement with 20% fetal bovine serum and a vitrification solution consisting of 15% ethylene glycol, 15% dimethyl sulfoxide and 0.5 mol/Lsucrose, then were plunged in liquid nitrogen directly and sealed in liquid nitrogen-filled cryovials. For thawing, the needles holding ovarian tissues were serially transferred into 1.0, 0.5, 0.25 mol/L sucrose solution and TCM-199 supplemented with 20% fetal bovine serum. In the slow-freezing group, pieces of human ovarian cortex fragments were placed in a 1.8-mL cryovial containing 1 mL of TCM-199 medium supplemented with 0.1 mol/L sucrose, 20% fetal bovine serum and 1.5 mol/L dimethyl sulfoxide, the cryovials were placed in the programmable freezer and cryopreserved by pre-set slow-cooling protocol. For thawing, the ovarian tissue strips were washed in a stepwise manner: 1.0 mol/L dimethyl sulfoxide + 0.1 mol/L sucrose, 0.5 mol/L dimethyl sulfoxide + 0.1 mol/L sucrose, 0.25 mol/L dimethyl sulfoxide + 0.1 mol/L sucrose and 0.1 mol/L sucrose. The frozen-thawed and fresh controlled human ovarian tissues were cultured in vitro. The expression of VEGF and CD34, as well as microvessel density, was analyzed by immunohistochemistry. RESULTS AND CONCLUSION: There was patchy and mild expression of VEGF in the stromal cells of all the three groups before and after culture. The expression of VEGF increased and reached peak value after culture for 2 days, began to decrease after culture for 4 days and further attenuated after culture for 6 days in all the three groups. Compared with slow-freezing group, the expression of VEGF in NIV group was closer to that in fresh control group. Microvessel density of all the three groups increased and reached peak value after culture for 2 days, and the microvessel density of fresh control group and NIV group was significantly higher than that of slow-freezing group (P < 0.05). The microvessel density of slow-freezing group after culture for 4 days and that of all the three groups after culture for 6 days significantly decreased compared with after culture for 2 days (P < 0.05). NIV is superior to slow-freezing to preserve stromal cells and extracellular matrix of human ovarian tissue, and plays less influence in VEGF expression and angiogenesis in human ovarian tissue. Related Articles | Metrics

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Meta analysis of soft tissue profile repair in patients treated with and without premolar extraction Liu Ze-ping, Chen Ting-ting, Li Gui-feng, Jiang Si-cong, Zou Shu-juan 2010, 14 (2):  237-239.  doi: 10.3969/j.issn.1673-8225.2010.02.011 Abstract ( 342 )   PDF (881KB) ( 522 )   Save OBJECTIVE: To analyze the impact of extraction and nonextraction in soft tissue profile repair. METHODS: Databases of CNKI, Wanfang, VIP Chinese Periodical Database, CBM, Medline (ovid) and Science direct had been searched for studies of soft tissue profile after orthodontic treatment with and without extractions published in recent decade, with orthodontics, corrective, tooth extraction, soft tissue profile as key words. Hand searching was also done to obtain any further information about the studies. Meta analysis was used to comprehensively quantitatively analyze the upper (lower) lip with esthetic plane and soft tissue plane before and after extraction and nonextraction treatment. According to homogeneity test, fixed effect model and random effect model were adopted to calculate weighted mean difference (WMD) and 95% confidence interval (95% CI). Revman 5.0 was used for data management. RESULTS: Five studies with 356 patients were included. Homogeneity test showed that lower lip-esthetic line and lower lip-soft tissue line I2 < 50%, without heterogeneity, so fixed effect model was adopted. Upper lip-esthetic line and upper lip-soft tissue line were more than 50%, indicating heterogeneity, so random effect model was adopted. Result of meta-analysis showed that, the WMD value and 95% CI of lower lip-esthetic line, lower lip-soft tissue line, and upper lip-soft tissue line has statistically significance. CONCLUSION: The extraction treatment significantly decreased the distance of lower lip-esthetic line, lower lip-soft tissue line, and upper lip-soft tissue line. Related Articles | Metrics

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Protective effect of basic fibroblast growth factor on renal tubular epithelial cell toxicity induced by gentamicin    Huang Rong-shi, Huang Ju-en, Cen Yan-hui, He Guo-zhen, Zhao Fei-lan, Fan Rong 2010, 14 (2):  240-244.  doi: 10.3969/j.issn.1673-8225.2010.02.012 Abstract ( 268 )   PDF (1397KB) ( 402 )   Save BACKGROUND: In vivo experiments have confirmed that fibroblast growth factor can effectively protect gentamicin-induced renal tubular epithelial cell injury, but the effect on the in vitro cultured cells is still rare. OBJECTIVE: To explore the mechanisms of basic fibroblast growth factor (bFGF) at different concentrations on preventing nephrotoxity mediated by genamicin on the primarily cultivated renal tubular epithelial cell models. METHODS: By use of enzyme and mesh screening, renal tubular epithelial cells were isolated from Kunming mice and purified, adjusting the cell concentration of 1×108/L, then cell suspension was moved to a 96-well culture plate and divided into different groups for culture: blank control group: normal culture; gentamicin group: 10, 30, 50 μL/hole (ie, 400, 1 200, 2 000 U/holes) are recorded as G1, G2, G3; bFGF group: 20, 50, 80 μL/hole (ie, 90, 225, 360 ng/hole) are recorded as B1, B2, B3; gentamicin plus bFGF intervention group: after adding bFGF 12 hours, then added gentamicin 12 hours, assigned into 9 dose subgroups, namely, G1B1, G1B2, G1B3, G2B1, G2B2, G2B3, G3B1, G3B2, G3B3, each subgroup contained four-hole complex. Cell morphology and quantity was observed. RESULTS AND CONCLUSION: Gentamicin showed a dose-dependent effect on the renal tubular epithelial cell injury, epithelial cells in the medium and high concentration groups exhibited shrinkage, rounded, swelling, poor adhesion, severely damaged cytoplasm and structural disorder. In the low concentration group, the number change of cells was not obvious, and fibroblasts began to appear; In the bFGF groups, cells were full, exhibited strong refraction, the cell number increased significantly, these manifestations were significant in 50 μL/hole concentration, and there was no significant difference compared with 80 μL/hore concentration; in case of gentamicin plus bFGF intervention, cells with low concentrations of gentamicin had no obvious damage to cells, which increased, the damaged cells collapse was reduced in the group of low concentration of gentamicin, cell shrinkage and poor adhesion were slightly relieved, high concentrations of bFGF intervention could yield to good cell morphology, but high concentrations of gentamicin caused cell swelling and necrosis of injury, which could not be improved by any concentrations of bFGF intervention. 50 μL/hole bFGF has antagonistic effect on the nephrotoxicity mediated by medium and low concentrations of gentamicin, but has no protection on high concentration of gentamicin-induced nephrotoxicity. Related Articles | Metrics

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Effect of mechano growth factor on radial fracture healing in rabbits Liao Bao-qiang, Deng Mo-yuan, Fu Ya, Wang Yuan-liang, Rao Quan-zhen 2010, 14 (2):  245-248.  doi: 10.3969/j.issn.1673-8225.2010.02.013 Abstract ( 268 )   PDF (1406KB) ( 591 )   Save BACKGROUND: Current research of mechano growth factor (MGF) mainly focuses on the muscles and nerve damage and repair, and it has been confirmed that MGF can promote muscle cell hypertrophy and nerve repair significantly. Regarding its role in fracture healing is unclear. OBJECTIVE: To investigate the effect of MGF on radial fracture healing in rabbits. METHODS: By using random digital table method, 12 New Zealand rabbits were divided into 3 groups: blank control group, low-dose MGF group and high-dose MGF group. The models with 5 mm bone defect were produced in the middle of the left radius in rabbits. At 3 days after the surgical operation, the defective areas were given 0.2 mL PBS or 0.2 mL MGF (0.36 and   0.72 g/L) injected into the ends of fracture areas, respectively, once per day for continuous 5 days. At 4, 6, 8 weeks after operation, X-ray photography was used to evaluate the healing of fracture, and the histological examinations were performed at the 8th week to observe the cell morphology at the fracture lesion. RESULTS AND CONCLUSION: At 1 day after operation, the activities of rabbits were reduced, with slightly reduced food intake, at 2 days they almost recovered to normal activities and diet. At 3 days, the surgical incision slightly swelled with a small amount of bleeding and without obvious signs of infection. All 12 rabbits entered the final analysis. X-rays showed that two fracture ends have basically combined in the high-dose MGF group at 4 weeks post-surgery, cortical bone was continuous and fracture lines were unclear. At 6 weeks, the bone medullary cavity almost run through and fully run through at 8 weeks. The healing time in the high-dose MGF group was remarkably shorter than that in blank control group and low-dose MGF group, the healing was in high quality. At 8 weeks after operation, a large number of osteoid tissues were observed in the blank control group, a small amount of woven bone formed, at a transition period from the fibrous bone callus to the bony bone callus; a large number of woven bone formed in the low-dose MGF group, at bony bone callus period; in the high-dose MGF group, a large number of woven bones converted into mature lamellar bone, at the callus rebuilding phase, which was consistent with imaging results. It is indicated that MGF can accelerate fracture healing significantly in a rabbit model and shows a dose-dependent manner in a certain range. Related Articles | Metrics

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Peripheral nerve transplantation with growth factors for acute spinal cord injury Hou Zhan-jiang, Yang Shuo, Meng Xiang-lin, Xia Hao-chen, Fan Chang-dong, Yan Jing-long 2010, 14 (2):  249-252.  doi: 10.3969/j.issn.1673-8225.2010.02.014 Abstract ( 261 )   PDF (1106KB) ( 360 )   Save BACKGROUND: At present the strategy of nerve regeneration and repairing are main promoting nerve intrinsic regeneration capacity and improving the micro-environment. Studies have shown a number of combined treatment which could promote the regeneration and growth of nerve axon. OBJECTIVE: To explore the feasibility and effect of rat spinal cord injury repaired by peripheral nerve combined growth factor. METHODS: Sixty healthy adult female SD rats were randomly divided into 4 groups: nerve graft group, nerve graft combined growth factor group, spinal cord transaction group and laminectomy group. Taking T9 as the center, a longitudinal incision was conducted in rat skin, revealing dural sac, spinal cord was transected and removed 3 mm, 2-cm segment of the eighth to tenth intercostal nerve was obtained from nerve graft group and nerve graft combined with growth factor group, autologous intercostal nerve was cross-transplanted into spinal defect (proximal white matter and distal gray matter, distal white matter and proximal gray matter) after pruning appropriately. The transplanted intercostal nerves were fixed with fibrin glue in nerve graft group, while those in nerve graft combined growth factor group were fixed with fibrin glue containing 2.1 mg/L acidic fibroblast growth factor, followed by dural suture. Stump of broken ends was done in spinal cord transection group, while laminectomy was performed in laminectomy group. RESULTS AND CONCLUSION: At 90 days post-surgery, somatosensory evoked potential (SEP) and motor evoked potential (MEP) were determined, the motor function of hind limbs was evaluated by the Basso. Beattie.Bresnahan (BBB) test at 70 days. Both SEP and MEP were led in the laminectomy group, but not lead in spinal cord transection group; in nerve graft group, 3 rats showed bilateral SEP, 4 led unilateral SEP, 4 led bilateral MEP, 3 led unilateral MEP; in nerve graft combined with growth factor group, 5 led bilateral SEP and 2 led unilateral SEP, 5 led bilateral MEP and 2 led unilateral MEP. The SEP and MEP latency and amplitude in the nerve graft group and nerve graft combined growth factor group were significantly superior to the spinal cord transection group (P < 0.01), autologous rib nerve graft group was better than nerve graft combined growth factor group (P < 0.01). In the laminectomy group, awake rats following anesthesia returned to normal exercise, rats in spinal cord transection group continued to extend limbs and rotated within 3 months, rats in other two groups recovered functions obviously 3 weeks post-surgery and gradually restored throughout the entire observation period. Nerve graft group and nerve graft combined growth factor group showed significantly increased BBB score compared with spinal cord transection were (P < 0.01), and the nerve graft combined growth factor group was superior to nerve graft group (P < 0.01). The peripheral nerve graft can promote the spinal function following spinal cord injury, while the nerve combined growth factor can better restore the function. Related Articles | Metrics

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Expression of type Ⅰ transforming growth factor beta receptor and type Ⅰ collagen in the peripheral and central area of human skin hypertrophic scar Geng Zhong-li, Dong Xiang-lin, Ma Bin-lin 2010, 14 (2):  253-257.  doi: 10.3969/j.issn.1673-8225.2010.02.015 Abstract ( 271 )   PDF (1648KB) ( 416 )   Save BACKGROUND: Many studies focus on transforming growth factor β (TGF β) and its receptors, however, the distribution of type Ⅰ TGF receptor (TGF-βR Ⅰ) in peripheral region of hypertrophic scars remain poorly understood.  OBJECTIVE: To determine the expression and distribution of TGF-βR Ⅰ and type Ⅰ collagen in the peripheral and central areas of human skin hypertrophic scar. METHODS: A total of 30 cases with human cutaneous scars admitted at the Department of Plastic Surgery, First Affiliated Hospital and Department of Mammary Gland, Head and Neck Surgery, Tumor Hospital of Xinjiang Medical University from 1999 to 2002, were selected, including 20 cases with hypertrophic scar and 10 cases with normal scars. A total of 180 scars were obtained from central and peripheral areas of scars as well as normal skin tissues. The protein contents of TGF-βR1 and type Ⅰ collagen was detected by immunohistochemistry. In addition, the immunostaining positive in these samples was analyzed by semiquantitative analysis. RESULTS AND CONCLUSION: Compared to non hypertrophic scar and normal skin tissues, the TGF-βR1 expression of hypertrophic scar was obvious greater with strong positive reaction. The TGF-β RⅠ content was 100% in peripheral region of hypertrophic scar, which was notably 20% greater than that of central area (P < 0.05). The content of type Ⅰ collagen was both 100% in peripheral and central areas. The differences of positive TGF-β  RⅠ and type Ⅰ collagen had no significance between peripheral and central areas of non hypertrophic scars (P > 0.05). There were few contents of TGF-β RⅠ and type Ⅰ collagen in normal skin tissues. The expression of TGF-β RⅠ is higher in peripheral than central areas of hypertrophic scar. Therefore, the peripheral area would be emphasized in the clinic work. Related Articles | Metrics

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Small interfering RNA targeting vascular endothelial growth factor gene inhibits proliferation of gallbladder cancer cells: An in vitro and in vivo experiment Qu Hua-wei, Zhang Yang-de, Chen Yu-xiang, Zhao Jin-feng, Liao Ming-mei, He Jian-tai 2010, 14 (2):  258-261.  doi: 10.3969/j.issn.1673-8225.2010.02.016 Abstract ( 305 )   PDF (1227KB) ( 354 )   Save BACKGROUND: Previous studies demonstrated that proliferation of cancer cells can be inhibited via RNA interference on the expression of vascular endothelial growth factor (VEGF). However, few studies report RNA interference on the expression of VEGF in gallbladder carcinoma. OBJECTIVE: To design and screen shRNA targeting VEGF, and to observe the effect of small interfering RNA targeting on proliferation of gallbladder cancer cells. METHODS: The VEGF-shRNA fragment was synthetized and connected with pCYU6/GFP/Neo-shRNA plasmid vector. shRNA was transfected into gallbladder cancer cells. The gallbladder carcinoma models of nude mice were prepared and randomly divided into blank control, negative control and experimental groups, with 6 animals in each group. shRNA was injected into tumor. Cell growth was detected by fluorescence microscope MTT. The RNA interference efficiency was examined by fluorescent quantitative RT-PCR. Changes of tumor volume were also observed. RESULTS AND CONCLUSION: Gallbladder cancer cells were shrunk with round shapes and a part of cells were dead after RNA interference on VEGF. shRNA-VEGF1 and shRNA-VEGF2 could significantly inhibit mRNA gene expression of VEGF, the inhibition ratio was 86% and 82%, respectively. The tumor volume of the experimental group was smaller than the other groups, with slowly growth (P < 0.05). No obvious changes were found in the blank control and negative control groups. The constructed hVEGF-shRNA vector markedly decreases VEGF gene expression and inhibits cellular proliferation, eventually, to treat gallbladder cancer. Related Articles | Metrics

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Effect of eucommia bark on osteopontin and osteoprotegerin expressions in rat bone marrow stem cells: Is there any difference between water extract and methanol extract? Zhang Yan-hong, Xie Huan-song, Xia Shu-lin, Zhao Chun, Tan Xiang-ling 2010, 14 (2):  262-266.  doi: 10.3969/j.issn.1673-8225.2010.02.017 Abstract ( 283 )   PDF (1332KB) ( 503 )   Save BACKGROUND: Previous studies demonstrated that eucommia bark can promote bone marrow stem cells (BMSCs) differentiated into osteoblasts, but relative mechanism is poorly understood.  OBJECTIVE: To investigate the effects of eucommia bark water/methanol extracts on expressions of osteopontin (OPN) and osteoprotegerin (OPG) in rat BMSCs. METHODS: Totally 2 g eucommia bark powder were added into water or methanol to 15 mL and oscillated for 1 hour at room temperature. After soaked overnight, both extracts were centrifuged at 15 000 r/min for 10 minutes. Water extract was obtained from supernatant in water soaked powder. In methanol soaked powder, methanol extracts was obtained by concentrated supernatant in vacuo and resolved using 15 mL water. Water and methanol extracts were then filtered by 0.22 μm membrane, and conserved at -20 ℃. Six SD rats, aged 2 months, were selected, and the 3rd passage of BMSCs were induced by water or methanol extracts with dilution of 1×10-2, 1×10-3, 1×10-4 and 1×10-5, respectively. PBS was added in the negative control group. All cells were cultured for 6 days. Expressions of OPN and OPG was measured by immunocytochemistry at 6 days with induction. The expression of OPN and OPG induced by water and methanol at 1×10-3 and 1×10-4 dilution was detected by RT-PCR. RESULTS AND CONCLUSION: Immunocytochemistrical results indicated that both water and methanol extracts of eucommia bark simulated OPN and OPG expression, in particular with dilution of 1×10-4. The methanol extracts had a stronger effect than water extract, but the expression of OPG did not change obviously. RT-PCR demonstrated that at the 3rd day of inducement, the level of OPN expression induced by water extract was higher than that of methanol extract, and no OPG expression was detected. Osteogenic differentiation of rat MSCs induced by eucommia bark water/methanol extracts relates to stimulating expression of OPN, which has no correlation to OPG. OPN expression induced by water extract is early than that of methanol extract.  Related Articles | Metrics

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Effect of tripterygium wilfkrdii Hook, ginseng and arotinoid ethyl ester on the sun-induced aging cultured fibroblasts of the human dermis Zhong Gui-shu, He Yuan-min, Liao Yong-mei　 2010, 14 (2):  267-271.  doi: 10.3969/j.issn.1673-8225.2010.02.018 Abstract ( 259 )   PDF (1516KB) ( 503 )   Save BACKGROUND: The expression of matrix metalloproteinases (MMPs) is enhanced in fibroblasts of aging skin; tripterine and ginsenoside Rd possess immunosuppression and anti-aging effects. Arotinoid ethyl ester can used to treat sun-induced aging skin, which accompanied by many side effects. OBJECTIVE: To discuss the regulation role of ginsenoside Rd, celastrol, arotinoid ethyl ester on MMP-1 and MMP-3 of the cultured fibroblasts of the human dermis in vitro. METHODS: The human dermis circumcision by surgical excision on newborn was obtained from Department of Surgery, Luzhou Medical College, and the informed consent obtained from patients. Culture of fibroblasts were as follow: ①No treatment in the normal control group. ②Radiated with 80 kJ/m2 ultraviolet and treated by 100 mg/L 8-methoxypsoralen in the positive control group. ③Treated by ultraviolet radiation+8-methoxypsoralen+arotinoid ethyl ester in the arotinoid ethyl ester group. ④Treated by ultraviolet radiation +8-methoxypsoralen+triptolide in the triptolide group. ⑤Treated by ultraviolet radiation+8-methoxypsoralen+ginsenoside Rd in the ginsenoside Rd group. The triptolide group was divided into 3 groups with doses of 10, 20, and 40 mg/L. The ginsenoside Rd group was divided into 3 groups with doses of 20, 50 and 100 mg/L. The immunohistochemistry staining of MMP-1 and MMP-3 of the fibroblasts in each group was observed. RESULTS and CONCLUSION: The expressions of MMP-1and MMP-3 were significantly increased in the positive control group, which had a significant difference to the normal control group (P < 0.05). Compared to the positive control group, the expression of MMP-1and MMP-3 significantly decreased in the treatment groups (P < 0.05); however, the differences among each treatment groups had no significance (P > 0.05). The results demonstrated that: ①Ginsenoside Rd and tripterine exhibit comparably effect on prevent and cure sun-induced aging skin. ②The wide ranges of effective concentration lead to little side effect to the body. ③The action mechanisms of ginsenoside Rd, celastrol and arotinoid ethyl ester in preventing and curing sun-induced aging skin is adjusting the expressions of MMP-1 and MMP-3. Related Articles | Metrics

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Establishment of a rat midpalatal suture expansion model Guo Jing, Zhang Qi-feng, Li Gui-feng, Liu Ze-ping, Zhou Jing, Zou Shu-juan 2010, 14 (2):  272-28-75.  doi: 10.3969/j.issn.1673-8225.2010.02.019 Abstract ( 328 )   PDF (1439KB) ( 329 )   Save BACKGROUND: Monkeys, dogs, pigs, rabbits and other large animals have been applied previously to prepare animal models of midpalatal suture expansion, but there are high cost, small sample size, difficult to obtain antibodies and other disadvantages. Wistar rats have wide heads to facilitate cavity operation, with low cost and high reproduction rate, as the midpalatal suture model, it is possible to overcome the above deficiencies. OBJECTIVE: To establish a rat model of midpalatal suture expansion, and to supply basement for further relative researches of animal models. METHODS: Twenty Wistar male rats of 5 weeks old, average weight of 65 g, were randomly divided into 2 groups, a experiment and a control, with 10 rats in each group. Rats in experimental group were placed on the expansion appliance, inserting into the diastema between the first and second molars, then stick to molar lingual using light-cured resin for retention. The rats in control group were sham operated, followed by one week of active expansion. After expansion, the midpalatal sections were observed by X-ray and light microscopy. RESULTS AND CONCLUSION: Maxillary X-ray film showed that midpalatal suture in experimental group was significantly widened, molar lead to cheek. Observed by light microscopy, partial oral side of midpalatal suture in the experimental group was obviously enlarged, mesenchymal cells were spindle, in the same direction to tension force. Below it, traumatic inflammatory response appeared, with a clear bleeding area. The midpalatal suture expansion model in rats is available, simple and reproducible. Related Articles | Metrics

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Establishment of rabbit femoral head necrosis models using liquid nitrogen refrigeration method: Is it feasible, ideal and confident? Dai Shuang-wu, Ding Shuai, Li Zhang-hua, Wu Mao-kui, Xi Jiang-ming, Meng Fan-sheng, Xiong Wen-hua 2010, 14 (2):  276-279.  doi: 10.3969/j.issn.1673-8225.2010.02.020 Abstract ( 360 )   PDF (1359KB) ( 460 )   Save BACKGROUND: The deficiency of perfect animal femoral head necrosis model limited its further investigation.  OBJECTIVE: To verify the feasibility of establishing rabbit femoral head necrosis models using liquid nitrogen refrigeration method, and to provide a foundation for subsequent research.  METHODS: A total of 20 adult, New Zealand, white rabbits were selected in the study. The round ligament of femur was not cut off and femoral head was not dislocated, and the exposed femoral head were quick frozen using cotton bud carrying liquid nitrogen for successive 25 times, with 10 s per time. The specimens were examined by gross anatomy, X-ray film, MRI and histological observation at day 3, 7 and weeks 2, 4, 6, and 8 after operation.  RESULTS AND CONCLUSION: The histological section showed that chondrocyte, osteocyte, and myeloid tissues presented necrosis in freezing and periphery at 3 days after model preparation, and the repair process appeared at 2 weeks after operation. The articular surface of femoral heads appeared collapse at 4 weeks after operation, and these changes became obvious at 6 weeks. The femoral head presented osteoarthritis-like disorder, with seriously collapsed articular surface at 8 weeks, and the contour of femoral head changed in 2 animals. The results demonstrated that without hip dislocation, rabbit femoral head necrosis models can be established successfully using liquid nitrogen refrigeration method. This method is simple, feasible, with high succeed rate, which can be used in subsequent research. Related Articles | Metrics

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Establishment of rabbit models of ischemic lumbar vertebrae adjacent to endplate: Feasibility of MRI and pathology verification Hou Chang-long, Yang Jian-yong, Zhuang Wen-quan, Tan Guo-sheng, Fan Hui-shuang, Mao Li-juan, Zhang Zhong-wei 2010, 14 (2):  280-284.  doi: 10.3969/j.issn.1673-8225.2010.02.021 Abstract ( 363 )   PDF (741KB) ( 472 )   Save BACKGROUND: Currently, the widely used intervertebral disc degeneration models are induced by altering intervertebral disc biomechanics, damaging intervertebral disc structure or changing hereditary features with genetic technique. All these methods are varies from natural duration of intervertebral disc degeneration. OBJECTIVE: To evaluate the feasibility of the establishment of rabbit model of ischemic lumbar vertebrae adjacent to endplate by percutaneous puncture followed by pingyangmycin injection. METHODS: A total of 46 New Zealand white rabbits were selected and two vertebraes were divided as experimental group (L5) and control group (L4) in every rabbit. Vertebrae adjacent to endplate was punctured. Pingyangmycin (2 g/L) 1 mL was injected into rabbits in the experimental group. And 1 mL normal sodium was injected into the control group. Lumbar artery angiography was performed in 4 rabbits before operation. Six rabbits were randomly performed MRI and then were executed for vertebral histology at weeks 1, 2, 3, 4, 5 and months 2, 3 after operation. Ischemic areas of L5 were measured by the MRI and histological section at week 4 after operation. RESULTS AND CONCLUSION: MRI and histology of control group had not specific changes. MRI had not significant signal intensity changes in the first 2 weeks in the experimental group. At week 3 after operation, it demonstrated slightly hyperintense signal on T2-weighted image (T2WI) and fat-suppression T2-weighted image (FS T2WI), while fat-suppression T1-weighted image (FS T1WI) was hypointense signal. The signal changed more obviously at week 4. Histology of experimental group had not specific changes in the first 2 weeks. From weeks 3-4, bone trabecula arranged confusedly and disorderly, with gradually decreased osteocyte and marrow haemocytes, while adipocytes increased and coalesced. Cartilage corpuscle of endplate decreased and architecture became disorder. But the anulus fibrosus and nucleus pulposus had no obviously changes. The intervertebral disk of the experimental group degenerated at week 5, and the ischemia of lumbar vertebrae still existed and intervertebral disk degenerated more obviously at months 2-3 after operation. There was significant positive correlation of ischemic areas of experimental group between MRI and histology at week 4 (r=0.965, P < 0.001). The rabbit model of ischemic lumbar vertebrae adjacent to endplate can be established successfully by percutaneous puncture vertebrae adjacent to endplate followed by pingyangmycin injection. The operation is minimally invasive, simple and reproducible, with high success rate. This is a fairly ideal animal model to study the degeneration of the lumbar spine and intervertebral disc. Related Articles | Metrics

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Dual-dissociation method to record whole-cell current of wistar rat neurons  Shen Pei-tong, Bi Ping, Li Gang 2010, 14 (2):  285-288.  doi: 10.3969/j.issn.1673-8225.2010.02.022 Abstract ( 307 )   PDF (478KB) ( 482 )   Save BACKGROUND: The main difficulties of whole-cell patch clamp are cell culture and whole-cell current recording. OBJECTIVE: To reduce the difficulty of patch-clamp experiment by combining acute cell isolation with current separation technique. METHODS: A total of 40 wistar rats aged 4-7 days, irrespective of genders, were selected. The pallium of wistar rats were cut into slices with 400-600 μm, rested for 1 hour in artificial cerebrospinal fluid with gas mixture, simultaneously put in oxygen. The brain tissues were placed into artificial cerebrospinal fluid containing 16 u/mL( type X ) and 2 u/mL(type XIV) proteinase, incubated for 60 minutes, and cleared the digestive enzyme. Under the whole-cell voltage clamp mode, the potential was hold at -80mV, depolarizing pulse stimulation from -60 mV to 60 mV, 10 mV step and 160 ms width. The total Tran membranes current was recorded, and then 70 mmol/LCsCl, 70mmol/L CsF was 11 μmol/L Na+ channel blockers tetrodotoxin, 30 mmol/L tetraethylammonium, 1 mmol/L 4-AP was successively added into extracellular fluid. The inward sodium current, transient outward potassium current and delayed rectifier potassium current was recorded, and then the result was analyzed using clampfit.  MAIN OUTCOME MEASURES: ①Cell morphological observation; ②Whole-cell current recording; ③Inward sodium current recording; ④Outward potassium current recording. RESULTS AND CONCLUSION: The cell had clearly three-dimensional structure, smooth surface and whole neurodendrite or neuraxon. The cell viability could maintained for 8-10 hours at 25 ℃ temperature. The added 11 tetrodotoxin in extracellular fluid could block sodium current. The outward potassium current could be blocked by tetraethylammonium with 30 mmol/L and 4-AP with 1 mmol/L. The cells harvested by modified rapid dissociation have good functions. Using current separation method, only specific blockers are needed, without extracellular fluid or electrode solution replacing, which can record the inward sodium current, transient outward potassium current, as well as delayed rectifier potassium current. This method can obviously improve the work efficiency than traditional one. Related Articles | Metrics

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Construction and identification of Bcl-x minigene and its mutated minigene Huang Bo, Wang Xiao-zhong, Wang Cai-wen, Li Jing, Zhang Hai-bin, Xiong Huo-mei, Xiao Yun 2010, 14 (2):  289-293.  doi: 10.3969/j.issn.1673-8225.2010.02.023 Abstract ( 305 )   PDF (1292KB) ( 365 )   Save BACKGROUND: It is not elucidated about its mechanism of the rising ratio of Bcl-xL/Bcl-xS linked closely with the development of tumors. The Bcl-x minigene is an important tool to study its mechanism of alternative splicing. OBJECTIVE: To construct the human apoptosis-related Bcl-x minigene and its mutated minigene, and to provide basis for studying the alternative splicing of Bcl-x gene. METHODS: At First, the fragment (P1P2) including exon 2 and part of 5’ sequence of intron 2 in bcl-x gene were amplified by PCR from human leukemia cell K562 genomic DNA, then directionally inserted into the eukaryotic expression vector pcDNA3.1(-), called pcDNA3.1(-)-p1p2; secondly, the fragment (P3P4) including 3’ sequence of intron 2 and part of exon 3 in bcl-x gene were amplified, then directionally inserted into the downstream of fragment p1p2, called pcDNA3.1(-)-p1p2-p3p4, it is pcDNA3.1(-)-bcl-x minigene. By sequencing and identifying, pcDNA3.1(-)-bcl-x minigene was introduced into HL-60 by transient transfection, so as to identify its expression by RT-PCR. In addition, pcDNA3.1(-)-bcl-x minigene was utilized as a template to construct its mutated minigene called pcDNA3.1(-)-bcl-x-CRCE1 (M) by inverse PCR. RESULTS AND CONCLUSION: Taking human leukemia cell K562 genome DNA as the template, P1 and P2 as primers, approximately 686-bp target fragment were obtained by PCR amplification; taking P3 and P4 as primers, approximately 178-bp target fragment could be obtained. Through XbaⅠ and Eco RⅠdouble enzyme digestion, Xba Ⅰ and Xho Ⅰ double enzyme digestion, the anticipated fragment could be obtained from minigene pcDNA3.1(-)-bcl-x, also from mutated minigene pcDNA3.1(-)-bcl-x-CRCE1(M) through Eco RI single enzyme digestion, the sequence result was correct. Results showed that we successfully construct the minigene and mutated minigene of human apoptotic-related gene bcl-x.  Related Articles | Metrics

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Autologous nucleus pulposus induces rat dorsal root ganglion damage without mechanical compression  Lu Zhi-dong, Chen Zhi-rong 2010, 14 (2):  294-297.  doi: 10.3969/j.issn.1673-8225.2010.02.024 Abstract ( 286 )   PDF (1074KB) ( 428 )   Save BACKGROUND: Resent studies demonstrate that sciatica is caused by inflammatory reaction of nucleus pulposus, and dorsal root ganglion (DRG) is in involved in this process; however, its pathophysiological changes remain poorly understood. OBJECTIVE: To explore pathogenesis of sciatica related to nucleus pulposus protrusion without mechanical compression. METHODS: Totally 24 male SD rats were randomly divided into the experimental group and control group, with 12 rats in each group. The coccygeal intervertebral discs of rats were opened to obtain 5 coccygeal nucleus pulposus, mixed with 50 μL physiological saline to prepare suspension. Rats in the experimental group were prepared non-compressive models by injecting suspension to the epidural cavum of rats. Meantime, the same volumes of physiological saline were injected into rats in the control group. The rat hind paw mechanical withdrawal thresholds were measured, pain-related behavior and dorsal root ganglion morphology were observed. RESULTS AND CONCLUSION: In the absence of mechanical pressure, disappearance of the nucleolus, degeneration of the nuclear membrane and significant hyperalgesia derived by auto-transplantation of nucleus pulposus to the epidural cavum of rats were observed. Dorsal root ganglion cells often had hyperemia, edema and intracytoplasmic vacuoles, with large and irregular nuclei. Prominent Nissl bodies were not seen. The Inflammatory reaction derived by autograft nucleus pulposus is an important factor in DRG injury and sciatica. Related Articles | Metrics

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Expression of interleukin 1 beta in the lateral meniscus of rabbits with anterior cruciate ligament rupture Li Guo-jun, Li Kang-hua, Zhang Shi-qing, Wang Xiao 2010, 14 (2):  298-301.  doi: 10.3969/j.issn.1673-8225.2010.02.025 Abstract ( 291 )   PDF (1432KB) ( 460 )   Save BACKGROUND: Previous studies demonstrate that interleukin 1β (IL-1β) plays a great role in the degeneration of articular cartilage in knee joints which was caused by the anterior cruciate lagement (ACL) rupture; however, the role of IL-1β in the degeneration of lateral meniscus remains poorly understood. OBJECTIVE: To investigate the changes and significance of the expression of IL-1β in the lateral meniscus of rabbits with ACL rupture. METHODS: A total of 48 male, New Zealand, white rabbits, were matched mode pairs and prepared for ACL rupture models. In the experimental side, the ateral meniscus of rabbits was cut down to prepare ACL rupture models, simultaneously, the control sides were exposed without cut down. A total of 12 rabbits were sacrificed under anesthesia at weeks 1, 3, 6 and 8 after model preparation. Degeneration scores of lateral meniscus and expression of IL-1β in lateral meniscus were performed. RESULTS AND CONCLUSION: With time prolonged, the scores of lateral meniscus degeneration were increased, which was higher in the experimental group than those in the control group at each time points (P < 0.05). The positive rate of IL-1β was greater in experimental group than that in the control group (P < 0.05). In addition, the positive rate of IL-1β in the experimental group was greater at weeks 3 and 6 than weeks 1 and 8 (P < 0.05), which was higher in week 8 than week 1 (P < 0.05). The results demonstrated that the highly expressed IL-1β in the experimental sides hints that IL-1β maybe a promotional factor which causes the degeneration of lateral meniscus. The decreasing of the expression of IL-1β is not a stopping signs of lateral meniscus degeneration. Related Articles | Metrics

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Transfection of mouse L-M (TK-) cells with Wnt3a and its effect on the subcellular distribution of beta-catenin Shang Yan-chang, Wang Shu-hui, Zhang Cheng, Xiong Fu, Li Yong, Liu Zheng-shan, Xu Yong-feng 2010, 14 (2):  302-305.  doi: 10.3969/j.issn.1673-8225.2010.02.026 Abstract ( 296 )   PDF (1190KB) ( 407 )   Save BACKGROUND: Wnt signaling pathway plays an important regulative role in the embryonic development processes. Accordingly, it is of great significance to establish the cell model of Wnt signaling pathway so as to conduct study on it. OBJECTIVE: To establish Wnt signaling pathway cell model by transfecting L-M (TK-) cells with Wnt3a eukaryotic expression plasmid, and to investigate the effect of canonical Wnt signal pathway on the β-catenin subcellular distribution. METHODS: The eukaryotic expression plasmid pgk-Wnt3a-pcDNA3.0 after amplification was digested by restriction endonuclease first. Then it was transfected together with the control plasmid pgk-neo-pcDNA3.0 into L-M (TK-) cells via lipofection, after which the cell colony was screened by G418 for amplification. RT-PCR was used for detecting the expression products and the indirect immunofluorescence assay for observing the effect of Wnt3a on the β-catenin subcellular localization of L-M (TK-) cells. RESULTS AND CONCLUSION: The Wnt3a plasmid was verified by endonuclease digestion to have produced the expected plasmids after amplification. According to the RT-PCR detection to the 10 stably-transfected cell colonies achieved by 3 weeks of G418 screening, it was seen, on the L-Wnt3a cDNA, a strip of bright band of 320 bp in length, which showed that the products of amplification were exactly the expected fragments and that the Wnt3a plasmid was expressed on mRNA transcriptional level after being transfected with L-M (TK-) cells. In contrast, no expected band was found on the cDNA of L-M (TK-) cells transfecting the control plasmid. In addition, the immunofluorescence assay detection showed that the protein expression of Wnt3a was found in the cytoplasm of the L-M(TK-) cells tranfecting Wnt3a plasmid, while for those transfecting the control plasmid, it was opposite. β-catenin, as showing by bright red fluorescence, was found to concentrate and enter into the nucleus of the L-M (TK-) cells transfecting Wnt3a plasmid, while for those transfecting the control plasmid, it was opposite. Cell model with continually activated Wnt signaling pathway is established. The stable expression of Wnt3a in L-M (TK-) cells transfected with pgk-Wnt3a-pcDNA3.0 is obtained. The expression of Wnt3a is able to promote the transfer of β-catenin from cytoplasms into nucleus in L-M (TK-) cells.  Related Articles | Metrics

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Clinical application and progress of tissue-engineered trachea  Bi Guang-liang, Ma An-de 2010, 14 (2):  309-312.  doi: 10.3969/j.issn.1673-8225.2010.02.028 Abstract ( 266 )   PDF (625KB) ( 489 )   Save BACKGROUND: Tissue-engineered trachea serving as a tracheal transplant, can not only reduce the expression of antigen cells, suppress immunoreaction, but also in advance conduct vascularization of blood vessels and regeneration culture of tissue cells in vitro. OBJECTIVE: To summarize the commonly used methods for tracheal transplantation, to introduce the latest clinical transplantation method of tissue-engineered trachea, and to provide reference evidence for carrying out such operations. METHODS: Taking tissue engineering, trachea, transplantation in English as search terms, Pubmed database from January 1990 to May 2009, ScienceDirect database from January 1990 to May 2009, Springer database from January 1990 to May 2009 was retrieved; Taking tissue engineering, trachea, transplantation in Chinese as search terms, CNKI database from January 1990 to May 2009 was researched. Literatures were limited to English and Chinese languages. Inclusive criteria: clinical studies and animal experiments related to the tracheal transplantation; exclusive criteria: duplicate documents. RESULTS AND CONCLUSION: Totally 270 literatures were screened out by computer, according to inclusion and exclusion criteria, 32 documents of which were involved for analysis. There is an increasing cases with irreversible injury of tracheal resulted from various reasons and in need of transplantation during clinical work, and the material source of tracheal transplantation is always the main problem to limited the clinical use. Constructing tissue-engineered trachea in vitro is considered to be a good way for repairing trachea. This paper will review the origin and development of tracheal transplantation and tissue-engineered trachea, summarize the common method of allotransplantation, autotransplantation and biomaterial transplantation, as well as the research of constructing bio-scaffold, choosing seed cells and clinical applied development of tissue-engineered trachea, and emphatically introduce the first case of successful artificial tracheal transplantation by combining the method of allotransplantation and culturing the seed cell without the immune cells of donor.  Related Articles | Metrics

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Research progress in the treatment of avascular necrosis of the femoral head with bone marrow mesenchymal stem cells transplantation   Qiu Wei, Meng Zeng-dong 2010, 14 (2):  313-316.  doi: 10.3969/j.issn.1673-8225.2010.02.029 Abstract ( 324 )   PDF (551KB) ( 516 )   Save BACKGROUND: Avascular necrosis of the femoral head occurs in the hip joints at high load-bearing areas, it is a special form of aseptic bone necrosis. Bone marrow-derived mesenchymal stem cells may have an excellent repair effect for the necrosis of femoral head. OBJECTIVE: To understand bone marrow mesenchymal stem cell transplantation for treatment of early avascular necrosis of the femoral head, and to lay a foundation for further study. METHODS: Taking avascular necrosis of femur head, femur head necrosis, osteonecrosis, bone marrow mesenchymal stem cells, mesenchymal stem cells, multipotent stem cells in English as search terms, Pubmed database from January 2000 to April 2009 was retrieved; Taking avascular necrosis and bone marrow mesenchymal stem cells in Chinese as search terms, CNKI database from January 1989 to April 2009 was searched. Literatures were limited to English and Chinese languages. Inclusive criteria: Study of the biological characteristics and functions of bone marrow mesenchymal stem cells, the etiology and pathology of avascular necrosis of femoral head, bone marrow mesenchymal stem cell transplantation for treatment of avascular necrosis; Exclusive criteria: duplication of documents. RESULTS AND CONCLUSION: More than 800 literatures were screened out by computer search, according to inclusive and exclusive criteria, 38 documents of which were involved for analysis. Marrow mesenchymal stem cells can differentiate into chondrocytes, osteocytes and dipocytes under suitable cultural condition. With the development of stem cell engineering, marrow mesenchymal stem cell transplantation becomes an effective method for repairing osteonecrosis. This article describes the cause and the pathology of avascular necrosis of the femoral head, analyzes the theory basis for the treatment of avascular necrosis of the femoral head with marrow mesenchymal stem cell, and verifies its effect through comparing both animal and clinical experiments of this aspect at home and abroad, with the hope of seeking a better treatment. Related Articles | Metrics

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Feasibility of predicting fracture risk with bone turnover markers and bone mineral density Lin Jian-ping, Song Shi-feng, Yao Lun-long 2010, 14 (2):  317-320.  doi: 10.3969/j.issn.1673-8225.2010.02.030 Abstract ( 269 )   PDF (644KB) ( 661 )   Save BACKGROUND: Although bone mineral density can diagnose osteoporosis, it cannot reflect the existing bone metabolism of the subjects; bone turnover markers cannot diagnose osteoporosis, but it can timely reflect the current bone turnover rate of the subjects. OBJECTIVE: To summarize osteoporotic fracture risk predicted by bone turnover markers and bone mineral density, and to theoretically analyze the feasibility of fracture healing after operation. METHODS: With bone turnover, biochemical marker, osteoporosis in English for the search terms, Pubmed database from January 1999 to January 2009 was retrieved; With bone turnover, bone metabolism, biochemical markers, bone mineral density, osteoporosis, fractures in Chinese for the search terms, CNKI database from January 1999 to January 2009 was searched. Literatures were limited to English and Chinese languages. Inclusive criteria: the article closely related to predict fracture risk with bone turnover markers and bone mineral density; Exclusive criteria: duplicated documents. RESULTS AND CONCLUSION: Totally 631 literatures were screened out by computer, according to inclusion and exclusion criteria, 31 documents of which were involved for analysis. Osteoporosis is one of the main causes of elderly disease and death. Early prediction of osteoporotic fracture risk has a great significance. This article describes the bone metabolism, the type of bone biochemical markers, bone mineral density measurement, multi-cell basic unit and the OPG-RANKL-RANK system in bone reconstruction after a long-term fixation; focuses on the use of bone turnover markers and bone mineral density to prognosticate the fracture risk in osteoporosis or after long-term fixation by domestic and foreign academician, and theoretically analyze the feasibility of predicting postoperative fracture healing. Related Articles | Metrics

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Research methods of gait kinematics and kinetics   Song Ya-wei, Sun Wen, Kou Heng-jing 2010, 14 (2):  321-324.  doi: 10.3969/j.issn.1673-8225.2010.02.031 Abstract ( 447 )   PDF (627KB) ( 1375 )   Save BACKGROUND: Kinematics studies the movement of the objects using a method of geometry, with strength and quality not taken into consideration; Kinetics major studies the force of people acting on objects and its relationship with the movement of objects. OBJECTIVE: Through an introduction to kinematics and kinetics research equipment, this study was aimed to lead to the current status of kinematics and kinetics, to provide for experimental basis for the research of new equipment and gait experiments. METHODS: Taking “gait, kinetics, kinematics” in English for the search terms, Pubmed database (1999-01/2009-06) was retrieved. Taking “gait, kinematics, kinetics” in Chinese for the search terms, CNKI database (1999-01/2009-06) was searched. Literatures were limited to English and Chinese languages. The kinematics and kinetics research equipment regarding the gait were included, while other kinematics and kinetics experiments were excluded, also outdated and repetitive researches were excluded. RESULTS AND CONCLUSION: Totally 2 367 literature were obtained from the computer screen, and 33 documents of them were involved for analysis according to the inclusion and exclusion criteria. In case of gait analysis method, at home and abroad scholars have carried out extensive studies although the content are quite different, they used identical instruments and methods in the testing of gait, in spite of photography or video recording were applied to calculate and measure kinematic parameters, the force platform was all used to record kinetic parameters. Also there are a small number of studies using the distribution of plantar pressure to measure more precise foot-ground pressure distribution, and then proceed to a comprehensive statistical analysis. Kinetics and kinematics gait analysis is applied not only in rehabilitation medicine, but also widely in athletics sports and the related footwear manufactures, the commonly used method is the plane fixed-point photograph and force platform analysis. Related Articles | Metrics

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Trainability and recontruction of ageing’s muscle function   Qu Hui-fang, Zhang Wen-zhong, Yan Wan-jun 2010, 14 (2):  325-328.  doi: 10.3969/j.issn.1673-8225.2010.02.032 Abstract ( 420 )   PDF (545KB) ( 583 )   Save BACKGROUND: In many parts of the body, muscle diastolic and contractive activities can promote the body's energy metabolism. Furthermore, the muscles also play an irreplaceable role in maintaining body posture and joint stability. OBJECTIVE: To summarize the rules of muscle function and aging changes, to search for research related to the trainability of old muscle function, and to provide references for muscular anti-aging research in field of sports medicine. METHODS: Taking senescence, aging, muscle function, exercise, training in English as the search terms, Pubmed database from January 2000 to May 2009 was retrieved; Taking aging, muscle function, training in Chinese as the search terms, CNKI database from January 2000 to May 2009 was searched. Literatures were limited to English and Chinese languages. Inclusive criteria: research closely related to the trainability of motor function and muscle metabolic function in the elderly; Exclusive criteria: obsolete and lack of credibility of the literature. RESULTS AND CONCLUSION: Totally 67 literatures were screened out by computer, according to inclusion and exclusion criteria, 31 documents of which were involved in the analysis. Based on the recent research on muscle function, the muscle function can be summarized into three aspects: motor function, metabolic function and maintenance function. The elderly muscle aging is mainly showed in these 3 aspects. How to reconstruct these functions through resistance training? The concerning data were summarized in this paper, to verify the trainability of ageing’s muscle motor and metabolism functions, but no study indicates that the maintenance function can be reconstructed, and deserves further studies of sports science.                              Related Articles | Metrics

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Theory study and medical application of real-time quantitative polymerase chain reaction Liu Xiao-rong, Zhang Li, Wang Yong-ping 2010, 14 (2):  329-332.  doi: 10.3969/j.issn.1673-8225.2010.02.033 Abstract ( 538 )   PDF (649KB) ( 1005 )   Save BACKGROUND: Real-time fluorescence quantitative polymerase chain reaction (PCR) refers to join the fluorescence groups into PCR reacting system, and to real-time monitor entire PCR process using the fluorescence signal accumulation, finally to make the quantitative analysis of the unknown template through the standard curve. OBJECTIVE: To study the theory of real-time fluorescence quantitative PCR and to explore its applications and progress in medicine. METHODS: With “real-time fluorescence quota PCR, theorem, application” in English for the search term, PubMed database was retrieved from January 2000 to December 2008. With “real-time fluorescence quantitative PCR, principle, application” in Chinese for the search term, Wanfang Database from January 2000 to December 2008, Tsinghua Tongfang Chinese database from January 2000 to December 2008 were was retrieved. Literatures were limited to English and Chinese languages. Cell factor and tumor resistance genes served as the evaluation index. The methodology of research on the real-time fluorescence quantitative PCR technology and medical applied research on real-time fluorescence quantitative PCR technology were included. While repetitive research and Meta analysis were excluded. RESULTS AND CONCLUSION: Because real-time fluorescence quantitative PCR technology has not only realized PCR develops from qualitative to quantitative levels, but also has strong specificity, high sensitivity, good duplication, accurate quantization, high automaticity, and entire blocking response compared with conventional PCR, thus it becomes the important tool in the molecular biology research. Real-time fluorescence quantitative PCR technique has been widely applied, such as mRNA expression, detections of DNA copy number and determination of mononucleotide polymorphism, as well as in the clinical medicine including accurate quantitative examination of mycobacterium tuberculosis, Type B and Type C hepatitis, AIDS virus, gonococcus, and chlamydia trachomatis. Its quantitative scope extremely extends, no need of gradient dilution, the specificity is stronger, overcomes the false positivity. Due to the traditional PCR technology cannot give the accurate quantization, it is greatly limited in the practical application. Therefore, the accurate quantization of the PCR product, particularly the dynamic monitoring of viral etiology, becomes the urgent need. Related Articles | Metrics

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Hypothesis of textural composite hydrate system IV: Study on hydration composite ways  Piao Dong-xu, Mao Li-jiang, Hu Yuan-jie, Chen Xiao-dong, Zhang Ying, Sun Rui-huan 2010, 14 (2):  333-336.  doi: 10.3969/j.issn.1673-8225.2010.02.034 Abstract ( 343 )   PDF (842KB) ( 457 )   Save BACKGROUND: The water molecules and biological macromolecules in human tissue may combine to form hydrate composite. Different types of macromolecules show different approaches to combine with water molecules. OBJECTIVE: To discuss the basic form of water molecules combined with biological macromolecules. METHODS: The theoretical hypothesis of texture composite hydration system was proposed. The relevant literatures were retrieved through the computer to discuss these issues. RESULTS AND CONCLUSION: According to the role of water in the hydrate composite, the composite ways can be divided into the following 4 groups. ①Surface composite: Hydrated membrane layers can form on the surface of globular protein or DNA molecules. The hydrated membrane layer shows that the speed of water molecular and hydrate density were reduced gradually from surface to inside. The hydrated layer makes the shape and physical state of the macromolecules stabilized, and it plays important role in forming of TCHS. ②Swelling composite: As a result of the strong hydration of glycosaminoglycans, the hydrated proteoglycan often swells and shows high viscosity and hydrogel character. The hydrated proteoglycan play some mechanical roles in connective tissue. For example, it can make the cartilage have a strong deformation-resistibility. ③Hydrophobic composite: Hydrophobic effect drives structural rearrangement, including the formation of bilayers (Bio-membrane). This hydrophilic-hydrophobic-hydrophilic “sandwich” structure is conducive to the self-stability in mechanics, combination with membrane protein and transmembrane transport. ④Slit composite: A lot of slit spaces with nano scale were existing inside and outside of the cells. Aqueous solution in the slit spaces shows some abnormal characters such as higher viscosity and lower ice point, because of the rearrangement of water molecules. Slit composite and surface composite have some universality, and it can be regarded the former as a special case of the latter. The classification way mentioned above has not any absolute connotations. However, the rational utilization of these hydration composite ways will be conducive to research the theory on textural composite hydrate system.    Related Articles | Metrics

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Biological property and clinical application of tissue engineered skin Tian Bao-xiang, Fan Hua, Liu Feng-bin, Wei Chun-lin 2010, 14 (2):  337-340.  doi: 10.3969/j.issn.1673-8225.2010.02.035 Abstract ( 357 )   PDF (511KB) ( 603 )   Save BACKGROUND:Tissue engineered skin has been used to repair defect skin, but it is not as good as to completely replace permanent skin. OBJECTIVE: To explore the biological characteristics and clinical application of various types of tissue engineered skin. METHODS: Pubmed data (1990-01/2005-12) was retrieved by computer with the key words of “tissue-engineered skin”. CNKI database (2002-01/2007-12) was retrieved with the same key words. RESULTS AND CONCLUSION: Totally 226 literatures were primarily selected. After reading titles and abstracts, studies addressing irrelative objective or repetitive content were excluded, and 11 literatures were included. The aim of skin tissue engineering is to create skin substitute that close to human skin in histology and functional characteristics. There were many mature tissue engineered skin in clinic, and various artificial skins were developed. Their structure was similar to human, but only had skin barrier function and lack of skin appendages, so they did not have complete skin function, and did not reach skin reestablishment. Ideal skin substitute should process cuticular layer and dermial layer. The dermial layer can reach rapid vascularization and nerve redistribution, can promote physiological wound repair. The cuticular layer can rapidly obtain full barrier and protection function, which can be fully integration with wound surface. A good dermial carrier has good histocompatibility, and a certain physical and chemical properties, such as strength, scratchability, elasticity, gas permeation, water vapor permeability. These can provide a good adherence between dermis and wound surface, offer an ideal fiber stent for cell growth, and finally promote autologous skin growth. Tissue engineered skin has good prospect, but there are many problems that should be solved in biocompatibility and skin function reestablishment.   References | Related Articles | Metrics

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Theoretical hypothesis and clinical practice of Bobath concept: A therapeutical basis for nervous tissues repairing and functional reconstruction Zhao Jian-le, Huang Xia 2010, 14 (2):  341-346.  doi: 10.3969/j.issn.1673-8225.2010.02.036 Abstract ( 422 )   PDF (626KB) ( 735 )   Save OBJECTIVE: A series of Bobath concept, Bobath theoretical hypothesis and clinical practice were summarized, to provide a better understanding of Bobath concept involved in nervous tissues repairing and functional reconstruction. METHODS: Articles concerning the Bobath treatment for stroke-related literature were retrieved in Highwire press, MEDLINE, and Google from 2001 to 2009, using the key words of “Bobath concept, stroke physical therapy”. Data of each study were extracted and analyzed following retrieval. Inclusion criteria: ①Bobath concept and its theoretical hypothesis. ②New development of Bobath concept in clinical practice. Exclusion criteria: Repetitive studies. The results of retrieved literature were analyzed. RESULTS: The theoretical basis of Bobath concept is based on present-day knowledge of motor control, motor learning, neural and muscle plasticity, and biomechanics. The latest theories and viewpoints are resourced from International Bobath Instructors Training Association and the British Bobath Tutors Association. They expounded on the Bobath concept from linking participation, activities and underlying impairments, organization of human behavior and motor control, the consequences of injury and dysfunction in the execution of movement, neural and muscle plasticity, motor learning and so. This article also discusses several important questions which meet in clinical practice. CONCLUSION: Bobath therapy, as a traditional method for stroke rehabilitation, is constantly absorbing the research results from current neuroscience, psychology and other disciplines, and blended with other treatments to promote the development of neuro-rehabilitation medicine. Related Articles | Metrics

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Correlation of peripheral blood T lymphocytes with the type of lumbar disc herniation and straight leg raising test Tian Peng, Ma Xin-long, Wang Tao, Ma Jian-xiong 2010, 14 (2):  347-350.  doi: 10.3969/j.issn.1673-8225.2010.02.037 Abstract ( 326 )   PDF (602KB) ( 355 )   Save OBJECTIVE: To observe peripheral T lymphocytes changes of patients with lumbar disc herniation, and investigate the relationship between the type of herniation, signs and T lymphocyte subsets. METHODS: Blood samples from 49 patients with single-level lumbar intervertebral disc herniation were collected from General Hospital of Tianjin Medical University from January to October 2008, including 26 males and 23 females. There were 22 cases of L4-5 and 27 cases of L5S1. In the straight leg raising (SLR) test before treatment, 34 cases were positive and 15 were negative. According to the position of intervertebral disc observed in operation, the patients were divided into ruptured disc herniation (RDH, n=30) and degenerative disc herniation (DDH, n=19) groups. In addition, blood samples from 20 healthy blood donors were collected as control group. T lymphocytes subsets were detected by flow cytometry. RESULTS: The levels of CD3+ , CD4+ , and CD4+ /CD8+ in RDH group were significantly higher than control group (P < 0.05), and the levels of CD8+ in RDH group were significantly lower than control group (P < 0.05). The levels of CD4+ and CD4+ /CD8+ in positive SLR test group were significantly higher than patients with negative results, but significantly lower levels of CD8+ was found in positive group than negative group (P < 0.05). The positive rate of SLR test in RDH group was significantly higher than that of SLR testing in DDH group. CONCLUSION: Changes of T lymphocyte subsets in peripheral blood were found after herniation of lumbar intervertebral disc. T lymphocytes mediated immune response may play an important role in the occurrence and development of signs in patients with herniated lumbar intervertebral disc. The SLR test may help to confirm that the disc herniation may cause nerve root impairment by mechanical loading or inflammatory stimulus. Related Articles | Metrics

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Changes of serum osteoprotegerin and bone mineral density in diabetic nephropathy patients Huang De-fang, Chen Gao-xiang, Zhang Song, Zhang Feng, Cao Hong 2010, 14 (2):  351-354.  doi: 10.3969/j.issn.1673-8225.2010.02.038 Abstract ( 287 )   PDF (540KB) ( 363 )   Save BACKGROUND: At present, the relationship between diabetic nephropathy and osteoprotegerin remains poorly understood. OBJECTIVE: To investigate the correlation between bone mineral density (BMD) and serum osteoprotegerin changes in patients with 2 type diabetic nephropathy. METHODS: Totally 104 patients with 2 type diabetes were divided following 5 groups according to glomerular filtration rate: simple diabetes, mild renal injury, moderate renal injury, severe renal injury, and renal failure groups. Additional 20 healthy people were selected as the control group. The level of serum osteoprotegerin was measured by ELISA. Meantime, levels of serum calcium, phosphonium, alkaline phosphatase, creatinine, urea nitrogen, and glycosylated hemoglobin were measured by the automatic biochemistry analyzer. The bone mineral density of entopic L 2-4 was determined by dual X-ray bone density equipment. The whole data was analyzed by multiple regression correlation analysis. RESULTS AND CONCLUSION: The level of serum osteoprotegerin in patients with diabetic nephropathy was obviously greater than that of the healthy people (P < 0.05), but BMD of the mild renal injury, moderate renal injury, severe renal injury, and renal failure groups was obviously lower than that of the healthy people (P < 0.05). Generally, the worse renal function accompanied by higher osteoprotegerin level, and lower BMD. There was a negative correlation between osteoprotegerin level and BMD in patients with diabetic nephropathy (r=-0.497, P < 0.01). However, the relationships between osteoprotegerin level and diabetic duration (r=0.566, P < 0.01), serum creatinine level (r=0.772, P < 0.01), serum urea nitrogen level (r=0.708, P < 0.01), serum phosphonium level (r=0.329, P < 0.01), or serum intact parathyroid hormone level (r=0.702, P < 0.01) were positive. Meantime, the serum phosphonium level had negative correlation to serum calcium level (r=-0.505, P < 0.01). It demonstrated that when the renal function got worse in diabetic nephropathy patients, the serum osteoprotegerin level was increasing accompanied by BMD decreasing. The osteoprotegerin level presents a negative correlation to BMD and serum calcium level, but positive to diabetic duration, serum creatinine, serum urea nitrogen, serum phosphonium and serum intact parathyroid hormone levels.  Related Articles | Metrics

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Comparative study on the physical fitness changing characteristics of adult males in Shanghai, Beijing and Guangdong Hu Shi-xing 2010, 14 (2):  355-359.  doi: 10.3969/j.issn.1673-8225.2010.02.039 Abstract ( 313 )   PDF (442KB) ( 430 )   Save OBJECTIVE: To compare and analyze the index characteristics relevant to the body shapes, physical functions and physical qualities of adult males in Shanghai, Beijing and Guangdong, based on the statistics of the second National Physical Fitness Monitoring in these three regions. METHODS: The physical fitness indexes were looked up from the “National Physical Fitness Monitoring Bulletin” issued by the National Physical Education Office and Physical Education Offices in Shanghai, Beijing and Guangdong. Then these indexes were divided by age groups of 20-24, 25-29, 30-34, 35-39, 40-44, 45-49, 50-54 and 55-59 years, to receive statistical processing and comparative analysis through Microsoft Excel. RESULTS: Body shape: ①The adult males in both Shanghai and Beijing were taller than those in Guangdong. ②The weight, Quetelet index, chest circumference and waist line indexes followed by ascending were Guangdong, Shanghai and Beijing. ③The body mass indexes in all age groups followed by ascending were Guangdong, Shanghai and Beijing. Physical functions: ①The diastolic pressure and vital capacity indexes followed by ascending were Beijing, Shanghai and Guangdong. ②For the adult males less than 30-34 ages, the highest step-test index was in Guangdong, and Shanghai came the second. Physical quality: ①The index of antexion with sitting position in all age groups in Shanghai were higher than in Beijing and Guangdong. ②The choice reaction time indexes of adult males in all age groups in Shanghai were the highest among the three regions. CONCLUSION: ①The statures of males in Shanghai, Beijing and Guangdong have been rising at a quick speed and will last a long time, and the rising speed in Guangzhou is relatively slow. ②The overweight phenomenon of adult males in Beijing and Shanghai tends to begin at a younger age, and the overweight range tends to increase with age. ③The physique indexes of adult males in Shanghai are better than those in Beijing and Guangdong, except the overweight index. Related Articles | Metrics

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Correlation between body composition and the components of metabolic syndrome in postmenopausal women  Jin Jing, Ruan Xiang-yan, Hua Lin 2010, 14 (2):  360-364.  doi: 10.3969/j.issn.1673-8225.2010.02.040 Abstract ( 308 )   PDF (711KB) ( 419 )   Save OBJECTIVE: To learn the prevalence of abnormal components of metabolic syndrome in postmenopausal women, and to explore the optimal body composition indices to predict the abnormal components of metabolic syndrome in postmenopausal women. METHODS: A total of 181 participants were recruited from postmenopausal women who came to Beijing Obstetrics and Gynecology Hospital for health examination between July 2006 and August 2007. They were 41-60 years of age, and there were 1-9 years since menopause. Their clinical and biochemical parameters were measured, including body height, body weight, waist circumference, hip circumference, sagittal abdominal diameter, blood pressure, follicle stimulating hormone, luteinizing hormone, estradiol, triglyceride, high density lipoprotein-cholesterol, fasting plasma glucose, fasting insulin and body fat. The prevalence of the abnormal components of metabolic syndrome in postmenopausal women was investigated. The correlations between body composition indices and the components of metabolic syndrome were analyzed by partial correlation analysis independent of age, years since postmenopause and the level of estradiol. The body composition indices to predict the abnormal components of metabolic syndrome in postmenopausal women were explored. RESULTS: ①The prevalence of central obesity (waist circumference ≥ 80 cm), hypertriglyceridemia (triglyceride ≥    1.7 mmol/L), low high density lipoprotein-cholesterol (high density lipoprotein-cholesterol < 1.29 mmol/L), hypertension (systolic pressure ≥ 130 mm Hg or diastolic pressure ≥ 85 mm Hg, 1 mm Hg=0.133 kPa) and high fasting glucose (fasting plasma glucose ≥ 5.6 mmol/L) was 48.1%, 31.5%, 16.6%, 38.7% and 50.3%, respectively. ②Through partial correlation analysis, body composition indices (body mass index, waist circumference, waist-hip ratio, sagittal abdominal diameter, and body fat) had a positive correlation with triglyceride, systolic pressure, diastolic pressure and fasting plasma glucose, while a negative correlation with high density lipoprotein-cholesterol, independent of age, years since postmenopause and the level of estradiol. ③Body composition indices and critical value of them used to predict the abnormal components of metabolic syndrome in postmenopausal women were as follows: hypertriglyceridemia: body mass index ≥ 24.845 kg/m2; low high density lipoprotein-cholesterol: sagittal abdominal diameter ≥ 18.85 cm; hypertension: body mass index ≥ 23.33 kg/m2; high fasting glucose: sagittal abdominal diameter ≥ 18.10 cm or body fat ≥ 36.515%. CONCLUSION: Central obesity and high fasting glucose are the most common metabolic disorders in postmenopausal women. Obesity is an independent risk factor of metabolic syndrome for postmenopausal women. Body composition indices and their critical values to predict the abnormal components of metabolic syndrome in postmenopausal women are found. Related Articles | Metrics

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From Minu3D cultural system to aldosterone-induced tubulogenesis Hu Kang-hong 2010, 14 (2):  365-369.  doi: 10.3969/j.issn.1673-8225.2010.02.041 Abstract ( 333 )   PDF (398KB) ( 458 )   Save The existing technical disadvantages of culture systems so far developed have limited the molecular biological study of tubulogenesis. Recently, a newly developed Minu3D system creates controllable growth conditions and allows the generation of tubules from renal stem/progenitor cells. It is for the first time indicated that aldosterone is able to induce the formation of tubules. Aldosterone may trigger some of mineralocorticoid receptor-mediated signaling pathways for the initiation and the regulation of the tubulogenesis. The implication of these studies toward unveiling tubulogenesis is discussed. Related Articles | Metrics

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Effects of berberine hydrochloride on periodontal tissues and cytokine expression in rats with experimental periodontitis　 Zhao Wei, Yu Zhan-hai, Gao Cheng-zhi 2010, 14 (2):  370-374.  doi: 10.3969/j.issn.1673-8225.2010.02.042 Abstract ( 322 )   PDF (383KB) ( 515 )   Save BACKGROUND: The broad-spectrum antibacterial action of berberine hydrochloride mainly contributes to recurrent aphtha, periapical periodontitis, radioactive mucositis and pericoronitis, however a little evidence support the action mechanism underlying periodontitis treatment.  OBJECTIVE: To determine the effects of berberine hydrochloride on the expressions of related cytokines in periodontal tissues of experimental periodontitis rats, to reveal and understand the action pathway of berberine hydrochloride on oral tissue repair. METHODS: Sixty Wistar rats weighing 160-200 g, aged 3 months Models, were involved in this study. Models of experimental periodontitis were established in rats through a use of local steel-wire ligation and systemic injection of prednisone acetate. Forty successfully established models were randomized into periodontitis model group (n=8) and periodontitis treatment group (n=32), at the same time, 10 normal rats served as control group. The treatment group of animals were fed with 0.06 g/kg berberine hydrochloride daily and medicated to death over the 1, 2, 3, 4 weekends (8 rats each). The model group was fed with isodose normal saline. The model group and normal control group were killed at the fourth weekend. Main observations: ①Oral gross observation and X-ray film examination; ②Pathological assay of periodontal tissues; ③Immunohistochemical SABC method was conducted to determine the expressions of tumor necrosis factor-α (TNF-α), bone gla protein (BGP), interleukin-1β (IL-1β), interleukin-6 (IL-6) in periodontal tissues in rats. RESULTS AND CONCLUSION: ①Following hormone injection, gum tissue exhibited erosion and pyorrhea in model group of rats; the above-mentioned symptoms were relieved in rats of treatment group; there was no abnormality in periodontal tissues of normal rats. X-ray examination revealed alveolar crest resorption and obvious interradicular shadow in the model group. ②Rats of model group showed obvious pathologic changes in periodontal tissues, the levels of TNF-α, IL-1β, IL-6 were significantly higher and the level of BGP was dramatically lower than those in normal group (P < 0.05); Treatment with berberine hydrochloride decreased the levels of TNF-α, IL-1β, IL-6 in periodontal tissues and increased the level of BGP compared with model group (P < 0.05). The periodontal tissues in groups treated with berberine hydrochloride exhibited pathological changes at inflammatory repair stage. Results showed that berberine hydrochloride inhibits the expressions of TNF-α, IL-1β, IL-6 in periodontal tissues in experiment rat models of periodontitis, and promotes the expression of BGP and repair of periodontal tissue. Related Articles | Metrics

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Motor coordination mode of transverse pelvic and thoracic rotations during walking in healthy pregnant women Wu Wen-hua, Onno G Meijer○, Huang Zi-jie, Liu Xian-xiang 2010, 14 (2):  375-360.  doi: 10.3969/j.issn.1673-8225.2010.02.043 Abstract ( 291 )   PDF (313KB) ( 558 )   Save BACKGROUND: During pregnancy, the female body undergoes a number of physiological and physical changes. One would expect that the demand on the musculoskeletal system will be considerable and there may be some changes of posture or gait during pregnancy. OBJECTIVE: To identify the coordination of axial rotations of pelvis and thorax during walking in healthy pregnant women, and to quantify the difference between normal female and pregnant women. METHODS: A volunteer sample of 12 maternal subjects with pregnancy between 20 and 34 weeks and 13 healthy nulliparous females served as controls. The comfortable velocity of walking, the axial rotations of pelvis and thorax, as well as coordination between these rotations were compared between healthy pregnant and healthy nulliparous women. The comfortable velocity and maximum velocity of walking were measured in two groups. Index of harmonicity of pelvic and thoracic rotations was calculated. The coordination between these rotations was characterised by Fourier phase relationship and coupling strength. RESULTS AND CONCLUSION: The comfortable walking velocity was significantly reduced in pregnant women. There was no significant difference with respect to Tempa Scale, index of harmonicity of transverse pelvic and thoracic rotations, phase relation and coupling strength between these rotations between the pregnant women and controls. No effect of pregnancy on the coordination between transverse pelvic and thoracic rotations was found. The pregnant women were able to adapt the changes caused by the pregnancy in terms of gait coordination. The pregnant women are suggested to reduce comfortable walking velocity in order to economise the energy cost. Related Articles | Metrics