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11 November 2012, Volume 16 Issue 46 Previous Issue    Next Issue

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Osteoarthritis suppression by delivery of the adenoviral vector-mediated nuclear factor kappa Bp65-specific small interfering RNA Chen Lian-xu, Yu Chang-long 2012, 16 (46):  8551-8555.  doi: 10.3969/j.issn.2095-4344.2012.46.001 Abstract ( 297 )   PDF (513KB) ( 445 )   Save BACKGROUND: Preliminary studies have demonstrated that recombinant adenovirus expressing nuclear factor κBp65 (NF-κBp65) specific small interfering RNA (siRNA) can suppress NF-κBp65 expression in the knee cartilage and synovium, decrease transcriptional activity of NF-κB and limit the levels of interleukin 1β and tumor necrosis factor α. OBJECTIVE: To observe the effect of adenoviral vector-mediated NF-κBp65-specific siRNA on experimental osteoarthritis in the rat knee. METHODS: Three-month aged male Sprague-Dawley rats were divided into four groups. The osteoarthritis model was induced by transection of the medial collateral ligament and partial medial meniscectomy in the rat knee. Then, 0.2 mL adenoviral vector-mediated NF-κBp65-specific siRNA and adenovirus were injected into the knee of NF-κBp65-specific siRNA and adenovirus groups, respectively. There was no treatment in the osteoarthritis group, no modeling in the normal group. RESULTS AND CONCLUSION: The scores on the knee cartilage and synovium were significantly increased in the osteoarthritis and adenovirus groups (P < 0.01), while the scores in the NF-κBp65-specific siRNA group were decreased dramatically (P < 0.01), but still higher as compared with the normal group (P < 0.01). Adenoviral vector-mediated NF-κBp65-specific siRNA can suppress the progression of early experimental osteoarthritis. Related Articles | Metrics

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Effect of estrogen on the bone deposition around implants in osteoporosis rats undergoing cigarette smoke inhalation Hou Yu-dong, Yu Ping-ping, Han Xiao-peng, Ren Guang-hui, Jia Ting-ting 2012, 16 (46):  8556-8560.  doi: 10.3969/j.issn.2095-4344.2012.46.002 Abstract ( 223 )   PDF (527KB) ( 511 )   Save BACKGROUND: Osteoporosis and smoking can increase dental implant failure rate significantly. OBJECTIVE: To investigate the effect of estrogen on bone deposition around implants in osteoporosis rats undergoing cigarette smoke inhalation. METHODS: Fifty 3-month-old female SD rats were randomly divided into five groups (n=10): sham operation group, ovariectomy group, ovariectomy+smoke inhalation group, ovariectomy+estrogen group and ovariectomy+smoke inhalation+estrogen group. Rats in all groups were subjected to ovariectomy except the sham operation group. After ovariectomy, the latter two groups were given cigarette smoke inhalation for 24 weeks. Then, screw-type titanium implants were placed into the right tibiae in rats at 12 weeks after ovariectomy or sham surgery. Next, rats in the ovariectomy+estrogen group and ovariectomy+smoke inhalation+estrogen group were injected with estradiol benzoate intramuscularly. After 24 weeks, before sacrificed, densitometry of the tibiae in rats was detected by X-ray test. After that the samples of dead rats were collected and the tibiae with implant were removed by hand-mill piece of bone implant for observation. RESULTS AND CONCLUSION: The densitometry and X-ray results demonstrated that estradiol could improve bone deposition around implants. Hard tissue slicing observations result showed that in ovariectomy+estrogen group, the implants obtained good osseointegration, besides, cancellous bone area combined with bone lamella was connected with the trabecular bone. In addition, the bonding bone lamella in ovariectomy+smoke inhalation+estrogen group was thicker than that in ovariectomy+smoke inhalation group. Moreover, the amount of trabecular bone was increased. These results suggest that estrogen can improve the bone deposition around implants to a certain extent and promote implant osseointegration in osteoporosis rats undergoing cigarette smoke inhalation. Related Articles | Metrics

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Caffeic acid inhibits osteoclast formation and expression of cathepsin K gene Tang Quan-yong, Jia Zhi-yu, Zhao Yun-zhuan, Chen Biao, Zhang Rui, Yang Wei, Zhang Ying-huai 2012, 16 (46):  8561-8565.  doi: 10.3969/j.issn.2095-4344.2012.46.003 Abstract ( 298 )   PDF (446KB) ( 433 )   Save BACKGROUND: To date, there are few researches on the effect of caffeic acid on the formation and differentiation of osteoclasts, and fewer researches on the mechanism of osteoclastic bone resorption. OBJECTIVE: To explore the mechanism of sweet potato extract caffeic acid inhibiting osteoclastic bone resorption by studying the effect of caffeic acid on the formation and differentiation of osteoclast and on the expression of cathepsin K gene. METHODS: Caffeic acid effects on formation and differentiation of osteoclasts in three osteoclast cell lines were observed and the inhibition effect of caffeic acid on the expression of cathepsin K gene was detected by reverse transcription-PCR. RESULTS AND CONCLUSION: Caffeic acid could directly inhibit the formation and differentiation of osteoclasts in three osteoclast cell lines. Caffeic acid had no influences on the expression of receptor activator of nuclear factor-kappa B ligand and receptor activator of nuclear factor-kappa B mRNA, but could markedly inhibit the expression of cathepsin K gene based on the semi-quantitative gene analysis of reverse transcription-PCR. Caffeic acid, an effective component of sweet potato extract, cannot only directly inhibit the formation and differentiation of osteoclast, but also markedly inhibit the expression of cathepsin K gene, which seems to be the mechanism of sweet potato extract inhibiting osteoclastic bone resorption. Related Articles | Metrics

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Miao Nationality’s Jiuxianluohanjiegu decoction promotes tibial shaft fracture healing Yang Yan-jun, Xiong Yi, Cheng Zhi-gang, Tang Liang-hua 2012, 16 (46):  8566-8570.  doi: 10.3969/j.issn.2095-4344.2012.46.004 Abstract ( 401 )   PDF (498KB) ( 460 )   Save BACKGROUND: Clinical researches have shown that Miao Nationality’s Jiuxianluohanjiegu decoction has the effect of promoting the blood circulation, remove blood stasis and obstruction, and reunion of bone, detumescence and analgesia, but the report on the effect in promoting the fracture healing is rare. OBJECTIVE: To observe the effect of Miao Nationality’s Jiuxianluohanjiegu decoction in promoting the rabbit tibial shaft fracture healing. METHODS: Using traditional Chinese medicine method of decocting, preparing experiment required concentration of decoction, millipore filtration standby. New Zealand rabbit white right tibialis anterior disruption caused by width 3 mm and depth 2 mm of the bone defect model, and the models were randomly divided into three groups; Jiegu decoction group, Sanqi control group, normal saline group. The rabbits in the three groups were gavaged with Miao Nationality’s Jiuxianluohanjiegu decoction, Sanqi powder and liquid and normal saline respectively. All the rabbits were administrated since fracture, and the whole blood viscosity, plasma viscosity, red blood cells deposited, blood calcium, serum phosphorus, alkaline phosphatase concentration, X-ray score and pathological changes were measured at 1, 7, 14, 21 and 28 days after administration. RESULTS AND CONCLUSION: Compared with Sanqi control group and normal saline group, the blood calcium, serum phosphorus and alkaline phosphatase concentration in the Jiegu decoction group were increased and reached to peak early, then gradually decreased (P < 0.05), while the hemorheology indicators were gradually decreased (P < 0.05). X-ray score half quantitative analysis showed that the fracture healing of the Jiegu decoction group was faster than that of the Sanqi control group and normal saline group, and the difference was significant (P < 0.05). Hematoxylin-eosin staining revealed that the callus formation speed and quality in the Jiegu decoction group were much higher than those in the other two groups. Miao Nationality’s Jiuxianluohanjiegu decoction can promote the callus formation and shorten the fracture healing time. Related Articles | Metrics

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Explant culture and identification of SD rat’s osteoblasts Li Juan1, 2, Shen Guang-hao, Cheng Guang, Luo Er-ping, Li Fei-jiang, Jing Da, Fei Zhou, Chen Xiao-yan 2012, 16 (46):  8571-8574.  doi: 10.3969/j.issn.2095-4344.2012.46.005 Abstract ( 376 )   PDF (512KB) ( 537 )   Save BACKGROUND: It is difficult to acquire sufficient osteoblasts with high purity. Therefore, it is imperative to master simple and fast extraction osteoblast method for culture and identification. OBJECTIVE: To culture in vitro and identify SD rat’s osteoblasts with explant culture. METHODS: The newborn (< 24 hours) SD rats were executed, and the skulls were taken out. Excess tissues around the skull were removed. The picked skull was cut off into 1 mm3 pieces. The osteoblasts were cultured by conventional method and tissue explant (modified method) as well as identified with morphology, alkaline phosphatase assay and Von Kossa staining methods. RESULTS AND CONCLUSION: The osteoblasts cultured by tissue explant had typical morphological characteristics; alkaline phosphatase was positive staining, and mineralization nodules formed after Von Kossa staining. Besides, the osteoblasts had unitary cell component which was typical, and the culture time was short, what’s more, the quantity, purity and density were coincident. These findings suggest that the osteoblast lineage acquired is stable, which provides a good plate for in vitro experiments. Related Articles | Metrics

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A mitochondrial PCR gene chip based study on the pathogenesis of osteoarthritis Pan Jian-ying, Shen Jun, Zhang Rong-kai, Li Zhi-fu, Cai Dao-zhang 2012, 16 (46):  8575-8582.  doi: 10.3969/j.issn.2095-4344.2012.46.006 Abstract ( 328 )   PDF (595KB) ( 559 )   Save BACKGROUND: Chondrocytes play a significant role in repairing damaged cartilage tissue as well as in maintaining the integrity of the cartilage. Mitochondria are involved in a large amount of biochemical processes, and mitochondrial impairment has a closed relationship with cell apoptosis, senescence and pathological process of osteoarthritis. OBJECTIVE: To detect the differential expression of mitochondrial genes by using gene chip based mitochondrial gene analyses. METHODS: Articular chondrocytes were collected from healthy people and osteoarthritis patients, then extracted and cultured followed by RNA isolation and quality assessment, mRNA isolation and strand cDNA synthesis. After all, real-time quantitative PCR was performed. RESULTS AND CONCLUSION: Among 84 mitochondrial genes, 18 genes were unambiguously identified as significantly altered in osteoarthritis: 15 of them (BBC3, BCL2, SLC25A37, etc.) were up-regulated at both fold changes and fold regulation > 2, and three of them (CPT1B, SLC25A16, SLC25A24) were down-regulated at fold change < 0.5 and fold regulation < 2. The grouping of 18 functional genes is as follows: membrane polarization & potential: BCL2, BCL2L1, TP53, UCP1, UCP3; mitochondrial transport: BCL2, BCL2L1, CPT1B, FXC1 (TIMM10B), MFN2, STARD3, TP53, UCP1, UCP3; small molecule transport: SLC25A16, SLC25A2, SLC25A24, SLC25A31, SLC25A37; targeting proteins to mitochondria: FXC1 (TIMM10B), MFN2; mitochondrion protein import: COX18, FXC1 (TIMM10B); inner membrane translocation: FXC1 (TIMM10B), TIMM17B; mitochondrial fission & fusion: COX18, MFN2; mitochondrial localization: MFN2; apoptotic genes: BBC3, BCL2, BCL2L1, SOD2, P53. These findings indicate that mitochondrial energy metabolism dysfunction occurs obviously in osteoarthritis chondrocytes. Related Articles | Metrics

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Gene expression profile in the synovium of patients with knee osteoarthritis in Yanbian area Ren Hong-ge, Li Lin, Li Zhen-hao, Sun Pei-lei, Cui Feng-de 2012, 16 (46):  8583-8588.  doi: 10.3969/j.issn.2095-4344.2012.46.007 Abstract ( 290 )   PDF (481KB) ( 508 )   Save BACKGROUND: The progression of primary osteoarthritis is the result of abnormal expression of various genes. We use mRNA gene chip technique to detect the synovial gene expression of knee osteoarthritis, which provides evidence for related pathological mechanism of knee osteoarthritis. OBJECTIVE: To compare the alteration of synovial gene expression in patients with knee osteoarthritis and normal people in Yanbian area, and to evaluate the potential osteoarthritis related discriminating genes and their roles in pathological mechanism. METHODS: This study included nine patients with knee osteoarthritis and three healthy controls. mRNA gene chip technique was used to analyze the gene expression profile of synovial tissue in these subjects and GO analysis was performed. RESULTS AND CONCLUSION: The results of gene expression in the synovial tissue of osteoarthritis patients and healthy controls group showed that 1 261 genes were differently expressed, including 451 up-regulated genes and 810 down-regulated genes. The GO analysis showed that the gene expression about signal transduction, regulation of transcription, chemotaxis, inflammatory response, and cell proliferation was different. Gene chip technique is effective for screening associated genes, and the pathological progression of osteoarthritis is a complex progression which many genes are involved. Related Articles | Metrics

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Guided bone regeneration repairs human mandibular bone defects: Histological observation Chen Hong-liang, Zhao Cheng-chu, Sun Yong, Zhao Feng, Zhong Ke 2012, 16 (46):  8589-8592.  doi: 10.3969/j.issn.2095-4344.2012.46.008 Abstract ( 332 )   PDF (443KB) ( 617 )   Save BACKGROUND: Osteogenesis effects of guided bone regeneration have been verified in basic research and animal experiments. However, histological observation of bone regeneration following repair of human mandibular bone defects using guided bone regeneration is not reported in China. OBJECTIVE: To study the osteogenesis effects of guided bone regeneration on the repair of human mandibular bone defects through histological observation. METHODS: Twenty-four defects from 19 mandibular bone defect patients were subjected to guided bone regeneration therapy. After 6 months, dental implantation was performed. New bone tissues in the surgical area were taken using a hollow drill for histological observation and osteogenesis evaluation. RESULTS AND CONCLUSION: Different developmental stages of bone formation were visible in new bone tissues. Lamellar bone tissues were mainly found near the root apex, while fibrous bone tissues containing osteoid tissue and cartilage bone formed at the dental crown. It indicated that there is a certain 6-month osteogenesis effect of guided bone regeneration, but the long-term effect needs further observation. Related Articles | Metrics

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Isolation and culture of canine articular chondrocytes in vitro Wang Chao-qiang,Tian Feng-de, Xie Hui, Wang Ben-jie, Liu Bao-yi, Zhao De-wei 2012, 16 (46):  8593-8598.  doi: 10.3969/j.issn.2095-4344.2012.46.009 Abstract ( 241 )   PDF (656KB) ( 434 )   Save BACKGROUND: Since Manning et al reported to isolate chondrocytes using trypsin and bacterial collagenase digestion in 1967, in vitro isolation and culture of chondrocytes has been widely studied, but there is still no unified standard. OBJECTIVE: To study the conditions for in vitro culture and isolation of chondrocytes and to explore a simple, feasible, and efficient experimental method to isolate, culture and proliferate canine chondrocytes in vitro. METHODS: The articular cartilage of 3-week-old puppies were isolated and digested with trypsin and type Ⅱ collagenase for harvesting chondrocytes in different conditions. We compared the number of cells cell survival rate obtained by different methods. Isolated cells were passaged and morphology of chondrocytes was observed. RESULTS AND CONCLUSION: The number and survival rate of chondrocytes was highest by using simple type Ⅱ collagenase digestion. Chondrocytes can be amplified through in vitro culture and maintain good cell morphology and phenotype, but can only be passaged less than five passages. Related Articles | Metrics

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Biological properties of rat articular chondrocytes cultured with autologous serum versus fetal bovine serum Fu Xin, Duan Xiao-ning, Zhang Ji-ying, Yu Chang-long 2012, 16 (46):  8599-8606.  doi: 10.3969/j.issn.2095-4344.2012.46.010 Abstract ( 284 )   PDF (743KB) ( 532 )   Save BACKGROUND: At present, fetal bovine serum is mainly use for chondrocytes culture. However, because the security of the chondrocytes culture using heterologous serum in clinical application has been questioned in recent years, the autologous serum for chondrocytes culture has been paid more and more attention. OBJECTIVE: To compare the difference in the biological properties between rabbit articular chondrocytes cultured with 10% autologous serum and 10% fetal bovine serum in vitro. METHODS: After the culture medium of rat autologous serum was prepared, rabbit articular chondrocytes were isolated from rabbits and seeded into the medium with 10% autologous serum and 10% fetal bovine serum, respectively for monolayer subcultivation. Then, passage1, 3 and 5 cells were chosen and cell morphological changes were observed by light microscope, meanwhile, growth curves were drawn to evaluate cell poliferation rate. Besides, biological properties of chondrocytes were observed by toluidine blue staining, typeⅠ and typeⅡ collagen immunohistochemical staining. In addition, the expression changes of typeⅠ, typeⅡcollagen, CD26 and CD44 were analyzed by flow cytometry. RESULTS AND CONCLUSION: (1)There was no significant difference in cell morphology between chondrocytes cultured with autologous serum and fetal bovine serum. (2)The cell growth rate of chondrocytes cultured with autologous serum was m ore rapid than that of chondrocytes cultured with fetal bovine serum. (3)Toluidine blue staining showed that both chondrocytes cultured with autologous serum and fetal bovine serum showed a passage-dependent reduction of staining. In passage 5 cells, the two groups almost had no different dying. For passage 1 and 3 cells, chondrocytes cultured with autologous serum showed deeper staining than fetal bovine serum. (4)Immunohistochemical staining and flow cytometry results showed that expression of type Ⅰ collagen was increased with passage, while type Ⅱ collagen expression was decreased with passage. Moreover, typeⅠcollagen expression of chondrocytes cultured with autologous serum was significantly lower than that chondrocytes cultured with fetal bovine serum at passage 3 (P < 0.05). (5)CD26 expression was increased at first and then decreased, while the expression of CD44 did not change with passage. These results suggest that compared with 10% fetal bovine serum, the growth rate of chondrocytes cultured with 10% autologous serum is more rapid, and has a better preservation of phenotype based on the results of different observations. Related Articles | Metrics

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Expression of type Ⅱ collagen and aggrecan in human articular cartilage tissues after blocking stromal cell derived factor-1/chemokine receptor 4 signal pathway in vitro Li Yan-lin, Wang Guo-liang, Cao Bin, He Chuan, Gao Gang, Chen Wen-dong, Ma Ke, Xu Peng, Yang Guang 2012, 16 (46):  8607-8610.  doi: 10.3969/j.issn.2095-4344.2012.46.011 Abstract ( 446 )   PDF (374KB) ( 393 )   Save BACKGROUND: Stromal cell derived factor-1(SDF-1)/chemokine receptor 4 (CXCR4) signal pathway plays a key role in the pathogenesis of osteoarthritis. OBJECTIVE: To explore the effect of SDF-1/CXCR4 signal pathway on the expression of type Ⅱ collagen and aggrecan mRNA of the synthesis in human articular cartilage after blocking the SDF-1/CXCR4 signal pathway with AMD3100. METHODS: The cartilage blocks from osteoarthritis patients who had total knee replacement and normal cartilage blocks from patients who had traumatic amputation were collected and divided into three groups: experimental group, experimental control group and blank control group. The three groups were cultured in the nutrient solution containing of SDF-1 and AMD3100, SDF-1 and MAB310, as well as SDF-1 only, respectively. RESULTS AND CONCLUSION: The expressions of type Ⅱ collagen and aggrecan mRNA, as well as type Ⅱ collagen content in the experimental group was higher than those in the experimental control group and blank control group (P < 0.05). These results suggest that SDF-1 can induce the degradation of type Ⅱ collagen and aggrecan thruogh the SDF-1/CXCR4 signaling pathway. Besides, AMD3100 can block SDF-1/CXCR4 signal pathway and reduce the degradation of type II collagen and aggrecan, and therefore slow down the degeneration of articular cartilage. But AMD3100 cannot recover type II collagen and aggrecan in osteoarthritis cartilage to normal levels Related Articles | Metrics

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Effect of extracorporeal shock wave on the proliferation of rabbit articular chondrocytes and type Ⅱ collagen expression Wang Ming-bo, Zhao Zhe, Yun Wen-ke 2012, 16 (46):  8611-8614.  doi: 10.3969/j.issn.2095-4344.2012.46.012 Abstract ( 400 )   PDF (395KB) ( 370 )   Save BACKGROUND: Injured articular cartilage has a very limited self-repair ability. Extracorporeal shock wave may provide a method that has high-quality repair of articular cartilage defects and can achieve good late outcomes. OBJECTIVE: To explore the effect of extracorporeal shock wave on the proliferation of rabbit articular chondrocytes and type Ⅱ collagen expression. METHODS: Normal rabbit knee joint chondrocytes were obtained by enzymatic digestion method. The cells were cultured, passaged, and then randomized into three experimental groups: A, B and C groups were given 0.5×105, 1.5×105 and 2.5×105 Pa extracorporeal shock wave intervention, respectively. The blank control received no intervention. RESULTS AND CONCLUSION: The cell growth curve in the group B at day 6 was higher than that in the other groups (P < 0.05). Enzyme-linked immunosorbent assay showed that compared with the other groups, the absorbance of type Ⅱ collagen in the group B was significantly increased (P < 0.05). Reverse transcription PCR indicated that type Ⅱ collagen expression was enhanced obviously in the group B, as compared with the other groups (P < 0.05). These results suggest that the extracorporeal shock wave stimulation of appropriate energy intensity and frequency can significantly promote chondrocyte proliferation and type Ⅱ collagen expression. Related Articles | Metrics

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Vitrification method for cryopreservation of the articular cartilage Song Hong-qiang, Xuan Yan-hua, Wu Ya-di, Qi Jian-hong 2012, 16 (46):  8615-8619.  doi: 10.3969/j.issn.2095-4344.2012.46.013 Abstract ( 340 )   PDF (493KB) ( 461 )   Save BACKGROUND: Articular cartilage allograft transplantation is one of effective methods for treatment of articular cartilage defects. However, the short preservation time of osteochondral allograft in vitro limits its clinical application OBJECTIVE: To discuss the feasibility and superiority of vitrification method on the viability of chondrocytes. METHODS: Osteochondral tissues were isolated from adult pigs, and cut into approximately 5 mm×6 mm (diameter×length) cylindrical osteochondral blocks. Fresh cartilage was taken as control. The osteochondral blocks was pretreated with 0.5 mol/L glycerol, 1 mol/L dimethyl sulfoxide, and 1 mol/L vitrification solution, respectively, and then cryopreserved for 8 weeks. Histochemistry staining and immunofluorescence staining were performed to observe and compare the changes in chondrocyte viability. RESULTS AND CONCLUSION: The viability of chondrocytes was 74.5% in the vitrification pretreatment group, significantly higher than the glycerol and dimethyl sulfoxide pretreatment groups. In addition, there was a small loss of cartilage matrix in the vitrification pretreatment group. In summary, the vitrification-cryopreservation method can greatly improve the viability of frozen chondrocytes. Related Articles | Metrics

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Primary culture, cryopreservation and resuscitation of human gingival fibroblasts Hui Min, Ni Ying, Zheng Jing, Liu Jian, Wang Shuai, Zhang Xiao-ming 2012, 16 (46):  8620-8624.  doi: 10.3969/j.issn.2095-4344.2012.46.014 Abstract ( 378 )   PDF (507KB) ( 512 )   Save BACKGROUND: Human gingival fibroblasts (HGFs) are the main cells in the connective tissue layers, which play an important role in many physiological and pathological processes. OBJECTIVE: To perform primary culture, identification, cryopreservation and resuscitation for HGFs. METHODS: First, the HGFs were cultured by the methods of tissue culture and identified by morphological and immunocytochemical analysis. Next, HGFs were frozen and resuscitated. And then their morphological changes were observed by the inverted microscope. RESULTS AND CONCLUSION: The success rate of HGFs from the primary cells was 86.7%, and the HGFs showed fusiform or spindle-shaped. The immunochemistry study indicated that vimentin showed positive staining; while cytokeratin showed negative staining, which originated from mesoblastma. The cryopreservation and resuscitation of HGFs were successful. The biological character of HGFs after second passage was similar to that of the original generation. These results suggest that the primary culture, cryopreservation and resuscitation of HGFs can be feasible by using the method of tissue culture. Related Articles | Metrics

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Effect of caffeine on rat tooth germ development Liu Shu-fa, Wang Xiao-wei, Wang Li-feng, Huo Dong-ying, Gu Xiao-ming 2012, 16 (46):  8625-8628.  doi: 10.3969/j.issn.2095-4344.2012.46.015 Abstract ( 325 )   PDF (510KB) ( 432 )   Save BACKGROUND: Coffee drinking during pregnancy can cause calcium malabsorption in pregnant women and affect fetal tooth development. OBJECTIVE: To observe the effect of caffeine on rat tooth germ development. METHODS: The Wistar pregnant rats were randomly divided into two groups. In the caffeine group, caffeine was added into the drinking water at 7 days after pregnancy; and in the control group, no caffeine was added into the drinking water. At 20 days after pregnancy, half of pregnant rats in each group were sacrificed to take the fetal rat, and hematoxylin-eosin staining was performed to observed the morphology of tooth germ; the rest half of the pregnant rats in the two groups were administrated till 20 days after delivery, the calcium level in the serum of rat pups was detected and the first molar of rat pups was observed by scanning electron microscope. RESULTS AND CONCLUSION: The calcium ion content in the serum of rat pups in the caffeine group was lower than that in the control group (P < 0.05). Hematoxylin-eosin staining showed that compared with the control group, the volume of the tooth germ tissue of the fetal rats in the same period in the caffeine group was reduced and the development was delayed. Electron microscopy results showed that the surface voids on the first molar of the rat pups in the caffeine group after acid etching was more than those in the control group. Caffeine application in the pregnant rats can inhibit the tooth germ development of rat pups, lead to the insufficiency of enamel matrix calcification, and also result in the intolerance etching of the tooth after development. Related Articles | Metrics

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mRNA expression of insulin-like growth factor-1 and its receptor in pathological scars Cai Hao, Tang Bing, Zhu Jia-yuag, Zhu Bin, Bi Liang-kuan 2012, 16 (46):  8629-8635.  doi: 10.3969/j.issn.2095-4344.2012.46.016 Abstract ( 303 )   PDF (569KB) ( 437 )   Save BACKGROUND: In many tumors, insulin-like growth factor 1 and its receptor express abnormally. Insulin-like growth factor is the mitogen and anti-apoptotic factor of a variety of cells, controlling the proliferation, differentiation and apoptosis of various cells. OBJECTIVE: To observe the expression of insulin-like growth factor-1 and insulin-like growth factor -1 receptor in pathological scar. METHODS: Surgical specimens of hypertrophic scars, keloids and normal skin were collected from the patients admitted in the Department of Burn Surgery, First Affiliated Hospital of Sun Yat-sen University. Fluorescent quantitative real-time PCR technique was used to detect the mRNA expression of insulin-like growth factor-1 and insulin-like growth factor -1 receptor in different tissues. RESULTS AND CONCLUSION: The mRNA expression of insulin-like growth factor-1 was higher in the hypertrophic scars and keloids than in the normal skin (P < 0.05). However, there was no difference in the mRNA expression of insulin-like growth factor-1 between the hypertrophic scars and keloids (P > 0.05). The expression of insulin-like growth factor-1 receptor in the hypertrophic scars was close to that in the normal skin (P > 0.05); while, the expression of insulin-like growth factor-1 receptor in the keloids was significantly higher than that in the surrounding tissues of keloids (P < 0.05). These findings indi Related Articles | Metrics

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Transforming growth factor-beta 2 effects on the expression of E-cadherin and alpha-smooth muscle actin in human lens epithelial cells Zhu Yan, Zhu Yu-guang, Zhong Ying-ying, Du Xiao-nan, Zhang Rong, Wang Jie 2012, 16 (46):  8636-8640.  doi: 10.3969/j.issn.2095-4344.2012.46.017 Abstract ( 349 )   PDF (427KB) ( 421 )   Save BACKGROUND: Studies have shown that mesenchymal transition in human lens epithelial cells is the main pathological change after cataract. Transforming growth factor-β2 can induce the mesenchymal transition. OBJECTIVE: To in vitro culture human lens epithelial cells and to investigate the effect of transforming growth factor-β2 on the expression of E-cadherin and α-smooth muscle actin in human lens epithelial cells. METHODS: First, human lens epithelial cells were cultured in vitro by tissue block method and passaged. Then, the 5th generation of human lens epithelial cells were collected and treated with transforming growth factor-β2 (100 ng/L) for inducement for 48 hours. Next, the expression of α-smooth muscle actin mRNA and E-cadherin mRNA was detected by reverse transcription-PCR, and the expression of the two proteins mentioned above were detected with Western blot method. RESULTS AND CONCLUSION: After human lens epithelial cells were treated with transforming growth factor-β2 for 48 hours, the expression of E-cadherin was decreased significantly; while the expression of α-smooth muscle actin was increased significantly. These results suggest that transforming growth factor-β2 can successfully induce mesenchymal transition in human lens epithelial cells, and the lens epithelial cells treated with transforming growth factor-β2 can be used as a cell model for mesenchymal transition. Related Articles | Metrics

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Effect of nerve growth factor pretreatment on phosphorylated Tau protein expression in hippocampal neurons in Alzheimer’s disease rats Ma Hui-ping, Lü Xin-rui, Shi Hong-yun, Shi Zhen-xia, Li Xin-chun, Li Zheng 2012, 16 (46):  8641-8646.  doi: 10.3969/j.issn.2095-4344.2012.46.018 Abstract ( 359 )   PDF (614KB) ( 452 )   Save BACKGROUND: Nerve growth factor (NGF) can promote the differentiation of cholinergic neuron, determine the growth of axons and involve in the regeneration and functional recovery of injured nerve. OBJECTIVE: To further validate the effects of NGF pretreatment on neurofibrillary tangles (NFT) and phosphorylated Tau protein expression in the hippocampal CA1 region of Alzheimer’s disease (AD)-like model rats. METHODS: Male Wistar rats (3-5 months old) were randomly divided into control group, AD-like model group and NGF pretreatment group. In the AD-like model group, AD-like rat model was established by injecting okadaic acid into the hippocampal CA1 region. In the NGF pretreatment group, NGF was injected into the lateral ventricle of the brain in rats before okadaic acid was injected for establishing the AD-like animal model. The rats of the control group were injected an equal volume of dimethyl sulfoxide using the same method. The behavior changes of rats were observed by Morris water maze. Then, the NFT in hippocampal CA1 region was detected by improved Bielschowsky staining. Besides, the changes of phosphorylated Tau protein expression in the hippocampal CA1 region were observed by immunohistochemical and western blot methods. RESULTS AND CONCLUSION: Learning disabilities and memory deficits in the AD-like model rats were found. Compared with the control group, in the model group, there were more NFTs; in addition, the phosphorylated Tau protein expression was increased in the hippocampus. However, the above symptoms in the rats of the NGF pretreated group were improved obviously. These results suggest that NGF pretreatment can significantly improve learning and memory capabilities of the AD-like model rats, inhibit NFT formation and decrease the expression of phosphorylated Tau protein. Related Articles | Metrics

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Effect of stress shielding of the Achilles tendon on transforming growth factor beta in rats Zhao Hua-kun, Yan Xiao-yan, Ma Yan-hong 2012, 16 (46):  8647-8650.  doi: 10.3969/j.issn.2095-4344.2012.46.019 Abstract ( 392 )   PDF (404KB) ( 395 )   Save BACKGROUND: The changes of cytokines induced by stress shielding in the tendon are closely related to tendon contracture. But the situation of transformation growth factor (TGF)-β remains unclear. OBJECTIVE: To investigate the effect of stress shielding on TGF-β concentration. METHODS: Twenty mature male SD rats were randomly divided into model and control groups. Rats in the model group were undergone stress shielding of the Achilles tendon in the left hind limb, and then rose for 2 and 4 weeks. Randomly selected 10 rats’ normal Achilles tendons in the right hind limb served as controls. The levels of TGF-β in the two groups were tested by detection of enzyme-linked immunosorbent assay. RESULTS AND CONCLUSION: The level of TGF-β in the model group was higher than that in the control groups (P < 0.01). There was no difference between the 2 weeks model group and 4 weeks model group (P > 0.05). These results suggest that after stress shielding of the Achilles tendon for 2 weeks, the level of TGF-β increases obviously and does not decrease at week 4, moreover, the anabolism of tendon is enhanced. Related Articles | Metrics

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Assessment and clinical analysis of female pelvic floor muscle Manrepa·Tuerxun, Li Yan, 　Gulina·Ababaikeli 2012, 16 (46):  8651-8656.  doi: 10.3969/j.issn.2095-4344.2012.46.020 Abstract ( 443 )   PDF (475KB) ( 830 )   Save BACKGROUND: Understanding the related factors of female pelvic floor disorders in Xinjiang multi-ethnic area and exploring the quantitative examination method which can relatively accurately reflect the pelvic floor function plays an important role in preventing female pelvic floor disorders. OBJECTIVE: To investigate factors related to female pelvic floor muscle strength and to analyze the relevance and significance of the pelvic floor muscle assessment with the symptoms and physical examination of pelvic floor dysfunction disease. METHODS: The questionnaire, physical examination, pelvic floor muscle strength testing data were collected from 210 cases of female patients who were in hospital during April 2010 to November 2010 at Department of gynecology, the First Affiliated Hospital of Xinjiang Medical University. Rank sum test was used to analyze the packet variables factors related to pelvic floor muscle strength, and than Spearman rank correlation analysis was carried out to make further analysis which has statistically significance data. RESULTS AND CONCLUSION: Age, body mass index, delivery times, menopause, symptoms of chronic constipation were negative correlated with pelvic floor muscle strength. The symptoms of stress urinary incontinence and physical examination of pelvic organ prolapse were negative correlated with the pelvic floor muscle strength. It is conductive to the early prevention and treatment of pelvic floor dysfunction disease by controlling the body weight and reducing the incidence of chronic constipation; well perimenopausal hormone replacement therapy; controlling the pelvic tissue damage of pregnant women and making the patients to pay more attention on their own pelvic floor muscle strength. Related Articles | Metrics

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Pressure distribution to prevent pressure ulcers under different supine positions Hou Xi-hong, Zhang Jian-guo, Xue Qiang, Liu He-rong 2012, 16 (46):  8657-8661.  doi: 10.3969/j.issn.2095-4344.2012.46.021 Abstract ( 401 )   PDF (518KB) ( 544 )   Save BACKGROUND: Pressure sore or pressure ulcer is a comprehensive complication in clinic. Although there are many anti-pressure ulcer materials and drugs, their effects are temporary rather than permanent. Therefore, to explore the human body pressure distribution reasonable to prevent pressure ulcer is of great consequence. OBJECTIVE: Through changing supine positions, to realize the different human body pressure distribution, and then to study the relationship between human body pressure distribution and supine positions so as to explore more reasonable pressure distribution in the human body. METHODS: Human body pressure distribution indexes were measured at different supine positions under the different air pressure of the mattress by using the pressure distribution measurement and analysis of the XSENSOR system. Then, the experimental data were collected and tidied through Excel and MATLAB software to compare and analyze the human body pressure distribution indexes under different supine positions using SPSS 16.0 statistical analysis software. t-test was done for pairwise comparisons of body pressure indexes. RESULTS AND CONCLUSION: P values for human body pressure distribution indexes under different supine positions were less than 0.05, indicating that changing supine positions exerts an important influence to human body pressure distribution. When the angle of the leg elevation-the back elevation was 10º-0º, the human body pressure distribution was more reasonable resulting in the biggest pressure of the whole body and the least heel pressure, which is the ideal human body pressure distribution. Related Articles | Metrics

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Stress distribution of molar uprighting periodontal ligament under combined force with micro-implant anchorage system Guo Dong-mei, Chang Shao-hai, Hu Ling-ling, Lu Ying-juan, Ye Yu-shan 2012, 16 (46):  8662-8666.  doi: 10.3969/j.issn.2095-4344.2012.46.022 Abstract ( 316 )   PDF (474KB) ( 461 )   Save BACKGROUND: Although mini-screws anchorage implant can provide strong anchorage, it cannot control the three-dimensional movement of the tooth. Tomas-pin can be fixed in a square archwire and designed into a cross, in order to control the three-dimensional movement of the tooth. And this research has not been reported. OBJECTIVE: To investigate the effects of combined force for maxillary second molar uprighting with micro-implant anchorage system. METHODS: Maxillary second molar and tipped mesiobuccally were selected to be the tooth sample, and maxillary which corresponds with sample tooth was also selected to be the alveolar bone sample. Multi-slice spiral CT scanning was taken for both of samples to get legible and accurate image information. A three-dimensional model of maxillary second molar with periodontal tissues was established by finite element software. Effects of embedding position of micro-implant, orientation of the loading force and loading force on the Von Mises stress of the periodontal tissue were investigated. RESULTS AND CONCLUSION: The maximum stress of the periodontal ligament was detected in the cervix of the maxillary second molar, and the stress was gradually decreased along with the apical. However, stress concentration could not be found in the apex of each root. The finite element model could be exhibited similarity in geometry and biomechanics, and could be used for the precisely biomechanics analysis. The bodily movement or tipping movement can be achieved with Tomas-pin. Related Articles | Metrics

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Small interfering RNA mediated silencing of FOXO3a influences the arsenic trioxide-inhibited proliferation of endothelial cells Li Hai-li, Liu Hong 2012, 16 (46):  8667-8670.  doi: 10.3969/j.issn.2095-4344.2012.46.023 Abstract ( 315 )   PDF (366KB) ( 469 )   Save BACKGROUND: Early studies have confirmed that As2O3 can promote FOXO3a factor expression in breast cancer cells and inhibit vascular endothelial growth factor expression in tumor cells. However, the effect of As2O3 on proliferation, FOXO3a and growth factors in vascular endothelial cells remains unclear. OBJECTIVE: To investigate the effect of silencing of FOXO3a using small RNA on inhibiting the As2O3-inhibited proliferation of human umbilical vein endothelial cells and angiogenesis. METHODS: There were four groups in this experiment: As2O3 group, control small interfering RNA group, FOXO3a small interfering RNA group and control group. (1)In As2O3 group, human umbilical vein endothelial cells were cultured in RPMI-1640 medium containing 4 μmol/L As2O3 after adherent. (2)In control small interfering RNA group, human umbilical vein endothelial cells were cultured in RPMI-1640 medium containing 4 μmol/L As2O3 after irrelevant sequence small interfering RNA was tranfected. (3)In FOXO3a small interfering RNA group, human umbilical vein endothelial cells were cultured in RPMI-1640 medium containing 4 μmol/L As2O3 after FOXO3a specific small interfering RNA was transfected into the cells. (4)In control group, a volume of PBS equal to the original volume of As2O3 served as a control, and human umbilical vein endothelial cells were cultured in the complete medium. After that, cell proliferation was detected by CCK-8 kit; the expressions of FOXO3a protein and vascular endothelial cell growth factor proteins in human umbilical vein endothelial cells were observed by immunocytochemical method. RESULTS AND CONCLUSION: Compared with the control group, the inhibitory effect of As2O3 on the proliferation of human umbilical vein endothelial cells was weakened by silencing of FOXO3a. Besides, As2O3 could promote FOXO3a protein expression and inhibit vascular endothelial cell expression. However, FOXO3a protein expression was inhibited obviously and protein expression of vascular endothelial growth factor was increased after FOXO3a using small interfering RNA. These results suggest that the transcription factor of FOXO3a can be an important strategy for the inhibition of tumor cell proliferation and angiogenesis. Related Articles | Metrics

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Expression of EphB3 receptor protein in injured spinal cord tissues of rats Wu Gang, Lü Gang, Zhi Xiao-dong 2012, 16 (46):  8671-8674.  doi: 10.3969/j.issn.2095-4344.2012.46.024 Abstract ( 233 )   PDF (371KB) ( 430 )   Save BACKGROUND: Eph receptors can regulate axon-mediated environment inhibiting axonal repair and regeneration. EphB3 is an extremely important member of the Eph family, therefore, to study the relationship between EphB3 and spinal cord injury will become the new direction of research at home and abroad. OBJECTIVE: To observe the expression of EphB3 receptor protein and gene in rats with spinal cord injury. METHODS: The spinal cord hemisection method was used to establish a rat model of spinal cord injury. Using reverse transcription-PCR and Western blot assay, we observed the mRNA and protein expressions of EphB3 in the spinal cord at 1, 2, 4 and 8 weeks after spinal cord injury in comparison with normal rats. RESULTS AND CONCLUSION: EphB3 gene and protein were highly expressed in injured spinal cord tissues, but they had no significant changes at 1, 2, 4 and 8 weeks after injury (P ≥ 0.05). In the spinal cord tissues of normal rats, the mRNA and protein expressions of EphB3 were very low. A significant difference in the mRNA and protein expressions was found between spinal cord injury rats and normal rats (P < 0.05). These findings indicate that spinal cord injury can result in the long-term, stable and high expression of EphB3 receptor protein. Related Articles | Metrics

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Ca2+ concentration changes in the spinal motoneurons after brachial plexus root rupture injury Sun Hong-bin, Wu Guang-zhi, Xing Wan-ying, Li Qiang, Cui Shu-sen 2012, 16 (46):  8675-8679.  doi: 10.3969/j.issn.2095-4344.2012.46.025 Abstract ( 410 )   PDF (424KB) ( 376 )   Save BACKGROUND: Peripheral nerve injuries can cause Ca2+ channel opening in the cell membrane and the extracellular Ca2+ internal flow, thereby resulting in intracellular calcium overload. OBJECTIVE: To observe free Ca2+ concentration changes in the spinal cord anterior horn motor neurons after brachial plexus root rupture injury. METHODS: Eighty-four healthy adult male Wister rats were randomly divided into three groups: sham operation group, control group and experimental group. Rats in the sham operation group were only subjected to expose the brachial plexus nerve without rupture. Rats in the control group had not treatment after brachial plexus nerve injury and those in the experimental group were intraperitoneally injected with 4 mg/(kg·d) verapamil. After the nerve was cut for 12 hours, 24 hours, 48 hours, 72 hours, 1 week, 2 weeks, and 4 weeks, four samples was collected from each group. RESULTS AND CONCLUSION: At the beginning of peripheral nerve injury, Ca2+ concentration in the spinal cord anterior horn motor neurons of the control and experimental groups was increased and reached a peak after 48 hours, but subsequently was decreased gradually. After 1 week, Ca2+ concentration of the experimental group basically returned to a normal level, but was still higher than that of the sham operation group. These results suggest that after brachial plexus root rupture injury, the L-type Ca2+channel on the nerve cell membrane is open, and Ca2+ internal flow enters into the cells, which leads to the increase in intracellular free Ca2+concentration. Moreover, L-type Ca2+channel can be blocked by verapamil, to decrease the Ca2+ internal flow and the number of apoptotic motor neurons. Related Articles | Metrics

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Two-dimensional gel electrophoresis analysis of opaque corneal tissue protein in B6-Co mutant mice Guan Cheng-qi, Jiang Kang-lun, Wu Liu-cheng, Ling Xue-ping, Shao Yi-xiang 2012, 16 (46):  8680-8684.  doi: 10.3969/j.issn.2095-4344.2012.46.026 Abstract ( 184 )   PDF (468KB) ( 630 )   Save BACKGROUND: The similarities and differences between opaque cornea of B6-Co mutant mice and normal cornea of normal B6 mice at the proteomic level have been studied, which can help to illuminate the mechanisms of human corneal opacity and develop new ways for studying gene function using the phenotype-driven approach. OBJECTIVE: To compare and analyze the similarities and differences of cornea proteomics between B6 mice and B6-Co mutant mice using two-dimensional gel electrophoresis method. METHODS: After the corneal tissue protein of B6 mice and B6-Co mutant mice was extracted respectively, the samples acquired were performed two-dimensional gel electrophoresis and gel staining. And then, the electropherograms were analyzed for comparing the similarities and differences of the protein between normal and opaque corneal tissues. RESULTS AND CONCLUSION: The extract of the corneal tissue protein, basic method and condition of two-dimensional gel electrophoresis were established successfully. The gel electropherogram was clear and the result showed that 13 protein spots were significantly downregulated and six protein spots were significantly upregulated in opaque corneal protein of B6-Co mutant mice. There was a significant difference in cornea proteomics between B6 mice and B6-Co mutant mice. These results suggest that gene mutation can lead to the gene up-, and down-regulation or silencing in related signal pathways. Related Articles | Metrics

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Effect of mechanical strain on hypertrophic scar formation and hydroxyproline content in mice Yang Heng-lian, Xiao Hu, Ran Li, Li Qiang 2012, 16 (46):  8685-8688.  doi: 10.3969/j.issn.2095-4344.2012.46.027 Abstract ( 314 )   PDF (399KB) ( 493 )   Save BACKGROUND: Mechanical strain plays a key role in cell proliferation, differentiation, apoptosis and gene expression. OBJECTIVE: To investigate mechanical strain effects on the formation of hypertrophic scar and hydroxyproline content in mice. METHODS: Four weeks old C57/BL mice were randomly divided into mechanical strain group and control group. The mice in the mechanical strain group were subjected to a 2-cm full-thickness skin incision, and then sutured with nylon wire. After stitches were taken out at day 4, 222-cm-mm stretching screw was fixed on the healed wound with nylon wire and expanded every other day. While stretching screw in the control group had no treatment. Six mice in each group were collected respectively at weeks 1, 2, 3, 4 and 5 after wounded, and then their hypertrophic scar thickness, cross-sectional area changes and hydroxyproline content in the hypertrophic scar tissues were observed. RESULTS AND CONCLUSION: Compared with the control group, scar hyperplasia in the mechanical strain group was more obvious, besides, the thickness, cross-sectional area of hypertrophic scars was bigger (P < 0.01). Hydroxyproline content in the hypertrophic scar tissues of the mechanical strain group was significantly higher than that in the control group at each time point (P < 0.01). These results suggest that mechanical strain can result in obvious scar hyperplasia in mice and a significant increase in hydroxyproline content. Related Articles | Metrics

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Effects of streptolysin O on increasing cell permeability Tang Xin-jie, Sheng Ling-ling, Zhang Qun, Xie Feng 2012, 16 (46):  8689-8692.  doi: 10.3969/j.issn.2095-4344.2012.46.028 Abstract ( 429 )   PDF (408KB) ( 569 )   Save BACKGROUND: Streptolysin O (SLO)-induced cell permeability is reversible to a certain extent, and the pores can be closed in the medium containing Ca2+. OBJECTIVE: To observe whether SLO could improve the permeability of pig fibroblasts and to observe the states of cells treated with SLO of different concentrations. METHODS: Pig fibroblasts were permeated with SLO solution of different concentrations (0, 230, 300 and 400 μg/L respectively). After incubation of 50 minutes, the cells were stained with PI to observe cell permeability. When adding the medium containing Ca2+, the cells were plated. Morphology and multiplication capacity of these cells were detected. RESULTS AND CONCLUSION: Along with increase of SLO solution concentration, the cell permeability improved slightly. When cells were treated with SLO, the cell morphology was not changed. 230 μg/L SLO had no influence on the capacity of cell proliferation, and however, 300 μg/L and 400 μg/L SLO reduced the proliferation capacity significantly appearing statistically difference when compared with 0 μg/L group. These findings indicate that 230 μg/L SLO could increase cell permeability with keeping the normal shape and proliferation ability of cells. Related Articles | Metrics

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Cellular repressor of E1A-stimulated genes inhibits apoptosis of human umbilical vein endothelial cells Yan Cheng-hui, Wang Na, Tao Jie, Li Yi, Wang Xiao-zeng, Han Ya-ling 2012, 16 (46):  8693-8697.  doi: 10.3969/j.issn.2095-4344.2012.46.029 Abstract ( 371 )   PDF (475KB) ( 369 )   Save BACKGROUND: Previous studies have demonstrated that there is a protective effect of cellular repressor of E1A-stimulated genes (CREG) against atherosclerosis through prevention of vascular smooth muscle cell apoptosis. However, the role of CREG in endothelial cells (ECs) apoptosis and its underlying signal mechanism is unknown. OBJECTIVE: To further illustrate the effect of CREG on the apoptosis of ECs. METHODS: The apoptosis of human umbilical vein endothelial cells (HUVECs) was induced by serum deprived culture, as well as the expression of CREG was detected by Western blot. TUNEL staining and caspase-3 activity assays were used to evaluate the apoptosis of HUVECS. Furthermore, gain- and loss-of-function analysis was used to reveal the bio-function of CREG in HUVEC apoptosis. RESULTS AND CONCLUSION: Western blot revealed that the apoptosis of ECs was increased with the decreasing of CREG expression. Moreover, gain- and loss-of-function identified that CREG could significantly inhibit ECs apoptosis after the overexpression, whereas the decrease of CREG overexpression could obviously increased and induced. Meanwhile, Western analysis demonstrated that the protective effect of CREG on ECs apoptosis might mainly mediated by activating PI3K/AKT signaling serum starvation pathway. These results suggest that CREG plays a critical role in protecting the vascular endothelium from apoptosis, and the protective effect of CREG against ECs apoptosis may be related with the transduction of PI3K/AKT signaling pathway. Related Articles | Metrics

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Effects of chemical pretreatment on hypoxic preconditioned mice Zhang Jian-jun, Sui Xin, Lü Guo-wei, Zhang Yan-bo, Shao Guo 2012, 16 (46):  8698-8702.  doi: 10.3969/j.issn.2095-4344.2012.46.030 Abstract ( 286 )   PDF (460KB) ( 433 )   Save BACKGROUND: Hypoxia-inducible factor-1 α (HIF-1α) regulates the expression of genes which increase resistance to hypoxia tolerance. CoCl2 is a chemical reagent that can stabilize HIF-1α. OBJECTIVE: To detect the effects of CoCl2 pretreatment on acute repetitive hypoxic exposure of mice. METHODS: Balb/c mice were randomly divided into chemical pretreatment group and normal group. At 3 hours before treatment, the mice in the two groups were respectively injected with CoCl2 and normal saline. After that, the mice in the two groups were subjected to hypoxic exposure for 0 run (normal control group), 1 run, and 4 runs, respectively. Subsequently, hippocampi of mice were removed immediately after the exposure for index detection. RESULTS AND CONCLUSION: The tolerance time after one-time hypoxic exposure in the chemical pretreatment group was higher than that in the control group, but there was no significant difference in four times hypoxic exposure between the chemical pretreatment and control groups. HIF-1 DNA binding activity of the four times hypoxic exposure subgroup in the control group was significantly higher than that of the one-time hypoxic exposure subgroup in the chemical pretreatment group and the normal control group. There was significant difference in each group. HIF-1 DNA binding activity of the one-time hypoxic exposure subgroup in the chemical pretreatment group was obviously higher than that in the normal group (P < 0.01). In the control group, erythropoietin and vascular endothelial growth factor mRNA level in the one-time hypoxic exposure and four times hypoxic exposure subgroups showed increased. Each group had significant difference (P < 0.05). There was no significant difference in erythropoietin and vascular endothelial growth factor mRNA levels among the chemical pretreatment group. These results suggest that CoCl2 chemical pretreatment can improve the tolerance time in hypoxic exposure, and thereby reduce induction of tolerance after hypoxic preconditioning and HIF-1 DNA binding activity. Related Articles | Metrics

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Preparation and purification of a great amount of monoclonal antibodies against the c-terminal of very low density lipoprotein receptor Yang Pu, Li Qi, Liu Zhi-guo, Tian Jun, Zong Yi-qiang, Qu Shen 2012, 16 (46):  8703-8708.  doi: 10.3969/j.issn.2095-4344.2012.46.031 Abstract ( 342 )   PDF (482KB) ( 415 )   Save BACKGROUND: Very low density lipoprotein receptor is regarded as a Swiss Army Knife receptor, and many researchers have studied this receptor. But there are only several kinds of commercial antibodies and the price is on the high side. OBJECTIVE: To get the monoclonal antibody against the c-terminal of very low density lipoprotein receptor for the future study. METHODS: In order to obtain the very low density lipoprotein receptor, hybridoma IgG 6A6 cells related with very low density lipoprotein receptor intracellular domain was injected into peritoneal of Balb/c mice. After 2 weeks, ascites was collected, and the target protein was preliminary purified by salting-out method (n-octanoic acid-saturated ammonium sulfate precipitation method) and further purified by affinity chromatography, and the monoclonal antibody related with very low density lipoprotein receptor intracellular domain was obtained. The relative molecular mass and purity were detected by sodium dodecyl sulfate polyacrylamide gel electrophoresis, and the activity was detected by enzyme-linked immunosorbent assay. RESULTS AND CONCLUSION: Western Blotting results showed that the antibody from the ascites directly can bind the very low density lipoprotein receptor protein, but the background was not clear. The purified antibody from the ascites through salting-out method (n-octanoic acid-saturated ammonium sulfate precipitation method) and then by protein A affinity chromatography can bind specifically with the very low density lipoprotein receptor. The pure and activity monoclonal antibodies can be obtained through purification by the salting-out method and Protein A Agarose Related Articles | Metrics

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MicroRNA studies on osteoblast differentiation Wang Zhen-heng, Wang Rui, Zhao Jian-ning 2012, 16 (46):  8709-8715.  doi: 10.3969/j.issn.2095-4344.2012.46.032 Abstract ( 291 )   PDF (535KB) ( 414 )   Save BACKGROUND: MicroRNAs act as key regulators in osteoblast differentiation which show a new insight for us into the osteoblast differentiation progress. OBJECTIVE: To analyze the regulation rules of microRNAs on related regulators and signaling in osteoblast differentiation. METHODS: A computer online search of CNKI database (2000-2011) and Web of Knowledge database (1995-2011) was performed by the first author using “osteoblast, signaling, regulators, microRNA” as the key words in Chinese and English, respectively, to retrieve relevant literatures about microRNA regulating osteoblast differentiation. Articles with stale content and duplicate literatures were excluded, and finally 37 literatures were included for analysis. RESULTS AND CONCLUSION: MicroRNAs play an important role in cell growth, cell differentiation, and cell death which are a novel regulation way to biology. MicroRNAs play critical rules on related regulators and signaling in osteoblast differentiation. The expression of several target proteins may be regulated by the same microRNA, and the expression of one target protein may be regulated by many microRNAs. The interactions between microRNAs and their targets must be considered when making a further research on the regulation of gene expression of microRNAs. Related Articles | Metrics

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Research advances in construction of tissue-engineered cartilage Liu Shang-lun, Dong Rui, Ma Yong 2012, 16 (46):  8716-8720.  doi: 10.3969/j.issn.2095-4344.2012.46.033 Abstract ( 242 )   PDF (638KB) ( 392 )   Save BACKGROUND: Tissue engineering method provides a new treatment for repair of articular cartilage defects, which has broad application prospects. OBJECTIVE: To explore the methods, researches and application prospects of the tissue-engineered cartilage construction. METHODS: A computer-based online search of China Journal Full-text Database and Web of Science was performed for articles and reviews in Chinese and English from 1991 to 2011. The key words were “tissue engineering, cartilage defects, seed cell, scaffolds” in Chinese and English. By Screening reading titles and abstracts, relevant literatures were included. Repetitive articles with poor quality were excluded. Finally, 30 articles were included. RESULTS AND CONCLUSION: Tissue-engineered cartilage with a variety of seed cells loaded composite scaffolds has made a great progress and shown a good role in repair of cartilage defects, but there are some specific problems to be solved in clinic. Related Articles | Metrics

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Brain injury with fracture can accelerate fracture healing and heterotopic ossification Liu Guan-hua, Zhang Liu, Liang Chun-yu 2012, 16 (46):  8721-8726.  doi: 10.3969/j.issn.2095-4344.2012.46.034 Abstract ( 295 )   PDF (568KB) ( 596 )   Save BACKGROUND: Clinical practices and experimental researches have found that brain injury with fracture can accelerate fracture healing. Besides, the hypertrophic callus and heterotopic ossification can be found. OBJECTIVE: To describe the research progresses and exploration direction of the effect of brain trauma on fracture healing. METHODS: PubMed, Springer Link, CNKI, Elsevier-Sdol databases were searched by computer for papers concerning the effect of brain injury on fracture healing as well as the relationship between brain injury and heterotopic ossification from 1990-01/2011-11. The key words for search were “traumatic brain injury, fracture healing, heterotopic ossification" in English and Chinese. Literatures of irrelevant and repetitive content were excluded, and 32 literatures were included for further study. RESULTS AND CONCLUSION: The blood-brain barrier damage leads to the increase of permeability when brain injuries. Certain factors which have osteogenic action can enter the systemic circulation through an open blood-brain barrier, and therefore, the expression levels of these factors in serum are significantly increased; meanwhile, the role of osteogenic factors in the fracture can promote callus formation and accelerate fracture healing. These findings suggest that the incidence of heterotopic ossification can be increased due to the coexistence of osteogenic factors and environment for a long time. Related Articles | Metrics

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Traumatic osteoarthritis chondrocytes damage and repair mechanism Zhang Wen-xian, Wang Xiao-yan, Feng Kang-hu, Yao Xing-zhang, Ji Jian-gang, Pu Jun 2012, 16 (46):  8727-8732.  doi: 10.3969/j.issn.2095-4344.2012.46.035 Abstract ( 280 )   PDF (613KB) ( 474 )   Save BACKGROUND: Although it has been confirmed in macro and micro level, the pathogenesis of osteoarthritis is still unclear, even self-contradictory and unable to form the more systematic, comprehensive, scientific theory system. OBJECTIVE: To review the progress in chondrocyte damage following traumatic osteoarthritis and repair mechanism. METHODS: A computer-based search of CNKI and the PubMed databases from January 1994 to October 2011 was performed for articles related to chondrocyte damage following traumatic osteoarthritis and repair mechanism. The keywords were “osteoarthritis, apoptosis, metalloprotease, free radical” in the title and abstract. Articles related to pathogenesis of traumatic osteoarthritis were selected, and articles published in the same field recently or in authorized journals were preferred. Totally 140 articles were checked, and finally 31 articles addressing periodontal local sustained release agent were reviewed. RESULTS AND CONCLUSION: Traumatic osteoarthritis occurs based on the macroscopic biomechanical changes, starting with cell transduction mechanism in cellular communication system, which inspires a programmed “destruction and repair” mechanism. Wherein, the chondrocyte apoptosis, catabolic enzymes, free radicals and related cytokine immune response may be the “axial” mechanism of traumatic osteoarthritis. Related Articles | Metrics

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Regulation mechanism underlying fatty acid oxidation during exercise Hu Yan, Niu Jie 2012, 16 (46):  8733-8740.  doi: 10.3969/j.issn.2095-4344.2012.46.036 Abstract ( 362 )   PDF (617KB) ( 409 )   Save BACKGROUND: The plasma fatty acid to the working muscle is primarily supplied from lipolysis of triacylglycerol stored in adipose tissue. The inability of fatty acid oxidation to support the energy demand during high intensity exercise can reflect a limitation in skeletal muscle to oxidize fatty acids. OBJECTIVE: To review findings of research on which related to the regulation mechanism about fatty acid oxidation during exercise, and to propose the problems that need to be solved on mechanism about fatty acid oxidation. METHODS: The PubMed database was used to search the articles on the regulation mechanism about fatty acid oxidation during exercise published from 1995 to 2012 with the key words of “exercise, fatty acid oxidation, intensity, carnitine, acetylcarnitine, mitochondria” in English. The repetitive contents were deleted. RESULTS AND CONCLUSION: A total of 94 articles were collected after the initial search, the repetitive contents were deleted, and finally 56 articles were classified and sorted. Fatty acid oxidation during exercise involves multiple possible regulatory steps, ranging from adipose tissue lipolysis to mitochondrial metabolism in skeletal muscle. It seems that the most attractive regulatory candidate for fatty acid oxidation is the muscle metabolite carnitine, which is essential in carnitine palmitoyl transferase-1 regulation and fatty acid oxidation. At high intensity exercise, the rapid glycolysis provides the mitochondria with excess Acetyl coenzyme A which buffered by free carnitine to form acetylcarnitine. Accordingly, a fall in muscle concentration of free carnitine may reduce carnitine palmitoyl transferase-1 activity, and thus the ability to transport fatty acid into the mitochondria, and therefore also the rate of fatty acid oxidation. In this way, rapid glycogen decomposition and glycolysis are suggested to have a major impact on inhibiting fatty acid oxidation. Related Articles | Metrics