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12 November 2013, Volume 17 Issue 46 Previous Issue    Next Issue

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Receptor activator of nuclear factor kappa-B ligand induces osteoclast precursor culture and differentiation Zhu Wei-ping, Shi Wei, Lin Lin, Li Zhong-he, Huang Jin 2013, 17 (46):  7981-7987.  doi: 10.3969/j.issn.2095-4344.2013.46.001 Abstract ( 330 )   PDF (708KB) ( 492 )   Save BACKGROUND: Previous studies have applied long-bone mechanical separation method to obtain osteoclasts, which are terminally differentiated cells and cannot further proliferate. Therefore a large number of osteoclasts can be harvested with bone marrow cells inducing culture method and RAW264.7 cells inducing culture method to meet clinical requirements. OBJECTIVE: To investigate the optimal method of receptor activator of nuclear factor kappa-B ligand (RANKL) induced osteoclast precursors to differentiate into mature osteoclasts. METHODS: After bone marrow cells were isolated from mouse, RANKL and macrophage colony stimulating factor were added into the medium together, or RAW264.7 cells were cultured with RANKL to induce osteoclasts. The osteoclast precursors were treated with different concentrations of RANKL to observe the number of generated osteoclasts and evaluate the dose-effect relationship between RANKL and osteoclastogenesis. Annexin V-FITC and propidium iodide staining were used for flow cytometry to analyze the mononuclear-macrophage apoptosis during osteoclastogenesis. RESULTS AND CONCLUSION: When 10 μg/L RANKL was used, the peak of osteoclastogenesis appeared at days 6-7; when the concentration of RANKL was up to 100 μg/L, the peak appeared at days 4-5. The number of new osteoclasts was dose-dependent on the RANKL concentration. 50 μg/L of RANKL was the turning point in the fitted curve from osteoclastogenesis and RANKL concentration. After the RANKL concentration was more than 150 μg/L, the number of osteoclasts slowed down obviously. RANKL can induce monocyte-macrophage to form osteoclasts and promote monocyte-macrophage apoptosis. The highest number of osteoclasts would be obtained to each unit of RANKL when monocyte-macrophage cells were cultured with 30 μg/L of RANKL in the same vaccination density (104/cm2). Experimental findings indicate that, RAW264.7 cells or bone marrow cells inducing culture methods are simple and feasible, the optimum cell seeding density was 104/cm2; the appropriate stimulation concentration of RANKL was 30-50 μg/L. Figures and Tables | References | Related Articles | Metrics

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Inhibitory effect of interleukin-1 receptor-associated kinase-4 silencing on mitogen-activated protein kinases expression in human osteoblast-like cells Yang Zi-bo, Huang Bao-ding, Xiang Shan-shan, Wu Pei-hui, Zhang Zhi-qi, Liao Wei-ming 2013, 17 (46):  7988-7993.  doi: 10.3969/j.issn.2095-4344.2013.46.002 Abstract ( 251 )   PDF (397KB) ( 328 )   Save BACKGROUND: The molecular mechanism of periprosthesis osteolysis is not yet completely clear. Periprosthetic osteolysis and absorption is the pathological and physiological process typical of artificial joint loosening. Interleukin-1 can affect bone resorption process through a mitogen-activated protein kinases (MAPK) signaling pathway. OBJECTIVE: To explore the effects of siRNA-induced interleukin-1 receptor-associated kinase-4 gene (IRAK-4) silence on MAPK expression in MG63 cells, which may provide experimental basis for treatment and prevention of periprosthesis osteolysis. METHODS: The siRNA sequences of the target gene, IRAK-4, were constructed and transferred into MG63 cells using Lipofectamine 2000. There were three groups: blank group=MG63 cells, control group = MG63 cells transfected with scrambled IRAK-4siRNA, and silence group=MG63 cells transfected with specific IRAK-4 siRNA. The protein level of extracellular regulated kinase (ERK), c-Jun N-terminal kinase (JNK) and p38 mitogen-  activated protein kinases (p38MAPK) were detected by western blot assay. RESULTS AND CONCLUSION: The expression of IRAK-4 mRNA and protein in the silence group was significantly decreased compared with the control group. Compared with the blank and control groups, 48 hours after the transfection, IRAK-4 gene silencing in MG63 cells decreased protein expression of p-JNK1/2P46, p-ERK1/2 and p-p38MAPK  (P < 0.05). IRAK-4 silencing inhibited ERK, JNK and p38MAPK expression in osteoblast-like cells. Figures and Tables | References | Related Articles | Metrics

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Local injection of simvastatin affected reconstruction of trabecular bone of condyles of femur of osteoporotic rats Li Yang, Feng Shi-qing, Yang Ning 2013, 17 (46):  7994-7999.  doi: 10.3969/j.issn.2095-4344.2013.46.003 Abstract ( 541 )   PDF (401KB) ( 541 )   Save BACKGROUND: Local injection with simvastatin can induce osteogenesis, significantly increase the bone mineral density and mechanical strength of femoral neck and femoral condyle of rats with osteoporosis and analyze effects of local injection of simvastatin on trabecular bone of the femoral condyle. OBJECTIVE: To explore the effects of local injection of simvastatin on trabecular bone of the femoral condyle of osteoporotic rats and provide experimental basis for the application of simvastatin in clinical topical treatment of osteoporosis. METHODS: Eighteen female Sprague-Dawley rats received bilateral ovariotomy at 3 months, and were used to produce rat models of osteoporosis. They were assigned into three groups. Experimental rats received 5 and 10 mg simvastatin by single injection into right femoral cavity. Control rats received blank vector. The rat models were sacrificed at 1 month after injection and specimens were collected. Right femoral condyles were taken out for bone histomorphometric analysis by Micro-CT. RESULTS AND CONCLUSION: One month post-injection, Micro-CT scanning results revealed that cortical bone thickness, trabecular bone density and connection rate were significantly better in the simvastatin group than those in the control group. Results indicated that single injection of small-dose simvastatin obviously promoted rebuilding of trabecular bone of condyles of femur, improved microstructure of skeleton, strengthened local skeleton, prevented and treated osteoporosis, and provided a further basis for the prevention and treatment of osteoporosis, especially for osteoporotic fractures. Figures and Tables | References | Related Articles | Metrics

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Expression of transforming growth factor-beta 1 in rat embryohic cochlea Li Cui-e, Shi Yan-li 2013, 17 (46):  8000-8003.  doi: 10.3969/j.issn.2095-4344.2013.46.004 Abstract ( 273 )   PDF (312KB) ( 308 )   Save BACKGROUND: Many researches on the development of cochlea have been made, but mainly depend on the pathological conditions and developmental deformity, while the researches on the development process of normal embryonic cochlea and expression of related factors are rare. OBJECTIVE: To investigate the expression of transforming growth factor β1 in rat embryonic cochlea. METHODS: Fifty-six Sprague Dawley rats were selected and the rats were pregnancy, and the embryos were obtained from the rats, trimmed the inner ear specimens under the anatomical microscope, and then the specimens were treated with dehydration, decalcification, directly paraffin embedding and slicing processing. The morphology evolution of inner cochlea was observed under light microscope after hematoxylin-eosin staining. The expression of transforming growth factor β1 in rat embryonic cochlea was observed by immunohistochemical streptavidin biotin-peroxidase complex method. RESULTS AND CONCLUSION: The development of rat inner ear began in ectoderm thickening area of E8 phase, otocyst and the emergency of the mesenchymal could be seen in E9 phase, otocyst and ossicular chain began to develop in E9.5 phase, formation of cochlear duct anlage could be seen in E12.5 phase, cochlea tube development was completed in E18.5 phase and Corti’s formation could be seen in E16 phase. The structure and function of the inner ear could be fully developed after birth. The expression of transforming growth factor β1 could be seen in E12.5-E19 phases, and the expression was changed from weak to strong and then weakened further, and strongest in E14.5 phase. This suggested that transforming growth factor β1 may be involved during the development of rat cochlear epithelium. Figures and Tables | References | Related Articles | Metrics

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Psychological stress affects the expressions of two kinds of cytokines in rat temporomandibular joint cartilage Huang Xu, Xiao Peng, Wang Yan, Zhang Hong-yu, Liu Hai-xia 2013, 17 (46):  8004-8011.  doi: 10.3969/j.issn.2095-4344.2013.46.005 Abstract ( 293 )   PDF (519KB) ( 348 )   Save BACKGROUND: Animal modeling has reported that psychological stress can lead temporomandibular joint disease in rats, and there will be pathological changes. OBJECTIVE: To observe the effects of psychological stress on the expressions of vascular endothelial growth factor and transforming growth factor-β1 in rat temporomandibular joint condylar chondrocytes METHODS: Forty-eight adult male Wistar rats were randomly divided into the experimental group and the control group, 6 rats in the experimental group and 12 rats in the control group. Thirty-six Wister male adult rats in the experimental group were subjected to the following stimulating factors, such as electroshock, keeping stillness, climate change and food tempting in order to make them under psychological stress. Then, the experimental group rats were executed at 3 weeks after stimulation (3 weeks stimulation group), 6 weeks after stimulation (6 weeks stimulation group) and 6 weeks normal feeding after 6 week stimulation (recovery group), 12 rats were sacrificed at each time point. The 12 rats in the control group were executed at the same time as the last experimental group. The temporomandibular joint specimens were taken to produce slices. RESULTS AND CONCLUSION: The vascular endothelial growth factor-positive rate of recovery group was significantly larger than that of the 3 weeks stimulation group and the control group (P < 0.05); transforming growth factor-β1 positive rate in the 6 weeks stimulation group was significantly greater than that in the control group  (P < 0.05). It indicated that vascular endothelial growth factor was related with the new blood capillaries repairing tissue, and transforming growth factor-β1 might play a role that stimulates bone resorption in psychological stress. Figures and Tables | References | Related Articles | Metrics

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Estrogen effects on interleukin-1 expression in alveolar bone remodeling of osteoporotic rats Wang Xiu-he, Wang Chang-qing, Zhu Yu-ping, Zhang Xiao-dong 2013, 17 (46):  8012-8017.  doi: 10.3969/j.issn.2095-4344.2013.46.006 Abstract ( 311 )   PDF (415KB) ( 504 )   Save BACKGROUND: Currently rats are the most frequently used animal for the models of osteoporosis and ovariectomized rat models have been widely applied due to ovariectomized female rats are similar to human body in bone mineral density changes and response after estrogen administration. OBJECTIVE: To establish rat models of osteoporotic tooth extraction wound healing, and to investigate the effect of estrogen on the interleukin-1 expression and distribution in the remodeling of osteoporotic alveolar bone.     METHODS: Sixty-five purebred female rats, 3 months old, were randomly divided into two groups: osteoporosis model group (n=40; ovariectomy under general anesthesia) and sham operation group (n=25; fat tissue around ovary was removed). After 8-week feeding, osteoporosis models were established and the left upper molar was pulled out under general anesthesia. Histomorphomeric parameters test was performed on the jaw bone. In osteoporosis model group, 15 rats were randomly selected to give subcutaneous injection of estradiol benzoate, as the estrogen treatment group. Immunohistochemical method was applied to observe the interleukin-1 expression in the remodeling of osteoporotic alveolar bone. RESULTS AND CONCLUSION: After ovariotomy, the amount of trabecula decreased and the medullary cavity of the bone became larger, the jaw bone intensity decreased. After administration of estrogen, the positive expression of interleukin-1 was reduced as compared with osteoporosis model group. Experimental findings indicate that, osteoporosis can be detected in Sprague-Dawley female rats aged 3 months at 8 weeks after ovariotomy, and administration of estrogen can obviously decrease interleukin-1 positive expression in the remodeling of osteoporotic alveolar bone. Figures and Tables | References | Related Articles | Metrics

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Effect of hyperbaric oxygen preconditioning on expression of Bcl-2 and Bax after spinal cord injury Zhang Yu-qiang, Li You, Cao Yang, Dong Ming-yan, Lü Gang 2013, 17 (46):  8018-8023.  doi: 10.3969/j.issn.2095-4344.2013.46.007 Abstract ( 461 )   PDF (321KB) ( 402 )   Save BACKGROUND: Studies have shown that hyperbaric oxygen preconditioning can reduce neuronal apoptosis and improve neurologic dysfunction after spinal cord injury. However, the mechanism of inhibiting apoptosis is still unclear. OBJECTIVE: To study the effect and significance of hyperbaric oxygen preconditioning on expression of Bcl-2 and Bax protein after spinal cord injury in rats. METHODS: Sixty-six male adult Sprague-Dawley rats were divided into sham operation group, model group, treatment group at random. Animals in the sham group were six, and animals in the model and treatment groups were 30, respectively. Animals in the treatment group received hyperbaric oxygen preconditioning at 2.5 MPa (100% O2), once for 1 hour and totally five times within 10 days before establishment of spinal cord injury models. In the sham operation group, only laminectomy was done. In the model group, a spinal cord injury model was prepared using Allen’s method.  RESULTS AND CONCLUSION: The results of immunochemistry indicated that there were few Bcl-2 and Bax-positive cells in the sham operation group; a number of Bcl-2 and Bax-positive cells were visible in the model and treatment groups. Bcl-2 and Bax expressions reached the peak at day 7 and 3, respectively, after injury. After treatment with hyperbaric oxygen preconditioning, Bcl-2 positive cells decreased significantly, while Bax-positive cells decreased significantly (P < 0.05). It indicates that the hyperbaric oxygen preconditioning can increase Bcl-2 expression but decrease Bax expression. Figures and Tables | References | Related Articles | Metrics

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Construction of human caudal type homeobox transcription factor 2 retrovoral vector and transfection of urothelium cells Lin Ming-en, Deng Bi-hua, Lü Yi-song, Rong Lu, Yao You-sheng 2013, 17 (46):  8024-8029.  doi: 10.3969/j.issn.2095-4344.2013.46.008 Abstract ( 581 )   PDF (392KB) ( 522 )   Save BACKGROUND: Caudal type homeobox transcription factor 2 plays an important role in the development of epithelium in digestive tract, especially the small intestine and colon.  OBJECTIVE: To construct the retroviral expression vector of pLNCX2-caudal type homeobox transcription factor 2, and to observe the effect of in vitro transfection of pLNCX2-caudal type homeobox transcription factor 2 in intestinal metaplasia. METHODS: Gene recombinant technology was employed to clone human caudal type homeobox transcription factor 2 gene to the retroviral expression vector of pLNCX2, then identified with enzyme digestion and sequencing and packed to the PA317 cells. The plasmids were transfect in urothelium cells, real-time PCR and western blot were used to detect the expressions of caudal type homeobox transcription factor 2, villin and liverintestin- cadherin at the protein and mRNA levels． RESULTS AND CONCLUSION: The retroviral vector pLNCX2-caudal type homeobox transcription factor 2 was successfully constructed. The levels of caudal type homeobox transcription factor 2 protein and mRNA expressions in bladder urothelium cell transfected with pLNCX2-caudal type homeobox transcription factor 2 were higher than that in control. And the over-expression of caudal type homeobox transcription factor 2 could up-regulate the levels of villin and liverintestin-cadherin expressions. Interestingly, specific changes of intestine-like cells were seen in the bladder urothelium cells transfected by pLNCX2-caudal type homeobox transcription factor 2. Over-expression of caudal type homeobox transcription factor 2 can activate the caudal type homeobox transcription factor 2 of urothelium cells and induce intestinal epithelial differentiation, thus inducing the development of cystitis glanduaris. Figures and Tables | References | Related Articles | Metrics

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Establishment of rat models of Parkinson’s disease by unilateral two-point injection with 6-hydroxydopamine Wang Yu-ling, Gao Hua, Yang Xin-ling 2013, 17 (46):  8030-8035.  doi: 10.3969/j.issn.2095-4344.2013.46.009 Abstract ( 545 )   PDF (358KB) ( 950 )   Save BACKGROUND: The establishment of animal models of Parkinson’s disease plays an important role in clinical and basic experiments of Parkinson’s disease, and its stability directly affected study results. OBJECTIVE: To evaluate the behavior and pathological changes of rats with Parkinson’s disease established by unilateral two-point injection with 6-hydroxydopamine. METHODS: A total of 62 Sprague-Dawley rats were randomly assigned into experimental group (n=50) and normal group (n=12). Using stereotatic technique, 6-hydroxydopamine was injected into the right substantia nigra compact part and midbrain ventral tegmental area to establish models of Parkinson’s disease in the experimental group. Behavioral changes of rats, tyrosine hydroxylase and dopamine content were observed. RESULTS AND CONCLUSION: At 2 weeks, after induction with apomorphine, left rotation speed > 7 r/min was observed in only 22 rats of the experimental group. No significant difference in the number of rotation between 2 and 4 weeks was detected (P > 0.05). Tyrosine hydroxylase and dopamine content significantly reduced in the model group. Results indicated that unilateral two-point injection of 6-hydroxydopamine could be used to successfully establish rat models of Parkinson’s disease with similar behaviors and pathologies. Figures and Tables | References | Related Articles | Metrics

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Establishment of an overtraining rat model on the treadmill Zhang Cai, Yang Hua-yuan 2013, 17 (46):  8036-8042.  doi: 10.3969/j.issn.2095-4344.2013.46.010 Abstract ( 836 )   PDF (294KB) ( 459 )   Save BACKGROUND: Overtraining is a series of functional disorder or pathological state induced by continuous fatigue accumulation because exercise load and body function are incommensurate to each other. At present, commonly used methods for establishing rat models of overtraining included treadmill, swimming and climbing rod, but treadmill is comparatively accepted in the world. OBJECTIVE: To establish the standard of overtraining rat model and to implement objective of model establishment by dynamically monitoring biochemical indexes and observing behavioral changes. METHODS: A total of 16 Sprague-Dawley rats were randomly assigned to model and blank control groups. The model group received movement training according to the plan. After adaptable feeding, training was performed, 6 days every week, with a rest of 1 day. Increasing intensity on treadmill was used. From the first week of training, the speed, gradient and running time were gradually increased. However, the blank control group was conventionally fed, without any training. RESULTS AND CONCLUSION: Behavior changes of the training rats were arisen after five weeks. Serum creatine kinase levels increased continuously in training process, and higher than basic levels at 5 weeks  (P < 0.01). Serum urea nitrogen levels persistently increased, and higher than basic levels at 3 weeks (P<0.05).    Hemoglobin and serum testosterone levels increased and then decreased, and significantly lower than basic levels at 8 weeks (P < 0.05). Behaviorally, overtraining appeared. Simultaneously, hemoglobin and serum testosterone levels were significantly lower than basic levels. Serum creatine kinase and serum urea nitrogen levels were significantly higher than basic levels. These results indicated that the body was in overtraining state. The standard of overtraining rat model was established in this study. The overtraining rat model was established according to the training program when the training was lasted for 8 weeks, the training speed was 30 m/min; every training time was 110 minutes, and the gradient was 15°. Figures and Tables | References | Related Articles | Metrics

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Time for establishing an animal model of acute high intraocular pressure Zhang Kun-ya, Qian Xiu-qing, Liu Zhi-cheng 2013, 17 (46):  8043-8048.  doi: 10.3969/j.issn.2095-4344.2013.46.011 Abstract ( 342 )   PDF (315KB) ( 554 )   Save BACKGROUND: An animal model of acute high intraocular pressure offers an easy and effective method to glaucoma research. Research results will be distorted if the animal model is not induced successfully. Nowadays, a successful animal model can only be identified by observational indexes. OBJECTIVE: To observe the induction time for an animal model of acute high intraocular pressure, and thereby to provide accurate and feasible method for identifying whether the animal model is induced successfully. METHODS: Acute high intraocular pressure animal models were induced by anterior chamber perfusion using communicating vessels-based device with a glass vessel filled with normal saline and 2-mm-long mercury. Liquid volume was perfused to rabbit eyes. A total of 10 animal models were prepared successfully at an intraocular pressure of 4.9 kPa. The experiment was complete till 30 minutes after the perfused liquid was not moved. At the certain intraocular pressure, the variation of fluid inflow over time was recorded, and the induction time for successful modeling was calculated. RESULTS AND CONCLUSION: The curve between fluid inflow into rabbit eyes (y) and induction time (t) could be fitted to logistic function,        while U, b0 and b1 are 957.44±313.22, 0.023±0.008 and 0.980±0.003, respectively. The fluid inflow to the eye ball did not change when the induction time was over (282.93±42.68) minutes, and the animal model of acute high intraocular pressure was established successfully. Figures and Tables | References | Related Articles | Metrics

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Radiation metabolomics of minimally invasive urine biomarkers for X-ray radiation exposure in mice Wang Min, Pan Xiao-jing, Liu Bin, Zhang Hong 2013, 17 (46):  8049-8055.  doi: 10.3969/j.issn.2095-4344.2013.46.012 Abstract ( 423 )   PDF (416KB) ( 466 )   Save BACKGROUND: Single hematology analysis can only reflect the body injury at a certain time point after radiation damage, but cannot reflect the longer-term cumulative status after radiation damage. OBJECTIVE: To identify the biomarkers in blood and urine in mice after radiation damage with metabolomics method based on nuclear magnetic resonance hydrogen spectrum. METHODS: Forty-eight mice were randomly divided into four groups and received 0 (sham radiation), 3, 9 and 27 Gy radiation. The blood samples were collected at 24 hours and 5 days after radiation. Another 36 mice were collected and divided into three groups and received 0 (sham radiation), 9 and 27 Gy radiation, then the urine samples were collected at 2 days before radiation and 5 days after radiation for 24 hours. The blood and urine samples were analyzed with proton nuclear magnetic resonance spectroscopy. RESULTS AND CONCLUSION: The content of aspartic aminotransferase and alanine aminotransferase in blood plasma maintained a stable level after 3- or 9-Gy X-ray radiation, but the level of alkaline phosphatase in blood plasma was increased significantly after 9-Gy radiation, which indicating that low-dose head radiation may cause increased radiation damage and repair. The level of total superoxide dismutase in blood plasma was significantly decreased at 5 days after radiation which indicating that head radiation in mice could cause systemic oxidative stress. Meanwhile, the N-hexamolglycine and -thymidine level in the urine samples was significantly increased after different doses X-ray radiation, which can be used as the radiation damage markers in urine samples after head radiation. The 3-hydroxy-2-methylbenzoic acid 3-O-sulfate level in urine samples was increased by 2.5 times after received 9-Gy radiation, which can be used as the specific markers of middle- and low-dose radiation damage; the level of taurine in the urine samples was increased by 20% after 27-Gy radiation, which can be used as the specific markers of high-dose radiation damage.  Figures and Tables | References | Related Articles | Metrics

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Cationic liposome-mediated enhanced green fluorescent protein plasmid transferred into skeletal muscle satellite cells Xu Zhi-feng, Li Jing-lai, Han Zhen, Feng Gang, Ren Ming-ming 2013, 17 (46):  8056-8061.  doi: 10.3969/j.issn.2095-4344.2013.46.013 Abstract ( 609 )   PDF (545KB) ( 464 )   Save BACKGROUND: Skeletal muscle satellite cells are totipotential stem cells with multi-directional differentiation potential, locate in skeletal muscle interstitium, have a certain tolerance to ischemia and hypoxia, and are important cells in stem cell engineering. OBJECTIVE: To establish a thrifty, convenient culture procedure and create a simple, efficient method to transfect skeletal muscle satellite cells, and investigate genetic expression after the transfection for cellular cardiomyoplasty. METHODS: Skeletal muscle satellite cells were isolated from rabbit thigh and cultured. Their growth curves were determined by CKK-8 method. Grouped by different proportions of the plasmid and liposome, skeletal muscle satellite cells were transfered by the enhanced green fluorescent protein plasmid based on liposome. After transfection, the efficiency and character of target genetic expression was determined. RESULTS AND CONCLUSION: Satellite cells were isolated, cultured and transfected successfully. In suitable ratio of plasmid and liposomes, the transfection efficiency reached up to above 35%. The target protein was expressed within   12 hours after transfection, reached maximum in 48-72 hours and decreased gradually after one week. The expression still could be observed two weeks latter. The enhanced green fluorescent protein plasmid conducted by cationic liposome could be transfered into skeletal muscle satellite cells efficiently. The transfection efficiency was correlated closely to the ratio of plasmid and lipofectamine. The change of target gene expression depended on time. Figures and Tables | References | Related Articles | Metrics

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Effect of small interfering RNA on gene expression of synovial cells in patients with rheumatoid arthritis Hou Chun-feng, Sun Min, Li Shu-jie, Zhao Jian-hong, Wang Ji-bo 2013, 17 (46):  8062-8068.  doi: 10.3969/j.issn.2095-4344.2013.46.014 Abstract ( 451 )   PDF (435KB) ( 455 )   Save BACKGROUND: The etiological factor for rheumatoid arthritis remains unclear, but the effects of nuclear factor-κB on the onset of rheumatoid arthritis have been gradually paid great attention by rheumatologists. OBJECTIVE: By using the RNA interference (RNAi) technique to block the signal pathway of nuclear factor-κB mRNA of human rheumatoid arthritis synovial cells, this study explored its application prospect in the treatment of rheumatoid arthritis. METHODS: The synovial cells were isolated, digested, and cultured for further use. In accordance with the design principle of small interfering RNA (siRNA), target sequences of siRNA of nuclear factor-κB were identified, and siRNA expression vector of nuclear factor-κB was synthesized and constructed. The four pGenesil-1/nuclear factor-κB siRNA expression vectors were transfected into the first passage of synovial cells that well grew. Blank and negative control groups were set. Cells at 12, 24, 48, 72 hours, 5 and 7 days after transfection were collected, and RNA was extracted. Intracellular nuclear factor-κB mRNA expression levels were measured, and siRNA  plasmid vector that could effectively inhibit nuclear factor-κB mRNA expression was screened out. RESULTS AND CONCLUSION: Nuclear factor-κB highly expressed in synovial cells after human rheumatoid arthritis. 3#pGenesil-1/nuclear factor-κB apparently suppressed nuclear factor-κB mRNA expression in synovial cells with human rheumatoid arthritis. RNAi technique blocked nuclear factor-κB mRNA expression. Therefore, the block of nuclear factor-κB signal pathway might be a good target for rheumatoid arthritis gene therapy. Figures and Tables | References | Related Articles | Metrics

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Optimal animal model of osteoarthritis He Ming-jiang, Zhang Hong-mei, Jing Lin, Tao Zhi-kun, Hu Rui 2013, 17 (46):  8069-8074.  doi: 10.3969/j.issn.2095-4344.2013.46.015 Abstract ( 655 )   PDF (376KB) ( 1028 )   Save BACKGROUND: There are a variety of methods for establishing osteoarthritis animal model, and different methods and different animals have their own characteristics. OBJECTIVE: To review the status of the research on osteoarthritis animal models. METHODS: The relevant articles to the osteoarthritis animal models were searched in Medline database from January 2002 to October 2011, with the key words of “animal model, osteoarthritis” in English by the computer. Similarly, Chinese Journal Full-text Database was retrieved for related articles published in Chinese from January 2002 to October 2011, with the key words of “animal model, osteoarthritis, research development” in Chinese. Totally 423 relevant articles were collected and 128 of them conformed the standard. At last 40 articles were included to review after full-text reading. RESULTS AND CONCLUSION: The artificial induced animal models are the principal means to establish animal models of osteoarthritis and they have been widely used in the study of osteoarthritis pathogenesis, drug efficacy, cartilage physiology and pathology. Spontaneous osteoarthritis animal models have great advantages in the study addressing the initial mechanism of osteoarthritis, the biochemical changes of articular cartilage and the comparison of prevention and cure effects. Transgenic animal models have great application prospects on studying cartilage repairing mechanisms and testing a gene to prevent, delay or reversal the morphological changes. Accordingly the choice of models should be based on research needs. Figures and Tables | References | Related Articles | Metrics

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Alzheimer’s disease transgenic animal models: How to get more similar pathological characteristics? Dong Xian-hui, Chai Xi-qing 2013, 17 (46):  8075-8082.  doi: 10.3969/j.issn.2095-4344.2013.46.016 Abstract ( 563 )   PDF (412KB) ( 2393 )   Save BACKGROUND: Alzheimer’s disease causes and pathogenesis remain unclear, which greatly restrict the screening of drugs. And the main reason is lack of suitable animal models. The developing transgenic animal technology allows studying the role of certain pathogenic gene in vivo, and has regarded the ideal animal models for Alzheimer’s disease. OBJECTIVE: To summarize the research advance of Alzheimer’s disease transgenic animal models. METHODS: Using “Alzheimer’s disease, transgenic mouse, animal model, dementia” in Chinese and English as the key words, the first author retrieved PubMed and CNKI databases published before July 2013. Finally, 41 articles were included in result analysis. RESULTS AND CONCLUSION: The etiology of Alzheimer’s disease is diverse, and genetic factor is one important factor. The existing transgenic animal models of Alzheimer’s disease include single genetically modified models, double genetically modified models and multiple transgenic models. Single transgenic animal models can make a kind of mutated exogenous gene integrate into the genomes of animals by using recombinant DNA technology. This kind of models can be applied to only study one specific pathological change of Alzheimer’s disease. Double transgenic animal models can make two kinds of mutated exogenous gene integrate into the genomes of animals and simultaneously transfect animals by using recombinant DNA technology. This kind of models is closer to the pathological changes of Alzheimer’s disease than single transgenic animal models, but still cannot simulate Alzheimer’s disease. Multiple genetically modified models are obtained with different transgenic mice hybridization or several genes transfection, which are most similar to clinical process and pathological features of Alzheimer’s disease. However, this kind of models may develop a decline in consanguinity. Each kind of animal model has their advantages and shortcomings, and a better transgenic animal model is urgently needed to completely simulate pathological characteristics of Alzheimer’s disease. Figures and Tables | References | Related Articles | Metrics

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Hip stress technical strength training and evaluation: Document comparison and logical analysis Ge Qi, Hong Tao, Wang Bing-cheng 2013, 17 (46):  8083-8089.  doi: 10.3969/j.issn.2095-4344.2013.46.017 Abstract ( 293 )   PDF (413KB) ( 453 )   Save BACKGROUND: At the beginning of the 20th century a strength training method in the backward direction appeared: hip stress strength training method. There are many articles focusing on the concept, function, effect and mechanism of this training method. So far, relevant research has formed a unique branch of special strength training research directions. OBJECTIVE: To conclude the concept, function, effect, features and mechanisms of hip stress technical strength training through the systematic analysis of literature concerning hip stress strength training. METHODS: CNKI (1998/2010) and Wanfang databases were searched by the first author for relevant articles. The keywords were “hip stress, strength training, trajectory control, exercise training, muscle strength, knee, physical fitness, bone” in Chinese and English. The obtained data were summarized using contrast method and logic analysis method. RESULTS AND CONCLUSION: Totally 90 papers were retrieved, and finally 34 papers were included. The results showed that the hip technical strength training method is the most effective training method currently to cultivate the running, jumping and throwing back pedaling force after hip extension forces. It has a new training idea and special training equipments, which can effectively solve many sports difficulties in strength training and  technical training and has the vital significance for the high-level sports training, skills assessment and sports teaching. Hip stress special technical strength training research has formed a special research branch of special strength trainings, which will play a more significant research role. Figures and Tables | References | Related Articles | Metrics

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Technology progress in the in vitro construction and culture of tissue-engineered bone Liu Yan-xiao, Bei Kang-sheng 2013, 17 (46):  8090-8095.  doi: 10.3969/j.issn.2095-4344.2013.46.018 Abstract ( 285 )   PDF (391KB) ( 377 )   Save BACKGROUND: The in vitro construction, maturation and differentiation of cell scaffold complexes into tissue- engineered bone is the necessary process of bone tissue engineering construction, but there are no uniform methods and standards. OBJECTIVE: To summarize the basic method and technology to build bone tissue engineering at present and to discuss the related development. METHODS: A compute-based online search was conducted on the PubMed database and CNKI database for the articles related to the in vitro construction and culture of bone tissue engineering bone from January 1997 to January 2013 with the key words of “bone tissue engineering, cell biological scaffold, cell inoculation, seeding density, culture in vitro, bioreactor” in English and Chinese. Finally, 44 articles were included according to the inclusion and exclusion criteria. RESULTS AND CONCLUSION: As the carrier of bone tissue engineering seed cells, the primary prerequisite of biological scaffold is sterile, because the sterile biological scaffold can be able to survive. Sterilization of biological scaffolds includes ultraviolet sterilization, 60Co γ-ray sterilization, soaking in ethanol with the volume fraction of 75%, autoclave method, and ethylene oxide sterilization. 60Co γ-ray sterilization is the common method in the biological scaffold sterilization. The inoculation density of seed cells is the key factors that influence the adhesion growth and proliferation of seed cells on the scaffolds. The adhesion between cells and scaffold materials will be affected by the affinity of scaffolds, cell adhesion and gravity, and other factors. The method for the inoculation of bone tissue engineering seed cells includes static inoculation and dynamic inoculation. Each construction method has its advantages and disadvantages. Overcomeing these disadvantages, forming a uniform construction method and fully clinical application are the direction of future development. Figures and Tables | References | Related Articles | Metrics

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Cell therapy of chronic wound healing Yan Long-zong, Chen Bin 2013, 17 (46):  8096-8101.  doi: 10.3969/j.issn.2095-4344.2013.46.019 Abstract ( 474 )   PDF (378KB) ( 775 )   Save BACKGROUND: The chronic wounds, also called non-heading wounds, can seriously affect the quality of life of patients and has brought heavy burden to patients, as well as health care professionals. Cell therapy is a new method for promoting wound healing and plays an important role in the repair of chronic wounds. OBJECTIVE: To summarize the progress of researches on the chronic wound healing mechanism and cell therapy, and to provide evidences for the clinical management of chronic wounds and relative basic researches. METHODS: A computer search of CNKI database from 2005 to 2012, PubMed database from 1995 to 2012 and Foreign Medical Journal Full-Text Service database from 2000 to 2012 was performed using “non-healing wounds, diabetic foot ulcer, wound healing, cell therapy” in Chinese and English as the key words to retrieve articles about chronic wound healing mechanism and the application of cell therapy. Totally, 42 articles meeting the inclusive criteria were included in result analysis。 RESULTS AND CONCLUSION: The wound healing is a complex biological process, involving multiple cell types, extracellular matrix and cytokine factors. The delayed healing of refractory wound seriously affects the quality of life of patients and has brought heavy economic burden to patients. At present, many methods have been employed to promote wound repair, such as local hyperbaric oxygen therapy, surgical treatment, herbal Chinese medicine, application of various growth factors, cell therapy and gene therapy. Cell therapy is noninvasive and those delivered cells can adapt to their environment, are able to release growth factors and cytokines, and more importantly, are able to deliver the growth factors for the wound healing process due to cell signaling capabilities. Currently, cells used for the treatment of chronic wound cells mainly include bone marrow stem cells, bone marrow-derived mesenchymal stem cells, cord blood stem cells, peripheral blood stem cells, epidermal stem cells, skin-derived progenitor cells, adipose stem cells, fibroblasts and platelets. Figures and Tables | References | Related Articles | Metrics

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Biological mechanism, clinical application and medical experiment of millimeter waves Luo Qing-lu 2013, 17 (46):  8102-8107.  doi: 10.3969/j.issn.2095-4344.2013.46.020 Abstract ( 736 )   PDF (444KB) ( 1005 )   Save BACKGROUND: Millimeter wave has been applied in clinic for about 20 years, while the mechanisms and clinical curative effect are still undefined and lacked of evidence based medicine.   OBJECTIVE: To review the clinical application and mechanism of millimeter wave in various diseases in the past 10 years. METHODS: A computer-based online search was performed in the CNKI database, China Standard database and Science Direct database from January1998 to December 2012 with the key words of “millimeter wave, biological mechanisms, experimental studies, clinical application” put in the title and abstract. The articles related to the biological effect of millimeter wave were included, and finally, 68 articles were included for review. RESULTS AND CONCLUSION: When the millimeter wave used in the human body, it can only go through the skin, but its energy can resonance with some molecules in the tissues and provide the treatment effects. Millimeter wave therapy has a good affinity for aqueous tissue, which can improve the metabolism and blood circulation of partial tissues, and can enhance the metabolic and pathological product absorption and excretory, thus can eliminate inflammatory, alleviate swell and relieve pain. Millimeter wave filed has comprehensive Figures and Tables | References | Related Articles | Metrics

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Meta-analysis on the effect of negative pressure therapy in body surface wound treatment Bai Ming, Zhao Ru, Wang Zhi, Long Xiao, Zeng Ang, Zhang Hai-lin, Wang Xiao-jun 2013, 17 (46):  8108-8115.  doi: 10.3969/j.issn.2095-4344.2013.46.021 Abstract ( 444 )   PDF (365KB) ( 420 )   Save BACKGROUND: Negative pressure wound therapy has been widely recognized, the currently published papers are limited in academic value and lack of scientific, objective, qualified index to confirm the therapy effectiveness. OBJECTIVE: To systemically evaluate the clinical effect of negative pressure wound therapy, provide more evidence for its clinical application, and guide clinical research. METHODS: Fifteen articles were screened out of peer-reviewed publications (Cochran library, Embase, PubMed-Medline and Chinese BioMedical Literature Database). Scientific data were collected and evaluated by two researchers. The data were statistically analyzed with RevMan software. RESULTS AND CONCLUSION: Only 15 random-controlled trials were finally preserved, including 10 as B-grade moderate bias risk and focused on the effect of negative pressure wound therapy on chronic wounds, and 5 as C-grade high bias risk and focused on the effect of negative pressure wound therapy on acute wounds. There were significant differences in the main outcome measures between negative pressure wound therapy and conventional wound therapy. As for chronic wound patients, no significant difference was observed in the operation-preparing period, reducing wound area, promoting wound granulation, and amputation rate between two therapies. As for acute wound patients, the differences were significant in the operation-preparing period, promoting wound granulation, wound infection rate, and cost materials between two therapies. However, no difference was significant in the healing of wound and hospitalization time. Our findings indicate that, negative pressure wound therapy is an effective means for both acute and chronic wounds, it can shorten operation-preparing period, promote wound granulation, and reduce amputation rate and infection rate, thus providing evidence for clinical application. The well-designed study is needed to develop high-quality random controlled trails. Figures and Tables | References | Related Articles | Metrics

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Stem cells in cartilage tissue engineering and influential factors Wang Yan 2013, 17 (46):  8116-8121.  doi: 10.3969/j.issn.2095-4344.2013.46.022 Abstract ( 278 )   PDF (397KB) ( 401 )   Save BACKGROUND: A variety of stem cells as seed cells for cartilage tissue engineering have been extensively studied. OBJECTIVE: To analyze the biological characteristics of different kinds of stem cells and their application in cartilage tissue engineering. METHODS: By retrieving recent studies on sources of stem cells and influential factors in cartilage tissue engineering, the authors focused on optimization of harvesting stem cells from different sources in bone tissue engineering, problems, and factors that affect cell growth, differentiation, reproduction and metabolism, which provide a theoretical basis for tissue-engineered cartilage repair. RESULTS AND CONCLUSION: A large number of experimental studies have confirmed that exogenous stem cells can be used as seed cells for cartilage tissue engineering. Influential factors relative to cartilage tissue engineering construction using seed cells include growth factors, cell scaffold, cell seeding density, oxygen concentration, stress and micro-gravity. However, tissue-engineered cartilage is still in the stage of laboratory research, and stem cells for cartilage tissue engineering involves many issues, such as stem cell transformation, in vitro culture of stem cells, how to prolong the survival of cells, reduce cell antigenicity and enhance the host’s immune tolerance, which are yet to be resolved. Figures and Tables | References | Related Articles | Metrics

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Tissue-engineered cartilage repair for sports-induced articular cartilage injury Zhang Lu-yao 2013, 17 (46):  8122-8127.  doi: 10.3969/j.issn.2095-4344.2013.46.023 Abstract ( 357 )   PDF (412KB) ( 357 )   Save BACKGROUND: The emergence and development of tissue engineering technology provides a new idea for reconstruction of joint functions after sports-induced articular cartilage injuries, and realizes complete regeneration of cartilage tissue. OBJECTIVE: To understand the basic principles of tissue engineering research, to analyze factors influencing tissue-engineered cartilage construction, and to explore the feasibility of tissue-engineered cartilage repair for sports-induced articular cartilage injury. METHODS: Through the retrieval of literatures on the construction and application of tissue-engineered cartilage, we analyzed the feasibility of new cartilage formation in vivo using tissue engineering technology, focused on tissue-engineered cartilage construction and its application in cartilage injury and repair, thereby providing a theoretical basis for tissue-engineered cartilage repair of sports-induced cartilage injuries. RESULTS AND CONCLUSION: Seed cells, scaffolds and in vitro culture environment constitute two elements of cartilage tissue engineering, which, as a whole, promote and restrict each other. The proper configuration of seed cells, scaffolds and in vitro culture environment is the key issue to be solved in the treatment of sports-induced cartilage injuries. Figures and Tables | References | Related Articles | Metrics

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Knee muscle function recovery of patients with knee osteoarthritis: Treatment and evaluation Li Yi,Yao Jian-feng, Wu Liang, Liang Xiao-jun, Chen Peng, Ma Yan 2013, 17 (46):  8128-8132.  doi: 10.3969/j.issn.2095-4344.2013.46.024 Abstract ( 339 )   PDF (345KB) ( 540 )   Save BACKGROUND: Muscle atrophy and weakness exist around the knee joint of patients with knee osteoarthritis, comprehensive treatment combined with training muscle power around knee joint can obviously relieve symptoms and restore muscle power around the knee joint. OBJECTIVE: To test the relationship between muscle function around the knee joint and the knee joint function with isokinetic muscle strength test before and after comprehensive treatment of knee osteoarthritis, and to investigate the pathological mechanisms of knee osteoarthritis in order to provide basis for the clinical rehabilitation and treatment.  METHODS: We screened out 30 subjects with unilateral knee osteoarthritis (stage Ⅱ and Ⅲ). Hospital for Special Surgery was used to evaluate the knee function, and visual analogue scale score was used to assess the patient pain. The knee muscle function of knee osteoarthritis patients was measured before treatment; the patients with knee osteoarthritis at stage Ⅱ and Ⅲ received comprehensive treatment for at least 1 month (active quadriceps isometric muscle strengthening exercises, oral celecoxib capsules combined with intra-articular injection of sodium hyaluronate), and knee muscle function was measured after treatment. Hospital for Special Surgery and the visual analogue scale score were evaluated during follow-up period. RESULTS AND CONCLUSION: Compared with the healthy side, the muscle power of the affected side of the patients with knee osteoarthritis was decreased when the flexion and extension muscle peak torque and work volume tested at 60 (°)/s and 180 (°)/s (P < 0.05). After comprehensive treatment, the Hospital for Special Surgery scores of the knee osteoarthritis patients were increased significantly (P < 0.05), while the visual analogue scale scores were decreased (P < 0.05), and the muscle power of the affected side was increased significantly (P < 0.05); there was no significant difference in the muscle power between affected side and healthy side after treatment (P > 0.05). There was no significant difference in the peak torque ratio of quadriceps and hamstring measured at 60 (°)/s in the knee osteoarthritis patients before and after treatment (P > 0.05), and there was significant difference measured at 180 (°)/s (P < 0.05). Knee osteoarthritis patients have decreased muscle strength around the knee joint, and the comprehensive treatment of muscle functional training combined with effective pain relief and nutrition joint drugs can significantly relieve symptoms, improve muscle strength and restore knee function Figures and Tables | References | Related Articles | Metrics

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Ten-year prospective study on adult obesity in Jiangsu province Li Sen, Huang Hui-ming, Xu Hao, Huang Jian-ya 2013, 17 (46):  8133-8140.  doi: 10.3969/j.issn.2095-4344.2013.46.025 Abstract ( 320 )   PDF (454KB) ( 410 )   Save BACKGROUND: Longitudinal study of changes in obesity is an important method to explore the etiology, which can provide scientific basis for preventing and controlling obesity. OBJECTIVE: To investigate the effect of age, observation period and birth cohort on the obesity prevalence of adults in Jiangsu province through the age-period-cohort analysis. METHODS: 20-69-year-old adults in Jiangsu province were collected as the research objects. The stratified cluster sampling method was used to collect the obese data in 2000-2010, and analyzed with SAS software. RESULTS AND CONCLUSION: Obesity prevalence of 1946-1950 birth cohort to 1976-1980 birth cohort was gradually increased (P < 0.05) from 2000 to 2010. Obesity prevalence from 1931-1935 birth cohort to 1941-1945 birth cohort was not significantly increased (P > 0.05). With the increasing age in each age group of over 25 years old, the risk of obesity was increased gradually. There were significant differences in the odds ratios between the baseline groups of 20-25 years old and the age groups of over 25 years (P < 0.05). Compared with the 1995-1999 birth cohort, the other periods had a slight influence on the obesity prevalence (P > 0.05). Compared with 1976-1980 birth cohort, there were no significant differences in the risks of obesity of the birth cohorts after 1951-1955 (P > 0.05). But the risk of obesity from 1946-1950 birth cohort to 1931-1935 birth cohort was gradually increased (P < 0.05). The 1936-1940 birth cohort ranked in the highest risk of obesity (odds ratio=2.93). With the increasing of the age, the risk of obesity was increased gradually; the impact of era changes on obesity was not significant since the 2000; obesity risks of those born in the different times were different; rural area will be the key area of obesity control in the future, and the key population of the excessive growth in obesity control was those aged less than 45 years old. Figures and Tables | References | Related Articles | Metrics