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11 June 2011, Volume 15 Issue 24 Previous Issue    Next Issue

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Point implantation of allograft chondrocytes for repairing articular cartilage defects Chen Yi-quan, Shen Hui-liang, Yong Yi-min, Hu Huai-jian 2011, 15 (24):  4371-4374.  doi: 10.3969/j.issn.1673-8225.2011.24.001 Abstract ( 83 )   PDF (1390KB) ( 344 )   Save BACKGROUND: Although many biological reconstruction can repair defects and rebuild articular surfaces, the function is inferior to normal cartilage. Transplantation of biodegradable polymer-coated cartilage cells may obtain real hyaline cartilage. OBJECTIVE: To observe effects of collagen-embedded chondrocytes by point implantation method on repairing articular cartilage defects. METHODS: Articular cartilage defects were made in the knee of New Zealand rabbits. The rabbits were randomly divided into three groups, treated with collagen gel-embedded chondrocytes, chondrocytes or no treatment. RESULTS AND CONCLUSION: Histological changes were observed at 2, 4, 12 and 24 weeks postoperatively. Hyaline cartilage was observed in rabbits following transplantation of collagen gel-embedded chondrocytes, while only fibrous tissues were observed in the defects in the other two groups. Moreover, mean histological and histochemical scores were greater in the rabbits treated with collagen gel-embedded chondrocytes compared with the other groups (P < 0.01). Results show that transplantation of collagen gel-embedded chondrocytes can obtain hyaline cartilage, and can be used for repairing large-area cartilage defects. Related Articles | Metrics

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Effect of the small molecular weight peptide on expression of osteoprotegerin and receptor activator of nuclear factor-kappa B ligand in MC3T3-E1 cells Wu Kun-peng, Yang Qiao-zhu, Ma Feng-yan, Dai Yong-wu, Liu Yi 2011, 15 (24):  4375-4378.  doi: 10.3969/j.issn.1673-8225.2011.24.002 Abstract ( 138 )   PDF (1079KB) ( 656 )   Save BACKGROUND: The in vivo experiment indicated that the small molecular weight peptide (KP) can treat osteoporosis in ovariectomized rat by increasing the level of calcium (Ca) and bone mineral density. The in vitro experiment indicated that KP can induce the proliferation, differentiation and mineralization of MC3T3-E1 cells by inhibiting nuclear factor-κ B p50 and p65. It is unknown whether KP affects the expression of osteoprotegerin (OPG) and receptor activator of nuclear factor-κB ligand (RANKL) in osteoblasts. OBJECTIVE: To investigate the effect of KP on the expression of OPG and RANKL in MC3T3-E1. METHODS: MC3T3-E1 were cultured with various concentrations of KP (0, 50 and 100 mg/L). Osteoblasts cultured without KP were assigned as control. Collected cells and analyzed the expression of OPG and RANKL with Western Blot after treating for 3, 6, 12, 18, 24, and 30 days. RESULTS AND CONCLUSION: After 50 mg/L and 100 mg/L KP treatment, the OPG expressions in MC3T3-E1 cells were gradually increased (P < 0.01), and KP had no effect on the expression of RANKL. Compared with control, KP obviously increased the OPG/RANKL ratio (P < 0.01). Therefore, we think that KP affects the system of OPG/RANKL and indirectly inhibits osteoclast number and functions by increasing the expression of OPG, but RANKL. Related Articles | Metrics

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A loading device for study of bone mechanobiology Xu Qiang, Zhang Chun-qiu, Guo Yong, Wu Han, Dong Xin 2011, 15 (24):  4379-4382.  doi: 10.3969/j.issn.1673-8225.2011.24.003 Abstract ( 96 )   PDF (609KB) ( 382 )   Save BACKGROUND: Articular cartilage cells respond to mechanical stimulation to maintain the morphological structure near cartilage of human body. A loading device can provide mechanical environment for studying mechanobiology.  OBJECTIVE: To construct a dual-frequency loading device according to the bionic principle of tissue engineering. METHODS: This mechanical environment was achieved by using dual-frequency loading method and implementing the coupling of low-frequency high-amplitude mechanical loading and high-frequency low-amplitude mechanical loading by controlling amplitude and frequency of cam and piezoelectric ceramics respectively, meanwhile, finite element method was used to make mechanics analysis on cartilage. RESULTS AND CONCLUSION: In the light of the principle of simulation, a dual-frequency loading device has been developed for bone mechanobiology study. Under dual-frequency loading condition, mechanical loading on the surface layer of cartilage was largest, followed by the middle layer, smallest on the deep layer; when high-frequency of 10 and 20 Hz superpositioning low-frequency of 1 and 2 Hz, the cartilage show different mechanical responses; but the difference between two response was little. The construction of the mechanical environment of the device may be helpful to tissue engineering development and clinical applications; in addition, it will be tested with biological experiment in future. Related Articles | Metrics

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Expression of bone morphogenetic protein-2 during the distraction process in midpalatal suture of rats Hu Hai-kun, Zhou Jing, Wang Yao, Li Jing, He Wu-lin, Zou Shu-juan 2011, 15 (24):  4383-4386.  doi: 10.3969/j.issn.1673-8225.2011.24.004 Abstract ( 104 )   PDF (766KB) ( 320 )   Save BACKGROUND: Orthodontists usually use palatal suture expansion to solve the problem of maxillofacial deformity. Some studies have shown that bone morphogenetic protein-2 (BMP-2) can induce bone and cartilage formation, promote the distraction osteogenesis in the process of bone remodeling under mechanical tension forces. However, underlying mechanisms of BMP-2 that drive bone formation during palatal suture expansion remain unknown. OBJECTIVE: To investigate the law of BMP-2 expression in mid-palatal suture remodeling after subjecting to tensional strain loading. METHODS: Wistar male rats of 5 weeks old, with average weight of 70-80 g, were randomly divided into 2 groups, separately, Rats in experimental group were placed on the expansion appliance, with opening loops with an initial force of 50 g applied to mid-palatal suture for periods of 1, 4 ,7 and 14 days. After expansion, the mid-palatal suture was observed by IHC staining and real time-PCR to investigate the expression of BMP-2 protein and mRNA during mid-palatal suture remodeling after suture expansion. RESULTS AND CONCLUSION: Sections with IHC staining revealed BMP-2 was detected in the osteoblast, osteocytes, chondrocytes, also in the connective tissue cells of suture. Coincident with IHC, the result of real-time PCR demonstrated time-differential expression of the mRNA that was significantly higher than that of controls at all-time points during the expansion suture remodeling. It is suggesting that BMP-2 was susceptible to mechanical tension forces and may play an important role in the remodeling of mid-palate suture. Related Articles | Metrics

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Preparation of rat femoral fracture model with an external fixator Zhao Zhen-yu, Shao Lin, Liu Jian-yu, Yang Da-ping, Hao Chen-guang, Zhao Hong-mei, Liu Guo-feng 2011, 15 (24):  4387-4390.  doi: 10.3969/j.issn.1673-8225.2011.24.005 Abstract ( 126 )   PDF (704KB) ( 630 )   Save BACKGROUND: The external fixator has strong anti-rotation capability, thus, it not affect local blood supply or binding joints, which is suitable for preparing bone fracture models.  OBJECTIVE: To design a new type animal model to study the fracture. METHODS: Twenty-four right SD rats were performed right femoral osteotomy and fixed by a newly developed external fixator. Gross observation, imaging observation and histological method were used to observe the femoral positioning and healing of osteotomized bone ends. RESULTS AND CONCLUSION: The external fixator maintained in the correct location of femur, with no signs of screw pull out, pinhole loosen, or fracture. Callus formation could be vague seen at 2 weeks after fixation. There were successive callus at 4 weeks, but the fracture end still clear. At 8 weeks after fixation, bone union could be found. The results demonstrated that external fixator utilized has sufficient stability for the management of fracture. This is a successful model of fracture in rats. Related Articles | Metrics

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Different types of rat cerebral ischemia models after middle cerebral artery occlusion Zhang Qian, Mao Shan-ping, Li Tao, Tan Jie 2011, 15 (24):  4391-4394.  doi: 10.3969/j.issn.1673-8225.2011.24.006 Abstract ( 91 )   PDF (1352KB) ( 514 )   Save BACKGROUND: Suture method caused transient middle cerebral artery occlusion (MCAO) is the commonly used method to study focal cerebral ischemia. However, differences exist in cerebral ischemia models may lead to deviation of the experimental results. OBJECTIVE: To analyze the types of cerebral ischemia in rats after MCAO method and its influential factors in different types. METHODS: Referring to Longa’s method, 166 rats were prepared for MCAO models and scanned by MRI at 24 hours after the surgery. The rats were divided to cortex infarction group, subcortical infarction group and non-infarction group according to the scan results, and to analyze the insertion depth of suture in the surgery. RESULTS AND CONCLUSION: The insertion depth were (19.9±0.9), (19.0±1.1) and (17.7±1.3) mm in the cortical infarction group, subcortical infarction group and non-infarction group, respectively. There was significant difference in the depth of suture insertion between the three groups of rats (P < 0.01). It is indicated that different types of cerebral infarction in rats may be due to the different insertion depths of suture. The insertion depth of suture is deeper into the blood vessel, and the risk of cortical infarction may be higher. Related Articles | Metrics

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Establishment of non-alcoholic fatty liver model in mice Wang Jun-jie, Fang Hui-long, Li Chun-wei, Chen Fu-chun,Tang Wei-jun, Jiang Xian-yong, Wu Hai-yan, Chen Dong-lin 2011, 15 (24):  4395-4399.  doi: 10.3969/j.issn.1673-8225.2011.24.007 Abstract ( 240 )   PDF (1376KB) ( 969 )   Save BACKGROUND: Methods for inducing non-alcoholic fatty liver model (NAFLM) should be improved. OBJECTIVE: To establish NAFLM in mice by high fat feeding and carbon tetrachloride. METHODS: Kunming mice were randomly divided into three groups: control, high fat, and high fat plus carbon tetrachloride. Liver morphological and pathological changes were observed at 2, 4, 6 and 8 weeks. Serum levels of total cholesterol (TC), triglyceride (TG), alanine transaminase (ALT) and aspartate transaminase (AST) and hepatic TC, TG changes were determined. RESULTS AND CONCLUSION: In high fat plus carbon tetrachloride group, inflammatory cell infiltration appeared in the mice liver after 2 weeks. There were a small amount of lipid droplets at 4 weeks, and steatosis emerged after 6 weeks. Hepatic fibrosis began to appear at 8 weeks. The serum levels of total TC, TG, ALT and AST, and hepatic TC, TG were significantly advanced in high fat plus carbon tetrachloride group compared with control group at 8 weeks (P < 0.05 or P < 0.01). The NAFLM can be established successfully by feeding with high fat food and injecting 5% carbon tetrachloride. Related Articles | Metrics

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Establishment of rat models of polysystic ovarian syndrome characterized by skeletal muscle insulin resistance Wang Kai-jun, Wang Luan, Ma Rui-xin, Zhao Wen-juan 2011, 15 (24):  4400-4404.  doi: 10.3969/j.issn.1673-8225.2011.24.008 Abstract ( 154 )   PDF (1668KB) ( 753 )   Save BACKGROUND: Studies have demonstrated that insulin resistance plays an important role in the occurrence and development of polycystic ovarian syndrome (PCOS). Establishing rat models of PCOS characterized by skeletal muscle insulin resistance is the basis for studying PCOS. OBJECTIVE: To establish the ideal rat models of PCOS characterized by skeletal muscle insulin resistance. METHODS: Female Sprague-Dawley rats, aged 8-9 weeks, were randomized to model group and control group with 20 animals in each group. Insulin and human chorionic gonadotrophin were administered to the model group rats which were fed with high fat diet and 50 g/L glucose solution. The rats in the control group were given normal diet and injected normal saline daily. After 6 weeks of treatment, the body weight was measured and then all of rats were sacrificed. Ovaries were taken out for histological observation. Serum concentrations of estradiol, testosterone, progestogen, follicle-stimulating hormone and luteinizing hormone were detected. Fasting blood glucose and insulin levels were measured for evaluating the insulin resistance. The expression of glucose transporter 4 in the skeletal muscle was detected by immunohistochemical staining. RESULTS AND CONCLUSION: The ovarian size in the model group was larger than that in the control group. The ovaries of the model group showed multiple follicular cysts. The fasting blood glucose, fasting insulin, testosterone and luteinizing hormone levels in the model group were respectively higher than those in the control group. While, the expression of glucose transporter 4 in PCOS rats was significantly lower than that of the control group. In model group, the intracytoplasmic granules of glucose transporter 4 were mostly located in the cytoplasm, however, in the control group, they were near the muscle cellullar membrane. Therefore, the administration of insulin and human chorionic gonadotrophin combined with high fat diet and 50 g/L glucose solution is an ideal method to establish the rat models of PCOS with significant muscle insulin resistance. Related Articles | Metrics

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Construction of rabbit models with thigh soft tissue injury and repair effect of traditional Chinese medicine Diechuangsan  Zhang Gui-fu, Wang Zhi-yi, Liu Dan-ping 2011, 15 (24):  4405-4408.  doi: 10.3969/j.issn.1673-8225.2011.24.009 Abstract ( 101 )   PDF (1474KB) ( 627 )   Save BACKGROUND: Traditional Chinese medicine Rheum palmatum L. and Cortex Phellodendri Chinensis have the property of activating blood circulation and removing stasis, as well as deswelling and alleviating pain, which obtain good outcomes in treating soft tissue injury. OBJECTIVE: To study the effect of Diechuangsan (DCS) on experimental soft tissue injury and prostaglandin E2 expression. METHODS: Totally 36 New Zealand big ear white rabbits were randomly divided into 3 groups, namely, DCS, model and control groups. Rabbits were established acute soft tissue injury models and treated by DCS at 30 minutes after model preparation, once per day. No medication was performed in the model and control groups. Prostaglandin E2 expression and hemorheology index changes were observed at 5 days after model preparation. The recovery of injured soft tissues was observed by hematoxylin-eosin staining and pathological score at 5 and 13 days after model preparation. RESULTS AND CONCLUSION: Compared with the model group, the pathological score of the DCS group was higher (P < 0.05), which was earlier close to the normal tissues. In addition, the prostaglandin E2 expression and blood viscosity of the DCS group was lower than that of the model group (P < 0.05). It is suggested that DCS can repair injured soft tissues via decreasing inflammatory mediators prostaglandin E2 and reducing injury animal blood viscosity. Related Articles | Metrics

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Effects of berberine on expression of hepatic peroxisome proliferator-activated receptors and its target genes in type 2 diabetic Chinese hamsters Liu Xu-han, Li Guo-sheng, Huang Lan, Zhu Hua, Liu Ya-li, Ma Chun-mei 2011, 15 (24):  4409-4414.  doi: 10.3969/j.issn.1673-8225.2011.24.010 Abstract ( 117 )   PDF (1611KB) ( 835 )   Save BACKGROUND: Although Berberine has been reported to treat type 2 diabetes, the underlying mechanisms of berberine on insulin resistance of type 2 diabetes, especially hepatic insulin resistance, remains not fully understood. OBJECTIVE: To study the effects of berberine on the expression of hepatic peroxisome proliferator-activated receptors (PPARs) and their target genes in type 2 diabetic Chinese hamsters. METHODS: The insulin-resistant and type 2 diabetic Chinese hamster models were induced by high-fat diet without or with low-dose streptozotocin. After the induction of models, the hamsters were randomly divided into normal control (standard food), insulin-resistant (high-fat diet), diabetic (high-fat diet and streptozotocin) and berberine-treated diabetic (high-fat diet and streptozotocin and berberine) groups. All groups were treated for 9 weeks. RESULTS AND CONCLUSION: Results of real-time quantitative PCR indicated that compared with normal control group, the expression of PPARα, PPARβ/δ, acyl-Coenzyme A oxidase (Acox), carnitine palmitoyltransferase 1 (Cpt1) and acetyl-Coenzyme A dehydrogenase, medium chain (Acadm) was decreased (P < 0.05) and the expression of sterol regulatory element binding factor 1 (SREBP1c), sterol regulatory element binding factor 2 (SREBP2), PPARγ, lipoprotein lipase (LPL), CD36/FA transporter (FAT/CD36) and adipocyte fatty acid-binding protein (ap2) was increased (P < 0.05) in the fatty liver of insulin-resistant and diabetic hamster groups. Berberine effectively improved insulin resistance, reversed the altered expression of PPARs and its target genes in diabetic hamsters. PPARs and its target genes involved in the therapeutic molecular mechanisms of berberine on fat-induced hepatic insulin resistance in type 2 diabetic hamsters. Related Articles | Metrics

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Role of autologous platelet-rich plasma on fracture healing in rabbits Zhou Hai-yang, Wang Chen, Geng Zhen 2011, 15 (24):  4415-4418.  doi: 10.3969/j.issn.1673-8225.2011.24.011 Abstract ( 136 )   PDF (1512KB) ( 595 )   Save BACKGROUND: Platelet-rich plasma (PRP) is often prepared with autalloploid and can be used to promote the fracture healing of flat bone and cancellous bone. OBJECTIVE: To prepare PRP with Landsberger’s method and to analyzes the therapeutic role of PRP on the fracture healing of radius in rabbits. METHODS: New Zealand rabbits were randomly divided into a control group and a PRP treated group. Unilateral open femoral fracture models were established in the lower 1/3 radius bone. PRP group received was treated with autologous PRP gel, while control group was only sutured after fracture modeling. Autologous PRP was prepared by Landsberger’s method and then implanted to the fracture site, followed by plaster immobilization of the fracture fragments. RESULTS AND CONCLUSION: The fracture specimens exhibited varying degrees of callus formation at 1, 2, 4, 6 weeks. X-ray score in the PRP group was significantly higher than that in control group at 4 weeks (P < 0.05). Immunohistochemistry showed that, type I collagen content in the PRP treatment group was significantly higher than that in the control groups for 4 weeks (P < 0.05). PRP accelerates bone fracture healing of rat femurs by increasing expression of type I collagen and the number of bone island. Related Articles | Metrics

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Establishment of rabbit models of corticosteroid-induced avascular necrosis of femoral head Wang Yuan-he, Zhang Cai-long, Tian Shao-qi, Sun Kang, Wang Cui 2011, 15 (24):  4419-4422.  doi: 10.3969/j.issn.1673-8225.2011.24.012 Abstract ( 132 )   PDF (430KB) ( 274 )   Save BACKGROUND: Hormone use has become the primary cause of steroid-induced avascular necrosis of femoral head (SANFH). OBJECTIVE: This study used a combination of injection of horse serum and a large dose of corticosteroid to develop a hormone-induced rabbit model of early avascular necrosis of femoral head (ANFH), and preliminary discussed the pathogenesis of ANFH. METHODS: New Zealand rabbits were randomly divided into three groups. Methylprednisolone with horse serum group: horse serum (10 mL/kg) was injected. Three weeks later, 6 mL/kg of horse serum was injected. Two weeks later, 45 mg/kg of methylprednisolone was daily injected for 5 consecutive days. Methylprednisolone group: 45 mg/kg of methylprednisolone was daily injected for 5 consecutive days. Control group: no treatment was given. Serum cholesterol and triacylglycerol levels were detected at 1, 3, 7 and 14 days before and after hormone injection. MRI and histopathological detection was done in femoral head at 2, 4 and 8 weeks after hormone injection. RESULTS AND CONCLUSION: The serum triglyceride and total cholesterol in methylprednisolone with horse serum group and methylprednisolone group were higher than control group at 1 and 3 days after hormone injection (P < 0.01). MRI results displayed abnormal signal in femoral head at 4 weeks in methylprednisolone with horse serum group, but in the methylprednisolone group at 8 weeks. Histological detection results exhibited that at 4 weeks, some trabeculae were broken into fragments, and the empty bone lacunae increased. At 8 weeks, the trabeculae showed thinning and broken. There were large amount of empty bone lacunae with bone cell atrophy and larger fat cells which were fused into bubbles. In methylprednisolone group, the level of necrosis was lighter than methylprednisolone with horse serum group during each period. Results suggest that hormone combined with horse serum can successfully prepare early-stage hormone-induced ANFH. Related Articles | Metrics

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Phosphorylated p38 mitogen-activated protein kinase and transforming growth factor beta-1 expression in degenerative versus normal disc tissues Yuan Ye, Zhao Jing, Zhao Jie, Li Yong-min, Wang Xu 2011, 15 (24):  4423-4426.  doi: 10.3969/j.issn.1673-8225.2011.24.013 Abstract ( 83 )   PDF (1279KB) ( 390 )   Save BACKGROUND: Studies show that p38 mitogen activated protein kinase (MAPK) signal pathway is highly correlated with inflammatory reactions during intervertebral disc degeneration. OBJECTIVE: To study the phosphorylated p38MAPK (p-p38MAPK) and transforming growth factor β1 (TGF-β1) expression in patients with intervertebral disc degeneration. METHODS: Degenerative disc tissues resected by surgery from 36 patients with lumbar disc herniation were selected and normal disc tissues from 10 cases of trauma were used as control group. The expression of p-p38MAPK and TGF-β1 was evaluated with immunohistochemistry method and analyzed using pathological imaging analysis system. RESULTS AND CONCLUSION: The positive rate of p-p38MAPK and TGF-β1 expression in degenerative disc tissues was greater than normal disc tissue (P < 0.05). Results demonstrated that p-p38MAPK and TGF-β1 are highly expressed in the process of disc degeneration. Related Articles | Metrics

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Construction of basic fibroblast growth factor gene fluorescent eukaryotic cell expression vector and inhibitory effect on hydrogen peroxide-induced apoptosis and related gene expression in vascular endothelial cells Xu Bin, Lin Gui-xian, Wu Xiao-ying, Mao Jian-wen 2011, 15 (24):  4427-4431.  doi: 10.3969/j.issn.1673-8225.2011.24.014 Abstract ( 97 )   PDF (801KB) ( 305 )   Save BACKGROUND: It has been verified that exogenous basic fibroblast growth factor (bFGF) has obvious inhibitory effect on apoptosis in vascular endothelial cells. OBJECTIVE: To construct a fluorescent eukaryotic cell expression vector carrying the gene of human bFGF, and to investigate its effect on apoptosis induced by hydrogen peroxide (H2O2) and related gene expression in vascular endothelial cells. METHODS: bFGF and GFP gene was subcloned into eukaryotic expression plasmid pcDNA3.1 by means of gene cloning to obtain pcDNA3.1-bFGF-GFP. pcDNA3.1-bFGF-GFP was transfected into umbilical vein endothelial cells by liposome mediating, RT-PCR and observing green fluorescence by fluorescence microscope were used to determine the expression of bFGF and GFP. Umbilical vein endothelial cells were divided into three groups: control group (transfected pcDNA3.1), H2O2 group (transfected pcDNA3.1+H2O2) and bFGF-transfected+H2O2 group (transfected pcDNA3.1-bFGF-GFP+H2O2), the apoptosis of umbilical vein endothelial cells was detected by flow cytometry, the expression of caspase-3 P17 subunit and bax protein were determined by Western blot. RESULTS AND CONCLUSION: pcDNA3.1-bFGF-GFP was successfully constructed, bFGF mRNA was increased significantly and specific green fluorescence was observed by fluorescence microscope after pcDNA3.1-bFGF-GFP transfected. Compared with the control group, apoptosis rate, the expression level of caspase-3 P17 subunit and bax protein significantly increased in H2O2 group (P < 0.01). Compared with H2O2 group, bFGF gene transferring significantly decreased apoptosis rate, down-regulated the expression of caspase-3 P17 subunit and bax protein (P < 0.01). bFGF gene transferring can inhibit the apoptosis induced by H2O2 in vascular endothelial cells, the underlying mechanism might be associated with regulation on the expression of bax protein and activity of caspase-3. Related Articles | Metrics

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Effect of polypeptide from Chlamys farreri on ultraviolet B radiation-induced HaCaT cell apoptosis: Tumor necrosis factor-alpha/tumor necrosis factor receptor 1 pathway Zheng Song-wen, Li Jin-lian, Wang Chun-bo, Han Yan-tao 2011, 15 (24):  4432-4436.  doi: 10.3969/j.issn.1673-8225.2011.24.015 Abstract ( 102 )   PDF (861KB) ( 350 )   Save BACKGROUND: Polypeptide from Chlamys farreri (PCF) exerts protective effects on ultraviolet B radiation-induced human keratinocyte cell line HaCaT cells through Fas pathway and nuclear factor-κB pathway. OBJECTIVE: To investigate HaCaT cell apoptosis after ultraviolet B radiation, and activation of tumor necrosis factor-α/ tumor necrosis factor receptor-1, and intervention of PCF. METHODS: Cell injury models were established by 20 mJ/cm2 ultraviolet B radiation on HaCaT cells for 0.5 hour. Low-dose, moderate-dose and high-dose drug groups, positive control group and inhibitor group were treated with 1.42, 2.84, 5.69 mmol/L PCF, 5.68 mmol/L vitamin C and 50 mg/L anti- tumor necrosis factor-α monoclonal antibody. RESULTS AND CONCLUSION: After ultraviolet B radiation, HaCaT cell apoptosis became more. Tumor necrosis factor-α, tumor necrosis factor receptor 1 mRNA and phosphorylated c-Jun N-terminal kinase expression increased. 1.42, 2.84 and 5.69 mmol/L PCF could reduce ultraviolet B radiation-induced tumor necrosis factor-α, tumor necrosis factor receptor 1 mRNA and phosphorylated c-Jun N-terminal kinase expression, and inhibit cell apoptosis, especially 5.69 mmol/L PCF, which was identical to the effects of 5.68 mmol/L vitamin C. 50 mg/L anti-tumor necrosis factor-α monoclonal antibody significantly decreased ultraviolet B radiation-induced phosphorylated c-Jun N-terminal kinase expression. These suggest that PCF inhibited ultraviolet B radiation-induced HaCaT cell apoptosis by tumor necrosis factor-α/tumor necrosis factor receptor 1 pathway. Related Articles | Metrics

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Effect of breviscapine on transforming growth factor beta 1 in human peritoneal mesothelial cells induced by peritoneal dialysis solution Du Fei, Zhang Ke-fei, Gao Yuan, Zhang Wen-hui, Yuan Li-ying 2011, 15 (24):  4437-4439.  doi: 10.3969/j.issn.1673-8225.2011.24.016 Abstract ( 78 )   PDF (538KB) ( 435 )   Save BACKGROUND: Based on the understanding of breviscapine effects and mechanism on inhibiting and delaying peritoneal failure, breviscapine is expected to be widely introduced to prolong peritoneal dialysis time, improve the quality of dialysis, and reduce dialysis failure rate so as to promote long-term effect of peritoneal dialysis in patients with end stage renal diseases. OBJECTIVE: To observe the effects of breviscapine on the secretion of transforming growth factor beta 1 (TGF-β1) and proliferation activity of human peritoneal mesothelial cells (HPMCs) cultured in vitro with peritoneal dialysis solution (PDS). METHODS: HPMCs were cultured in vitro and divided into five groups: control group, PDS group, 5 µmol/L breviscapine group, 10 µmol/L breviscapine group, and 20 µmol/L breviscapine group. The levels of TGF-β1 and proliferation of HPMCs in supernatant were detected and compared. RESULTS AND CONCLUSION: Significantly increased levels of TGF-β1 and decreased cell proliferation activity were observed in the PDS group. The level of TGF-β1 in the PDS group was higher than that in the 5 µmol/L breviscapine group (P < 0.05), and significantly higher than that in the 10 µmol/L breviscapine group and 20 µmol/L breviscapine group (P < 0.01). The proliferation activity in the PDS group was lower than that in the 5 µmol/L breviscapine group (P < 0.05), and significantly lower than that in the 10 µmol/L breviscapine group and 20 µmol/L breviscapine group (P < 0.01). The results indicated that breviscapine can reduce the secretion of TGF-β1 in cultured HPMCs. Also it can restore the reduced cell proliferation activity caused by PDS． Related Articles | Metrics

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Oxygen concentration variation influences the severity of retinopathy and vascular endothelial growth factor expression in newborn rats Wei Er-xia, Ke Xiao-yun, Chen Hui, Liu Zi-bin, Wu Wei 2011, 15 (24):  4440-4443.  doi: 10.3969/j.issn.1673-8225.2011.24.017 Abstract ( 141 )   PDF (1344KB) ( 508 )   Save BACKGROUND: Oxygen plays an important role in premature retinal neovascularization, but its scope of oxygen and the specific mechanism of retinopathy are not clear. OBJECTIVE: To study the effects of different concentrations of oxygen on neovascularization in oxygen-induced retinopathy of newborn rats. METHODS: Forty newly born rats were divided into four groups: air group (10 rats) was raised in air room. Fluctuation group 1, 2, 3 (10 rats) was exposed to cycles of 50%-20%, 40%-10%, 50%-10% for 14 days, respectively. Retinas were dissected and stained by adenosine-diphosphatase (ADPase) histochemistry for assessment of intraretinal vascular development and preretinal angiogenic vessel growth. Expressions of vascular endothelial growth factor (VEGF) were studied by Western-blot. RESULTS AND CONCLUSION: Retina flat-mounts presented increasing neovascular tufts, severity of retinal neovascularization and the number of vascular nuclei extending from the internal limiting membrane into the vitreous, as well as VEGF-expression in fluctuation group 2 and fluctuation group 3. There were no obviously neovascularization in the air and fluctuation 1 groups. This study suggested that a small change in blood oxygen profile could trigger a disproportionate increase in subsequent neovascularization, accompanied by changes of retinal VEGF level. VEGF promote retinal neovascularization. The over-expression of VEGF plays a synergistic role during the formation of neovascularization. Partial pressure of inspiratory oxygen was positively correlated with the retinopathy. Hypoxia may be more important for retinal neovascularization on the occurrence of retinopathy than hyperoxia. Related Articles | Metrics

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Expression of platelet-derived growth factor-D in pathogenesis of unilateral ureteral obstruction during renal interstitial fibrosis Zhang Qian-qian1, Sun Jian-ping, Gao Yan-xia, Dong Hui, Zhou Li-min, Liu Hong-yan, Xiao Yu-cui 2011, 15 (24):  4444-4447.  doi: 10.3969/j.issn.1673-8225.2011.24.018 Abstract ( 99 )   PDF (1428KB) ( 412 )   Save BACKGROUND: The platelet-derived growth factors (PDGF) are highly expressed in the diseased renal, and induce renal interstitial fibrosis through tubulointerstitial cells proliferation, phenotypic modulation and inflammatory cell infiltration. OBJECTIVE: To observe expression of the PDGF-D in kidney tissues in the process of renal interstitial fibrosis. METHODS: Totally 60 SD rats were randomly divided into the model and sham-surgery groups. In the model group, rats were constructed unilateral ureteral obstruction models by left ureteral ligation. Animals were sacrificed at 3, 7, 14, 21 and 28 days after operation, and the expression and distribution of the PDGF-D in the kidney tissues were detected by immunohistochemistry, and the expression levels of PDGF-D mRNA in tissue samples were determined by real-time RT-PCR. RESULTS AND CONCLUSION: There were few PDGF-D expressed in glomerular mesangial cells and vascular smooth muscle cells in the sham-surgery group, but highly expressed in the model group, which were focally observed in interstitial cells in areas of tubulointerstitial fibrosis, and increased with time prolonged. The mRNA level of PDGF-D in the model group was obviously greater than that of the sham-surgery group (P < 0.05). These observations suggest that PDGF-D plays an important role in the pathogenesis of unilateral ureteral obstruction during renal interstitial fibrosis. Related Articles | Metrics

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Effect of pilose antler polypeptides on chondrocytes proliferation in knee osteoarthritis Xiu Zhong-biao, Jiang Zhi-hao, Sun Lei 2011, 15 (24):  4448-4452.  doi: 10.3969/j.issn.1673-8225.2011.24.019 Abstract ( 96 )   PDF (1562KB) ( 506 )   Save BACKGROUND: Experimental studies on chondrocytes culture in vitro confirm that pilose antler polypeptide (PAP) has mitogenic activity and can stimulate the proliferation of cartilage cells. OBJECTIVE: To investigate the regulatory effect of PAP on related cytokines and chondrocytes proliferation in experimental osteoarthritis process. METHODS: Totally 80 New Zealand White rabbits were randomly divided into 3 groups: normal group (n=4), sham operation group (n=4) and model group (n=72).Normal group was not given any treatment. Sham operation group was sutured after incising the medial aspect of the left knee. And model group was surgically induced into osteoarthritis models. After successful modeling, the rabbits of the model group were further divided into 2 groups: PAP group and normal saline control group, 32 rabbits in each group. PAP group received 0.5 mL intra-articular injection of PAP dilution liquid once daily for 30 days while normal saline control group received 0.5 mL intra-articular injection of physiological saline. On days 1,7,15 and 30 after intervention, articular cartilage samples and synovial fluid were collected respectively. The morphological changes of articular cartilage were observed under optical microscope and the structural change of chondrocytes was observed by transmission electron microscopy. The levels of interleukin-1β (IL-1β), tumor necrosis factor-α (TNF-α) and transforming growth factor β1 (TGF-β1 ) in synovial fluid were detected by enzyme-linked immunosorbent assay (ELISA). The expression levels of PCNA were detected by the method of immunohistochemistry and the cell proliferation index was analyzed. RESULTS AND CONCLUSION: At the same period, the levels of IL-1β and TNF-α in synovia fluid of PAP group were lower than those of normal saline control group, but the level of TGF-β1 was higher in the former group, which were statistically significant   (P < 0.05). The expression of proliferating cell nuclear antigen in artcular cartilage of PAP group was more than that of normal saline control group and the cell proliferation index was significantly higher, which had statistical difference. These results suggest that PAP can decrease the levels of IL-1β, TNF-α, increase the levels of TGF-β1 in synovial fluid and promote the proliferation of chondrocytes in experimental osteoarthritis process. Related Articles | Metrics

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Effect of natural and recombinant hirudin on superoxide dismutase, malonaldehyde and endothelin in rat random skin flap congestion models Guo Ying-xin, Yin Guo-qian, Li Jia-quan 2011, 15 (24):  4453-4456.  doi: 10.3969/j.issn.1673-8225.2011.24.020 Abstract ( 111 )   PDF (1070KB) ( 290 )   Save BACKGROUND: Most scholars believed that the effect of hirudin can improve the survival rate of congestion flap based on its antithromotic and antithrombin activity. However, whether it has anti-inflammatory and antioxidant effects remains unclear. The disparity between recombinant hirudin and recombinant hirudin needs to be explored. OBJECTIVE: To investigate the protective effect of natural hirudin and recombinant hirudin on random skin flap survival in rats vein congestion models.  METHODS: Thirty Wistar rats were employed to establish animal model of vein congestion. Dorsal random skin flap was prepared on each animal, 10 cm×3 cm in size. The flaps were divided randomly into 3 groups. 5 U natural hirudin, 5 U recombinant hirudin or physiological saline was injected at 1.5 cm and 3.0 cm at distal part of flap immediately, 3 and 5 days after operation. The 7 days flap survival, morphological changes of flap and superoxide dismutase (SOD), endothelin (ET) and malonaldehyde (MDA) levels were measured RESULTS AND CONCLUSION: The flap survival of the natural and recombinant hirudin groups were greater than that of the control group, with increased SOD level and decreased ET and MDA levels. In particular, the effect was more manifest in the natural hirudin group. The findings demonstrated that hirudin can improve the flap survival rate and SOD content but reduce the rate of flap necrosis, MDA, and ET levels. Related Articles | Metrics

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Effect of ginsenoside Rd on collagen expression in liver fat-storing cells Qiao Shi-wen, Wang Ji-feng, Huang Qin, Zhu Zong-gui, Zhang Zhi-gui 2011, 15 (24):  4457-4460.  doi: 10.3969/j.issn.1673-8225.2011.24.021 Abstract ( 95 )   PDF (1118KB) ( 283 )   Save BACKGROUND: Collagens produced by liver fat-storing cells are the directly reasons for cirrhosis, thus, regulating capacity of fat-storing cells producing collagens can prevent cirrhosis.   OBJECTIVE: To explore effect of ginsenoside Rd on expression of type Ⅰ and Ⅲ collagens in liver fat-storing cells in vitro. METHODS: The liver fat-storing cells were cultured in vitro and interfered with ginsenoside Rd. The expression of type Ⅰ and Ⅲ collagens was detected by immunohistochemistry and immunofluorescent. RESULTS AND CONCLUSION: Compared with the control group, the positive/negative-area and the absorbance of the experimental group were obviously decreased (P < 0.05), suggesting that ginsenoside Rd regulates the expression of type Ⅰ and Ⅲ collagens in liver fat-storing cells in vitro, which affects liver fat-storing cells producing collagens. Related Articles | Metrics

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High glucose induces podocyte apoptosis and constitutive photomorphogenic 1 expression changes Wang Xiao-lin, Gan Hua, Du Xiao-gang 2011, 15 (24):  4461-4464.  doi: 10.3969/j.issn.1673-8225.2011.24.022 Abstract ( 108 )   PDF (1242KB) ( 353 )   Save BACKGROUND: Studies have shown that constitutive photomorphogenic 1 (COP1) is involved in cell apoptosis. OBJECTIVE: To observe the effects of high glucose exposure on expression of in vitro cultured podocyte apoptosis and COP1 expression. METHODS: Mouse podocytes of an immortalized cell line were treated with different concentrations of glucose. Apoptosis index and mortality index at different culture time were evaluated by flow cytometry to screen best dose-effect glucose concentration (30 mmol/L), which was used to treat high-glucose group. Control group and mannitol group (30 mmol/L) were set. RESULTS AND CONCLUSION: Glucose induced podocyte apoptosis and death in time- and dose-dependent manner in certain concentration (P < 0.05). With increasing glucose concentration, mortality index was significantly elevated, but apoptosis index remained unchanged. Compared with control group, apoptosis index was significantly increased in high glucose group (P < 0.05), but COP1 mRNA and protein expression was significantly decreased (P < 0.05). Results showed that changes in COP1 expression may play a critical role in pathogenesis of high glucose induced podocyte apoptosis. Related Articles | Metrics

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Effects of pretreatment with heat stress on free radical metabolism in skeletal muscle of rats after eccentric exercise Shan Jin-song, Ren Qiu-jun, Gao Qian-jin 2011, 15 (24):  4465-4467.  doi: 10.3969/j.issn.1673-8225.2011.24.023 Abstract ( 118 )   PDF (1156KB) ( 420 )   Save BACKGROUND: It has been demonstrated that pre-heat stress can elevate the ability of muscle against injury, but the precise mechanism is unclear. OBJECTIVE: To observe the effect of pre-heat stress on superoxide dismutase activities and malondialdehyde content in rat skeletal muscle after eccentric exercise. METHODS: Male Wistar rats were randomly divided into control group, eccentric exercise group, heat stress+eccentric exercise group. The temperature of heat stress was 43 ℃ for about 35 minutes. Large load intermittence eccentric exercise was performed at -16° down slope treadmill at 26.8 m/min for 5 minutes, with an interval of 1 minute, totally for 10 groups. Rat gastrocnemius muscle was obtained at 1 hour before exercise, 1, 24 and 48 hours after exercise separately. Rat malondialdehyde content was determined by thiobarbituricacid method. Superoxide dismutase activities were measured by xanthine oxidase method. RESULTS AND CONCLUSION: Compared with control group, malondialdehyde content was significantly increased in rat gastrocnemius muscle in eccentric exercise group (P < 0.05), and gradually increased over time after exercise. Superoxide dismutase activities were significantly decreased over time after exercise (P < 0.05). Compared with eccentric exercise group, superoxide dismutase activities were significantly increased (P < 0.05), but malondialdehyde content (P < 0.05) was significantly decreased in rat gastrocnemius muscle of heat stress+eccentric exercise group. These indicate that heat stress can enhance superoxide dismutase activities, reduce malondialdehyde content, and protect skeletal muscle from eccentric exercise induced injury. Related Articles | Metrics

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Role of pro-apoptotic protein Bim and PI3K/Akt signal-transduction pathway mechanism in hypothermia-induced cardiomyocytes damage in neonatal rats Bao Xiao-ming, Cheng Xiao-shu, Huang Xiao, Wang Yao-sheng, Li Ju-xiang, Hong Kui 2011, 15 (24):  4468-4472.  doi: 10.3969/j.issn.1673-8225.2011.24.024 Abstract ( 143 )   PDF (1620KB) ( 312 )   Save BACKGROUND: Cold stimulation can result in cardiovascular disease or aggravate cardiovascular disease. It is significant to research the occurrence patterns and molecular mechanism for preventing and treating cardiovascular diseases.  OBJECTIVE: To explore the effect of hypothermia on the cardiomyoctyes apoptosis and the level of lactate dehydrogenase (LDH) activity, as well as the role of pro- apoptosis protein Bim and PI3K/Akt signal-transduction pathway. METHODS: Cardiomyoctyes were exposed to 4℃ cold environment for 1 hour, then cultivating in 37 ℃ CO2 incubator for 0, 4, 8, 12, 16, and 24 hours. The degree of apoptosis was measured by flow cytometer by Annexin V-FITC/PI; LDH activity was determined with automatic biochemistry analyzer. Expressions of Bim were determined by Western blotting. All the changes were observed in the nutrient medium after adding PI3K inhibator LY294002. RESULTS AND CONCLUSION: After hypothermia treatment, the cell apoptosis was significantly increased, the survival rate decreased, and the LDH activity and Bim expression increased (P < 0.05), which reached the peak at 24 hours. After adding LY29004, cell apoptosis increased notably, the LDH activity and Bim expression increased, but p-PI3K expression were declined (P < 0.05). It suggested that hypothermia can induce cardiomyoctyes apoptosis. The PI3K/Aktsignal-transduction pathway is likely to participate in Bim expressions. Related Articles | Metrics

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High altitude hypoxia on the ultrastructure of heart and lung tissue Zhang Chun-ai, Li Wen-hua 2011, 15 (24):  4473-4476.  doi: 10.3969/j.issn.1673-8225.2011.24.025 Abstract ( 76 )   PDF (665KB) ( 532 )   Save BACKGROUND: Studies show that, the body inevitably will be injured to different degrees upon the rapid access to high altitude, among them damage to heart and lung is apparent. OBJECTIVE: To evaluate the influence of hypoxic acclimatization on the ultrastructure of rat heart and lung. METHODS: SD rats were randomly divided into 4 groups, namely the plateau hypoxia 1, 3, 30 days, and a control group. The hypoxic groups of rats were brought from the Xi'an (elevation 5 m), to Golmud, Qinghai (elevation 2 700 m; 1 day), to the Naqu (elevation 4 500 m; time-consuming 3 and 30 days). The cardiopulmonary specimens were observed for the histological changes. RESULTS AND CONCLUSION: At acute hypoxia 1 and 3 days, the microstructure and ultrastructure of lung tissues exhibited apparent interstitial pulmonary edema and alveolar pulmonary edema; rat ventricular myocytes showed varying degrees of cloudy swelling, vacuolar degeneration, necrosis and interstitial dissolved edema by the heart tissue observations under light microscopy; electron microscope displayed that, the mitochondria swelling, sarcoplasmic reticulum, myofibril dissolution, intracellular and extracellular edema in cardiac muscle cells. These changes in the right ventricular wall at hypoxia 3 days were more apparent compared with those in left ventricular wall. The interstitial edema after hypoxic acclimatization was significantly reduced at 30 days. High altitude acute hypoxia may induce pulmonary interstitial edema and alveolar pulmonary edema, as well as right ventricular-based heart damage. The heart and lung tissue lesions significantly reduced after high altitude hypoxic acclimatization. Related Articles | Metrics

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Effects of soybean trypsin inhibitor on glucose tolerance and hepatic glycogen in alloxan diabetic mice Guo Rui-hua, Wang Hui-li, Zhai Yi-min, Hu Yan, Jin Xiao-dong 2011, 15 (24):  4477-4480.  doi: 10.3969/j.issn.1673-8225.2011.24.026 Abstract ( 121 )   PDF (667KB) ( 380 )   Save BACKGROUND: Previous studies found soybean trypsin inhibitor (SBTI) may have effect on treating diabetes mellitus and regulating insulin disturbance. OBJECTIVE: To study the effects of SBTI on blood glucose, glucose tolerance, and hepatic glycogen in alloxan diabetic mice. METHODS: Alloxan diabetic mice models were prepared by intraperitoneal injection of alloxan, and 32 mice with successful model preparation were randomly divided into 4 groups, additional 8 mice were served as control. After model preparation, mice in the SBTI-L and SBTI-H groups were lavaged with 0.27 and 0.80 mg/kg SBTI solution, those in the glibenclamide group was lavaged with 0.16 g/kg glibenclamide. Saline was lavaged in the model and control groups. The medication lasted for 2 weeks. The blood glucose and glucose tolerance of mice were measured by using glucose oxidase method, and the content of hepatic glycogen was determined by the improved anthrone method. RESULTS AND CONCLUSION: Compared with the model group, the blood glucose of glibenclamide, SBTI-L, and SBTI-H groups was significantly decreased (P < 0.05), and the glucose tolerance rise lower (P < 0.05), but the hepatic glycogen increased obviously (P < 0.05). The SBTI-L and SBTI-H groups showed faster effect and longer effectiveness than the glibenclamide group, especially more manifest in the SBTI-H group. Experimental results show that SBTI can significantly decrease blood glucose, improve glucose tolerance, and increase the content of hepatic glycogen. Related Articles | Metrics

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Isokinetic test of flexor and extensor strength in primary student knees Ji Hua-juan, Wei Meng-tian, Wang Li-xia 2011, 15 (24):  4481-4484.  doi: 10.3969/j.issn.1673-8225.2011.24.027 Abstract ( 101 )   PDF (722KB) ( 334 )   Save BACKGROUND: Previous evaluation for lower limb strength in primary students are limited for physical quality measurement, such as standing long jump or jump touching, which lack of quantitative studies.  OBJECTIVE: To learn the characteristic of development in muscle strength in primary students using isokinetic testing system. METHODS: The flexion and extension strength of knee joints in 54 healthy primary students aged 9-12 (25 boys and 29 girls) were measured by using Kinitech Isokinetic testing system with 3 angular velocities: 60 (°)/s, 120 (°)/s and 240 (°)/s. RESULTS AND CONCLUSION: The peak torque (PT) of primary students was decreased with angular velocities increasing; The PT value of left side at speed of 60 (°)/s and 120 (°)/s, and right side at speed of speed of 60(°)/s was lower than that of the extensors in both genders (P < 0.01 or P < 0.05). At the same speed, the PT value of flexor and extensor in left side and flexor in the right side of boys were obviously higher than that of girls at 240 (°)/s (P < 0.01 or P < 0.05). Related Articles | Metrics

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Effect of treadmill exercise on the collagen, matrix metalloproteinases-1 and tissue inhibitors of metalloproteinase-1 mRNA in rat’s Achilles tendon Li Min, Jin Li 2011, 15 (24):  4485-4488.  doi: 10.3969/j.issn.1673-8225.2011.24.028 Abstract ( 101 )   PDF (637KB) ( 365 )   Save BACKGROUND: Collagen is the main protein component of tendon tissue. Metalloproteinases are a large family of enzymes capable of degrading all of the tendon matrix components. OBJECTIVE: To investigate the effects of 12-week treadmill running on content of collagen and the expression of procollagen I, matrix metalloproteinases-1 (MMP-1) and tissue inhibitors of metalloproteinase-1 of rat Achilles tendon. METHODS: A total of 20 male Sprague-Dawley rats were randomly assigned to control and exercise groups. At 1 week, exercise was performed for 20 minutes daily, at the increased speed from 12 m/min to 20 m/min. The treadmill speed was kept at 20 m/min. At 2 weeks, the gradient was 5%, and time was 30 minutes. At 3-12 weeks, the gradient was 10%, and the time was 40 minutes, for totall 12 weeks. In control group, the animals were fed normally. RESULTS AND CONCLUSION: 24 hours after the last exercise, the rat Achilles tendon was obtained. Compared with control group, the expression of procollagen Iα1, MMP-1 and its inhibitor 1 mRNA was significantly increased (P < 0.05 or P < 0.01) in the exercise group, and the hydroxyproline content was increased (P > 0.05). These results show that long-term treadmill running can increase the synthesis and degradation of collagen in achilles tendon. Related Articles | Metrics

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Construction and identification of small interfering RNA expression plasmid target to differentiated embryo-chondrocyte expressed gene 1   Wei Yan, Jia Yan-fei, Zheng Yan, Ma Xiao-li, Wang Yun-shan 2011, 15 (24):  4489-4492.  doi: 10.3969/j.issn.1673-8225.2011.24.029 Abstract ( 133 )   PDF (609KB) ( 349 )   Save BACKGROUND: Differentiated embryo-chondrocyte expressed gene 1 (DEC1) is a basic helix-loop-helix transcription factor, which is closely associated with some malignant cancers. OBJECTIVE: To construct small interfering RNA (siRNA) expression plasmid target to DEC1.   METHODS: The mRNA sequence of DEC1 gene was searched from NCBI. Utilize of Katahdin siRNA technology, DEC1-siRNA oligonucletides were inserted into pGreenPuro™ shRNA Cloning and Expression Lentivector, after annealing, then transformed into JM-109. The recombinant plasmid was identified by Agarose gel electrophoresis analysis, ultraviolet spectrophotometer analysis, PCR and DNA sequencing.   RESULTS AND CONCLUSION: The recombinant plasmid pGreenPuro™ shRNA Cloning and Expression Lentivector-DEC1 was obtained by connecting 25 bp segment containing DEC1 sequence to pGreenPuro™ shRNA Cloning and Expression Lentivector. Agarose gel electrophorsis analysis and ultraviolet spectrophotometer analysis confirmed that plasmid DNA had higher purity. DNA sequencing showed Targeting siRNA oligonucleotides were correctly inserted into the eukaryotic expression vector pGreenPuro™ shRNA Cloning and Expression Lentivector without base mutation. The interference vector pGreenPuro™ shRNA cloning and expression lentivector-DEC1 was successfully constructed. Related Articles | Metrics

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Preparation of DNA Ladder Wang Li, Dong Wei-hua, Zhang Jun-he, Wang Tian-yun 2011, 15 (24):  4493-4496.  doi: 10.3969/j.issn.1673-8225.2011.24.030 Abstract ( 152 )   PDF (877KB) ( 318 )   Save BACKGROUND: DNA molecular weight standard (DNA Ladder) is one of necessary reagents in molecular biological laboratory. It can correctly measure the length of DNA fragments and serve as the DNA molecular weight standard. Now, there are two methods to prepare the DNA Ladder, one is PCR technique, the other is through DNA digestion by restriction enzyme. The two methods all have some merits and drawbacks. The first one can produce standard bands, but is difficult to obtain large bands. The second method is to prepare DNA Ladder restriction digestion of plasmid or phage DNA. In previous study, we have amplified 100-500 bp DNA fragments by using PCR technique, then purified the PCR product and prepared DNA Ladder. The bands of the prepared DNA Ladder were clear, higher definition and could be equal with that of commercial products. It could be used in the molecular biology experiments.  OBJECTIVE: To prepare DNA molecular weight standard control. METHODS: A specific plasmid pUC-DNA for PCR amplification was constructed, according to the sequence of pUC-DNA. Primers were designed to amplify 100-1 000 bp DNA fragments followed by primer5.0. 100-1 000 b p series DNA fragments were amplified and electrophoresised in 2% agarose gels. After identification by PCR recovery products and DNA sequencing, the DNA sequences were analyzed according to the pUC-DNA and the homology analysis was compared analyzed by blast. The PCR products were extracted by phenol/chloroform, precipitated with ethanol and mixed them proportionally. RESULTS AND CONCLUSION: 100-1 000 bp DNA fragments were successfully amplified by PCR technique, and the size of the bands were the same as the expected. The purified bands were clear. The 100-1000 bp DNA ladder was prepared in our laboratory. The method is convenient and the size of the prepared DNA ladder is standard, it could be used in the molecular biology experiments. Related Articles | Metrics

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Clinical application and progress of human body composition measurement methods Yan Dan, Ruan Xiang-yan 2011, 15 (24):  4499-4502.  doi: 10.3969/j.issn.1673-8225.2011.24.032 Abstract ( 140 )   PDF (673KB) ( 1580 )   Save BACKGROUND: Body composition always be partitioned into fat mass and fat-free mass, the qualities and distribution of those two are closely related to the health situation of all human. There is an important relationship between imbalance of body composition and diseases. OBJECTIVE: Overview and contrast the body composition measurement methods. Then give a review about the progress of human body composition measurement methods in clinical application. METHODS: Documents about body composition measurement methods and clinical application were searched from CNKI, PubMed and Elsevier, which published during 1961 to 2010. The progress of human body composition measurement methods in clinical application was summarized.  RESULTS AND CONCLUSION: Totally 36 manuscripts were included. The imbalance of body composition is the source of many diseases. The measurement of body composition is related to evaluate the health situation of human. Currently, there are many body composition measurement methods, but each of the methods has its own advantages and defects, at the same time, as more and more new methods are continually appearance. Related Articles | Metrics

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Artificial dermis combined with autologous thin skin transplantation for repair of soft tissue defects Fang Hong-song, Wang Hu, Gan Jing-yue, Wang Wei, Li Bin-bin 2011, 15 (24):  4503-4506.  doi: 10.3969/j.issn.1673-8225.2011.24.033 Abstract ( 149 )   PDF (489KB) ( 881 )   Save BACKGROUND: Multiple skin transplantations are needed when repair soft tissue defects by skin grafting. Artificial dermis is rare used in clinic, and the experience is insufficient. OBJECTIVE: To evaluate curative effect of artificial dermis for repair soft tissue defects.  METHODS: A total of 22 cases underwent artificial dermis with autologous thin skin transplantation were selected, including 6 cases with bone exposure, 2 with tendon exposure, 3 with resection of skin tumor, and 11 with others. Debridement and artificial dermis transplantation was performed firstly, followed by split thickness autoskin transplantation when the local granulation tissue grew well and the exposed tendon or bone tissue was covered by dermis-like tissues. The area of taking the skin, site of injury, operating performance, sealing, side effects were observed. The clinical and overall results were evaluated. RESULTS AND CONCLUSION: All cases had good coverage of wounds and graft survival. 18.50±4.27 days were needed to secondly skin grafting. One case performed a second surgery due to infection, 2 cases with automatic epithelization did not need secondly skin transplantation. Until to 3 months follow-up, epidermal growth and appearance of injury site are all good without scar in 21 cases. One case had scar and unhealthy appearance because of infection, but epidermal growth was also good. The area of taking the skin had no pigmentation and pigment prolapse, no serious scar in 20 cases, with 15.35±4.67 days to epithelialization. Repair of skin and soft tissue defects with artificial dermis has high quality of wound healing, minor injury to the area of taking site, and take longer time to secondly skin grafting. The overall clinical effect is good. Related Articles | Metrics

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Orthopedic ballistic shock wave and herbal fumigation therapy for treatment of overuse injury of soft tissue Liu Bo, Liu Hui, Zhao Wei-xia, Wu Sa 2011, 15 (24):  4507-4510.  doi: 10.3969/j.issn.1673-8225.2011.24.034 Abstract ( 130 )   PDF (738KB) ( 760 )   Save BACKGROUND: Orthopedic ballistic shock wave is commonly used to cure the overuse injury of soft tissues, but this therapy combined with herbal fumigation simultaneously is rarely reported. OBJECTIVE: To observe the clinical effects of the orthopedic ballistic shock wave combined with Chinese medicinal herbal fumigation on overuse injury of soft tissues. METHODS: A total of 912 cases with soft tissue overuse injury were treated with the orthopedic ballistic shock wave and Chinese medicinal herbal fumigation. The pain intensity of trigger point was assessed by visual analogue scale. RESULTS AND CONCLUSION: The intensity of pain decreased obviously after one weak treatment, but there were no significant changes of visual analogue scale in the second week. Two weeks later, the intensity of pain decreased significantly compared with before treatment (P < 0.05). Compared with tennis elbow and tenovaginitis of flexor digitorum, the visual analogue scale of calcanodynia decreased slowly (P < 0.05). The therapy of orthopedic ballistic shock wave and herbal fumigation is significantly efficient for overuse injury of soft tissues. Related Articles | Metrics

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Genetic variations of bone morphogenetic protein 7 associated with obesity in Xinjiang Uygur populations Yan Zhi-tao, Li Nan-fang, Guo Yan-ying, Yao Xiao-guang, Wang Hong-mei, Hu Jun-li 2011, 15 (24):  4511-4515.  doi: 10.3969/j.issn.1673-8225.2011.24.035 Abstract ( 130 )   PDF (629KB) ( 281 )   Save BACKGROUND: Studies show that bone morphogenetic protein 7 (BMP-7) gene maps to chromosome 20q13, a susceptibility locus for obesity. BMP-7 represents a strong biological and positional candidate for a susceptibility factor for obesity. OBJECTIVE: To investigate the association between the genetic variations of functional region in BMP-7 gene and obesity in Xinjiang Uygur populations. METHODS: A case-control study was conducted based on epidemiological investigation. 1063 Uygur populations were selected. The exons, segmental introns and the promoter regions of BMP-7 gene were sequenced in 48 Uygur obese patients (male/female: 24/24). Representative variations were selected according to the minor allele frequency and linkage disequilibrium and genotyped using the TaqMan polymerase chain reaction method in 1063 Uygur individuals, a relatively isolated general population in a relatively homogeneous environment and a case-control study was conducted to analyze the association between genetic variations of BMP-7 gene and obesity. RSSULTS AND CONCLUSION: Five novel and eight known variations in the BMP-7 gene were identified. Two genotype distributions, rs60254222 and rs17480735, followed Hardy-Weinberg equilibrium. There were significant differences in AA, AG, GG genotype distribution frequency of rs6025422 variation between obese and control groups in old adults (32.2%, 55.7%, 12.2% vs. 23.8%, 48.3%, 27.9%; P < 0.01)．There were significant differences in means of body mass index, waist circumference and waist-to-hip ratio were significantly decreased among individuals with AA, AG and GG genotypes of rs60254222 in old adults (P < 0.05). Logistic regression analysis showed that GG genotype of rs60254222 variation might be a obesity protective factor in old adults (OR = 0.578, 95% confidence interval 0.401-0.833, P = 0.003). Results showed that rs60254222 polymorphism in BMP-7 gene may be associated with obesity in Uygur populations. Related Articles | Metrics

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Effect of low-intensity ultrasound on washing open injury Zhang Huan-bo, Wei Wei, Zheng Hong-yu 2011, 15 (24):  4516-4518.  doi: 10.3969/j.issn.1673-8225.2011.24.036 Abstract ( 115 )   PDF (491KB) ( 465 )   Save BACKGROUND: Infection rate of open injury is high. OBJECTIVE: To compare the effects of low-intensity ultrasound and traditional method on washing open wounds. METHODS: Clinical data of 84 patients with open wounds were collected. Bacterial eradication and wound healing of 42 patients irrigated by low-intensity ultrasound were observed and compared with 42 patients (control group) undergoing regular flushing. RESULTS AND CONCLUSION: The wound tissue bacterial clearance rate and A level union rate at 2 hours after debridement were significantly greater in observation group compared with control group (P < 0.01, P < 0.05). The area of wound healing at 5 days was greater in observation group compared with control group, and the average healing time was significantly shortened than the control group (P < 0.01). Results shown that low-intensity ultrasound wound washing machine can effectively wash the wound with simple operation and promote wound healing. Related Articles | Metrics

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Marine algae polysaccharide derivant in treatment of osteoporosis in rats Si Yan-li, Li Dong-xia, Liu You-cai, Qian Zhi-bin 2011, 15 (24):  4519-4521.  doi: 10.3969/j.issn.1673-8225.2011.24.037 Abstract ( 97 )   PDF (326KB) ( 325 )   Save BACKGROUND: We have previously reported that marine algae polysaccharide derivant can significantly promote bone cell growth. OBJECTIVE: To observe the effect of marin algae polysaccharide derivant in treatment of osteoporosis. METHODS: Sixty female Wistar rats were randomly divided into control group, treatment group and model group. There were 20 rats in each group. The rats in treatment group and model group were respectively treated with retinoic acid to induce osteoporosis. The rats in treatment group were gavaged the marine algae polysaccharide derivant 10 mg/kg, and the rats in model group were gavaged the glucose 10 mg/kg orally for 14 days. Changes of rat femur bone histological examination and histomorphometry parameters were observed. RESULTS AND CONCLUSION: The mean Trabecular Number (Tb.N), the mean Trabecular Thickness (Tb.Th) and the percent Trabecular Area (Tb.Ar%) were significantly decreased in the model group compared with control group. The mean trabecular separation (Tb.Sp) was greatly increased. After intake of marine algae polysaccharide derivant, Tb.N, Tb.Th and Tb.Ar% of the treatment group were significantly more than that of the model group. The Tb.Sp was obviously reduced. These indicate that marine algae polysaccharide derivant can increase bone mass and have a therapeutic and preventional effect on the osteoporosis. Related Articles | Metrics

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Decoy receptor 1 expression in the intervertebral discs Yang Ji-kun, Zhang Guo-qing, Chen Bo-hua 2011, 15 (24):  4522-4524.  doi: 10.3969/j.issn.1673-8225.2011.24.038 Abstract ( 95 )   PDF (288KB) ( 273 )   Save BACKGROUND: Decoy receptor 1 (DcR1), which acts as decoy receptors, is resistant to apoptosis induction by related apoptosis inducing ligand (TRAIL). OBJECTIVE: To investigate the expression of the DcR1 (TRAIL-R3) protein in human herniated and normal lumbar intervertebral discs (IVD). METHODS: The expression and distribution of DcR1 proteins were assessed by using immunohistochemistry in 20 herniated lumbar IVD and 8 normal lumbar IVD tissues samples following discoidectomy from January to September 2010. RESULTS AND CONCLUSION: Percentage of nucleus pulposus and anulus fibrous cells with DCR1 expression was obviously more in the herniated group than in normal IVD group. The current results indicate that the expression of DCR1 rises after herniation. Related Articles | Metrics

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Effect of cathepsin L/G on venous vascular wall in traumatic deep vein thrombosis rat models Li Wen, Hu Ji-hong, Li Xing-guo, Li Hong-kun, Zhang Yu-bing, Zhao Xue-ling, Wang Bing 2011, 15 (24):  4525-4529.  doi: 10.3969/j.issn.1673-8225.2011.24.039 Abstract ( 122 )   PDF (474KB) ( 535 )   Save BACKGROUND: At present, the basic molecular etiological mechanism and core regulatory network of deep vein thrombosis (DVT) remains uncertain, and there is not an ideal measure for early diagnosis of DVT. OBJECTIVE: To study the underlying impact of cathepsin L/G in DVT rat model. METHODS: DVT rat models (n = 50) were established by clamping both femoral vein in three different positions within 3 seconds with mosquito forceps and fixing with cast. According to different observation phases and biological situations of the femoral vein thrombosis, model rats were divided into thrombogenesis group, pre-thrombogenesis group and non-thrombogenesis group. An additional 10 normal rats served as control group. Femoral vein was obtained at corresponding time points to exact total RNA. After a gene chip-based screening, the data of gene expression were further dissected by real-time PCR.  RESULTS AND CONCLUSUON: Gene chip hybridization analysis results demonstrated that differential expression of cathepsin L/G gene was significant among groups, and the expression was greatest in the thrombogenesis group, followed by pre-thrombogenesis and non-thrombogenesis groups, which was significantly greater than the control group (P < 0.05). Real-time PCR analysis results were consistent with gene chip hybridization analysis results. These indicate that DVT is associated with an increase in expression of cathepsin L/G in local venous vascular wall, and they may be candidate molecular markers for early diagnosis of deep vein thrombosis. Related Articles | Metrics

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Mechanotransduction in osteoblast and osteocyte regulation Zhang Shu, Cao Xin-sheng, Wang Bing 2011, 15 (24):  4530-4536.  doi: 10.3969/j.issn.1673-8225.2011.24.040 Abstract ( 134 )   PDF (501KB) ( 572 )   Save BACKGROUND: One of the basic functions of bone is its adaptation to mechanical loading environment. Bone cells are the mechanosensitive cells in bone tissue. However, the mechanisms by which mechanical signals are transduced to chemical signals that influence bone growth and metabolism remain unidentified.  OBJECTIVE: To understand the mechanotransduction pathways in osteoblasts and osteocytes, and to provide a theoretical basis for further study. METHODS: PubMed database was retrieved by computer with key words of “osteoblast, osteocyte, bone cells, mechanical stress”. According to inclusion criteria, 69 articles were included to summarize the transduction of mechanical signals of bone cells. RESULTS AND CONCLUSION: One of the basic functions of bone is its adaptation to mechanical loading environment. Bone cells are mechanosensitive cells. However, how the transduction of mechanical signals of cells realizes and how regulates skeleton remain poorly understood. Studies confirmed that due to the construction features and cell location of skeleton, osteoblasts and osteocytes are the most important mechanosensitive cells in bone tissue. The process of mechanotransduction can be divided into four distinct steps: ① mechanocoupling; ② biochemical coupling; ③ signal transmission; ④ effector response of bone cells. Through these four steps, the loads acting on the bones are transduced into biochemical signals, and then change the function of bone cells, finally induce the changes of bone structures to adapt the mechanical environment. The regulatory mechanisms of mechanical signals in bone marrow mesenchymal stem cells require further investigation.  Related Articles | Metrics

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Relationship of transforming growth factor beta 1 with bone turnover markers and bone mineral density Pi Yin-zhen, Liao Er-yuan 2011, 15 (24):  4537-4540.  doi: 10.3969/j.issn.1673-8225.2011.24.041 Abstract ( 168 )   PDF (447KB) ( 326 )   Save BACKGROUND: Transforming growth factor-β1(TGF-β1) is an important cytokine in the regulation of bone remodeling. Whether it can be used as a sensitive marker to bone turnover is incompletely understood. OBJECTIVE: To investigate the relationship of TGF-β1 with bone formation and resorption markers, and bone mineral density (BMD) at the anteroposterior lumbar spine. METHODS: Totally 663 healthy women from Changsha were analyzed, aged 20-80 years. Levels of TGF-β1, serum bone alkaline phosphatase (sBAP) and serum C-terminal cross-linked telopeptides (sCTx) were measured by ELISA, at the same time, the BMD of anteroposterior lumbar spine was measured by dual-energy x-ray absorptiometry. The associations between TGF-β1 and other indexes were analyzed.   RESULTS AND CONCLUSION: The TGF-β1 levels reached a peak in the 30-39 and 40-49 years groups, which had a negative correlation with age, but no correlation with body mass index (BMI). After corrected for BMI, TGF-β1 was negatively correlated to sBAP and sCTx, but was positively correlated with BMD of spine after correction of BMI or age. TGF-β1 can act as a sensitive cytokine to reflect the changes of bone turnover. Related Articles | Metrics

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Effects of antler powder on treatment of corticosteroid-induced avascular necrosis of the femoral head in rats Pharhat Baktursun, Peng Hao, Li Bin-bin, Huang Lei 2011, 15 (24):  4541-4544.  doi: 10.3969/j.issn.1673-8225.2011.24.042 Abstract ( 112 )   PDF (364KB) ( 379 )   Save BACKGROUND: Long-term utilization of steroid hormone easily induces avascular necrosis of the femoral head (ANFH). Cornu Cervi Pantotrichum possesses bone growth factor and can promote wound healing and tissue repairing, which may has effect on treatment of ANFH.   OBJECTIVE: To investigate therapeutic effect of antler powder on ANFH induced by corticosteroids in rats. METHODS: Totally 42 male wistar rats were randomly divided into the blank control, model and high-, moderate- and low-dose treatment groups. Exception those in the blank control group, all rats were intragluteally injected with dexamethasone at 30 mg/kg twice per week for 6 weeks to induce ANFH. Then the rats were treated with antler powder by oral gavage at 200, 400 and    800 mg/kg dosage, once per day for 60 days. Same volume of physiological saline was gavaged in the blank control and model groups. Serum lipid level and bone mineral density (BMD) of femur were measured, pathological change and expression of vascular endothelial growth factor (VEGF) were investigated. RESULTS AND CONCLUSION: Steroid hormone intervention resulted in obviously ANFH in rats. After treatment with antler powder, the degree of necrosis was significant reduced. Compared with the model group, BMD were increased, total cholesterol, triacylglycerol, and low density lipoprotein levels were decreased, and the VEGF expression increased in the treatment groups. The results suggest that antler powder has a positive curative effect on ANFH by promoting bone formation, fat metabolism and increasing VEGF expression. Related Articles | Metrics

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Influence of effective ingredients of Chinese kidney-tonifying drugs on rat bone healing and hemorheology Zhang Di, Jia Zhi-jie, Tian Yong-li, Xu Zhi-yu 2011, 15 (24):  4545-4548.  doi: 10.3969/j.issn.1673-8225.2011.24.043 Abstract ( 111 )   PDF (411KB) ( 398 )   Save BACKGROUND: Chinese kidney-tonifying herb is commonly used in orthopedics department. However, individual effects of different ingredients remain poorly understood. OBJECTIVE: To observe influence of effective ingredients of Chinese kidney-tonifying drugs on bone healing and hemorheology of rats with bone injury. METHODS: Rat model of fracture of shaft of the femur was established, and treated by intragastric administration separately with extractive rhizoma drynariae total flavonoids, epimedium total flavonoids, dodder total flavonoids, naringin, quercetin and hesperidin. Blood and bone samples were harvested after 21 days to detect the extent of bone healing and hemorheology indexes. RESULTS AND CONCLUSION: The effective constituents of different Chinese kidney-tonifying drugs were beneficial to bone healing, and the effect of rhizoma drynariae total flavonoids and epimedium total flavonoids was better than the other drugs. Rhizoma drynariae total flavonoids and epimedium total flavonoids effectively inhibited blood viscosity under low shear, and reduced erythrocyte aggregation index and the aggregation and adhesion of platelet (P < 0.05), but it had no evident effects on erythrocyte deformation. This shows that the effective ingredient of the Chinese kidney-tonifying drugs can promote bone healing and exert certain effects on hemorheology. In particular, rhizoma drynariae total flavonoids exhibits the best effect. Related Articles | Metrics

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Prokaryotic expression of antibiotic peptide Cecropin A gene and identification of expression products Chen Lei 2011, 15 (24):  4549-4552.  doi: 10.3969/j.issn.1673-8225.2011.24.044 Abstract ( 168 )   PDF (343KB) ( 437 )   Save BACKGROUND: Cecropins are a kind of micromolecule protein with antibacterial activity. Eukaryotic cell-expressed or artificially synthesized Cecropins is characterized by low efficiency and high cost. OBJECTIVE: To clone and express an antibacterial peptide gene of Musca domestica Cecropin A, and to identify recombinant expression product. METHODS: Mature Musca domestica Cecropin A encoding nucleotide sequence was searched from the GenBank and amplified by RT-PCR. The gene of Musca domestica Cecropin A was cloned into prokaryotic expression vector pET32a and fused with gene of Thioredoxin (Trx) and expressed in E.coli BL2l (DE3). After induction by isopropyl-β-D-thiogalactoside, the sera of the immunized rabbits were collected after rabbits were immunized with the hemolymph of housefly larvae. Recombinant protein was identified by western blot analysis and N-[Tris(hydroxymethyl)methyl]glycine-sodium dodecylsulfate-polyacrylamide gel electrophoresis. RESULTS AND CONCLUSION: After induction by isopropyl-β-D-thiogalactoside, E.coli BL21 expressed mature Cecropin. Rabbit anti- housefly larvae sera, N-[Tris(hydroxymethyl)methyl]glycine-sodium dodecylsulfate-polyacrylamide gel electrophoresis and western blot analysis results confirmed that expression products were mature Cecropin. These suggest that prokaryotic expression system can be utilized to obtain natural mature Cecropin. Related Articles | Metrics

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Construction and identification of RNAi expressing vector specific for leucine-rich repeats and immunoglobulin-like domains 1 gene and selection of stably transfected cell clone Liu Hong-chao, Wang Bao-feng, Xie Rui-fan, Cai Ming-jun, Guo Dong-sheng, Lei Ting 2011, 15 (24):  4553-4556.  doi: 10.3969/j.issn.1673-8225.2011.24.045 Abstract ( 108 )   PDF (363KB) ( 246 )   Save BACKGROUND: Leucine-rich repeats and immunoglobulin-like domains 1 (LRIG1) gene showed low expression in glioma cells. LRIG1 gene overexpression significantly enhanced LRIG1 mRNA and protein expression and inhibited its biological behavior. However, very few researchs are reported from the stand-point of inhibition of LRIG1 gene expression. OBJECTIVE: To construct specific RNA interference plasmids for LRIG1, establish stably transfected human glioma GL15 cell line, and observe its effect on expression of target gene LRIG1. METHODS: Designed and synthesized two shRNAs (named LRIG1-shRNA1 and LRIG1-shRNA2) specific for LRIG1 mRNA according to the GenBank, and one scrambled shRNA sequence as negative control, named pGenesil2-negative shRNA. The shRNA was inserted into pGenesil2 vector and sequenced. The recombinant vectors were transformed into E. coli. Picked up the positive clones and extracted the plasmids, which were transfected into GL15 cells by Metafectine. G418 was applied to select the stably transfected cell clones. Western Blotting was performed to examine the LRIG1 protein level. RESULTS AND CONCLUSION: The recombinant plasmids which contain shRNA were analyzed by restriction endonuclease digestion and DNA sequence, and it was proved that the fragment was inserted into the expected sites. Compared with the negative control group, the level of LRIG1 protein expression in pGenesil2-LRIG1-shRNA1(LRIG11) transfected cells and in pGenesil2-LRIG1-shRNA2(LRIG12) transfected cells was decreased by 47.9% (P < 0.01) and 32.8% (P > 0.05). The results confirmed that RNAi expressing vector specific for LRIG1 gene (pGenesil2-LRIG1-shRNA1) was successfully constructed, and the stable cell clones transfected with the shRNA expression vector showed inhibition of the expression of LRIG1 in glioma cell line GL15. Related Articles | Metrics

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Efficient purification and label of anti-human beta 2-microglobulin light chain monoclonal antibody Ruan Guang-ping, Yao Xiang, Pang Rong-qing, Wang Xing-ming, Dai Ying, Pan Xing-hua 2011, 15 (24):  4557-4560.  doi: 10.3969/j.issn.1673-8225.2011.24.046 Abstract ( 83 )   PDF (368KB) ( 283 )   Save BACKGROUND: The inoculation of hybridoma cell strain onto mouse abdominal cavity may obtain ascites containing mass antibody. Previous method to purify monoclonal antibody in ascites is complex and difficult to operate. OBJECTIVE: To prepare, purify and label anti-human leukocyte antigen (HLA)-I class molecule light chain monoclonal antibody, and to detect the expression of tumor cell surface HLA-I class molecules. METHODS: Hybridoma cells were inoculated onto the mouse abdominal cavity. Ascites containing anti-human light chain beta2-microglobulin antibody were obtained and purified with the modified caprylic acid-ammonium sulfate method. The purified monoclonal antibody was labeled with fluorescein isothiocyanate to detect peripheral blood mononuclear cells, T2 cells expressing blank HLA-A2 molecule and K562 cells surface HLA-I class molecules. The expression of HLA-I class molecules was determined by using flow cytometry and fluorescent microscopy. RESULTS AND CONCLUSION: The purified anti-human light chain beta2-microglobulin-fluorescein isothiocyanate monoclonal antibodies accounted for 96% purity. Flow cytometry results showed that, the HLA-I class molecules were highly expressed in peripheral blood mononuclear cells surface, lowly expressed in T2 cells, and not expressed in K562 cells surface. It is a simple and convenient method to purify ascites with the modified caprylic acid-ammonium sulfate method, and according prepare anti-human light chain beta2-microglobulin-fluorescein isothiocyanate. This method is effective to distinguish the levels of HLA-I class molecules expressed in various cells. Related Articles | Metrics