Objective To observe the changes in the expression of renal tissue TNF-α , NF-κB and the interrelation to renal cell apoptosis, and their influences of Inula Britannica(an inhibitor of inflammatory signal pathway) in exhausted swimming rats, and to investigate the role of inflammatory signal pathway. Methods Forty-eight male Wistar rats were randomly divided into three groups: control group (CN, n=8), exhaustive swimming group (ES, n=24) and Inula Britannica group (IB, n=16). The rats of CN were quiet without swimming. The rats of ES swam to exhaustion and were sacrificed at immediately (ESI, n=8), 6 hour (ES 6 h, n=8) and 24 hour (ES 24 h, n=8) after exhanstiing swimming. The rats of IB group took orally Inula Britannica at the dose of 25 ml/kg body weight at 24 h before swimming and then swam to exhaustive state. The rats of IB group were sacrificed at 6 hour (IB 6 h, n=8) and 24 hour (IB 24 h, n=8) after exhaustiing swimming. The renal cell apoptosis was measured by the method of terminal-deoxynucleotidyl transferase mediated d-UTP nick end labeling (TUNEL). The expression of TNF-α in renal tissue was examined by immunohistochemistry. The changes of NF-κB in renal tissue were measured by flow cytometry and immunnhistochemistry. The interrelation between TNF-α and NF-κB was analyzed by Pearson method, and the interrelation between TNF-α, NF-κB and renal tissue cell apoptosis was analyzed by Spearman method. Resulls The number of renal tissue apoptotic cells was increased progressively from ESI to ES 24 h rats (P <0.05). Immunohistochemistry staining showed that the positive expressions of renal tissue TNF-α and NF-κB were increased progressively at 0 h (0.136±0.009, 0.129±0.011), 6 h (0.171±0.011, 0.166± 0.009) and 24 h (0.229±0.008, 0.218±0.019) after exhaustiing swimming in ES compared with control group (0.109±0.010, 0.095±0.010) ( all P<0.05). The similar changes of renal tissue NF-κB was also revcalved by flow cytometry. The expression of TNF-α was positively correldted with NF-κB (r=0.955, P<0.01 ), and renal cell apoptosis was also positively correlated with TNF-α and NF-κB (r =0.953, r=0.939, P<0.01) in ES rats. Pretreatment with Inula Britannica, inhibited the up-regulation of expressions of renal tissue TNF-α (6 h:0.142±0.012, 24 h:0.130±0.010) and NF-κB (6 h:0.138±0.010, 24 h:0.136±0.011 ) induced by exhausting swimming. Conclusion Overtraining can induce the up-regulating expressions of renal tissue TNF-α and NF-κB, and Inula Britannica can partly counter the above changes in exhaustied swimming rats, which may be one important mechanisms of overtraining-induced renal tissue cell apoptosis and the anti- apoptosis effect of Inula Britannica.

Objective To explore remote effect of decalcification bone matrix (DBM) in treatment of delayed union and nonunion of long bone fractures. Methods From 1986, 96 cases of delayed union and nonunion of long bone fractures were treated with DBM. Of all, 38 cases of nonunion of long bone were treated with surgical implantation, 37 delayed union and 21 nonunion with percutaneous injection. Results All 96 cases were followed up for 4-16 years (average 7.5 years). Of 37 cases of delayed union treated with percutaneous injection, 35 attained bone union but 2 resulted in nonunion of tibial fractures with a union rate of 94.6%. Of 21 cases of bone nonunion treated with percutaneous injection, 17 got bone union but 4 did nonunion (3 tibial nonunion and 1 humeral nonunion) with a union rate of 80.1% and union time for 3-8 months (average 4.5 months). Of 38 cases treated with implantation of DBM, 36 had bone union but 2 nonunion at 1/3 inferior to the tibia with a union rate of 94.7%. Conclusions Either percutaneous injection or surgical implantation of DBM can attain satisfactory effect even to autograft. After prepared by dehydration, degrease, decalcification and irradiation disinfection, DBM is characterized by safer transplantation and less immunity than other bone transplantations and can be preserved for a long time so as to be applied both in the peacetime and the wartime.

Objective To study the expressions of apoptosis related genes Bcl-2 and Bax proteins in rat's renal tissue and the relationship between the ratio of Bcl-2/Bax and renal cell apoptosis induced by overtraining,and observe the effect of anisodamine on the expression of genes Bcl-2 and Bax in exhausted rats.Methods The animal model of acute kidney injury induced by exhausting swimming was reproduced.Forty eight male Wistar rats were randomly assigned into sedentary control group(CN,n=8),exhausting swimming group(immediate,6 hours and 24 hours after exhausting swimming,ESI,ES6h and ES24h,8 each),Anisodamine group(6 hours and 24 hours after exhausting swimming,AD6h and AD24h,8 each).The expressions of Bcl-2 and Bax were detected by immunohistochemical staining and image analyzer.The correlation between the ratio of Bcl-2/Bax and renal cell apoptosis was analyzed.Results The analysis showed that the expression of Bax increased,and of Bcl-2 decreased,the ratio of Bax/Bcl-2 increased remarkably in ESI group compared with CN group(P

Objective To observe the expression of DR5 in the ocular tissues of uveitis rats induced by endotoxin,and study the relationship between the apoptosis of inflammatory cells and expression of TRAIL / DR5.Methods SD rats were randomly divided into 3 groups:Blank control group,normal saline injection group and endotoxin injection group.The endotoxin injection group was injected with lipopolysaccharide into the rat posterior foot pad to make endotoxin-induced uveitis animal model.There were no operations in the blank control group,and the subgroups were divided into 6 hours,12 hours,24 hours and 48 hours groups according to the time of injection.The ultrastructural changes of inflammatory cells and endothelial cells in iris capillaries were observed by transmission electron microscopy (TEM).The expression of DtR5 protein on inflammatory cells at different time after endotoxin induction was detected by SABC method.Results TEM showed that the microvilli of the capillary endothelial cells in the iris tissue of the blank control group and saline injection group had more obvious vesicles with no obvious abnormal structure and shape.The number of swallowed vesicles in the capillary endothelial cells injected with endotoxin was decreased at 6 hours group,and the number of vesicles in the infiltrating neutrophils and lymphocytes decreased.Neutrophils and lymphocytes appear chromatin condensation,vacuolar changes in the expression of apoptosis.Immunohistochemistry showed that the DR5 protein was negative in the iridocular epithelium of the blank control group and saline injection group.In the endotoxin injection group,the DR5 protein was weakly colored in the iris pigment epithelium and appeared on the inflammatory cells.The number of staining and the intensity of coloring in the 24 hours group were significantly higher than those in the 6 hours group,and the color density was 0.085 9 ± 0.019 6,there were statistical differences compared with 6 hours group,12 hours group and 48 hours group (all P ＜ 0.05).Conclusion TRAIL and its receptor DR5 may be involved in the apoptosis of inflammatory cells in endotoxin-induced uveitis.

Adult chondrocytes had been used as seed cells in the previous tissue engineering; however, they possess the weaknesses including the limited proliferative capability in vitro and the liability to aging after amplification. Bone marrow stromal cells (BMSCs) are multipotent cells, which can differentiate into osteoblasts, chondrocytes, and adipocytes. It is of great importance to study the regulation and control of BMSC directed induction because directed differentiation is required in the tissue engineering. During the BMSC differentiation towards chondrocytes, serious kinds of biological inducing factors participate in precise induction as signal factors. The physical factors, such as biomechanical strength and ultrasound, have been shown to be involved in the regulation of BMSC differentiation towards chondrocytes. In terms of tissue repair, apart from biological factors which play an important role in the formation of cartilage tissue, the chondrocyte microenvironment in vivo is indispensable. Bioreactor is a kind of device intended for in vitro tissue culture that incubates the cells or tissues taken from living bodies in simulated physiological environment in vivo. On the basis of original cell culture, the present bioreactors apply biomechanical stimulation to simulate the stressed environment of articular cartilage in vivo.

Objective To observe the expression changes of renal tissue Bax,Bcl-2 and caspase-3,to wxamine the correlation between the ratio of Bax to Bcl-2,caspase-3 and renal tubular cells apoptosis,and to investigate the role of caspase-related signal pathway.Methods Forty-eight male Wistar rats were randomly divided into three groups: control (CN,n=8),exhaustive swimming (ES,n=24) and inula britannica (IB,n=16) group.The rats of CN were quiet without swimming.The rats of ES swam to exhaustive state and were sacrificed at immediately(ESI),6 hour (ES 6 h) and 24 hour (ES 24 h) after exhaustive swimming respectively.The rats of IB took orally inala britannica at the dose of 25 ml/kg body weight at 24 h before swimming and then swam to exhaustive state.The rats of IB group were sacrificed at 6 hour (IB 6 h) and 24 hour (IB 24 h)after exhaustive swimming.The animal model of overtraining-induced acute kidney injury was developed by exhaustive swimming.The renal cell apoptosis was measured by the method of TUNEL.The expressions of Bax,Bcl-2,caspase-3 in renal tissue were observed by immunohistochemistry.The expression of caspase-3 protein was examined by Western blotting.The correlation between the ratio of Bax to Bcl-2 and caspase-3 was analysed by Pearson method,and the correlation between the ratio of Bax to Bcl-2,caspase-3 and renal tubular cell apoptosis was analysed by Spearman method.Results The number of renal tubular apeptotic cells was increased progressively in ESI to ES 24 h rats by TUNEL (P＜0.05).Immunohistochemistry staining showed that the ratio of Bax to Bcl-2 and caspase-3 in renal tubular cells were increased progressively at 0 h,6 h and 24 h after exhaustive swimming compared with control group (P＜0.05).The change of renal tissue caspase-3 was also revealved by Western blotting analysis.The ratio of Bax to Bcl-2 and caspase-3 in renal tubular cell was correlated positively (r=0.865,P＜0.05),The ratio of Bax to Bcl-2,and caspase-3 was also correlated positively to renal tubular cell apoptosis (r=0.674,r=0.837,P＜0.05) in ES rats.Pretreatment with inula britannica inhibited the up-regulation of the ratio of Bax to Bcl-2,caspase-3 and cell apoptosis in renal tubular cell induced by exhaustive swimming.Conclusion Overtraining can induce renal tubular cells apoptosis through activating caspase-related signal pathway by impairing the balance of Bax and Bcl-2,which may be one of the important molecular mechanisms of overtraining-induceed renal tubular cells apoptosis.

This study is to investigate the effect of downregulation histone deacetylases 1 (HDAC1) gene by the technology of RNA interference on the differentiation of HL-60 cells line. The optimal segment targeting HDAC1 gene was designed and transfected into HL-60 cells by Lipofectamine 2000. The HDAC1 mRNA and protein level were detected by RT-PCR and Western blotting. The morphologic change of HL-60 cells was detected by an optical microscope with Wright-Giemsa. Cell differentiation was tested by NBT reduction assay. Expression of CD13, CD33 and CD14 was measured by flow cytometry. The results indicated that HDAC1 mRNA and protein were markedly suppressed by the siRNA targeting HDAC1 in a concentration-dependent manner. HDAC1 siRNA promoted cell differentiation. HL-60 cells became more mature in morphology after transfected to HDAC1 siRNA at a concentration of 30-60 nmol x L(-1) for 24 h. NBT reduction ability of HDAC1 siRNA with 30 nmol x L(-1) was 0.31 +/- 0.09, compared with negative control (0.20 +/- 0.02) (t = -3.1, P < 0.01), and with 60 nmol x L(-1) was 0.25 +/- 0.02 in comparison with negative control (0.21 +/- 0.04) (t = -2.12, P < 0.05). But it has no change in HDAC1 siRNA > or = 120 nmol x L(-1). After transfection with 60 nmol x L(-1) HDAC1 siRNA to HL-60 cells, the expression of CD13 was (96.50 +/- 0.70)% in compared to siRNA-NC (3.39 +/- 0.68) % (t = 164.9, P < 0.000 5), CD33 was (66.73 +/- 0.50) % in compared to siRNA-NC (96.80 +/- 1.70) % (t = 43.4, P < 0.000 5). CD14 was (0.53 +/- 0.00) % by comparison with siRNA-NC (0.49 +/- 0.02) % (t = -0.97, P > 0.1). HDAC1 siRNA promoted cell differentiation in indicated concentration. HDAC1 might be one of the targets of gene therapy for leukemia.

Carbonic anhydrase(CA) is one of a group of enzyme that contains zinc. They exist widely in the mammals. CA has at least 14 species of isoenzyme. Among them,CAIX is located at the downstream of VHL tumor suppressor gene,and it can be activated through HIF-1 pathway. CAIX primarily exists in the mucous membrane of gastrointestinal tract in normal human tissues and its expression is low. However,higher expression of CAIX can be detected in renal cell carcinoma,cervical carcinoma and lung cancer,it may be closely related with the occurrence and development of the tumor. In addition,we can predict the prognosis of tumor patients by analysis of the expression of CAIX,thus,it is considered that CAIX is a tumor specifi c antigen. Due to its highly specifi c expression in renal cell carcinoma,CAIX will become the potential target antigen of tumor vaccine as well as the target gene for the treatment of RCC. The application of CAIX shows high promise in RCC target therapy. Hence,it is defi nitely important for us to investigate the expression and regulation of the CAIX.

Objective To verify the authors' previous cDNA micro-array results and to further investigate the moleculer mechanisms of gastric cancer. Methods Quantitative real-time RT-PCR was employed to detect the expressions of three S100A calcium-binding genes in 22 fresh surgical samples of gastric tumor tissue and non-cancerous mucosa from the same patients. Results The transcription level of S100A2 in primary cancer lesion was elevated in 80% of samples when compared with matching non-neoplastic mucosa (P=0.018) and the average up-regulation level was 10.78 fold. 55% of cancer lesions showed higher transcription level of S100A4 than their adjacent non-neoplastic mucosa, the average up-regulation level was 2.31 fold. S100A6 transcription level was higher in 74% (P=0.01) of primary cancer lesion with an 2.25 fold up-regulation than the adjacent non-neoplastic mucosa. After rectified by ?_2-microglobulin, the relative expression levels of S100A2, S100A4 and S100A6 were 2.83?10~ -4 , 6.44?10~ -2 and 0.41, respectively. According to the Spearman correlation coefficient analysis there were significant positive correlations between S100A2 and S100A4, and S100A2 and S1006 (P value were 0.00 and 0.017, respectively). Conclusion The changes in S100A2 and S100A6 genes may be an early event in a majority of gastric cancer patients, while S100A4 may be associated with the infiltration of gastric cancer. Further study on the three genes might be helpful for understanding the nature of gastric carcinoma.

Objective To study the expression of S100A4 in gastric cancer and normal gastric tissue, and analyze its correlation with the clinico-pathological features and prognosis of gastric cancer. Methods Real-time quantitative reverse transcription PCR (real time qRT-PCR) was used to detect the S100A4 expression in 20 fresh surgical samples of gastric cancer and normal gastric tissue as controls. The microarray of gastric cancer tissue was established for the analysis of the S100A4 expression immunohistochemically in 208 gastric cancer tissue and isogeneic normal gastric mucosa and lymph node with metastasis. Results The S100A4 expression was increased in 55% (11/20) of gastric cancer samples with an average of 2.31 fold up-regulation of that of the normal mucosa. Patients with lymph node metastasis showed a higher percentage of elevated S100A4 transcription than those without metastasis (P=0.024). As displayed by immunohistochemistry, the positive rate of S100A4 in non-neoplastic mucosa, primary tumor and lymph node with metastasis was 9.4%, 28.1% and 32.2%, respectively (P≤0.01). A higher percentage of elevated S100A4 expression was shown in patients in advanced stage than in patients in early stage (P=0.004). In primary tumor lesions, the S100A4 expression correlated significantly with the depth of invasion (P=0.003) and poorer prognosis (P=0.034). S100A4 expression in lymph node with metastasis was also associated with poor outcome (P=0.002). Multifactorial Cox's regression analysis showed that TNM stage (P=0.029) and the expression levels of S100A4 (P=0.024) in lymph node were independent influence factors for prognosis. Conclusions Expression of S100A4 may be a late event which is associated with the progression and prognosis of gastric cancer. The analysis of S100A4 expression in lymph-node metastasis is helpful in judging the prognosis of gastric cancer.