Objective To compare expression of extracellular proteins of virulent H37Rv and attenuated H37Ra in order to search differential proteins,to provide a train of thought for studing M.TB toxicity further.Methods Extracellular proteins were extracted from H37Rv and H37Ra which were inoculated and cultured on Suton's medium for three weeks.The first dimensional electrophoresis was performed on immobilized pH gradient rod gels(pH 3～10).Then the proteins in the rod gels were separated using SDS-PAGE gels.The silver-stained gels were dried and scanned with image scanner.The 2D image analysis was performed with image Master 2D Elite 3 10.Results The most protein spots deriving from extracellular proteins of H37Rv and H37Ra strains were in acidic range.In the basic range(pI more than 9 0),the number of protein spots belong to extracellular proteins of H37Rv and H37Ra was few.Three protein spots belong to low molecular range in H37Rv strain.However,absent in H37Ra strain.Conclusions Two-dimensional gel electrophoresis is useful to separate protein in Mycobacterium tuberculosis.

Objective To evaluate the prospects of recombinant 38000 protein of Mycobacterium tuberculosis in tuberculosis epidemic investigation and subunit vaccine preparation.Methods Physicochemical characteristics of recombinant 38000 protein was detected by P I, peptide-mapping analysis and circular dichroism,guinea pig skin test,MTT stain,and peripheral blood macrophage phagocytosis were used to investigate the roles of recombinant 38000 protein in the cell immunity.Results Recombinant 38000 protein was acidic protein,its P I, was 4 67.The number of alkaline amino acid correspond with theoretic number;The secondary structure of recombinant 38000 protein was composed of ?-helix(32 6%),?-turn(31 6%) and random coil(35 8%) Recombinant 38000 protein could induce DTH in guinea pig sensitized by Mycobacterium tuberculosis Recombinant 38000 protein enhanced phagocytosis of macrophage in mice . PBMC from 30 8% healthy donors and 25% tuberculosis patients were stimulated by the recombinant 38000 protein.Conclusion Recombinant 38000 protein may be used as diagnostic reagent and as an candidate in development of subunit vaccine.

Objective To investingate the role of inflammation in coronary atherosclerosis and the relationship between the inflammatory factors and coronary risk score. Methods 56 patients with acute coronary syndrome were studied, among them 26 cases were diagnosed as acute myocardial infarction (AMI) and 30 unstable angina pectoris (UAP). The study group was compared with a control group of 30 cases who were identified as normal by coronary angiography. The concentrations of serum sICAM-1 and hsCRP were determined by ELISA assay. The coronary risk score was recorded in patients with UAP. Results Serum sICAM-1 levels were significantly elevated in patients with AMI or UAP compared with that of control group, while higher hsCRP level was observed only in the patients with AMI compared with those with UAP and the control group. By linear regression analysis, only serum sICAM-1 levels were correlated with coronary risk score (r=0.445, P

Aim To explore the influence of atorvastatin on the ultramicrostructure of the membrane surface of the rabbit endothelial cells in rabbit atherosclerosis(AS)in the nanometer level.Methods A total of 44 male New Zealand white rabbits were randomly divided into 3 groups:control group consisting of 12 rabbits,AS group consisting of 16 rabbits and atorvastatin group consisting of 32 rabbits.By the end of 2nd,6th week 6~8 rabbits of each group were sacrificed and the middle segments of thoracic aortas were obtained to be observed with atomic force microscope.Results The control group vascular endothelial cells(VECs)were fusiform in shape and aligned regularly.Their size were about 11.96 ?m?3.72 ?m and their macroaxis were in parallel with the direction of hemokinesis.VECs in the atherosclerotic group were in deformity and bigger than those of the control group.They aligned irregularly and their volumes changed to be swelled.The membrane protein of VECs in the control group was composed of many round and elliptical eminences,which were almost in the same size.and with distinct boundary lines.The membrane protein of VECs in the atherosclerosis group was composed of many irregular eminences in different size.It was vague among the eminences in which there were many holes.But the VECs of atorvastatin group were better than those of atherosclerosis group.The ultramicrostructure of the membrane surface of the atorvastatin group VECs was obviously improved.Meanwhile,the mean roughness(Ra)of membrane protein of three groups was compared.The Ra of the atherosclerosis group was significantly higher than that of the control group and the atorvastatin group(P

Objective To study the correlation of expression of DNA topoisomerase Ⅱ alpha (TOP2A)with expressions of human epidermal growth factor receptor 2 (HER2)and phosphatase and tensin homolog (PTEN)and gene mutation of phosphatidylinositol 3-kinase (PI3K)in breast cancer so as to provide reference for prognosis of the cancer and evaluation of drug efficiency.Methods This study enrolled totally 96 breast cancer patients. Tumor specimens were resected.The gene expressions of TOP2A,HER2 and PTEN were analyzed using branched DNA-liquid-chip,and PI3K gene mutation was detected by xTAG-liquid-chip.Correlations between gene expressions and gene mutation were further explored by Spearman correlation analysis so as to clarify the relationship between TOP2A and HER2 signaling pathway gene.Results Co-expression of TOP2A and HER2 was strong,and TOP2A tended to be highly expressed in the presence of high expression of HER2 (P =0.01).The expression of PTEN was not significantly correlated with the expression of TOP2A,whereas the mutation of PI3K had a positive association with the high expression of TOP2A (P =0.004).Conclusion Anthracycline drug resistance factor TOP2A may be related to the critical factors of HER2 signaling pathway,suggesting that HER2 expression and PI3K mutation may be key factors in regulation of TOP2A expression,which would provide important evidence for chemotherapeutic resistance.

Objective To investigate the growth inhibition effect of evodiamine (Evo) on renal carcinoma 786-0 cells and to explore its molecular mechanism. Methods After treated with Evo, methyl thiazolyl tetrazolium ( MTT) assay was used to detect the vitality of 786-0 cells, flow cytometry was employed to examine the cell cycle distribution in 786-0 cells, and immunoblotting was utilized to determine the expression levels of target proteins related to cell cycle progression. Results Evo remarkably inhibited 786-0 cells vitality in dose-dependent manner. Cell cycle analysis indicated that 786-0 cells were arrested in G2/M phase followed by Evo treatment. Furthermore, the results of immunoblotting showed that Evo up-regulated the protein expression levels of P53, P21 and its downstream target gene CyclinB1 in 786-0 cells. Conclusion Evo treatment can induce 786-0 cell cycle G2/M arrest, and its underlying mechanism might be dependent on the P53/P21 signal pathway.

Objective To evaluate the frequency of pncA gene mutations in pyrazinamide-resistant (PZA-resistant)Mycobacterium tuberculosis(M.TB)isolates.Methods The isolates were tested for PZA susceptibility with absolute concentration method.The pncA gene was amplified and the products were cloned into T-vector,followed with sequencing.Results In all the 36 M.TB isolates,there were 25 PZA-resistant strains and 11 PZA-susceptible strains.Mutations of the pncA gene were found in 10 PZA-resistant isolates,four of which belonging to high PZA-resistant strains.The wild type pncA sequence was present in 3 PZA-susceptible strains, synonymous mutations of pncA gene were detected in two PZA-susceptible strains.Additionally,11 PZA-resistant strains and 2 PZA-susceptible ones showed putative regulatory mutations in the upsteam of pncA gene.Condusions The mutation of the pncA gene can cause the PZA resistance.However.there ale other drug resistance mechanism except this mutation.

Objective To study the effects of MF59 in combination with heat-killed BCG ( hBCG) as adjuvant on the immunogenicity of Mycobacterium tuberculosis fusion protein PstS1-LEP.Methods BALB/c mice were divided into six groups from group 1 through group 6.They were immunized with PstS1-LEP+MF59 ( group 1 ) , PstS1-LEP+MF59/hBCG ( group 2 ) , PstS1-LEP+hBCG ( group 3 ) , MF59 ( group 4 ) , PstS1-LEP (group 5) and hBCG (group 6) for three times at intervals of two weeks , respectively.The mice were sac-rificed two weeks after the last immunization .The serum samples were collected for antibodies detection .The splenic lymphocytes and peritoneal macrophages were isolated and cultured with PstS 1-LEP.Indirect ELISA and sandwich ELISA were used to detect PstS 1-LEP-specific antibodies and cytokines in the supernatants of culture , respectively.Results The level of IFN-γ, IL-1β, IgG, IgG1 and IgG2a in group 1 were higher than those in groups 4, 5 and 6 (P<0.05).The level of IL-2 and IL-4 in group 1 were higher than those in groups 4 and 6 (P<0.05).The level of IFN-γ, IL-1β, IL-12, IgG, IgG1 and IgG2a in group 2 were higher than those in groups 4, 5 and 6 (P<0.05).The level of IL-2 was higher in group 2 than that in groups 4 and 6 (P<0.05). The level of IL-4 in group 3 was higher than that in group 4 ( P=0.05 ) .The level of IL-1βin group 3 were higher than that in groups 4 and 5 ( P<0.05 ) .The level of IgG was higher in group 3 than that in groups 4 and 6 (P<0.05).IgG1 level in group C was up-regulated in comparison with that in groups 4, 5 and 6 (P<0.05 ) .Conclusion hBCG as PstS1-LEP adjuvant induces a shift towards Th 2-type immune response , while MF59 induces Th1/Th2-type immune response.The combination of MF59 and hBCG inhibits the secretion of IL-4 by spleen lymphocytes , but enhances the secretion of IL-12 by macrophage .

BACKGROUND: Several studies have demonstrated that many American women who are at high risk of developing osteoporosis have higher levels of serum phosphorus. This indicates that some substances which can lower the serum level of phosphorus will supply a new and effective method to prevent and treat osteoporosis.OBJECTIVE: To observe the influences of porcine bone protein on bone mineral density (BMD) and serum levels of calcium and phosphorus in a rat model of osteoporosis.METHODS: Wistar rat models of osteoporosis were established by intramuscular injection of dexamethasone. Rat models were randomly divided into physiological saline, Jiegu Qili tablet, 50, 100, 200 mg/kg porcine bone protein groups. Rats that did not receive any treatments served as normal controls. After 12 weeks of treatment, serum was collected and serum levels of phosphorus and calcium were determined by biochemistry method. At the same time, tibia sections were made to determine tibial DMD by QDR-400 dual energy X-ray absorptiometry and to observe tibia marrow cavity by hematoxylin-eosin staining.RESULTS AND CONCLUSION: There was no significant difference in serum level of calcium among groups (P＞0.05).Compared with the physiological saline group, serum level of phosphorus in the 50, 100, 200 mg/kg porcine bone protein groups was significantly decreased (P ＜ 0.05). BMD was significantly higher in the 50, 100, 200 mg/kg porcine bone protein, Jiegu Qili tablet groups than in the physiological saline group (P ＜ 0.05). The tibia marrow cavity was smallest in the normal control group and largest in the physiological saline group. The tibia marrow cavity was larger in the 50, 100, 200 mg/kg porcine bone protein,Jiegu Qili tablet groups than in the physiological saline group. These results indicate that porcine bone protein cannot change the serum level of calcium, but it lowers serum level of phosphorus, and increases BMD, in a rat model of osteoporosis. However, the dose-dependent effect of porcine bone protein was not observed within the present experimental dosage. In addition, porcine bone protein can also reduce the marrow cavity of the tibia of rats with osteoporosis.

Objective To explore the risk factors for clinical anastomotic leakage after resection of rectal cancer in China. Methods By meta-analysis we made a comprehensive analysis of the risk factors for clinical anastomotic leakage after resection of rectal cancer based on 19 articles published in China between January 1999 and January 2009. Results The anastomotic leakage rate was higher in the patients aged 60 years old and above than in those younger, with the combined odds ratio (OR) value being 0.50 (95% CI: 0.33-0.76) (P<0.01). The incidence rate was higher in the male patients than in the female ones, with the combined OR value being 2.17 (95% CI: 1.38-3.42) (P<0.01). The incidence rate in the patients with the distance of tumor from the lower margin to anal verge being 7cm and shorter was higher than that with longer distance, with the combined OR value being 1.79 (95% CI: 1.37-2.35) (P<0.01). The incidence rate in the patients who had received radiotherapy preoperatively was higher than that in those who had not, with the combined OR value of 3.66 (95% CI: 2.19-6.09) (P<0.01). The incidence rate in the patients who had received stapler anastomosis was higher than that in the patients who had received manual anastomosis, with the combined OR value being 0.70 (95% CI: 0.47-1.05), but there was no significant difference between them (P>0.05). The incidence rate was higher in the patients with diabetes mellitus than in the healthy ones, with the combined OR value being 3.16 (95% CI: 2.27-4.39) (P<0.01). The incidence rate was lower in the patients with Dukes A and B stages than in those with Dukes C and D stages, with the combined OR value being 0.61 (95% CI: 0.45-0.83) (P<0.01). The incidence rate in the patients with high malignance degree in clinicopathological types was higher than that with low malignance degree, with the combined OR value being 2.17 (95% CI: 1.38-3.42) (P<0.01). The incidence rate was lower in the patients who had received preventive colostomy than in those who had not, with the combined OR value being 0.39 (95% CI: 0.14-1.05), but there was no significant difference between them (P>0.05). The incidence rate was higher in the patients who had got selective operation than in those who had got emergency operation, with the combined OR value being 0.27 (95% CI: 0.13-0.56). Conclusion The risk factors of anastomotic leakage after resection of rectal cancer are as follows: 60 years old and above, male patients, diabetes mellitus, preoperative neo-adjuvant radiotherapy, the distance of tumor from the lower margin to the anal verge being shorter than 7cm, Dukes C and D stages, high malignance degree in clinicopathological types, and emergency operation.