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.

Objective To investigate the efficacy and safety of tirofiban for emergency interventional therapy in patients with acute ST segment elevation myocardial infarction.Methods The 96 acute ST segment elevation myocardial infarction patients having accepted emergency percutaneous coronary intervention (PCI) were divided into treatment group and control group according to the treatment method with 48 cases each.The treatment group was given conventional standard treatment combined with tirofiban treatment [tirofiban intracoronary injection 10 μ g/kg and then intravenous 0.15 μ g/(kg· min) for 24-48 h].The control group was given conventional standard treatment only.The postoperative 60 min electrocardiogram ST segment recovery,the coronary blood flow TIMI grade,the major adverse cardiac events (angina,myocardial infarction,heart failure,death) and postoperative bleeding in 4 weeks were compared between the two groups.Results The patients having postoperative 60 min electrocardiogram ST segment recovery in treatment group was 45 cases (93.8％,45/48),in control group was 35 cases (72.9％,35/48),and there was statistical difference (P ＜ 0.05).The incidence rate of infarction vascular TIMI grade ≥ 3 grade flow in treatment group was 95.8％ (46/48),in control group was 75.0％ (36/48),and there was statistical difference (P ＜ 0.05).The incidence rate of the major adverse cardiac events 4 weeks in treatment group was significantly lower than that in control group,and there was statistical difference (P ＜ 0.05).There was no statistical difference in the incidence rate of bleeding between the two groups (P ＞ 0.05).Conclusion In patients with acute ST segment elevation myocardial infarction,the emergency interventional therapy with tirofiban is efficacy and safety.

Fructus Bruceae, the dry and ripe fruits of Brucea ja-vanica( L. ) Merr. ( Simaroubaceae) , has been used for dysenter-y, tumor, malaria and furunculosis treatment, and topical appli-cation for warts and corns. Quassinoids are the main chemical constituents of Fructus Bruceae, which have been proved to have anti-tumor, anti-inflammatory, insecticidal, anti-viral, anti-bac-terial and hypoglycemic activities, and so on. In view of the ex-tensive pharmacological activities of Fructus Bruceae, and better development and utilization of its medicinal value, the advances of quassinoids in Fructus Bruceae and their pharmacological ac-tivities are reviewed in this paper.

Objective To investigate the killing effect of ethacrynic acid (EA) on lung cancer A549 cells derived spheres and explore the underlying mechanism.Methods A549 spheres were cultured in serum-free medium,and the protein expression of CD133,SOX2,EpCAM and ABCG2 was detected by Western blotting.MTT assay was used to evaluate the cell viability of A549 spheres and A549 cells after treated by 1,2,5,10 and 20 mg/mL cisplatin (DDP) for 48 h.The activity of glutathione S-transferase (GST) was measured by colorimetric method after A549 spheres were treated with 10,50,100 and 200 μmol/L EA,respectively.Flow cytometry,Western blotting,real-time PCR and luciferase assay were used to analyze the levels of cellular reactive oxygen species (ROS),formation of A549 spheres,mRNA and protein expression levels of β-catenin,Sox2 and ABCG2,and promoter activity of β-catenin upon 200 μmol/L EA treated cells for 48 h.A549 sphere was infected with β-catenin adenovirus for 24 h,followed by 200 μmol/L EA treatment (in presence or absence of 5 mg/mL DDP) for 24 h.The expression of β-catenin,Sox2 and ABCG2 at mRNA and protein levels was detected by real-time PCR and Western blotting,and cell growth of A549 spheres was evaluated by MTT assay.Results The A549 spheres,with high expression of tumor stem cells markers CD133,SOX2,EpCAM and drug resistance related molecule ABCG2,and resistance to DDP at different doses,were successfully derived.After 200 μmol/L EA had treated A549 sphere for 48 h,the levels of ROS were significantly increased (P ＜ 0.05),and the mRNA and protein levels of β-catenin,Sox2 and ABCG2,and promoter activity of β-catenin were notably decreased (P ＜ 0.05).The treatment of 200 μmol/L EA enhanced the inhibitory effect on proliferation and the promoting effect on apoptosis in A549 spheres induced by 5 mg/mL DDP (P ＜ 0.05).Up-regulation of β-catenin by adenoviral infection partly reversed the effects of 200 μmol/L EA on suppressing the expression levels of β-catenin,Sox2 and ABCG2,compared to the spheres infected with blank adenovirus.Additionally,β-catenin over-expression significantly remitted the inhibitory effect of 200 μmol/L EA and 5 mg/mL DDP on the proliferation in A549 spheres.Conclusion EA exerts inhibitory effect on the proliferation and stemness of A549 spheres through suppressing GST activity and β-catenin expression,and then promotes cell apoptosis.EA might be a novel drug in treatment of lung cancer and cancer stem cells.

Total RNA was isolated from Trichomonas vaginalis and Tv-Sir2-like cDNA was amplified by RT-PCR and cloned into pGEM-T Easy plasmid. A fragment of Tv-Sir2-like cDNA was subcloned into the expression vector pET-41b and expressed in E.coli BL21 with induction of IPTG. The full-length of Tv-Sir2-like cDNA was cloned and sequenced. The prokaryotic expression system of pET-41b/Tv-Sir2-like was constructed. The fusion protein of Tv-Sir2-like was expressed in E.coli BL21, occupying 30% of the total bacterial protein after being induced by IPTG for 5 h. SDS-PAGE analysis showed that the fusion protein was about Mr 59 000. The recombinant protein of Tv-Sir2-like is efficiently expressed in E.coli BL21.

Objective To investigate the effects of valsartan on the expression of connective tissue growth factor(CTGF) in rat glomerular mesangial cells incubated with high concentration of glucose.Methods We used high concentration glucose and valsartan to stimulate the cultured rat glomerular mesangial cells in vitro. The protein expressions of CTGF and the activation of P38 mitogen-activated protein kinase(P38 MAPK) and cAMP response element binding protein 1(CREB1) were tested by Western blot. CTGF and fibronectin(FN) mRNA were measured by reverse transcription and polymerase chain reaction (RT-PCR). The protein synthesis of lamanin (LN) and type IV collagen in the supernatants of the GMCs were detected by radioimmunoassay. Results Compared with low glucose control group,the expression of CTGF,p-P38 MAPK,p-CREB1,CTGF mRNA,FN mRNA,LN and type IV collagen in the supernatants were significantly increased in GMCs incubated with high concentration of glucose medium. The expression levels of CTGF,p-P38 MAPK,p-CREB1,CTGF mRNA and FN mRNA were significantlylower in the valsartan group than those in the high concentration glucose group. The concentrations of LN and type IV collagen in the supernatantsin the valsartan group were also lower than those in the high concentration glucose group. Conclusion Valsartan can inhibit expression of CTGF and ECM proteins in rat glomerular mesangial cells incubated with high concentration of glucose,partly by regulating the phosphorylation of P38 MAPK and CREB1.

Objective To study the relationship between the mutation of the inverted repeat (IR) gene in the multiple transferable resistant (mtr) system and multiple antibiotic resistance of Neisseria gonorrhoeae. Methods The antimicrobial susceptibilities of isolated strains were tested. An agar plate dilution method was used to determine the minimum inhibitory concentrations. The target genes were amplified by PCR and subjected to sequencing. Results No mutation was found in the IR gene of either of 2 sensitive or 5 penicillin-resistant Neisseria gonorrhoeas strains. Among the 17 multiple-antibiotic-resistant strains, a strain with both azithromycin- and penicillin-resistance had T/A and T/A insertions, and another had A/T deletion. Conclusion Mutations in the IR gene of the mtr system of Neisseria gonorrhoeae might result in multiple antibiotic resistance.

Objective To discuss the expressions and clinicopathologic significances of insulin-like growth factor Ⅱ mRNA binding protein 3 (IMP3)in squamous intraepithelial lesion (SIL)and cervical squa-mous cell carcinoma (CSCC)before and after the therapy of radiation and chemotherapy.Methods The expressions of IMP3 in 80 cases of CSCC,90 cases of SIL (60 cases of HSIL,30 cases of LSIL)and 30 cases of cervicitis were detected by immunobistochemistry.The relations between IMP3 and clinicopathological cha-racteristics of CSCC were analyzed.Results The expression rates of IMP3 in CSCC,HSIL,LSIL and cervici-tis were 86.3%(69 /80),78.3%(47 /60),33.3%(10 /30)and 0(0 /30),and the difference among the four groups was statistically signicant (χ2 =87.01,P <0.01).The positive expression rate of IMP3 declined by radiation or chemotherapy (60.0% vs.85.0%,χ2 =5.79,P =0.013).The expression of IMP3 was related with lymph node metastasis (χ2 =3.97,P =0.046),differentiated degree (χ2 =5.95,P =0.018),clinical stage (χ2 =5.82,P =0.016)and invasion depth (χ2 =5.73,P =0.017).There was nothing to do with age (χ2 =0.11,P =0.745).Conclusion IMP3 expresses excessively in CSCC,and is associated with pathologi-cal grade and invasion progress.Radio-chemotherapy can reduce the expression of IMP3.

AIM:To investigate the effect of Sonic Hedgehog ( Shh) signaling blockade on the growth of hema-tocarcinoma cells and underlying mechanisms.METHODS: The expression of Shh signaling molecules in hematocarci-noma cell lines BEL-7402, Huh7 and HepG2 was detected by RT-PCR.The cell viability was detected by MTT assay.The cell cycle and apoptosis were analyzed by flow cytometry.The expression of apoptosis-related proteins was determined by Western blot.RESULTS:Shh signaling molecules were all expressed in BEL-7402, Huh7 and HepG2 cells.The mRNA expression of Patched ( Ptch) , Gli1 and Gli2 was down-regulated by anti-Shh antibody.Blockade of Shh signaling pathway inhibited the proliferation of hepatocarcinoma cells with increasing cells in G0/G1 phase and induced the apoptosis of hepa-tocarcinoma cells.Treatment with anti-Shh antibody down-regulated the protein expression of pro-caspase-3, pro-caspase-8 and pro-caspase-9, while up-regulated the protein levels of cleaved caspase-3, cleaved caspase-8 and cleaved caspase-9 in BEL-7402 cells.CONCLUSION:Blockade of Shh signaling pathway inhibits the growth of hepatocarcinoma at different levels by cell cycle arrest and inducing apoptosis of hematocarcinoma cells.

Objective To investigate the renal histopathological changes of over training induced acute renal injury in human being.Methods Eight patients treated in our hospital admitted overtraining were observed retrospectively about their clinical and pathological data,including clinical features,laboratory tests and pathological examinations.Results Eight patients with acute kidney injury after overtraining,manifested as urine occult blood positive in 2 cases,3 cases of urinary protein,urinary occult blood and urine protein were positive in 3 cases.Five cases of renal dysfunction,manifested as creatinine,urea nitrogen,uric acid significantly increased; renal ultrasound non-specific changes,manifested as increased echogenicity of the cortex.2 cases of renal pathology glomerular ischemic; Two cases of renal interstitial mild edema,five cases of inflammatory cell infiltration; Three cases of renal tubular epithelial vacuolar degeneration,2 cases of tubular atrophy,4 cases of renal tubular epithelial brush border loss,see intraluminal protein casts can be seen,1 case of calcium deposition.Conclusion The acute renal injury can be induced by overtraining.Kidney pathology ischemic is the most important change and renal tubular show most sensitive features of ischemic.In addition,inflammatory response and striated muscle damage were also induced because of overtraining.