Objective:To study the cytotoxic effects on tumor cell lines Hela,BGC823,MCF-7,L342,H9101,D6 induced by Fas ligand and anti-human DR5 monoclonal antibodies(Anti-DR5 mAb) and the underlying mechanism.Methods:Fas/DR5 mRNA were detected by RT-PCR. Cytotoxicity exerted by FasL/Anti-DR5 mAb on tumor cell lines was measured by MTT colorimetry and the induced apoptosis was determined by agarose gel electrophoresis.Results:The expression of DR5 on BGC823 and Hela cells were higher and DR5 didn’t express in D6. The expression of Fas on H9101 and L342 were higher and Fas didn’t express in D6. Cell line H9101 and L342 were sensitive to Anti-DR5 mAb and FasL in a dose dependent manner; Cell line MCF-7 and BGC823 were sensitive to FasL but were partially sensitive to Anti-DR5 mAb; Cell line Hela was partially sensitive to FasL but was sensitive to Anti-DR5 mAb; Cell line D6 was insensitive to two apoptosis inductions.Conclusion:Apoptosis induced by Fas ligand and Anti-DR5 mAb vary among tumor cell lines. The underlying mechanism may be relevant to Fas/DR5 mRNA expression,the release of Caspase-8 and Bcl-2.

Objective To explore the influence of acute myocardial infarction (AMI) patients with coronary lesions to the ST segment resolution (STR) and clinical prognosis after percutaneous coronary intervention (PCI).Methods 380 patients with AMI were detected twice each hour for the situation of STR,before and after coronary angiography and PCI.According to the STR,the patients were divided into STR (≥ 50％) group and no STR (＜ 50％) group.The relationship between STR and clinical prognosis of hospitalization or follow up were analyzed.Results The STR rate of patients with coronary multivessel disease or proximal lesions after PCI was lower than the rate of patients with coronary single vessel disease or distal lesions (P ＜ 0.05).The incidence rate of major cardiac events and LVEDd during hospitalization in patients with proximal coronary lesions was obviously higher than patients with distal lesions,while LVEF was lower than distal lesions(P ＜0.01).It had no significant differences in LVEF,LVEDd between coronary multivessl disease and single vessel disease during follow up (P ＜ 0.01).Conclusion It has significant relationship between coronary multivessel disease,proximal coronary,STR resolution in patients with AMI after PCI and the prognosis during hospitalization and follow up.

Objective To explore the relationship between ST-segment level,T-wave early inversion and patients' left ventricular function,prognosis specifically for ST-segment elevation acute myocardial infarction patients,which underwent emergency PCI.Methods 62 patients (M:40,F:22 ) with acute myocardial infarction were randomly divided into groups as follows:according to whether one hour ST segment's changes more than 50％ after PCI,they were divided into group A and B:group A (PCI after one hour ST segment ≥50％ ),group B (PCI ST segment after one hour ＜50％ ).According to whether 24-hour T wave inversion after PCI,they were divided into group C and D:group C (24-hour T-wave inversion),group D (24 hours without T wave inversion).After six months,the different LVEF betweeen group A and B,C and D was observed.Results Of group A and group C,after 6 months,the left ventricular function recovery was better than that of group B and group D ( P ＜ 0.05 ).Conclusion The analysis of ST segment and T wave are simple,practical and effective methods to predict myocardial reperfusion,especially for PCI therapy in patients with acute myocardial infarction.

Objective To study the mode and clinical implications of onset of spontaneous tosade de pointes in the congenital long QT syndrome. Methods We reviewed electrocardiograms (ECGs) of 55 patients with congenital QT syndrome for syncope. Documentation of the onset of tosade de pointes was available for 16 patients. All these patients had "definitive long QT syndrome" by accepted clinical and ECG criteria. Results One hundren and forty-nine runs of tosade de pointes were documented in 16 patients,of whom,there were 130 runs of pause-dependent tosade de pointes. Conclusion Our results show that the pause-dependent tosade de pointes,which has been recognized as a hallmark of tosade de pointes in the acquired long QT syndrome,plays a major role in the genesis of tosade de pointes in the congenital long QT syndrome.

AIM:Tostudywhe ther theangiotens in-(1-7)[Ang-(1-7)]/Mas receptor axis protects cardio-myocytes against high glucose (HG)-induced injury by inhibiting nuclear factor-κB (NF-κB) pathway.METHODS:The cell viability was measured by CCK-8 assay.The intracellular levels of reactive oxygen species ( ROS) were detected by DCFH-DA staining .The number of apoptotic cells was tested by Hoechst 33258 nuclear staining .Mitochondrial membrane potential ( MMP) was examined by JC-1 staining.The levels of NF-κB p65 subunit and cleaved caspase-3 protein were de-termined by Western blotting.RESULTS: Treatment of H9c2 cardiac cells with 35 mmol/L glucose (HG) for 30, 60, 90, 120 and 150 min significantly enhanced the levels of phosphorated ( p) NF-κB p65, peaking at 60 min.Co-treatment of the cells with 1 μmol/L Ang-(1-7) and HG for 60 min attenuated the up-regulation of p-NF-κB p65 induced by HG. Co-treatment of the cells with Ang-(1-7) at concentrations of 0.1~30μmol/L and HG for 24 h inhibited HG-induced cy-totoxicity, evidenced by an increase in cell viability .On the other hand, 1 μmol/L Ang-(1-7) ameliorated HG-induced apoptosis, oxidative stress and mitochondrial damage , indicated by decreases in the number of apoptotic cells , cleaved caspase-3 level, ROS generation and MMP loss .However, the above cardioprotective effects of Ang-(1-7) were markedly blocked by A-779, an antagonist of Ang-(1-7) receptor (Mas receptor).Similarly, co-treatment of H9c2 cardiac cells with 100 μmol/L PDTC ( an inhibitor of NF-κB) and HG for 24 h also obviously reduced the above injuries induced by HG.CONCLUSION:Ang-(1-7)/Mas receptor axis prevents the cardiomyocytes from the HG-induced injury by inhibiting NF-κB pathway .

Aim To investigate the role of ATP-sensi-tive potassium channels-Akt pathway in exogenous hy-drogen sulfide( H2 S) inhibiting the high glucose( HG)-induced injury in H9c2 cardiac cells. Methods The expression level of Akt protein was tested by Western blot assay. The cell viability was measured by cell counter kit-8(CCK-8 assay). The number of apoptotic cells was tested by Hoechst 33258 nuclear staining fol-lowed by photofluorography. The intracellular levels of reactive oxygen species ( ROS ) were detected by DCFH-DA staining followed by photofluorography. Mi-tochondrial membrane potential ( MMP ) was examined by JC-1 staining followed by photofluorography. Results H9c2 cells were treated with 35 mmol·L-1 glucose (high glucose, HG) for 0 ~24 h respectively. After treating for 3 h, the expression level of phosphorated ( p )-Akt protein began to be obviously reduced, the maximum reduced expression level was observed after the cells were exposed to HG for 24 h. Pretreatment of the cells with 50 μmol · L-1 pinacidil ( Pin, a KATP channel opener) or 400 μmol·L-1 NaHS( a donor of H2 S) prior to exposure to HG considerably blocked the down regulation of p-Akt expression level induced by HG. However, pretreatment with 1 mmol · L-1 KATP channel blocker glibenclamide( Gli) obviously attenua-ted the inhibitory effect of NaHS on HG-induced down-regulation of p-Akt expression level. On the other hand, the protective effects of NaHS against the HG-induced cardiomyocyte injury were markedly blocked by 30 μmol·L-1 Ly294002(an inhibitor of Akt), as indicated by the decrease in cell viability and MMP dissipation as well as the increases in the number of apoptotic cells and ROS generation. Conclution KATP channels-Akt pathway mediates the protective effect of H2 S against the HG-induced cardiac injury.

[ ABSTRACT] AIM:To investigate the role of ATP-sensitive potassium ( KATP ) channels in the inhibitory effect of hydrogen sulfide ( H2 S) on high glucose ( HG)-induced inflammation mediated by necroptosis in H 9c2 cardiac cells. METHODS:The expression levels of receptor-interacting protein 3 ( RIP3; an indicator of necroptosis ) and cyclooxyge-nase-2 (COX-2) were determined by Western blot.The levels of interleukin-1β(IL-1β) and tumor necrosis factor-α( TNF-α) were detected by ELISA .RESULTS:After H9c2 cardiac cells were treated with 35 mmol/L glucose ( HG) for 24 h, the expression of RIP3 was significantly increased .Pre-treatment of the cells with 100 μmol/L diazoxide ( DZ; a KATP channel opener) or 400 μmol/L NaHS (a donor of H2S) for 30 min considerably blocked the up-regulation of RIP3 induced by HG.Moreover, pre-treatment of the cells with 100 μmol/L 5-hydroxydecanoic acid (5-HD; a KATP channel
blocker) attenuated the inhibitory effect of NaHS on HG-induced up-regulation of RIP3.On the other hand, co-treatment of the cells with 100μmol/L necrostatin-1 ( a specific inhibitor of necroptosis ) or pre-treatment of the cells with 100 μmol/L DZ or 400 μmol/L NaHS attenuated HG-induced inflammatory responses , evidenced by decreases in the expression of COX-2 and secretion levels of IL-1βand TNF-α.However , pre-treatment of the cells with 100 μmol/L 5-HD significantly attenuated the above anti-inflammatory effects of NaHS.CONCLUSION:KATP channels play an important role in the inhib-itory effect of H2 S on HG-induced inflammation mediated by necroptosis in H 9c2 cardiac cells.

Objective: To construct the expressing vector of the extracellular domain of death receptor 5 (DR5), express it E.coli, identify the purified DR5 protein, and study its biological activity. Methods: The extracellular domain of DR5(eDR5) was assembled by overlapping PCR. The expression vector pET-22b(+)/ DR5 was constructed and transformed into E.coli BL21(DE3). The expression of eDR5 protein was induced by IPTG and purified by Ni 2+ -affinity chromatographic column. The purity and specificities were detected by SDS-PAGE and ELISA, respectively. The blocking effects of purified eDR5 on FMU1.5-induced apoptosis of U343, U373 cells were observed. Results: The extracellular domain of DR5 was obtained by overlapping PCR. The eDR5 protein was expressed in both supernatants and inclusion bodies with a yield more than 30% of total bacterial proteins. The purity of eDR5 was more than 95% and the yield reached 9 mg/ml. The result of ELISA showed the purified protein was eDR5. Purified eDR5 partially blocked the apoptosis of U343 cells induced by FMU1.5 and TRAIL. Conclusion: The successful construction, expression, and purification of the extracellular domain of DR5 protein lays a foundation for further study of DR5 function.

Objective To explore the clinical outcome of improved technique of Gamma nail in the treatment of intertrochanteric hip fracture of the elderly patients. Methods From March 2002 to October 2006.39 patients with intertrochanteric hip fracture were operated by improved technique of Gam-ma nail.There were 18 males and 21 females at average age of75.7 years(67_98 years).There were 6 patients with type A1 fracture,24 with type A2 fracture and 9 with type A3 fracture according to AO/ASIF classification.Of all.36 patients(92.3%)had osteoporosis.The operation improvements included the following points:(1)The patients were placed at the lateral decubitus position with the fractured limb on the uppermost,with flexion of knee and hip of 60°.The normal hip and knee were flexed as possible.(2)One-off indirect traction reduction was used after general anesthesia. no requirement of continuous mechanical traction.(3)C-arm image intensifier was employed to obtain normal and lateral projections.Results Of all,35 patients were followed up for a mean period of3 years and 2 months, ranging from 6 months to 5 years and 2 months.Operation data showed incision length of(4.3±1.2)cm,mean opera-tion time of(46±10)minutes,intraoperative bleeding volume of(65±26)ml and intraoperative X-ray exposure of(3.0±2.1)times.Postoperative recovery data showed survival in one-year follow up,with ambulation time of(10.5±3.6)days and fracture union time of(10.9±2.1)weeks.Mean Parker's score wag(6.9±3.2)points 6 months after operation. Conclusions Improved technique of Gamma nail can shorten operation duration,reduce operative trauma and bleeding,reduce X-ray exposure and im-prove success rate of surgery.as facilitates early pest-operative recovery and reduces the perioperative mortality rate of the elderly.

AIM:To study whether hydrogen sulfide (H2S) protects H9c2 cardiomyocytes against high glucose ( HG)-induced injury by inhibiting necroptosis .METHODS:The protein levels of RIP3 ( an indicator of necroptosis ) and cleaved caspase-3 were determined by Western blot .The cell viability was measured by CCK-8 assay.The intracellular le-vels of reactive oxygen species (ROS) were detected by 2’, 7’-dichlorfluorescein diacetate staining followed by photofluo-rography.Mitochondrial membrane potential (MMP) was examined by rhodamine 123 staining followed by photofluorogra-phy.The number of apoptotic cells was observed by Hoechst 33258 nuclear staining followed by photofluorography .RE-SULTS:After the H9c2 cells were treated with HG (35 mmol/L glucose) for 0~24 h, the protein expression of RIP3 in the H9c2 cells was significantly increased at 3 h, 6 h, 9 h, 12 h and 24 h, reaching the maximum level at 24 h.Pretreat-ment of the cells with 400μmol/L NaHS (a donor of H2S) or co-treatment of the cells with necrostatin-1 (Nec-1;a speci-fic inhibitor of necroptosis) considerably blocked the up-regulation of RIP3 protein induced by HG.Moreover, pretreatment with NaHS or co-treatment with Nec-1 obviously inhibited HG-induced injuries , leading to an increase in the cell viability , and decreases in the generation of ROS and MMP loss .On the other hand , pretreatment with NaHS also reduced the num-ber of apoptotic cells and the protein level of cleaved caspase-3 in the HG-treated H9c2 cardiomyocytes .CONCLUSION:H2 S protects H9c2 cardiomyocytes against HG-induced injury by inhibiting necroptosis .