Objective To investingate the effect of 5-Aza2'-deoxycytidine(5-Aza-CdR)on cell growth and to explore the possibility of re-expression of the hypermethylated and silenced RUNX3 gene in hepatocarcinoma cell line HepG2.Methods The change in expression of the tumor suppressor gene RUNX3 mRNA in cultured HepG2 cells was observed by RT-PCR before and after 5-Aza-CdR treatment.Activity of cell growth was observed by MTT assay and colony-forming test.The cell cycle was analyzed by flow cytometry.Apoptotic morphology was observed by transmitting electron microscopy.Results The gene was reactivated by two different doses of 5-Aza-CdR treatment in HepG2 cell without expressing RUNX3.The hepatocarcinoma cell line treated with 5-Aza-CdR displayed a slowed growth rate in contrast to the control group.The colony formation rate of HepG2 cell treated with 5-Aza-CdR decreased dramatically(P

[ ABSTRACT] AIM:To construct a prokaryotic expression plasmid to produce recombinant human tumor necrosis factor-related apoptosis-inducing ligand ( TRAIL) and to verify the biological activity of TRAIL.METHODS: The pro-karyotic expression plasmid pET-28a (+)-TRAIL114-281 was constructed.Human soluble TRAIL was obtained through opti-mized inducing protein expression and purification conditions.The biological activity of TRAIL was verified by CCK-8 as-say.The apoptosis-inducing effect of TRAIL alone and/or in combination with proteasome inhibitor bortezomib ( Velcade, PS-341) on the tumor cell lines H460 ( TRAIL-sensitive) and K562 ( TRAIL-resistance) for 24 h was determined.The ap-optotic rates of the cells were analyzed by flow cytometry with Annexin V-FITC/PI staining.The activities of caspase-8,-9 and -3 in the cells were detected by colorimetric method.The protein expression of Bax, Bcl-2 and cFLIP was measured by Western blot.The expression of DR4 and DR5 in the H460 cells and K562 cells after treated with bortezomib for 24 h was detected by flow cytometry.RESULTS:The recombinant human soluble TRAIL protein with stable bioactivity was success-fully acquired, which induced apoptosis in H460 cells and K562 cells.After treatment with different concentrations of TRAIL, the apoptotic rate of H460 cells was significantly increased with the increase in the concentration of TRAIL ( P<0.05), but the apoptotic rate of K562 cells was not affected by the increasing TRAIL concentration.Apoptotic rate in com-bination group was obviously higher than that in single group ( P<0.05 ) .In the process of apoptosis, the activities of caspase-8,-9 and -3 in H460 cells and K562 cells were both increased.The expression of Bcl-2 and cFLIP in treatment groups ( especially the combination group) was decreased compared with control group.No significant change of the Bax expression level was observed.The expression of DR4 and DR5 in the H460 cells and K562 cells was significantly up-regu-lated after treated with bortezomib ( P<0.05 ) .CONCLUSION: Bortezomib combined with recombinant human soluble TRAIL synergistically induces apoptosis in tumor cell lines H460 and K562 through initiating intrinsic apoptotic pathways by up-regulating death receptors DR4 and DR5, and reducing the expression of antiapoptotic proteins Bcl-2 and cFLIP.

Objective: To evaluate the compensation level of the Urban Employee Basic Medical Insurance ( UEBMI ) and Urban Resident Basic Medical Insurance ( URBMI ) in Jiangsu province . Methods: Take catastrophic health expense as the bottom line of compensation ratio for the basic medical insurance, the benefit of hospitalization expenses of those joining the insurance as the actual line of compensation ratio, and quartile division was used to comparatively analyze the differences between the bottom line of compensation ratio and the actual line of compensation ratio. Results: Take 10%as the critical value of catastrophic health expense, the actual line of compensation level is higher than the bottom line of compensation level in UEBMI, and there is reverse in URBMI. Conclusion: In some extent, the compensation level of UEBMI has relieved the economic burden of the poor jointed group because of sickness, while limited compensation level of UEBMI is in need of further improvement.

To discover factors affecting the inpatient’ actual reimbursement rate of urban resident basic medical insurance ( URBMI ) . Methods: Using the method of correlation analysis and multiple stepwise regression modeling to identify the influencing factors. Results: The per capita funding criteria, rate of inpatient out of pocket payment over resident annual per capita disposable income and per capita hospitalization rate have significant effect on the actual reimbursement rate of URBMI. Conclusion: It is needed to establish a sustainable steady financing mechanisms for URBMI, improve the evaluation system of reimbursement policy and gradually raise the level of actual reimbursement.

Objective To study the financial inputs into community health School of Medicine and Health Management, TONGJI Medical College of HUAZHONG[service (CHS) institutions by entities of different ownerships, by comparison of their investment, housing and human resources for policy recommendations.Methods Choose 6 cities in the East, Central and West of China through stratified random sampling. Collect data by literature review, in-depth interview, focus group discussion and questionnaire survey. Analyze quantitative data through EPI data 3.02 and SPSS 13.0.Results The institutions owned by the government and hospitals were advantageous in financial investment and housing; institutions under social organizations or individuals were inferior with development supports (P=0.00). Conclusion The community health service principle of government dominance and social involvement should be advocated, with equal opportunities for community service providers of various ownership. Emphasis should be placed on betterment of policies and measures, specific budget subsidy, as well as performance appraisal and service supervision. These measures aim at encouraging non-government providers to play greater role in community health service.

OBJECTIVE To observe the therapeutic effectiveness and safety of the indigenous teicoplanin in intensive care unit(ICU) patients with severe infections of Gram-positive(G+) cocci.METHODS Sixty patients with severe Gram-positive bacteria infection received teicoplanin which was produced by Xinchang Pharmaceutical Factory of Zhejiang Medicine Co,Ltd.by infusion for 7-14 d.Dosage was adjusted according to the creatinine clearance rate.Maximum temperature,APACHEⅡ score,indicators of inflammatory response,liver and kidney function,microbiologic examination and adverse reactions were observed before treatment and the end of 7-14 day treatment.RESULTS Seventy-two strains of Gram-positive cocci were isolated from 60 patients,in which meticillin-resistant Staphylococcus aureus(MRSA),meticillin-resistant S.epidermidis(MRSE),Enterococcus faecium,E.faecalis,S.haemolyticus and E.gallinarum accounted for 58.33%,8.33%,16.67%,5.56%,8.33% and 2.78%,respectively.After treatment,59 strains of Gram-positive cocci were eliminated while 13 strains were not eliminated and bacteriological eliminated rate(calculated by strains) was 86.1%;In this group,twenty-eight patients were cured while twenty-two cases were markedly improved.The total bacteriological eliminated rate was 81.94%.The incidence of adverse reactions was low and only a slight increase or decrease in platelet count was observed which didn′t require special treatment interventions.CONCLUSIONS Indigenous teicoplanin has certain therapeutic effectiveness and is safe for severe Gram-positive bacteria infected patients,particularly for septic patients with organ damage in ICU.In patients highly suspected with meticillin-resistant staphylococci or enterococci infection,teicoplanin may be a choice for initial empirical treatment.

Objective To observe critical patients with thrombocytopenia,and exam their coagulation function,so as to diagnose disseminated intravascular coagulation(DIC)in the early stage.Methods Totally 56 critical patients complicated with thrombocytopenia were enrolled in the investigation;complete set of DIC tests were performed.A total of 26 cases were diagnosed as DIC according to the criteria from the International Society of Thrombosis Haemostasis(ISTH).The sensitivity,specificity,accuracy,likelihood ratio,predictive value of prothrombin time(PT),activated partial thromboplastin time(APTT),thrombin time(TT),fibrinogen(Fg),antithrombin Ⅲ and bleeding signs(including petechia and ecchymosia)were calculated by applying fourfold table.Results The specificity of PT prolongation and Fg reduction was 87.0%;the sensitivity of ATⅢ decrease,and each of PT prolongation,Fg reduction,and bleeding(including petechia and ecchymosis)were as high as 96.0%.Conclusion Multi-markers based on thrombocytopenia are likely to improve sensitivity and specificity of DIC diagnosis.

Objective To investigate whether spinal cord is involved in the analgesic effect of propofol.Methods Fifteen adult SD rats of both sexes weighing 195～223g were randomly divided into three group of five animals each: control group received normal saline intraperitoneally(ip), fentanyl group received fentanyl 0 1mg/kg and propofol group propofol 100mg/kg ip 2 min later 4% formalin 150?l was injected subcutaneously into the planta region of right hindpaw 1h after formalin injection animals of all three groups were anesthetized with pentobarbital sodium ip After induction of anesthesia chest was open and 100 ml of normal saline was infused via left ventricle then followed by 4% formalin infusion for fixation of tissue, 90 min later spinal cord, L 3 5 sect, was removed for determination of c fos expression in spinal cord using fos immunohistochemistry technique Results In control group in less than 10 s after formalin intraplantar injection the animals were agitated, restless, lame and paw licking In fentanyl group and propofol group the righting reflex was suppressed for (19 4?7 8) min and (7 2?1 5)min and no pain response was seen during this period When righting reflex recovered the pain response was much lighter than that in the control group Formalin stimulation induced c fos expression was seen only in the ipsilateral spinal cord Both fentanyl and propofol significantly suppressed c fos expression evoked by formalin stimulation The number of fos like immunoreactivity neurons(FLIN) decreased by 57 8% and 36 3% respectively(P

Objective To study the 2244G→A, 2299 A→G single nucleotide polymorphism (SNP) in the 5' regulatory regions of Toll-like receptor 4 (TLR4) in patients with Gram negative bacteria infection in Shenzhen locality, and to discuss the occurrence, course and prognosis of patients with sepsis. Method Polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) method was used to detect the genotype of TLR4. After the whole blood DNA of patient was extracted and PCR was amplified, the products were 500bp and 599 bp, and were cut by endonuclease Mae Ⅱ and Sph Ⅰ respectively to determine the SNP 2244G→A and 2299 A→G in TLR4. These two kinds of allele frequencies were statistically calculated in all patients. In the meantime, the incidence of septic shock, average hospitalized days, cost and prognosis of all patients were recorded. Statistical analysis was performed with SPSS version 16 software. ANOVA was used for comparison among multiple groups, and t -test and Sighed rank test were used for paired comparison. Results The 2299 and 2244 sites in the 5' regulatory regions of TLR4 gene of patients with Gram negative bacteria infection in Shenzhen locality had various degrees of changes in single nucleotide. Compared with the documented data from Chinese people in general, there was a significant difference in 2299A→G genotype frequency in residents of Shenzhen locality ( P < 0.05). But there were no statistically significant difference in mortality, incidence of septic shock, average days of ICU stay or ICU cost between TLR4 SNP positive and negative groups of patients. Conclusions There is a wide range of genetic variation in the 2299 and 2244 sites in the 5' regulatory regions of TLR4 among citizens of Shenzhen locality with unique distribution. The 2299A→G genotype frequency probably has differences in distribution and population. The pathogenesis and the prognostic factors of sepsis are complicated, whereas the gene polymorphism may be just one of the factors affecting the prognosis of patients with Gram negative bacteria infection.

Obesity often results when regular caloric intake exceeds the energy expenditure. Hypothalamus is the center that regulates feeding and energy balance inside the body, especially the lateral hypothalamic area (LHA) and ventromedial nucleus (VMN). Acupuncture-moxibustion has achieved significant progress in treating simple obesity, and the relevant studies have revealed the central action mechanism underlying the treatment. By reviewing the literatures associated with the mechanism studies on acupuncture-moxibustion for simple obesity, this article is going to provide clinical and scientific guidance for the treatment of simple obesity with acupuncture-moxibustion.