Objective:To construct a prokaryotic expression plasmid containing HCVF gene,and purify the recombinant fusion protein in E.coli system. Methods: F gene of Hepatitis C virus was amplified by PCR method from plasmid H/FL(containing full length cDNA sequence of HCV 1a subtype),cloned into pET32a(+)vector,and then transformed into E.coli JM109. After identified by restriction diges- tion and DNA sequencing,recombinant plasmid was transformed into E.coli BL21 and induced with IPTG. The fusion protein trxA- F was further confirmed by Western blot analysis and purified by affinity chromatography method. Results: Restriction digestion and PCR screening showed that HCV F gene was cloned into pET32a(+)successfully. After BL21 was transformed with recombinant vector pET32a (+)- HCVF and induced with 0.5mmol/L IPTG,a 35.4kDprotein band was found bySDS- PAGE. And this recombinant protein showed a highly specific and strong reaction with anti- His monoclonal antibody by Western blot analysis. Conclusion: The prokaryotic expression plasmid pET32a(+)- HCVF was successfully constructed,which will be helpful for further research.

Apoptosis is important to the development of diseases. Research recently indicates that c-Jun N-terminal kinase (JNK) and cysteinyl aspartate-specific protease (caspase) play key roles in apoptosis and affect the development of diseases. This article is to introduce the function and relationship between JNK and caspase in apoptosis during the process of diseases.

Along with the increase of the population having malignant tumors with time,the effective treatment strategy is far behind the clinical needs. There are significant individual variation in terms of treatment response and toxic profile with the same chemotherapy regimen. Therefore,the research about the occurrence and the individualized treatment of malignant tumor is one of the current hot topics. cytochrome P450(CYPs) is part of I metabolism enzymatic system,and also is the most rich in content,the most widespread in distribution and has the broadest substrate spectrum in nature. This article reviews the relationship between cytochrome P450 and the occurrence and treatment of malignant tumors.

Objective To investigate whether MEG3 involved in the development of retinoblastoma by down-regulating the expression of P53 protein.Methods The MEG3 expression of retinoblastoma tissues and corresponding non-tumor tissues were detected by quantitative real-time PCR (qRT-PCR).Retinoblastoma cell lines SO-RB50 or HXO-RB44 were transfected with pcDNA-MEG3 or siRNA-MEG3,after which cell apoptosis was tested by flow cytometry and P53 protein expression was tested by Western blot.Results MEG3 expression of retinoblastoma tissues was significantly reduced compared with corresponding non-tumor tissues(P =0.014).MEG3 level was significantly increased in pcDNA-MEG3 transfected SO-RB50 cells (P =0.002) and significantly decreased in siRNA-MEG3 transfected HXO-RB44 cells (P =0.004).Flow cytometry showed that the SO-RB50 cells apoptosis was significantly increased with the MEG3 over-expression(P ＜ 0.05),as well as the HXO-RB44 cells apoptosis was significantly decreased with the MEG3 knockdown(P ＜ 0.05),compared with the control group,respectively.Furthermore,Western blot showed that P53 protein level was significantly increased after SO-RB50 transfected with pcDNA-MEG3 (P ＜ 0.05),while significantly decreased after HXO-RB44 transfected with siRNA-MEG3 (P ＜ 0.05),compared with the control group,respectively.Conclusion MEG3 is down-regulated in retinoblastoma,affect the development of retinoblastoma,and may induce the retinoblastoma cell apoptosis by promoting the expression of P53 protein.

Objective To evaluate the role of syndecan-4 (SDC-4) in inflammatory responses of rats with ventilator-induced lung injury (VILI).Methods Twenty-four healthy male wild type Sprague-Dawley rats and 24 male SDC-4 gene knockout Sprague-Dawley rats,aged 2-3 months,weighing 200-220 g,were assigned into 2 groups (n =12 each) using a random number table:control group (group C)and VILI group.The animals were anesthetized with pentobarbital sodium and tracheostomized.The rats kept spontaneous breathing in group C.The rats were mechanically ventilated for 4 h in group VILI.Blood samples were taken from the femoral artery at the end of mechanical ventilation for detection of arterial oxygen partial pressure (PaO2).The animals were sacrificed after blood sampling.The left lung was lavaged,and the broncho-alveolar lavage fluid (BALF) was collected for determination of the tumor necrosis factor-α (TNF-α),interleukin-1beta (IL-1β) and IL-18 concentrations (by enzyme-linked immunosorbent assay) and total protein concentrations (by BCA assay).The right lungs were removed for determination of the wet to dry weight ratio (W/D ratio) and expression of TNF-α,IL-1β and IL-18 mRNA in lung tissues (by real-time polymerase chain reaction) and for examination of the pathological changes (with a light microscope).The lung injury scores were recorded.Results Compared with group C of wild type rats,PaO2 was significantly decreased,W/D ratio and lung injury scores were increased,the concentrations of total protein,TNF-α,IL-1β and IL-18 in BALF were increased,and the expression of TNF-α,IL-1β and IL-18 mRNA in lung tissues was up-regulated in group VILI of wild type rats (P＜0.05),and no significant change was found in the variables mentioned above in group C of gene knockout rats (P＞0.05).Compared with group C of gene knockout rats,PaO2 was significantly decreased,W/D ratio and lung injury scores were increased,the concentrations of total protein,TNF-α,IL-1β and IL-18 in BALF were increased,and the expression of TNF-α,IL-1β and IL-18 mRNA in lung tissues was up-regulated in group VILI of gene knockout rats (P＜0.05).Compared with group VILI of wild type rats,PaO2 was significantly decreased,W/D ratio and lung injury scores were increased,the concentrations of total protein,TNF-α,IL-1β and IL-18 in BALF were increased,and the expression of TNF-α,IL-1β and IL-18 mRNA in lung tissues was up-regulated in group VILI of gene knockout rats (P＜0.05).Conclusion SDC-4 can inhibit inflammatory responses of rats with VILI and is involved in the endogenous protective mechanism.

Objective: To strengthen the education of ethics,honor and humiliation,and humanities for the working staff in community healthcare service,so as to better serve the community residents.Method: To strengthen the education of ethics,honor and humiliation,and humanities for the working staff in community healthcare service,and check the outcome.Conclusion: It is necessary to recognize the humanistic function of community healthcare service,strengthen practice and the ability of healthcare service to improve residents' trust and satisfaction of community healthcare service.

Objective To investigate the clinical efficacy of heat-sensitive point moxibustion in treating functional dyspepsia. Methods Fifty-six patients with functional dyspepsia were randomly allocated to treatment and control groups, 28 cases each. The treatment group received heat-sensitive point moxibustion and the control group took domperidone tablets. The symptoms were scored and plasma motilin was measured in the two groups before and after treatment. The clinical therapeutic effects were compared between the two groups.Results The total efficacy rate was 89.3% in the treatment group and 82.1% in the control group; there was a statistically significant difference between the two groups (P<0.05). There were statistically significant pre-/post-treatment differences in the symptom score and plasma motilin in the two groups (P<0.05). There were statistically significant post-treatment differences in the symptom score and plasma motilin between the treatment and control groups (P<0.05).Conclusion Heat-sensitive point moxibustion is an effective way to treat functional dyspepsia.

Objective To investigate relationship between Ala 54 Thr polymorphism in the intestinal fatty acid-binding protein (I-FABP) and plasma lipids in old people.Methods We collected fasting blood samples of selected groups,then we extracted DNA from whole blood,and detected lipids from serum.The 54A/T I-FABP genotypes were analyzed in 72 Han Chinese elderly by polymerase chain reaction (PCR) and DNA restriction enzyming.Total cholesterol (TC),triglyceride (TG),high density lipoprotein (HDL-C) and low-density lipoprotein (LDL-C) were detected.Genotype subgroups were Thr54 (-) and Thr54 (+) groups (36 cases for each).Results Compared with Thr54 (-) genotype group,Thr54 (+) genotype group had higher levels of plasma TC[(4.50±0.73) mmol/L vs.(5.48±0.49)mmol/L],TG[(1.08±0.48) mmol/L vs.(2.02±0.53)mmol/L],LDL-C [(3.10±0.44) mmol/L vs.(3.50±0.66)mmol/L],and had lower level of HDL-C [(1.14±0.25) mmol/L vs.(0.96±0.23)mmol/L](t=-6.67,-7.84,-3.03,3.05,all P＜0.05).

Objective To explore the relationship between FABP2 54 gene polymorphism and serum lipid levels in different ethnic patients with coronary heart disease.Methods We collected fasting blood from participants,then extracted DNA from whole blood,and detected lipids from serum.We performed 54A/T FABP2 genotyping for participants in the case and the control groups by polymerase chain reaction (PCR) and DNA restriction enzyme digestion techniques.Results ( 1 ) In Hans population with coronary heart disease,the allelic frequency of point in 54T gene was 0.542,and 54A gene was 0.458 ; in urban Mongolian with coronary heart disease,the allelic frequency of point in 54T gene was 0.708,and that of 54A gene was 0.292.The allelic frequency of point in 54T and 54A genes were 0.284 and 0.716 respectively in the Han population control group and 0.353 and 0.647 respectively in the normal urban Mongolian group.Compared with the control groups,mutant 54T allele frequency in the case groups of Han and urban Mongolian was significantly higher ( Han:x2 =14.967,P ＜ 0.05 ; Mongolian:x2 =28.083,P ＜ 0.05 ).Compared with the Han Chinese population with coronary heart disease,the mutant 54T allele frequency in urban Mongolian with coronary heart disease was significantly (x2 =7.111,P ＜ 0.05 ).There was no significant difference in mutant 54T allele frequency between the Mongolian control group and the Han control group ( x2 =3.392,P ＞ 0.05 ).( 2 ) Patients with coronary heart disease who carry Thr54 ( + ) had a significantly higher level of fasting plasma TG [ CC Thr54 (-):( 1.89 ± 0.57 ) mmol/L,CC Thr54 ( + ):( 3.92 ± 1.63 ) mmol/L; CM Thr54 (-):(2.23 ± 0.13 ) mmol/L,CM Thr54 ( + ):(4.03 ± 1.14) mmol/L; Hans:P =0.001,Mongolian:P =0.035 ],and LDL-C [ CC Thr54 (-):( 3.09 ± 0.92) mmol/L,CC Thr54 ( + ):(4.05 ± 1.14 ) mmol/L ; CM Thr54 (-):( 4.26 ± 0.08 ) mmol/L,CM Thr54 ( + ):( 5.10 ± 0.56 ) mmol/L; Hans:P =0.025,Mongolian:P =0.045 ] than those with Thr54 (-).Conclusion (1) Threre is FABP2 gene polymorphism in urban Mongolian and Han population.Gene mutation frequency is in accordance with Hardy-Weinberg law of genetic equilibrium,suggesting the sample group representative.(2) FABP2 polymorphism may have a certain contribution to dyslipidemia in patients with coronary heart disease.FABP2 gene polymorphism may be associated with coronary heart disease.Individuals of Mongolian with FABP2 Thr54( + ) has a higher risk of coronary heart disease.