Objective To investigate the effects of hypoxia-inducible factor-1α (HIF-1α)expression on pathogenesis of retinopathy of prematurity (ROP) and to find new target for gene therapy.Methods After liposome-mediated small interference RNA (siRNA) transfection into rat retinal endothelial cells,the cells were cultured in medium with CoCl2-induced hypoxic condition.Expression of HIF-1α mRNA was determined by fluorenscence quantitative reverse transcription-polymerase chain reaction(RT-PCR),HIF-1α protein expression was detected by Western Blot after cocultured for 8 hours.Cell proliferation was measured with 3-(4,5)-dimethylthiazol (-2-yl)-2,5-diphenyltetrazolium bromide(MTT) assay after cocultured for 24 hours.Difference between groups was compared with independent samples t test.Results Rat retinal vascular endothelial cells were successfully transfected with siRNA.Fluorescence quantitative RT-PCR results showed that at 48 hours of transfection,the expression of HIF-1α mRNA in the interference group of siRNA1,siRNA2 and siRNA4 were 0.1620 ± 0.0147,0.2034 ± 0.0251 and 0.3049 ± 0.0165,which were 16.20％,20.34 ％ and 30.49％ of blank control group (1.0000±0.0344),and were lower than that of negative control group (0.8334±0.0242) (t=16.786,8.953 and 4.087,P＜0.05 respectively).Western Blot results showed that HIF-1α protein expression was significantly inhibited by siRNA1(0.4956 ± 0.0421 ) and siRNA2 (0.6544 ± 1.0032) comparing with blank control group (3.5105 ±0.4084) and negative control group (3.4019 ± 1.0677) (t =6.861,2.893,4.567 and 5.072,P＜0.05 respectively).As for cellular proliferation activity,(49.5±2.9) ％ and (67.4±1.2) ％ of cells growth inhibition were observed after transfection with siRNA1 and siRNA2,which were higher than those of negative control group [(15.7±1.5) ％ ] (t=2.786 and 6.904,P＜0.05).Conclusions The synthetic HIF-1α siRNA could effectively inhibit the expression of HIF-1α gene and reduce cell proliferation in rat retinal endothelial cells under hypoxic condition.RNA interference technology targeting HIF-1α might become a new strategy for gene therapy of ROP.

Objective To evaluate the ablation zone and temperature changes in the liver caused by laser ablation in vitro and in vivo.Methods Single needle single point laser ablation was performed in vitro with porcine liver using 5 W and 7 W-power and in vivo with rabbit liver using 5 W-power.All energy outputs were 1 800 J.The ablation zone and temperature changes were evaluated.Results Coagulation caused by laser ablation was divided into a black carbonized area and a brown white necrotic area from inside to out,respectively.The mean in vitro ablation range obtained by 5 W and 7 W-power laser ablation was 1.9 cm × 1.2 cm × 1.2 cm and 2.5 cm × 1.5 cm × 1.5 cm,respectively.The mean in vitro black carbonized range obtained by 5 W and 7 W-power laser ablation was 1.1 cm × 0.5 cm × 0.5 cm and 1.4 cm × 0.7 cm ×0.7 cm,respectively.The average maximum temperature using 5 W-power was 243.57 ℃,62.47 ℃ and 34.27 ℃ in the ablation center,5 mm from the center,and 10 mm from the center,respectively.For in vivo liver experimentation,the average maximum temperature using 5 W-power was 62.26 ℃ and 44.35 ℃5 mm and 10 mm from the center,respectively.Conclusion Laser ablation can cause coagulation necrosis of the liver tissue in vitro and in vivo,and the ablation range increased with increasing power.Laser ablation may have potential use in the ablation of tumor localized in critical sites.

Objective: To investigate the effect of zinc supplementation on expression of MT-1 and MT-2 mRNA in placenta and small intestine of pregnant rats. Method: Eighteen healthy pregnant SD rats were randomly divided into two groups,normal control (ZC) and zinc supplement group (ZS). Rats in both groups were fed the same normal diet. Rats in ZC group drank deionized water and those in ZS group drank the water supplemented with 1.26 mmol/L Zn. At gestational 9.5d and 17.5d, serum zinc was determined by atomic absorption spectrophotometer and the expression of MT-1 and MT-2 mRNA of placenta and small intestine was examined by RT-PCR. Results: Serum zinc levels at gestational 9.5d and 17.5d were higher in ZS group. Relative expression of both MT-1 and MT-2 mRNA in placenta, MT-1 mRNA at gestational 17.5d and MT-2 mRNA at each time point were higher in ZS group. At gestational 9.5d, there was no difference between two groups in expression of MT-1 mRNA. Conclusion: The expression of MT-1 and MT-2 mRNA in placenta and small intestine was up-regulated by dietary zinc supplementation during pregnancy.

Objective:To investigate the effect of zinc supplementation on the expression profile of zinc transporters mRNA in placenta and small intestine of pregnant rats.Method:Eighteen pregnant SD rats were randomly divided into normal control(ZC)and zinc supplement group(ZS).Rats in ZC group drank deionized distilled water while those in ZS group drank water supplemented with zinc of 1.26 mmol/L.Placenta and small intestine were taken at gestational day 9.5 and 17.5,respectively.The expression of ZnT1,2,5 and 6 mRNA was examined by RT-PCR.Results:At gestational D9.5,the expression of placental ZnT1,2 and small intestinal ZnT1,2,6 mRNA was up-regulated,and placental ZnT5 mRNA down-regulated by dietary zinc supplementation.Dietary zinc intake had no effect on the expression of placental ZnT6 and small intestinal ZT5 mRNA.At gestational D17.5,the expression of placental ZnT5 and 6 mRNA was up-regulated by dietary zinc supplementation,and dietary zinc had no effect on the expression of placental and small intestinal ZnT1,2 mRNA.The expression of ZnT5 mRNA at gestational D17.5 in both groups was not detectable.Conclusion:Dietary zinc supplementation during pregnancy has significant effect on the expression profile of ZnT 1,2,5 and 6mRNA in placenta and small intestine of pregnant rats.

Objective To investigate the association between catechol-O-methyltransferase (COMT) Val158Met polymorphism and the memery function of schizophrenia treated with Aripiprazole.Methods Schizophrenic patients were diagnosed in according to Diagnostic and Statistical Manual of Mental Disorders-IV (DSM-IV).A total of 78 Chinese Han subjects were involved in this study.The polymerase chain reaction-restriction fragment length polymorphism(PCR-RFLP) was used to identify COMT genotypes.The Chinese revised version of Wechsler Memory Scale (WMS-RC) was used twice to evaluate the memory function of patients treated with Aripiprazole alone before and after drug treatment for 8 weeks.ANOVA,rank sum test and t-test were used to statistical analysis.Results There was statistical difference of cumulative memory among the three groups (F=12.371,P=0.002),and Met/Met genotype showed much poorer cumulative memory than Val/Met and Val/Val genotypes (Z=-3.168,-2.641 ; P=0.002,0.008,respectively) Memory function was significantly improved after patients been treated with Aripiprazole except the associative memory(P＜0.05).Met/Met genotype showed more improvement of cumulative memory than other genotypes after Aripiprazole treatment (t=4.203,P＜0.01 ; t=3.024,P ＜0.01,respectively).Conclusions There may be a relationship between COMT Val158Met polymorphism and the memory function of schizophrenia in Aripiprazole treatment.Met/Met genotype of schizophrenia showed worst cumulative memory but best responsiveness in Aripiprazole treatment.

Objective:To study the mechanism of moxibustion therapy on diabetic peripheral neuropathy for the peripheral neuroprotection.Methods:The DPN model was induced by intraperitoneal injection with streptozotocin (STZ).The rats were given moxibustion at the acupoint Yishu (Extra) and the acupoint Zusanli (ST 36).The treatment was carried out once a day and 15 minutes per acupoint,lasting for 56 d in total.The clinical effect of moxibustion was evaluated by detecting blood sugar,urine sugar,body weight and dietary intakes,as well as nerve conduction velocity with neuroelectrophysiological method.The structure variation of sciatic nerve was observed by HE staining and light microscopy,and the level of NGF in the sciatic nerve Was determined by ELISA.Results:Compared with the model group,the plasma glucose was significantly lower in the moxibustion group (P＜0.01),with significantly faster nerve conduction velocity (P＜0.01),more notably changes in pathological appearance (P＜0.01) and higher level of nerve growth factor (NGF)(P＜0.01).Conclusion:Moxibustion could improve the symptom and signs of peripheral neuropathy in rat models with DPN,which may relate to the increased NGF and enhanced peripheral nerve protection.

Aim To isolate, purify, and chemically characterize a polysaccharide showing inhibition of the six-?-helix bundle formation of HIV-1 envelope glycoprotein gp41 from cultured broth of a streptomyces sp.strain. Methods Ethanol was used to precipitate polysaccharides and macromolecules from the broth.Proteins in the precipitate were removed by sevage method.Purification was carried out by DEAE-Cellulose and sephadex G-25 column chromatography. The chemical structure of the polysaccharide was determined with the combined application of HPLC,UV,IR and 1H-NMR spectroscopy, and the methods of periodate oxidation and Smith degradation,etc. Activity of anti-HIV-1 ~gp41 six-?-helix bundle formation was assayed withsandwich ELISA method.Results The purified polysaccharide,designated as SMP for Streptomyces polysaccharide,is neutral with a molecular weight of approximately 4855 Daltons. Sugar analysis showed SMP contains glucose and fructose residues in an approximate molar ratio of 22∶1 (10.96 to 0.48). The glycosidic linkages were estimated to be (1→4)-?-D-pyranoside as its main chain, and 1→6 linkage was attached to the main chain. Activity analysis revealed SMP markedly inhibited the six-?-helix bundle formation of HIV-1 glycoprotein gp41 and the IC_~50 was (145.48?7.25) mg?L~-1 .Conclusion Streptomyces polysaccharide SMP showing inhibition of the six-?-helix bundle formation of HIV-1 envelope glycoprotein gp41 was isolated and purified.

Objective: To investigate the awareness of air pollution protection knowledge and its influencing factors among primary school students in Shennongjia Forest District, Hubei Province, so as to provide insights into targeting implementation of health education on air pollution protection knowledge. Methods: Students in Grade 3 to 5 in Shennongjia Shiyan primary school were enrolled by stratified cluster sampling method, and students' demographic features and awareness of air pollution protection knowledge were investigated using the Investigation on the Effects of Air Pollution Health Protection of Pupils (Volume A). Factors affecting the awareness of air pollution protection knowledge among primary school students were identified using a multivariable logistic regression model. Results: A total of 897 questionnaires were allocated, and 877 valid questionnaires were recovered, with an effective rate of 97.77%. The respondents included 446 men (50.86%) and 431 women (49.14%), 301 third grade students (34.32%), 284 fourth grade students (32.38%), and 292 fifth grade students (33.30%), and had a mean age of (10.32±0.93) years. The overall awareness of air pollution protection was 55.76%, and the awareness rates of basic concepts, basic knowledge, and basic behaviors and skills were 42.99%, 53.48% and 57.24%, respectively. Multivariable logistic regression analysis identified age (OR=1.453, 95%CI: 1.053-2.005), living with parents (OR=2.638, 95%CI: 1.571-4.429), mother's educational level (below primary school, OR=0.270, 95%CI: 0.084-0.862; primary school, OR=0.169, 95%CI: 0.069-0.416; junior high school, OR=0.309, 95%CI: 0.138-0.691; high school, OR=0.352, 95%CI: 0.160-0.773) and average annual family income (50 000 to 100 000 Yuan, OR=1.629, 95%CI: 1.162-2.282; 100 000 to 150 000 Yuan, OR=1.802, 95%CI: 1.101-2.948; ≥150 000 Yuan, OR=1.939, 95%CI: 1.065-3.529) as factors affecting the awareness of air pollution protection knowledge among primary school students. Conclusion The awareness of air pollution protection knowledge is 55.76% among primary school students in Shennongjia Forest District, and is affected by age, mother's educational level, average annual family income and living with parents.

OBJECTIVEThis paper aims to investigate the impact of anthracene addition on microbial community in agricultural soil irrigated with tap water or reclaimed wastewater.

METHODSThe changes of microbial community were characterized by terminal restriction fragment length polymorphism in combination with 16S rRNA gene clone library analysis.

RESULTSA significant change in microbial community composition was observed during the biodegradation of anthracene, with dominantly enriched members from the genus Methylophilus.

CONCLUSIONThis work might be useful for developing techniques for the isolation of novel putative PAH degrader.

Anthracenes ; analysis ; Bacteria ; classification ; genetics ; isolation & purification ; Biodegradation, Environmental ; DNA, Bacterial ; genetics ; isolation & purification ; Hazardous Waste ; Phylogeny ; Polymorphism, Restriction Fragment Length ; RNA, Ribosomal, 16S ; genetics ; Soil Microbiology ; Soil Pollutants ; analysis ; Waste Disposal, Fluid ; methods