Background and purpose:miR-22 has been reported to be down-regulated in gastric cancer, lung cancer, colorectal cancer, and breast cancer. However, its expression in glioma was still poorly known. This study aimed to explicit whether miR-22 suppresses cell proliferation by targeting MTDH, thus to reveal molecular mechanism that miR-22 functions as a tumor suppressor in glioma. Methods:Quantitative real-time polymerase chain reaction (qRT-PCR) was conducted for detecting the expression of miR-22 in gliomas and normal brain tissues. MTDH 3’UTR-luciferase vector was constructed and dual-luciferase reporter gene assay was employed to examine the effect of miR-22 on luciferase activity. U251 cells were transfected with miR-22 mimics, and MTDH siRNA as for postive control, then Western blot was performed to detect the expressions of MTDH protein. The proliferation ability of U251 cells was evaluated by MTT assay. Results:miR-22 was down-regulated in glioma tissues. Glioma patients with relatively high expression of miR-22 showed lower mortality compared with low expression of miR-22 by using Kaplan-Meier survival curves. We demonstrated miR-22 could bind to the 3’ untranslated region (UTR) of MTDH and inhibited the luciferase activity. Western blot showed that the expression of MTDH protein was inhibited by restored miR-22 or siR MTDH in U251 cells. Overexpression of miR-22 or siR MTDH inhibited the proliferation of U251 cells. Conclusion:miR-22 suppresses cell proliferation by targeting MTDH in glioma.

Objective To construct cDNA entry library and cDNA expression library of Armillifer agkistrodontis (A.) nymphs and make a preliminary immunoscreening for the cDNA expression library.Methods The nymphs were collected from the Kunming mice infected experimentally with A.agkistrodontis eggs and the total RNA were extracted from the nymphs using TRIzol Reagent.After purifying the mRNA,the synthesized cDNAs were cloned into the donor vector pDONR222 by BP reaction of Gateway technology and the recombinants were transformed into the DH10B cells by electroporation,the cDNA entry library was obtained.Next,the expression vector pDEST17 was ligated with entry clones by LR reaction,and the recombinants were transformed into the BL21 (DE3) cells.Hence,the cDNA expression library was constructed.Then,the expression library was immunoscreened with the mixed sera of mice infected with A.agkistrodontis,and the insertions of positive clones were sequenced.After that,the open reading frame(ORF) of positive slone sequence,the homology of the screened genes and their encoded proteins were analyzed by Finder and BLAST (basic local alignment search tool) program of National Center of Biotechnology Information(NCBI),and the discovered new genes were submitted into the GenBank.Besides,the physico-chemical properties,secondary structure and B cell epitopes of encoded proteins were also analyzed by bioinformatics software.Results The average titer and total clones of the cDNA entry library were 1.45 × 105 CFU/ml(colony-forming unit,CFU) and 1.74 × 106 CFU,respectively,and the range of fragment length of the inserted cDNA was between 0.2-4.0 kb,with an average of 1.4 kb.The total clones of cDNA expression library were 1.00 × 105 CFU,and the fragment length of the inserted cDNA was between 0.3-2.2 kb,with an average of 1.0 kb.Five positive clones,coded S1,S5,A1,D1 and F1,respectively,were obtained through preliminary immunoscreening.The sequence and homology of the five positive clones were sequenced and analyzed by BLAST program.No significant similarities were found in pentastomida species,which meant that they were all novel genes of A.agkistrodontis.The gene sequences were submitted to GenBank,with the accession number from JQ180451 to JQ180455.Also,results obtained by bioinformatics software showed that the predictive encoding proteins were all potential to be valuable recombinant diagnostic antigens.Conclusions The cDNA library of A.agkistrodontis nymphs is successfully constructed,and five new genes of A.agkistrodontis are discovered.The establishment of cDNA library and the discovery of the new genes will lay a foundation for further studying the gene functions and screening the immunodiagnostic antigens.

Objective To determine the effect of mild hypothermia on neonatal piglet cardiac hemodynamic function after hypoxia-ischemia (HI).Method Twenty five 7-day-old piglets were used for hypoxic ischemic brain damage (HIBD) model by the method of temporary occlusion of the bilateral carotid arteries and followed by mechanical ventilation with low concentration of oxygen (FiO_2=6%) for 30 minutes.The piglets were randomly divided into three groups:group A (normothermia with body temperature to 39℃,n=9),group B (body temperature to 36℃for 72 hours,n=8),and group C (body temperature to 34℃for 72 hours,n=8).Mild hypothermia was initiated at 4 hours after HI,the systolic and diastole function were evaluated by Doppler echocardiography at pre-HI,post-Hi 4 hours and post-HI 72 hours.Results There were no significant differences in left ventrieular ejection time/left ventrieular ejection time (LPEP/LVEF),right ventricular ejection acceleration time/right ventricular ejection time (RACT/RVET) and CO at post-HI with hypothermia 72 hours in three groups,but the heart rate decreased in B and group C group.Compared with nonnothermia,mild hypothermia treatment showed no significant differences in MAP,LPEP/LVET,RACT/RVET,CO,SV at post-HI with hypothermia 72 hours.Conclusions Body temperature decreased by 3～5℃for 72 hours will not aggravate hemodynamic abnormity.

Objective:To explore the biological function of miR-132 in ovarian cancer and the target. Methods: 22 cases ovarian cancer tissue and non-tumor tissue adjacent were collected,the expression of miR-132 in tumor tissue and non-tumor tissue, normal ovarian epithelial cells and ovarian cancer cell were detected by RT-PCR. The normal ovarian epithelial cells which the expression of miR-132 maximum or minimum were chosen, and they were divided into two groups, respectively with transfection of negative control plasmid ( NC) and miR-132 mimic plasmid. The expression of miR-132 after transfection was detected by RT-PCR,the cell proliferation and cell apoptosis were detected by CCK-8 method and flow cytometry instrument respectively,the expression of Ezrin protein was detected by Western blot. Results:The expression of miR-132 in tumor tissue was significantly lower than the tumor tissue adjacent,the expression of miR-132 in ovarian cancer cell lines was significantly lower than normal ovarian epithelial cells, the differences were statistically significant (P<0. 05). The SKOV3 cell lines was chosed for gene transfection,compared with NC group, transfection with miR-132 mimic plasmid could significantly reduce cell proliferation, increase cell apoptosis, the difference had statistical significance ( P<0. 05 ) . Western blot results showed that up-regulation miR-132 significantly increased the Ezrin protein expression in ovarian cancer SKOV3 cells ( P<0. 05 ) . Conclusion: In ovarian cancer, miR-132;inhibits proliferation and induces apoptosis of ovarian cancer via Ezrin,it may be a tumor suppressor gene.

Objective To explore the effect of comprehensive intervention technique in neuroses nursing.Methods 48 inpatient neuroses were randomly divided into observation group(drug treatment and comprehensive intervention technique)and control group(drug treatment),using self-rating anxiety scale(SAS)and self-rating depression scale(SDS)to evaluate clinical symptoms before and after 2 months treatment.Results Before treatment.SAS and SDS scores between two patient groups had no statistic difference(P>0.05),but after 2 months treatment.the observation group had lower SAS and SDS scores than the control group,the difference were obvious(P<0.05).Conclusions Comprehensive intervention technique was useful in neuroses nursing which can ameliorate depression and anxiety emotions,improve illness recover.

Objective To explore the relationship of vascular endothelial growth factor(VEGF),IFN-? and IL-4 and effect of Bude-sonide on their in asthmatic rats.Methods Thirty-six male SD rats were randomly divided into 3 groups:asthma group,Budesonide-treatment group and control group.On the first day of the experiment and the 8th day,the rat models of the asthma group and Budesonid treatment group were allergized by the OVA/Al(OH)3 through intraperitoneal injection,respectively.And starting from the 15th day,they were challenged by the OVA through atomization for 2 weeks.Control group was allergized and challenged by NS atomization.Budesonid treatment group was interfered in Budesonide inhalation before suscitation in 0.5 h.After 12 h the same inhal done was again in Budesonide group.Twenty-four hours after the last challenge,the rats in 3 groups were sacrificed,and blood and bronchoalveolar lavage fluid(BALF) were collected.The concentrations of IL-4,IFN-? and VEGF in serum and BALF were measured by enzyme-linked immunosorbent assay.Results The concentrations of IL-4 in serum and BALF in asthma group and Budesonide treatment group were significantly increased than those in control group(P

Apyrase activities in some tissues and cells, such as peripheral vascular endothelial cells, have been reported, but these in endocardium endothelial cells have not been reported. The present study was to characterise the properties of bovine endocardium endothelial cells (BEEC)-associated apyrase. Apyrase activity was assayed by inorganic phosphate release, which could be inhibited concentration-dependently by NaN3, an apyrase inhibitor. NaF (20 mmol/L), another inhibitor of apyrase, also markedly inhibited the activity. EDTA or EGTA (1 mmol/L) could also inhibit the activity completely. However, the inhibitor for Na+/K+-ATPase, ouabain (3 mmol/L) did not affect the enzyme activity. BEEC apyrase activity was dependent on divalent cations (Ca2+ or Mg2+) and pH value.

Retrospective cohort study.

Retrospective cohort study.

Objective To evaluate the outcome of health education program on drowning prevention among primary and secondary school children in rural areas. Methods A township was selected and all the students from grade 3 to 5, grade 7 to 8, and grade 10 to 11 were selected to take part in the program. Twelve intervention measures on natural water safety and drowning prevention were carried out for one year. Information was collected using the same questionnaire before and after the intervention program. Results One year after the intervention was carried out, children's knowledge on drowning prevention improved significantly (13.21, 95% CI: 12.51-13.90) , and a positive effect was also noticed among boys (12.77, 95%CI: 11.77-13.77), girls (13.80, 95%CI: 12.82-14.78),and among primary school children (15.51,95%CI: 14.30-16.72), senior high school children (10.78,95%CI: 9.50-12.05) and junior high school children (12.77,95%CI: 11.84-13.71). Overall rates on risk behaviors dropped from 41.4% to 32.2% (by 22.2%) including 15.6% for boys, 35.2% for girls and 13.8%, 29.3%, 26.3% for primary school children, senior high school children, junior high school children, respectively. The incidence rates for non-fatal drowning decreased by 58.9% (from 5.6% to 2.3%). The person-times for treatment on sight, in emergency settings, in outpatient clinic or in the hospitals had a reduction from 399, 78, 36 to 175, 32, 14, respectively. Conclusion Health education program could improve children's perception on water safety, and reduce their risk behaviors as well as on the incidence of non-fatal drowning in the rural areas.