Objective To investigate the inhibitive effects of biosynthesized short hairpin RNA(shRNA)on the duplication of Influenza A virus(IAV)in human bronchial epithelium(HBE)cells.Methods NP-shRNA and PA-shRNA targeting to the highly conservative sequences of IAV nucleoprotein(NP)and acidic RNA polymerase(PA)were designed and biosynthesized by in vitro transcription,and then were inserted into the plasmid pGenSil-1.After sequencing analysis,the recombinant plasmids NP-shRNA and/or PA-shRNA were transduced into HBE cells followed by infected with IAV A/PR/8/34 H1N1(A/PR8)virus.The cytopathogenic effect(CPE)of the HBE cells,hemagglutination assay(HA)and 50% tissue culture infective dose(TCID50)titer of A/PR8 in the culture supernatants were determined respectively.The mRNA and protein expressions of NP and PA were detected by RT-PCR and Western blotting respectively,and the results were compared in order to evaluate the inhibitive efficiency of NP-shRNA and/or PA-shRNA to A/PR8 duplication in HBE cells.Results The content of NP-shRNA and PA-shRNA biosynthesized by in vitro transcription were respectively 59.4 nmol and 50.6 nmol in each 100 ?l.The purity was more than 2.0 and the sequences were verified to be correct after sequencing analysis.The CPE of HBE cells and the virus titer in the culture supernatants of cells transfected with NP-shRNA,PA-shRNA or NP-shRNA+PA-shRNA were markedly lower than those of the control groups.In 3 h after 1 TCID50 A/PR8 infection,the mRNA level of NP in NP-shRNA group,the mRNA level of PA in PA-shRNA group and those in NP-shRNA+PA-shRNA group were 41.7%,43.4%,68.5% and 73.7% respectively,lower than those of the control group.At 48 h after 1 TCID50 A/PR8 infection,NP synthesis were 92.3%,84.0% and 91.7% respectively of those of the control group.Conclusion Anti-IAV NP-shRNA and PA-shRNA are successfully prepared by in vitro transcription.Both of those markedly inhibit the reproduction of IAV A/PR8 and produce a favorable protective effect on HBE cells.

Objective To investigate the role of P21 protein and XIAP on apoptosis-inducing effect by mevastatin against A549 cell line and its mechanisms. Methods The cells proliferation was detected by MTT assay. The cell cycle distribution and apoptosis induction were evaluated with flow cytometer and electron microscopy. The mRNA expression of P21 and XIAP was measured with reverse transcription-polymerase chain reaction.The protein expression of P21 and XIAP was tested with flow cytometer and Western Bolt respectively. Results Mevastatin inhibited A549 cell survival in a time-dependent and concentration-depended manner. Flow cytometry showed that mevastatin induced G0/G1 phase arrest and caused apoptosis. Mevastatin did not affect P21 expression at both mRNA and total protein level.Concomitantly,P21 protein localized on cellular membrane decreased.Both mRNA and protein expression in XIAP were down regulated by mevastatin. All these effects were reversed by mevalonate.Conclusion Mevastatin can inhibite proliferation,induce apoptosis and interfere with cell progression of A549 through mevalonate pathway. Its mechanisms are explained by decreasing the expression of XIAP mRNA and protern.Targeting HMG-CoA reductase,Mevastatin blocks the isoprenylation of P21 protein which affects its anchorage on the cellular membrane.

Objective To observe the effects of health promotion on tumor recurrence,quality of life and self feeling evaluation of patients with superficial bladder carcinoma.Methods 104 patients with superficial bladder carcinoma treated in our hospital from March 2002 to March 2005 were divided to two groups randomly.The patients in one group were treated by bladder irrigation with Bacille Calmette Guerin (BCG)simply,and those in the other group were treated by bladder irrigation and health promotion.Tumor recurrence,quality of life and self feeling evaluation between the two groups were compared.Results All patients were foUowed up for 7.3-36.0 months(mean 28.5 months).17 patients(15/51)in the simple BCG irrigation group had tumor recurrence in 0-12 months(2 patients),13-24 months(6)and 25- 36months(9)respectively.The recurrence rate in the first year was 3.9%.in the second year 11.8% and in the third year 17.6%.Eleven(11/53)patients had in the BCG irrigation and health promotion group.in 0-12 months(2),13-24 months(7),and 25-36 months(2)respectively.The recurrence rate in the first year was 3.8%.in the second year 13.2%.and in the third year 3.8%.There were no significant differences in the recurrence rates of the first and second year between the two groups(P>0.05).but there was significant difieFence in the recurrence rate of the third year between the two group(Y2=5.29,P< 0.05).There were no significant difierences in self feeling SCL-90 evaluation and quality evaluation of life after the first discharging between the two groups(P>0.05).After 3-year follow up,the life quality of the patients in the BCG irrigation and health promotion group was improved obviously compared with the simple BCG group(X2=6.47,P<0.05),and the bad negative emotion was diminished obviously compared with the simple BCG group(P<0.05).Conclusion Health promotion after operation on the patients with superficial bladder carcinoma can decrease the long-term tumor recurrence rate obviously,and improve the quality of life and self feeling significantly.

Aim To investigate the effect taurine on ischemia-induced neuron apoptosis and its mechanisms. Methods Focal cerebral ischemia was induced by middle cerebral artery occlusion (MCAO). After 6 h permanent MCAO, mRNA expressions of Caspase-3/8 of rat brain were detected with real-time quantitative reverse transcriptase-polymerase chain reaction (real-time QRT-PCR). Oxygen-glucose deprivation (OGD) was used to induce an in vitro ischemia model. Primarily cultured cortical neurons were exposed to 4 h OGD. Intracellular calcium concentrations([Ca~(2+)]_i) was measured with fluorescent Ca~(2+) sensitive probe fura 2 acetoxylmethyl ester (Fura 2/AM). Neuronal apoptosis was assayed with flow cytometry of FITC-annexinV/propidium iodide binding 24 h after OGD. Results mRNA expressions of Caspase-3/8 were upregulated in MCAO model. [Ca~(2+)]_i and neuronal apoptosis were markedly increased in OGD model. Taurine pretreatment reduced the upregulation of mRNA expression of Caspase-3/8 in vivo and ameliorated calcium overload and neuronal apoptosis in vitro. Conclusion Taurine has neuroprotective effect against ischema-induced neuronal apoptosis. This is partly due to its effects on Caspase-3/8 and [Ca~(2+)]_i

To support the ongoing health reform in China,Chongqing municipal government made the decision in 2008 "Building Healthy Chongqing" ,and pinpointed for the first time regional development as a key goal.In line with this situation,the hospital proposed such hypothesis changes as from "disease hypothesis to "health hypothesis",from "illness oriented" to "capability oriented",and from "shaping development" to "adaptive development",as well as from one-sided efforts of medical staff to minimize casualty to joint efforts with patients to safeguard life and health.Focusing on these four strategic shifts,the hospital works in developing its "technical attraction,service capacity,innovation capacity,and emergency assurance capability,and management soft power",aiming at building a sustainable hospital in its development.

Objective To identify the significance of expression and phosphorylation of protein kinase R(PKR) and nuclear factor NF-κB p65 in cervical lesions, and the effect of high-risk human papilloma virus(hsHPV) on expression and phosphorylation of R(PKR) and NF-κB p65. Methods A total of 67 patients with cervical cancer, 149 patients with cervi-cal intraepithelial neoplasia (CINⅠ-Ⅲ) and 15 normal control were included in this study. The expression levels of PKR, phosphorylated PKR (p-PKR), NF-κB p65 and phosphorylated NF-κB p65 (p-NF-κB p65) were detected by immunohisto-chemical SP method in three groups. Results The positive expression rates of PKR and p-PKR in cytoplasm were signifi-cantly lower in hsHPV positive group than those in hsHPV negative group (27.2% and 11.0% vs 41.1% and 21.1%,χ2 =4.858 and 4.371,P＜0.05). The positive expression rates of NF-κB p65 and p-NF-κB p65 in cytoplasm and nucleus were significantly higher in hsHPV positive group than those in hsHPV negative group (46.3%, 25.7%, 22.8% and 12.5% vs 32.6%, 14.7%, 11.6%and 4.2%,χ2=4.345,4.048,4.729 and 4.650 respectively,P＜0.05). The positive expression rates of PKR in kytoplasm and karyon were significantly lower in NF-κB p65 (+) group than those in NF-κB p65 (-) group (25.5%vs 38.0%and 20.4%vs 36.3%,χ2=3.898 and 4.396 respectively, P＜0.05). The positive expression rate of PKR in kyto-plasm was significantly lower in p-NF-κB p65 (+) group than those in p-NF-κB p65 (-) group (19.0%vs 36.0%,χ2=4.462, P＜0.05). Conclusion hsHPV may inhibit the expression and phosphorylation of PKR but promote the expression and phosphorylation of NF-κB p65. The expression and phosphorylation of NF-κB p65 may inhibit the expression of PKR. Regu-lating effects of three may be associated with the generation and progression of cervical cancer.

Objective To determine the taxonomic position of medically important gamasid mites and to compare numerical taxonomy with the traditional systematics in the classification of gamasid mites. Methods Based on hierarchical cluster analysis, numerical taxonomy was applied to study fifty-seven species of medically important gamasid mites. Results The results of cluster analysis based on squared euclidean distance showed that Hirstionyssus Fonseca and Echinonyssus Hirst should be separated from Laelapidae Berlese and formed an independent family. The taxonomic position of the remaining species remains the same as those in the traditional systematics. Conclusion Numerical taxonomy can objectively reflect the taxonomic position of the medically important gamasid mites. The result of classification by numerical taxonomy is consistent with traditional systematics for gamasid mites.

Objective To investigate the expression of VCAM-1 and E-selectin in vessel walls of the patients with thromboangiitis obliterans(TAO).Methods The vessel tissues were taken from 18 cases of amputation operation due to TAO and 16 cases of amputation operation due to non-vascular diseases.The expression of VCAM-1 and E-selectin were assessed by the immunohistochemical method.Results The positive expression rate of VCAM-1 in vessel wall of the patients with TAO was 77.8%(14/18),which of E-selectin was 72.2%(13/18);both of them were obviously higher than 6.3%(1/16) and 6.3%(1/16) in the control group.The positive rate of VCAM-1 and E-selectin had no correlation with the age,smoking amount,smoking time and drug use (P>0.05).Conclusion The expression of VCAM-1 and E-selectin have close correlation with the occurrence and development of TAO.Detecting the expression of VCAM-1 and E-selectin can serve as a diagnostic indicator of TAO.

Objective To investigate the relationship between evidence-based nursing tactics and quality of life of breast cancer patients so as to provide the reference for the best nursing strategy .Methods 87 patients who were diagnosed with breast cancer received surgical treatment were randomly divided into experimental group (44 cases) and control group (43 cases) .Patients in control group were given general nursing ,while those in experimental group were given evidence-based nursing tactics on the ba-sis of general nursing .Both groups were asked to fill in the quality of life questionnaire after 4 months breast cancer treatment and before treatment .Results The quality of life in experimental group was significantly improved after treatment compared with that of before treatment(P< 0 .05) ,the quality of life in the experimental group received evidence-based nursing tactics is significantly better than that of in control group (P< 0 .05) .Conclusion Evidence-based nursing tactics can improve the quality of life in breast cancer patients .

AIM:To explore the effect on stability of hyperforin with different medicinal herbs and the changes of hyperforin content in herb combinations with Hypericum perforatum L.(HP).METHODS:Several common antidepressant medicinal herbs were selected separately to match with HP.The herbal samples consisted of HP group,HP with Curcuma wenyujin Y.H.Chen et C.Ling group,HP with Curcuma longa.L.group,HP with Epimedium brevicornum Maxim group and HP with Wuling powder(WY)group.The contents of hyperforin with each group were determined by reverse phase HPLC.RESULTS:The rates of extraction and retention of hyperforin in HP with Curcuma wenyujin Y.H.Chen et C.Ling group and HP with Curcuma longa.L.group were significantly lower than that of HP group.There were no obvious differences in the extraction rates of the HP group,HP with Epimedium brevicornum Maxim group and HP with WY group.The retention rate of hyperforin in HP group with WY powder was higher than that of HP group.The retention rate of hyperforin in HP with Curcuma wenyujin Y.H.Chen et C.Ling group,was lower than that of HP group.CONCLUSION:The results show that compatibility of HP with traditional Chinese medicine have effects on the stability of active herbal component.