Objective To investigate the correlation of platelet activating factor(PAF)with cortisol(Cor) ,as well as their rela-tionship with patients after sepsis prognosis .Methods 102 patients with sepsis admitted in the ICU of Chongqing Emergency Med-ical Center from April 2012 to August 2013 were enrolled into sepsis group ,and 40 cases of volunteers served as control group .All the patients were divided into survival group and non-survival group ,according to their prognosis during hospitalization .Plasma PAF and Cor level in control group ,survival group and non-survival group were compared with each other ,and acute physiology and chronic health evaluationⅡ(APACHEⅡ)in survival group and non-survival group were compared .The correlations of plasma PAF with Cor level in control group ,survival group and non-survival group were analyzed ,respectively .Results Plasma PAF and Cor level in survival group and non-survival group were significantly higher than that in control group(P<0 .05) .Plasma PAF ,Cor lev-el and APACHEⅡ scores were higher in non-survival group than that in survival group(P<0 .05) .The correlation of plasma PAF with Cor level in control group was not significant(P>0 .05) ,while the plasma PAF level positively correlated with Cor level in survival group and non-survival group(P<0 .05) .Curves of receiver operating characteristics(ROC)showed that PAF ,Cor and A-PACHEⅡ score could be used as predictors of mortality during hospitalization(Area=0 .708 ,0 .715 ,0 .787) .Conclusion The plas-ma PAF level positively correlates with Cor level in patient with sepsis .PAF ,Cor and APACHEⅡ score have certain guiding signif-icance for the assessment of prognosis during hospitalization in patients with sepsis .

Objective To explore the effects and mechanism of S100A4 gene silence on invasion of human thyroid cancer cell. Methods After thyroid cancer cell ARO was transfected by S100A4 small interfering RNA (siRNA), mRNA and protein level of S100A4 and matrix metalloproteinase 2 (MMP-2) were determined by real time RT-PCR and Western blot respectively. The anchorage-independent growth was examined by colony formation assay in soft agar, and invasion ability was evaluated by boyden chamber model. Results The level of mRNA and protein of S100A4 was significantly inhibited in ARO cancer cells transfected by S100A4 siRNA.Transfection with S100A4 siRNA could inhibit anchorage-independent growth and invasion ability of thyroid cancer cell ARO in a dose-dependent manner. mRNA and protein expression of MMP-2 were down-regulated by S100A4 siRNA. Conclusion S100A4 siRNA can inhibit the invasion of thyroid cancer cell through down-regulation of MMP-2.

This paper identified misunderstandings of the measures taken in China to overcome the problems incurred by off-label uses (namely reliance on drug makers to modify their medicine specifications,on medical institutions to enhance their regulation and management of off-label uses,and on informed consent to avoid risks).Based on such findings,the paper named defects found with such measures,and puts forward feasible ideas and methods to make such off-label uses legitimate and reasonable.These include clarification of the legal status of the medicine specifications,encouraging authoritative guidance for off-label uses,and determining the subject of evaluation to approve the off-label uses as reasonable.Such efforts aim at helping off-label uses out of the legal difficulties.

AIM:To introduce the concept of bridging study and its strategies in clinical trials for new drug application.METHODS:The concept of bridging study proposed in the ICH E5 guideline was introduced,with a case using bridging strategies in the new drug applications(NDAs)approved by the regulatory authority in Japan.The concrete mode and the development of bridging studies in Asia were summarized.RESULTS:With the application of the ICH E5,some countries and regions have successfully used the bridging strategy in the new drug applications.The bridging strategy is becoming a common and practical basis for the decision making of marketing approvals of new drugs in the Asia-pacific country.CONCLUSION:The currently bridging studies in Asia will play an important role in the extrapolation of foreign clinical data in new drug application.Using bridging study is very helpful in judging ethnic differences of drugs,reducing duplication of clinical trails,as well as shortening clinical development periods.

Objective To investigate the X ray radiation protective measure during interventional procedure.Methods The X ray radiation dose at 1,2,3 meters from the X ray tube in front of and in the near of lead glass,plumbic suit were measured in 101 cases interventional procedure.The data were analyzed and evaluated.Results The shielding efficacy was 95% by the lead glass and plumbic suit double protection,the X ray radiation was reduced markedly.The X ray radiation attenuation was swift with the distance increasing.The X ray radiation attenuation was 53.8% from 1 meter to 2 meter,and 81% to 3 meter.The X ray radiation received by medical personnel increased significantly with the fluoroscopy and subtraction time adding.Conclusion The X ray radiation protection should be carried out from shielding protection,distance protection and radiation time reducing;That can reduce the damage of radiation to keep the operator healthy.

Objective To study the impact of TDGF-1 gene silience by small interfering RNA(siRNA)on the invasion and migration of human breast cancer cell. Methods 3 siRNA fragments were designed according to the characteristic of TDGF-1 gene sequence and the most appropriate siRNA was selected by fluorescence real-time quantitative RT-PCR method. After the human breast cancer cell line MDA-MB-468 was transfected by the selected TDGF-1 siRNA, mRNA and protein of TDGF-1 were determined by real time quantitative RT-PCR and western blot respectively. The migration and invasion ability of the cancer cell were evaluated by wound-healing assay and Boyden chamber model respectively. Results siRNA could down-regulate the level of mRNA and protein of TDGF-1 in a dose-and time-dependent manner. In vitro experiment showed that TDGF-1 siRNA transfection can effectively inhibit the clonal growth, invasion and migration of breast cancer cell in a dose-dependent manner. Conclusions TDGF-1 gene may play an important role in the migration and invasion of human breast cancer cells. siRNA transfection can inhibit the invasion of human breast cancer cells.

Objective To study effects of polo-like kinase-1(PLK1)small interfering RNA(siRNA)on proliferation and apoptosis of undifferentiated human thyroid cancer cells.Methods 5 PLK1 siRNA(S1,S2,S3,S4 and S5)were constructed and used to transfect human thyroid cancer cell line ARO.RT-PCR was employed to pick out the most effective siRNA,which was then used to transfect ARO cell.RT-PCR and western blot were used to detect PLK1 expression in thyroid cancer cells,which were divided into different groups.MTT assay was performed to examine the effects of PLK1 siRNA on thyroid cancer cells in all groups.Apoptosis of thyroid cancer cells was observed by caspase-3 activity and TUNEL.Results All the 5 siRNA down-regulated PLK1 mRNA expression.among which S4 showed the best effect.S4 transfection could obviously inhibit proliferation of thyroid cancer cells in dose and time dependent manner.Compared with control groups,caspase-3 activity of cancer cells in s4 transfeeted group increased significantly.The effect of S4 transfection was dose and time dependent.TUNEL results showed apoptosis of cancer cells transfected by S4 siRNA was obvious and apoptosis of cells was dose-dependent.Conclusions PLK1 may play an important role in proliferation of undifferentiated thyroid carcinoma.PLK1 siRNA transfection can inhibit proliferation of throid cancer cell through apoptosis induction.

The World Health Organization (WHO) standard assay for determining antibody level is the rapid fluorescent focus inhibition test (RFFIT) and is used to determine the degree of immunity after vaccination against rabies.To compare the difference in RFFIT results between the laboratories of The National Institute of Infectious Disease in Japan (NIID) and the Chinese Centre for Disease Control (CCDC) as well the influence of the choice of standard serum (STD) for the detection,the two laboratories detection methods were simultaneously manipulated by RFFIT.The reference serums used in NIID and the WHO standard serum used in CCDC were compared in the same RFFIT detection to determine the titer of four sera samples C1,S1,S2 and S4 in parallel,and the titers of the detected sera samples were calculated using the standard formula for neutralizing antibody titer.No significant difference was found in RFFIT methods from the two laboratories and the RFFIT testing procedures of the two laboratories have good consistency.However,different titers were obtained with the tentative internal standard serum (TI-STD) produced by adjusting to 2.0 IU of WHO standard serum in NIID and the WHO STD.The titer determined with the TI-STD was higher than that determined with WHO STD,This difference appears to be significant and requires further investigation.

To provide a foundation for the development of rapid and specific methods for the diagnosis of rabies virus infection,anti-rabies virus monoclonal antibodies were prepared and rabies virus nucleoprotein and human rabies virus vaccine strain (PV strain) were used as immunogens to immunize 6-8 week old female BALB/c mice.Spleen cells and SP2/0 myeloma cells were fused according to conventional methods:the monoclonal cell strains obtained were selected using the indirect immunofluorescence test; this was followed by preparation of monoclonal antibody ascitic fluid; and finally,systematic identification of subclass,specificity and sensitivity was carried out.Two high potency and specific monoclonal antibodies against rabies virus were obtained and named 3B12 and 4A12,with ascitic fluid titers of 1∶8000 and 1∶10000,respectively.Both belonged to the IgG2a subclass.These strains secrete potent,stable and specific anti-rabies virus monoclonal antibodies,which makes them well suited for the development of rabies diagnosis reagents.

Objective:To investigate the effects of cyclic tensile stress on the function and degeneration of nucleus pulposus cells.Methods:The human primary nucleus pulposus cells were isolated and cultured. The cyclic tensile stress (100 000 μ?, 10% tensile strain, 0.1 Hz, 8 640 cycles) was loaded on the cells for 24 h. The proliferation of the cells was examined by MTT method. The cell cycle and apoptosis were detected through flow cytometry. Gene expression profile chip was used to detect the differentially expressed genes between the tensile stress group and control group. The function of these gene was analyzed by bioinformatics. The expression of inflammatory related factors, TGF-β, matrix degrading enzymes and extracellular matrix molecules were examined by qRT-PCR.Results:The cyclic tensile stress significantly promoted proliferation and cell cycle of nucleus pulposus cells. The cell percentage of S phase ( t=5.336, P<0.05) and G2/M phase ( t=7.288, P<0.01) was significantly different between the tensile stress group and control group. The cyclic tensile stress inhibited apoptosis of nucleus pulposus cells (8.56%±0.48% vs 10.63%±0.32%, t=4.474, P<0.05). A total of 866 differentially expressed genes were detected. Gene ontology analysis showed the roles of these genes in cells including focal adhesion, extractable matrix, membrane raft, condensed chrome kinetochore, cytoskeleton, etc. The cyclic tensile stress significantly affected the mRNA expression of inflammatory related factors, TGF-β genes, matrix proteinase and extracellular matrix molecules. Compared with the control group, the mRNA expression of inflammatory related factors IL15 ( t=5.379, P<0.05), IGF1 ( t=5.454, P<0.05) and IGFBP7 ( t=13.57, P<0.01) were significantly decreased in the tensile stress group; The mRNA expression of TGF-β genes TGFB1 ( t=6.931, P<0.05), TGFB2 ( t= 15.56, P<0.01) and TGFB3 ( t=7.744, P<0.05) were significantly increased in the tensile stress group; The mRNA expression of matrix proteinase ADAMTS3 ( t=5.241, P<0.05) and MMP19 ( t=24.72, P<0.01) were significantly decreased, and TIMP3 ( t=8.472, P<0.01) increased in the tensile stress group; The mRNA expression of extracellular matrix molecules COL2A1 ( t=5.871, P<0.05), FLRT2 ( t=5.216, P<0.05) and FN1 ( t=4.289, P<0.05) were significantly increased. Conclusion:The cyclic tensile stress promoted cell cycle and proliferation and inhibited apoptosis of nucleus pulposus cells. The cyclic tensile stress may affect the function and degeneration of nucleus pulposus cells by regulating the expression of inflammatory related factors, TGF-β, matrix degradation enzymes and ECM molecules.