Objective To investigate and analyze the effect of serum micro inflammatory and nutritional indexes of psychological intervention combined with enalapril maleate tablets on the treatment of the patients with maintenance hemodialysis. Methods 100 patients with chronic renal failure in Jiande the first people's hospital from February 2015 to August 2016 were selected as the subjects, and randomly divided into the control group and the experimental group. The control group and the experimental group were given enalapril maleate tablets, at the same time, the experimental group were received psychological intervention. The changes of serum micro inflammation indexes and nutritional indexes in the the two groups were compared and analyzed. Results The HsCRP, TNF alpha levels in the experimental group decreased significantly than those in the control group, the differences had statistical significant (P<0.05). The levels of prealbumin and plasma albumin in the experimental group were significantly higher than those in the control group (P<0.05). After treatment, the SAS score in the experimental group was (42.3±5.2) points, and the SDS score was (40.1±5.9) points. The SAS score in the control group was (60.9±9.2) points, and the SDS score was (59.1±7.4) points. The scores of depression and anxiety in the experimental group were significantly lower than those in the control group (P<0.05). Conclusion Psychological intervention combined with enalapril maleate tablets used in maintenance hemodialysis patients, which can significantly improve patients' serum inflammation index, restore the nutritional status of patients, eliminate the negative emotion of patients, with further clinical promotion and application significance.

Abstract] Objective To establish an assay for the detection of porcine parvovirus ( PPV) and to verify its application for monitoring cells used for production.Methods A pair of primers and one probe were designed according to the conserved sequence encoding non-structural protein 1 (NS1).Based on the designed primers, a real-time fluorescent quantitative PCR assay for the detection of PPV was developed. Several parameters including the linearity, precision, minimum detection limit and anti-interference of the established assay were evaluated.A stock of PPV strains was prepared by infecting swine testicle ( ST) cells with PPV strains.An assay for the detection of PPV infection was developed by using ST cells as sensitive cells.A combined ST cell infection-PCR test was developed by combining the ST cell infection assay with the real-time fluorescent quantitative PCR assay.The sensitivity of ST cell infection-PCR test was analyzed.The cell samples used for production of biological products were detected by using the established assay.Results The real-time fluorescent quantitative PCR assay was specific for the detection of PPV without cross-reaction to other species of parvovirus virus, SV40 virus and other porcine viruses.The linear range of the assay was 1×109-1×104 copies/μl with a R2 value more than 0.98.The sensitivity of the real-time quantitative PCR assay was 1×104 copies/μl.Both of the intra-and inter-coefficient of variation (CV) were less than 5%in Ct values.The intra-and inter-CV in copies of detection were 5%-15% and 30%-40% respectively.The minimum detection limit of the real-time fluorescent quantitative PCR assay was 1CCID50/ml.The PPV strains were detected in cell samples with no interference.The sensitivity of ST cells infection-PCR test was 0.01CCID50/ml.All of the 22 cell samples were negative for PPV by using the real-time fluorescent quanti-tative PCR assay.Conclusion The real-time fluorescent quantitative PCR and the ST cell infection-PCR test for the detection of PPV in cells were established successfully.The application of the two assays was conducive to further enhance the safety of using cells for production and therapy.

Objective:To provide the basic data for the further revision of mycoplasma test method described in Chinese Pharma-copoeia and some references for the operation standardization of drug mycoplasma test. Methods:Two incubation conditions,namely aerobic conditions and microaerophilic conditions,were compared with respect to the growth status of mycoplasma in liquid and solid media. Results:The growth of mycoplasma was obvious difference between the two incubation conditions,and the microaerophilic con-ditions were better than the aerobic conditions. Conclusion:The microaerophilic conditions can be used in the incubation of mycoplas-ma test,which should be defined and standardized in the future Chinese Pharmacopoeia.

Objective:To investigate the growth-promoting properties and applicability of the mycoplasma test culture media pre-scribed in Chinese pharmacopoeia, and provide reference for the standardization of the drug mycoplasma test method. Methods: Re-spectively using four quantitative detection methods including sensitivity, degree of color change, colony counts and colony diameter, and mycoplasma media widely used in the world as the reference media, the growth-promoting properties of 4 batches of mycoplasma broth and 4 batches of arginine mycoplasma broth from four domestic manufacturers were investigated. Results:The results of sensitivity assay and absorbance detection showed that all the media inoculated with below 100 CFU test microorganisms exhibited visible color change. Furthermore, the results of color change degree and colony diameter showed that there were significant differences among the media products from different manufacturers(P<0. 01). Conclusion:Mycoplasma broth and arginine mycoplasma broth both can sup-port the growth of below 100 CFU test microorganisms. Due to the difference in the growth-promoting properties among the media prod-ucts from different manufacturers, the drug mycoplasma test workers should use more sensitive methods to examine the applicability of the media.

Objective To investigate the effect of aspirin on the endometrium of patients with severe intrauterine adhesions after electrical resection.Methods 130 patients with severe intrauterine adhesions after electrical resection from August 2013 to August 2015 in our hospital were selected and randomly divided into the observation group and the control group ( 65 cases in each group ); The control group were given progestogen sequential therapy, while the observation group were given aspirin for treatment on basis of the control group; All the two groups were treated for three menstrual cycles; Before and after treatment indexes including uterine endometrial thickness, ovulation of the uterine artery blood flow index artery pulsatility index in ultrasound (PI), resistance index (RI), endometrial blood flow parameters[vascular index (VI), flow index (FI) and vascular blood flow index (VFI)] were recorded and compared, as well as menstruation and uterine cavity shape 3 menstrual cycles after treatment;All the two groups were followed up for one year, and one year pregnancy rates in the two groups were recorded.Results After treatment, the total efficiency in the observation group was 92.3% significantly higher than 69.2% in the control group ( P <0.05 ) .After treatment, in the two groups endometrial thickness, PI, RI, VI, FI and VFI were significantly improved compared with before treatment (P<0.05), but the observation group improved more obviously (P<0.05).Within one years in the observation group the pregnancy rate was 57.1%higher than that in the control group 26.5%(P<0.05) .Conlcusions Application of aspirin can significantly improve the endometrial thickness on the basis of estrogen and progesterone sequential therapy , increase the endometrial blood flow and improve the pregnancy rate, which mechanism may be related to promoting endometrial repair and improving blood perfusion of the uterus.

Objective:To establish a podocyte cell injury model induced by puromycin aminonucleoside(PAN),an in vitro model for studying the role of podocytes,especial the slit diaphragm molecules in proteinuria at the cellular and molecular levels.Methods:MPC5 were treated for 24 and 48 hours by 15,45 and 75 mg/L PAN,respectively.The podocyte molecular behavior during podocyte injury was evaluated:the apoptotic podocyte cells were revealed with FITC-Annexin V and Propidium Iodide(PI) assay and the proliferative podocyte cells detected with MTT assay after PAN treatment.The distribution of Nephrin and Podocin was revealed with indirect-immnofluorescent staining under confocal microscope.The distribution of F-actin was revealed with direct-immnofluorescent staining under microscope.Results:The percentage of apoptotic podocyte cells was increased in a dose-and time-dependent manner after PAN treatment.In PAN-treated group,the apoptosis was obviously increased at hour 48,the PAN-45 treated group was 33.48%?14.55% and PAN-75 treated group 38.01%?12.13% vs the control group 6.38%?0.50%(P

Problem-based learning(PBL)plays an improtant role in fostering the ability of problem-solving,creative thinking and active learning of medical undergraduates.Based on the present clinical education resources,how to implement PBL at the largest advantages is the main issue we should concern.

Objective To establish an assay for the detection of Torque teno sus virus ( TTSuV) strains and to analyze its preliminary application to biologics.Methods Primers and probe were designed according to the conserved sequences.A fluorescent PCR assay for the detection of TTSuV strains was estab-lished.Several parameters including the specificity, linearity, accuracy, sensitivity and anti-interference of the established assay were verified.The fluorescent PCR assay was performed to detect the samples of por-cine blood, cell substrate and rotavirus vaccine.The porcine blood samples positive for TTSuV strains were further genotyped.Results The established fluorescent PCR assay was confirmed to have high specificity as no cross-reactions with parvovirus virus of various species, SV40 and porcine circovirus strains were detec-ted.The linear range of the assay was 1×109-1×103 copies/μl for TTSuV1 genotype and 1×109-1×102 cop-ies/μl for TTSuV2 genotype with a R2 value more than 0.993.The sensitivity of the fluorescent PCR assay was 1×103 copies/μl for TTSuV1 genotype and 1×102 copies/μl for TTSuV2 genotype.The intra-and inter-CVs were both less than 7%in Ct values and less than 25% and 45% respectively in copies.No interfer-ence was found in the detection of TTSuV nucleic acids from cell samples.8 out of 20 porcine blood samples were positive for TTSuV strains, among which one sample was positive for TTSuV1 genotype, four samples were positive for TTSuV2 genotype and the rest were positive for both TTSuV1 and TTSuV2 genotypes.Com-pared with the reference strain, strains genotyped as TTSuV1 and TTSuV2 were respectively shared 98%-99%and 98%homologies in sequences.All of the cell substrate and rotavirus vaccine samples detected by the fluorescent PCR assay were negative for TTSuV strains.Conclusion The fluorescent PCR assay for the detection of TTSuV was established successfully, the application of which would further improve the safety of biologics.

Objective To conduct extensive quality control tests on Madin-Darby Canine Kidney ( MDCK) cells used for the production of influenza vaccine .Methods Tests for characteristics , extraneous agents, endogenous agents and tumorigenicity were performed on MDCK cells according to Chinese Pharma -copeia Book III .Cell lysate and DNA of MDCK cells were tested for oncogenicity in the light of new interna -tional requirements .Results The MDCK cells extracted from canis were adherent cells with an epithelial morphology, whose average number of chromosome was 80±1.No bacteria, fungi and mycoplasma contami-nation were detected . The detection for extraneous and endogenous virus showed that there was no nonspecific virus causing cytopathic effect , hemadsorption , hemagglutination or animal death .Tests for re-verse transcriptase , bovine viruses and canine viruses were all negative .Each nude mouse was injected with 107 viable cells to observe their tumorigenicity .Twelve weeks after cell injection , no node was found at the injection site and in large organs by gross anatomy .There was no significant difference between test group and negative control group .The test for tumorigenicity of viable cells was negative .Cell lysate and cellular DNA collected from equivalent amount of cells were respectively injected into nude mice , and no node forma-tion was found.There was no significant difference between the test cells and negative controls in pathology indicating that the tested MDCK cells were non-oncogenic .Conclusion It showed the possibility of using MDCK cells for the production of influenza vaccine .

Objective To optimize and compare four in vitro cytotoxicity assays in order to find a relatively rapid assay that can replace the traditional 51Cr release assay to evaluate the cytotoxicity of immune cells prepared for immunotherapy.Methods Four assays including BATDA, CAM (calcein acetoxymethyl ester), CytoTox-Glo and PKH were optimized and used to measure the in vitro cytotoxicity of NK-92 cells to K562 cell line.Intra-and inter-assay reproducibility of these assays and their correlation with 51Cr release assay were analyzed.Results After optimization, all of the four cytotoxicity assays showed good correlation with effector to target (E/T) ratio in a certain range.Compared with the other three assays, CytoTox-Glo assay showed obvious hook effect at a high E/T ratio of 40∶1.BATDA assay could detect the significant cytotoxicity of NK-92 cells to K562 cells after incubating them for one hour and that was the shortest time taken by the four assays to detect in vitro cytotoxicity.Both CAM and PKH assays took about four hours and CytoTox-Glo assay took six to eight hours to detect the significant cytotoxic potency.All of the four assays, especially BATDA and CAM assays, showed good intra-and inter-assay reproducibility.Among these assays, BATDA assay showed the best correlation with the traditional 51Cr release assay.BATDA assay, as compared with the other three assays, could be used to detect the cytotoxicity of Caov3 cells, an adherent cell line, and showed good results in evaluating the cytotoxic potency of autologous primary NK cells and CD19-CAR T cells.Conclusion Compared with the other three assays, BATDA assay shows the best linear correlation with 51Cr release assay and has the advantages of time-saving and good reproducibility.Besides, it is a better assay for detecting the cytotoxicity of adherent cells.BATDA assay is a promising substitute for 51Cr release assay in evaluating the in vitro cytotoxic potency of NK cells and other immune cells.