Background Rhodopsin (RHO) gene is the most common disease gene for autosomal dominant retinitis pigmentosa (adRP),one of the main pathogenesis is that misfolded mutant RHO proteins accumulate in the endoplasmic reticulum and cause endoplasmic reticulum stress (ERS).Objective This study aimed to determine the genetic basis for a consanguineous Chinese Han adRP family.Methods This study procedure complied with Helsinki Declaration.All participants in the family were investigated under the informed consent.Regular ocular examination was performed on the patients in this family.Next-generation sequencing (NGS) was carried out to screen the mutations in 189 genes associated with hereditary retinal diseases (HRDs).After being analyzed and filtered,variations detected by NGS were validated by Sanger sequencing and evaluating of pathogenicity.The wild-type RHOWT and mutant RHOP53Rwere cloned into the vector pEGFP-N1.Then the two plasmids were transfected into adult retinal pigmentosa epithelium cell line(ARPE19) and human embryo kidney 293 line (HEK293) to observe the location of rhodopsin-GFP fusion protein in cells,and the expression of ERS related protein XBP1 in the cells was detected by quantitative-PCR and Western blot.Results This family included 5 generations with the typical adRP characteristics.Genetic analysis identified a heterozygous variation,p.P53R in RHO gene,which was fully cosegregated in the family.Wild-type RHOWT-GFP fusion proteins showed the green fluorescence on the endoplasmic reticulum and cytomembrane,but the misfolded mutant RHO-GFP fusion protein gathered only in endoplasmic reticulum.Compared to wild-type RHOWT,the XBP1 was activated and increased by (1.28 ±0.09) fold.The introns of 26 bases in XBP1 mRNA were removed in the HEK293 cells with mutant RHO-GFP fusion protein,and the expression of XBP1 was stronger in the HEK293 cells with mutant RHO-GFP than that in HEK293 cells with wild type RHO-GFP and cells with blank pEGFP-N1 plasmid.Conclusions Heterozygous variant RHO p.P53R is very likely the pathogenical mutation in the adRP family.The RHOP53R mutant rhodopsin protein can not be delivery effectively from the endoplasmic reticulum to the cell membrane,and these proteins accumulate in the endoplasmic reticulum,which causes ERS.

In constructing the system of medical functional teaching and enhancing the quality and ability of students,we adjust the content and method of teaching in practice,and try to find students'problems in study through questionnaire survey and promote teaching reform and disciplines construction.

Objective:To review the inactivation effect of high-voltage pulsed electric field on microorganism to provid theoretical basis for the further research and sterilization application of high-voltage pulsed electric field. Methods:The combination of high-volt-age pulse electric field, microbiology and pulse electric field as the keywords, the research results of high-voltage pulsed electric field on microbial inactivation in PubMed, CNKI, VIP Chinese journal full text database and Wanfang database during 2010 and 2015 were retrieved, summarized and reviewed. Results:As a non-thermal sterilization technology, high-voltage pulsed electric field not only had the characteristics of short sterilization time, narrow increasing extent of temperature and low energy consumption, but also could keep the original flavor of food. Conclusion:With the development of related technology, high-voltage pulsed electric field is expected to re-alize industrialization.

Objective To assess the efficacy and safety of BioLiver Ⅰ bioartificial liver support system (BALSS) on the treatment of two types of acute liver failure (ALF) canine models. Methods The drug induced ALF canine model was established by multiple subcutaneous injections of acetaminophen. Another model was operated to resect 80 percent of the liver tissue. The primary hepatocytes were separated from the swine and cultured within the BALSS. The ALF models were treated by BioLiver ⅠBALSS for 6 hours. The changes of physiological, biochemical and histological parameters were observed before and after the treatment. Results The canines developed ALF 48 hours after injections of acetaminophen, the established rate of the model was 63 3%. While the other ALF canine model developed 24 hours after 80 percent liver resection and the established rate was 84 2%. Using our modified enzymatic digestion method, the yield of hepatocytes was (1 0～3 0)?10 10 per swine with high viability. BioLiver ⅠBALSS treatment resulted in beneficial effects on blood biochemical parameters. The pathological lesions of the liver were repaired. BALSS treatment was harmless to other organs. The ALF canines in drug group survived longer than in operation group. Conclusion This type of BALSS can provide safe and efficacious liver function support in the two types of ALF canine models, and it may be as an hepaful and important therapy to ALF

AIM: To study the electrophysiological effects of nitric oxide (NO) on the pacemaker cells in guinea-pig left ventricular outflow tract under the condition of ischemia/reperfusion ( I/R) .METHODS:The spontaneous slow action potentials of guinea-pig left ventricular outflow tract were recorded by conventional intracellular microelectrode technique.The effects of NO donor sodium nitroprusside ( SNP) on the spontaneous slow action potentials under normal or I/R condition were investigated.RESULTS:SNP at concentrations of 1, 10 and 100μmol/L but not 1000μmol/L signif-icantly increased velocity of diastolic depolarization ( VDD) and rate of pacemaker firing ( RPF) , SNP at concentrations of 1, 10, 100 and 1 000μmol/L notably increased maximal diastolic potential (MDP), amplitude of action potential (APA) and maximal rate of depolarization ( Vmax ) , shortened 50%and 90%of the duration ( APD50 and APD90 ) .In ischemia 10 min group, VDD and RPF were significantly decreased, APA and Vmax were notably increased, and APD50 and APD90 were markedly lengthened compared with control group.In reperfusion 10 min group, VDD and RPF were significantly in-creased, MDP and APA were notably decreased, and APD50 and APD90 were markedly shortened compared with I 10 min group.In reperfusin 10 min group, the pacemaker activity was always irregular.In reperfusion 10 min group, the parame-ters of spontaneous slow action potentials restored to the levels of control group except for VDD and Vmax .In 1, 10 and 100 μmol/L SNP+R groups, VDD and RPF were significantly increased than in ischemia 10 min group .In 1, 10, 100 and 1 000 μmol/L SNP+R groups, APA, APD50 and APD90 were restored to the levels of control group.CONCLUSION:SNP significantly increases the spontaneous activity of left ventricular outflow tract, and relieves the effects of I/R on the sponta-neous slow action potential markedly.

Objective:To explore the ameliorative effect and machanism of MSCs conditioned medium on the ovarian granulosa cells damage induced by triptolide.Methods: Cell Counting Kit 8 assay was used to examine the cell vitality of KGNs with the treatment of triptolide.The mixed enzyme digestion method were used for the isolation of human umbilical cord mesenchymal stem cells (MSCs),and flow cytometry was used for the subsequent immunotype identification.MSCs conditioned medium was collected ,and Cell Counting Kit 8 assay and PI staining was used to analyse the effect of MSCs conditioned medium on the cell vitality and cell cycle distri -bution of triptolide-damaged KGN.Real-time PCR method was used to examine the expression of cell cycle related gene CDKN1A.Results:Triptolide can inhibit KGN cell growth with the inhibition of cell vitality and cell cycle of KGN.MSCs conditioned medium did not influence the proliferation and cell cycle of normal KGN , but improved the triptolide-induced vitality inhibition and extent of S-phase arrest, and inhibit the abnormal up-regulation of CDKN1A in KGN.Conclusion: MSCs conditioned medium ameliorated the KGN cell damage induced by triptolide.

Objective To establish the analysis criteria of nucleoplasmic bridges (NPB) and a dose-response curve of NPB frequencies in human peripheral blood lymphocytes irradiated by 60Co γ-rays.Methods Human peripheral blood samples were collected from three healthy males,and were irradiated with 0,1,2,3,4,5 and 6 Gy of 60Co γ-rays.A cytokinesis-block micronucleus assay was carried out to analyze NPBs and micronuclei (MN) in binucleated cells.Results NPBs in binucleated cells at each dose level of γ-ray was conformed to the Poisson distribution.The dose-response curve of the γ-ray-induced NPB frequencies in human peripheral lymphocytes followed the linear-quadratic model y =(1.39 × 10-3)x2 + (4.94 × 10-3)x (R2 =0.981,P ＜ 0.05).Conclusions The dose-response curve of NPB frequencies in human peripheral blood lymphocytes induced by 0-6 Gy 60Co γ-rays was established.

Objective To investigate the status of knowledge and performance on Chinese Heart Failure Diagnosis and Treatment Guideline (2014 version) in general practitioners of Shanghai Pudong communities.Methods The survey was conducted from April to June in 2014 with a self-designed questionnaire.Total 390 general practitioners (GPs) in Pudong New Area were selected by cluster sampling method.The contents of questionnaire included:diagnosis and differential diagnosis,drug therapy,non drug therapy of chronic heart failure.Result Total 385 questionnaires were retrieved with a response rate of 98.7％ (385/390).The results showed that in aspect of diagnosis and differential diagnosis,373 (96.9％) Gps made the diagnosis based on history and physical examination,171 (44.4％)Gps never used BNP or NTPro-BNP tests,280 (72.7％)GPs did not know how to identify systolic or diastolic heart failure,86 (22.3％)Gps made the differential diagnosis according to the EF value.In aspects of drug therapy,the rate of beta blockers use was 10％-30％ in 284 (73.8％) Gps,149 (38.7％) Gps did not use beta blockers because of not knowing the contraindications,289 (75.1％) Gps used a maximum dose of betaloc for 25-50 mg,no one used 101-200 mg,242 (62.9％)Gps did not know the target dose of betaloc,the rate of ACEI/ARB use was 10％-30％ in 330 (85.7％) Gps,258 (67.0％) Gps would increase the dose but not knowing the target dose.The main reason for not using the target dose of Betaloc and ACEI/ARB was not knowing the dose.In aspect of non-drug therapy:240 (62.3％)Gps never heard of cardiac resynchronization therapy (CRT) and 271 (70.4％)Gps never heard of implantable cardioverter defibrillator (ICD).The senior rank GPs grasped the guideline much better than Gps with primary and intermediate professional ranks.Conclusion General practitioners in community health centers should further study the guideline of heart failure,particularly need to strengthen the knowledge and ability of drug therapy.

Objective:To construct GM-CSF and EB virus LMP2A fusion gene recombinant adenovirus and identificate it , using recombinant adenovirus to do immunologic study.Methods:With RT-PCR and Western blot to detect GC 2A gene expression of the recombinant adenovirus , with ELISA to analysis antibodies of vAd-GC2A stimulation in murine body , lactate dehydrogenase assay to detect the mouse cellular immunity effect , thereby the recombinant adenovirus immune effect was initially analysis.T-test method was used to do preliminary analysis and evaluation of the immune effect of recombinant adenovirus .Results:GC2A fragment amplified by PCR, the size was 1 961 bp and it was the expected.The results showed, the recombinant adenovirus vAd-GC2A proteins can be recognized by sera antibody of GM-CSF ( LMP2A); the results of ELISA showed that vAd-GC2A can stimulate the generation of antibodies.EBV-positive tumor cell killing rate (%) by CTL induced recombinant adenovirus was (66.7 ±6.9)%, at the same time the adenovirus type 5 wild strains and PBS were (24.3 ±2.5)%and(32.4 ±3.1)%only(P≤0.05).Conclusion:Fusion gene has been successfully constructed , vAd-GC2A in mice have humoral and cellular immune function.The data suggest that the vAd-GC2A should play a potent role in preventing the positive EB virus tumor.

Randomized controlled trials (RCT) is the source of the raw data of evidence-based medicine. Blind method is adopted in most of the high-quality RCT. Sham acupuncture is the main form of blinded in acupuncture clinical trial. In order to improve the quality of acupuncture clinical trail, based on the necessity of sham acupuncture in clinical research, the current situation as well as the existing problems of sham acupuncture, suggestions were put forward from the aspects of new way and new designation method which can be adopted as reference, and factors which have to be considered during the process of implementing. Various subjective and objective factors involving in the process of trial should be considered, and used of the current international standards, try to be quantification, and carry out strict quality monitoring.
Acupuncture Therapy ; standards ; Biomedical Research ; standards ; Evidence-Based Medicine ; Humans ; Randomized Controlled Trials as Topic ; standards