In order to use retroviral-mediated gene transfer technology in clinical application, retroviral vector must be of high titer and free of detectable replication-competent retroviruses (RCR). The aim of this study was to optimize methods of defective retroviral vector production. Study was conducted using a LXSN vector inserted with human tumor necrosis factor-? gene and an amphotropic retrovirus packaging cell line-PA317. The results indicated that viral titer was influenced by volume of medium and concentration of fetal calf serum. Inactivation of retroviral vector was greater at 37癈 than at 32癈. In experiment of transfection of PA317 and transduction of 3T3, integration of retroviral vector into genome of packaging cells and target cells, and free of RCR were detected by polymerase chain reaction analysis. Viral vector with high titer and free of RCR is able to use in clinical trial

Objective To explore the clinical characteristics and risk factors of strokeassociated pneumonia (SAP) in patients with acute cerebral hemorrhage.Methods A total of 375 patients with acute cerebral hemorrhage were selected from the department of neurology and neurosurgery during January 2013 to December 2015 in our hospital.According to the incidence of SAP,they were divided into the observation group (complicated with SAP,n =79) and control group (not complicated with SAP,n =296).Clinical data were collected,and clinical characteristics and related risk factors of SAP complicated with acute cerebral hemorrhage were analyzed.Results Among all the 79 SAP patients in observation group,there were 38 cases with gram-negative bacterial infections,25 cases with gram-positive bacterial infections,16 cases with mixed infections.The SAP incidence in patients with massive hemorrhage was higher than that in patients with nonmasive hemorrhage (x2 =11.301,P＜ 0.01),and was higher in patients with cerebellum,brainstem,ventricle,thalamus and multifoeal hemorrhage than that in patients with basal ganglion and brain lobe hemorrhage(x2 =4.023,P＜0.05).The hospitalization days of the observation group was longer than that of the control group [(32.7 ± 16.2) versus (17.3 ± 6.7),t=2.93,P＜ 0.01].The mortality of the observation group was higher than that of the control group (24.1％ versus 3.7％),(x2 =8.720,P＜ 0.01).Multivariate Logistic regression analysis showed that age≥ 65 (OR =4.87),underlying lung diseases (OR =5.30),bulbar paralysis (OR =7.39),disorder of consciousness (OR=4.11),NIHSS score ＞ 4 (OR =3.96),invasive airway operations (OR=3.78),gastric tube (OR =4.37),H2-receptor blocking agents application (OR =2.09) were independent risk factors for SAP in acute intraerebral hemorrhage patients.Conclusions Gram-negative bacteria are the main pathogens of SAP in patients with acute cerebral hemorrhage.The patients complicated with SAP after acute cerebral hemorrhage have poor prognosis including prolonged hospitalization period and higher mortality.SAP in acute intraerebral hemorrhage patients is closely related to the following factors:age≥65,underlying lung diseases,bulbar paralysis,disorder of consciousness,NIHSS score ＞ 4,invasive airway operations,gastric tube,H2-receptor blocking agents.

Objective To investigate the levels of mieropartieles originated from platelet (PMP),endothelium (EMP),and tissue factor-bearing microparticles (TF+ MP) in diabetes mellitus and to analyze its relationship with diabetic angiopathy.Methods PMP,EMP or TF+ MP were measured in 106 cases of diabetes mellitus and 50 controls by flow eytometry.The differences of EMP between groups of diabetic macrovascular disease and diabetic microvascular disease were determined.Results The levels of EMP in diabetic patients were higher than that in the control(164.20±128.88 vs 63.81±40.84,P<0.05).Diabetic cases with complication showed higher expression level of EMP than those without complications(184.12±152.77,188.21±149.55 vs 138.53±99.87,both P<0.05).However,no distinct increase was observed in PMP and TF+ MP level in diabetes groups compared with control group.Conclusions Endothelial dysfunction,may contribute to the increased level of EMP in patients with diabetes,especially those complicated with vascular diseases.EMP level may be used to evaluate the status of endothelial function and the development of diabetic angiopathy.

Objective To explore the correlations of soluble thrombomodulin (sTM) and high sensitive C reactive protein(hs-CRP) levels with the hypercoagulable state and indicators of predicting thrombosis in children with primary nephrotic syndrome(PNS).Methods Sixty-four PNS patients who had primary clinical diagnosis or recurrence after hormone withdrawal for 6 months or more hospitalized in Department of Urology,the Second Children's Hospital of Tianjin from January 2010 to January 2014 were selected as PNS group.Thirty patients with inguinal hernia undergoing elective surgery hospitalized in Department of Surgery,the Second Children's Hospital of Tianjin from January 2010 to January 2014 were selected as control group.All the selected patients were extracted for venous blood samples,and the double-antibody sandwich enzyme-linked immunosorbent assay was used to detect plasma sTM levels,while immunoturbidimetric assay was used to detect plasma hs-CRP levels;the indicators of predicting thrombosis as blood lipid and fibrinogen(Fb) etc.were simultaneously detected.Inspection results were compared and the correlation was analyzed by using statistical methods.Results The level of plasma sTM in children with PNS was significantly higher than that in the control group[(1.63 ±0.68) μg/L vs.(0.30 ±0.24) μg/L],and the difference was statistically significant between the 2 groups (t =12.545,P ＜ 0.05).The level of plasma sTM in PNS group with urine protein negative was significantly lower than that before hormone treatment[(1.05 ± 0.56) μg/L vs.(1.63 ± 0.68) μg/L],and there was a statistically significant difference(t =6.298,P ＜ 0.05).The level of plasma sTM in children with PNS had positive correlation with 24 h urinary protein,low density lipoprotein,apolipoprotein B,lipoprotein a and Fb (r =0.379,0.203,0.184,0.177,0.224,all P ＜ 0.05).The level of plasma hs-CRP in children with PNS was significantly higher than that in the control group [(1.9 ± 1.7) mg/L vs.(0.7 ± 0.6) mg/L],and the difference was statistically significant between the 2 groups (t =3.487,P ＜ 0.05).In PNS group,the level of plasma hs-CRP with urine protein negative was significandy lower than that before hormone treatment[(0.4 ±0.3) mg/L vs.(1.9 ± 1.7) mg/L],and there was a statistically significant difference (t =5.473,P ＜ 0.05).The level of plasma hs-CRP in children with PNS was negatively correlated with plasma albumin (r =-0.194,P ＜ 0.05),but positively correlated with Fb (r =0.257,P ＜ 0.01).Plasma sTM and hs-CRP levels in children with PNS were positively correlated (r =0.395,P ＜ 0.05).Conclusions sTM involves in the formation process of PNS hypercoagulable state possibly through dual influencing by blood coagulation process and blood lipids.sTM may be used as one of the reference indicators for PNS activity and prediction of thrombosis.Plasma hs-CRP reflects the micro-inflammatory state in children with PNS,and involves in the formation of hypercoagulability by effecting blood coagulation process,and should be used as indicators for monitoring the hypercoagulability of active PNS.

Objective To investigate the signaling mechanisms underlying up-regulation of nontypeable Haemophilus influenzae(NTHi)-induced MUC5AC mucin expression. Methods The expression of MUC 5AC at mRNA level was measured by reverse transcriptase-polymerase chain reaction (RT-PCR)and real-time fluorescent quantitative PCR.Immunoprecipitation and Western blot were performed tO examine the phosphorylation of p38 mitogen-activated protein kinase(p38MAPK)and epidermal growth factor receptor(EGFR)or the effect of dominant negative mutant of EGFR on the phosphorylation of p38MAPK in HM3 cells treated with NTHi.Luciferase assay was also performed to examine the effect of p38MAPK and EGFR inhibitors or dominant negative mutant of EGFR on NTHi-induced MUC5AC expression at transcription level.Results NTHi induced MUC5AC mucin expression at both mRNA and transcription levels.Phosphorylation of p38MAPK and EGFR were observed in HM3 cells treated with NTHi.Either SB203580,a specific inhibitor for p38MAPK or AGl478,a specific inhibitor for EGFR.inhibited NTHi-induced MUC5AC up-regulation at transcription level. Furthermore,Overexpressing dominant negative mutant of EGFR also inhibited NTHi-induced MUC5AC upregulation at transcription 1evel in a dos-dependent manner.EGFR inhibitor reduced NTHi-induced p38MAPK phosphorylation in HM3 cells.Conclusion Bacterium NTHi up-regulates MUC5AC mucin transcription via EGFR-p38MAPK signaling pathway.

Objective To investigate the association of the polymorphism of the N-acetyltransferase 2 (NAT2 )gene with isoniazid-induced hepatotoxicity and anti-tuberculous treatment efficacy in Chinese Han patients with tuberculosis(TB).Methods A total of 108 TB patients who received initial anti-TB treatment were followed up prospectively.A polymerase chain reaction direct sequencing approach was used to detect genetic polymorphisms of the NAT2 gene.Associations between NAT2 genotype and isoniazid-induced hepatitis/early treatment were analyzed.Chi-square test was used for statistical analysis. Results Among the 108 TB patients, intermediate-acetylators (IA ) was the most frequent NAT2 genotype with the proportion of 54.63%(59/108).The proportion of rapid-acetylators(RA)was 33.33%(36/108),slow-acetylators (SA)was 10.19%(11/108)and super-rapid acetylators was 1 .85 % (2/108). Among the 20 patients who developed drug-induced hepatitis,2 were RA,5 were SA and 13 were IA. Regarding NAT2 genotype,RA patients had a lower incidence of hepatotoxicity (OR =0.176,95 %CI :0.038-0.809,P =0.014)and SA patients were more likely to developed drug-induced hepatic injury (OR=4.556,95 %CI :1 .231 -16.854,P =0.044 ).Statistical analysis revealed that the frequency of variant diplotypes,NAT2*4/*6A (OR=7.741 ,95 %CI :2.653-22.586,P <0.01 )and NAT2 *6A/*6A (OR=15 .353,95 %CI :1 .506 -156.552,P =0.020)were significantly increased in TB patients with hepatotoxicity.NAT2 *4/*4 was less likely to developed hepatic injury (OR =0.176,95 %CI :0.038-0.809,P =0.014).Among the 58 culture-positive patients,12(31 .03%)were persistent culture positive after 2 months standard therapy.Early treatment failure was observed with significantly higher incidence rate in RA than other genotypes (OR = 7.200, 95 % CI :1 .794-28.900, P = 0.008). Conclusions In Chinese Han TB patients,IA is the most frequent NAT2 genotype.The SA status of NAT2 is a risk factor of isoniazid-induced hepatotoxicity.The diplotype of NAT2 *6A has clearly high risk of isoniazid-induced hepatotoxicity.In contrast,NAT2 * 4/* 4 is protective diplotype.RA is associated with early treatment failure in culture-positive patients.

Objective The advantages of the treating allergic rhinitis (AR) by sphenopalatine ganglion stimulation with acupuncture were evaluated.Methods Databases including CBMDisk, CNKI, WanFang, VIP, Cochrane Library, PubMed, ProQuest, ChiCTR, ISRCTN, ClinicalTrials.gov and CENTRAL were searched from the beginning of database established to Jan 2015. All issues from Jan 2004 to Jan 2015 published onjournals Chinese Acupuncture & Moxibustion,Shanghai Journal of Acupuncture and Moxibustion, Acupuncture Research,Journal of Clinical Acupuncture and Moxibustion,Chinese Journal of Integrated Traditional and Western Medicine and Chinese Journal of Otorhinolaryngology in Integrative Medicine were searched by hand at meantime. All data were extracted based on the inclusive and exclusive criteria which was pre-designed, the Revman5.3 was applied for meta-analysis, and the studies qualities were analyzed by grade score.Results 118 articles were collected, 7 studies that involving 1 230 patients met the inclusive criteria. The result indicated that the sphenopalatine ganglion stimulation with acupuncture as the main treatment of AR showed the better total response rate compared to conventional drugs, theOR(95%CI) was 3.22(1.81 - 5.75); however the change of total symptom score had no statistical significant difference, the MD(95%CI) was 0.69 (-0.56 - 1.93), the change of IgE had no statistical significant difference,theSMD(95%CI) was -0.07 (-0.97 - 0.83).Conclusion The main treatment on AR by sphenopalatine ganglion stimulation with acupuncture may has better efficacy than western medicine. But due to the methodological biases existed in most studies, future high-quality RCTs were needed to be included into Meta-analysis to test today’s study conclusion.

ObjectiveTo evaluate the application of a whole blood interferon-γ (IFN-γ) release assay QuantiFERON-TB Gold In Tube (QFT-GIT) in the diagnosis of tuberculous pleural effusion.Methods IFN-γ released by specific T cells stimulated by early secreted antigenic target 6 × 103protein (ESAT-6),culture filtrate protein 10 (CFP -10) and TB7.7 were measured by QFT-GIT test in 44 tuberculous pleural effusion patients and 16 non-tuberculous pleurisy controls.The IFN-γ release level between groups was compared by Mann-Whitmey test.ResultsThe positive rates of QFT-GIT in patients with tuberculous pleural effusion and non tuberculous pleurisy were 95.5％ and 12.5％,respectively.The sensitivity,specificity,positive predictive value and negative predictive value of QFT-GIT were 95.6％,87.5％,95.6％ and 87.5％,respectively.The antigen-specific IFN-γ release level in the patients with tuberculous pleural effusion was significantly higher than that in non-tuberculous pleurisy controls (P＜0.01).Conclusions The whole blood INF-γ release assay QFT-GIT is a sensitive and specific assay for detecting pleural tuberculosis infection.It could be a useful diagnostic tool for the diagnosis of tuberculous pleural effusion in China.

Objective To investigate the prevalence of tuberculosis among silicosis patients and silica exposure patients,and to analysis the risk factors of tuberculosis among these population.Methods A total of 1 227 silica exposure patients from Wenling,Zhejiang were enrolled in this field study.Basic demographic information was collected and chest X-ray was taken for each patient.Sputum was collected for Mycobacterium tuberculosis culture and strain identification. In univariate analysis,t test was performed for continuous variables andχ2 test for categorical variables.In multivariate analysis,the odds ratio (OR )was calculated along with a 95 % confidence interval (CI )by binary Logistic regression. Results A total of 1 204 silica exposure patients had full basic information and 99.8% were male patients with mean age of (59.4 ± 6.8 )years.The patients in phase 0 + to phase Ⅲ were 172 (14.3%),255 (21 .2%),160 (13.3%)and 617 (51 .2%),respectively.The tuberculosis prevalence rate was about 7.3% among these population.The risk factors for tuberculosis including phase Ⅱ silicosis (OR =2.96, 95 %CI :1 .05 -8.32,P =0.04)and phase Ⅲ silicosis (OR=3.88,95 %CI :1 .58-9.56,P <0.01),and contacting with tuberculosis patients (OR=4.14,95 %CI :1 .91 -8.98,P <0.01).Patients complicated with tuberculosis lacked specific symptoms,but fever and weight loss were more frequent.Conclusion Tuberculosis is highly prevalent in silicotic patients,especially in patients with phase Ⅱ/Ⅲ silicosis and in patients with tuberculosis contact history.

Objective:To investigate the effects of modified radical prostatectomy via an extraperitoneal approach on urinary control and sexual function in patients with prostate cancer.Methods:Fifty-six patients with stable prostate cancer who received treatment in Deqing People's Hospital between March 2015 and March 2018 were included in this study. They were randomly divided into observation and control groups ( n = 28/group). The observation group was subjected to modified radical prostatectomy via an extraperitoneal approach. The control group underwent standard laparoscopic surgery. Clinical efficacy and the effects of modified radical prostatectomy via an extraperitoneal approach on urinary control and sexual function were compared between the two groups. Results:Amount of blood loss and postoperative drainage were (125.39 ± 11.12) mL and (65.39 ± 10.12) mL in the observation group, and (224.79 ± 14.01) mL and (104.79 ± 15.01) mL in the control group. There were no significant differences in amount of blood loss and postoperative drainage between the two groups ( t = 18.83, 15.67, both P < 0.05). At 1, 3 and 6 months after surgery, the percentage of patients who had urinary control recovery in the observation group was 53.57% (15/28), 78.57% (22/28), 98.21% (27/28), respectively, which were significantly higher than those in the control group [21.43% (6/28), 35.71% (10/28), 67.86% (19/28), χ2 = 4.12, 7.21, 5.01, all P < 0.05]. At 1, 3 and 6 months after surgery, the score of erectile function recovery in the observation group was (15.98 ± 0.28) points, (15.99 ± 0.72) points, and (18.91 ± 0.48) points, which were significantly higher than those in the control group [(17.11 ± 0.34) points, (13.11 ± 0.48) points, (13.41 ± 0.39) points, t = 3.01, 12.89, 15.78, all P < 0.05]. Conclusion:Modified radical prostatectomy via an extraperitoneal approach can improve postoperative urinary control and sexual dysfunction.