Objective To establish a simple and rapid new method for DNA extraction of FTA bloodstain samples.Methods Genomic DNA was extracted from FTA bloodstains of 1.2mm diameter by FTA-DNA direct extraction and FTA routine method respectively,and their genotypes were analyzed using ABI IdentifilerTM kit in 10?l and 25?l of reaction volume respectively.Results For 25?l of reaction volume,all DNA extracted by two different methods was successfully genotyped.For 10?l of reaction volume,however,the typing success rate of DNA extracted by FTA routine method was significantly lower than those by FTA-DNA direct extraction procedure.Using FTA routine method,the value of RFU ranged from 100 to 2000,and the peak imbalance result from preferential amplification of the smaller allele was a common phenomenon.Moreover,allelic dropout occurred in approximately nineteen percent of samples,and this was not obviously improved even if performed by automatic DNA workstation.However,using FTA-DNA direct extraction procedure,the typing results were similar to those in 25?l of reaction volume,and better results can be obtained using automatic DNA workstation.Conclusion The FTA-DNA direct extraction method is simple and rapid,and can be used to automatic establishment of DNA database with FTA bloodstains.

Objective To investigate the gene expressions of IL-1 and TNF-? in bronchoalveolar lavage fluid(BALF) and lung tissues in mice with pulmonary fibrosis(PF) at various periods and to study the effects of cytokines on pulmonary fibrosis.Methods Forty male mice were randomly divided into two groups:experimental group(n=30) and control group(n=10),and poured respectively by bleomycin solutions and saline and killed on the 3rd,7th,14th,28th and 56th day.RT-PCR method was used to analyse the mRNA expressions of IL-1 and TNF-?.Results ① The expressions of IL-1 mRNA and TNF-? mRNA in BALF in experimental group were higher than those in control group(P

AIM: To further investigate the immunologic mechanism of myasthenia gravis(MG). METHODS: Acetylcholine receptor antibodies(AchRab) and presynaptic membrane antibodies (PsMab) was determined by ELISA in 285 cases of MG patients. Tumor necrosis factor- alpha(TNF-?) and RBC-C3b receptor rosette rate(RBC-C3bRR)and RBC-IC rosette rate (RBC-ICR)was determined in 202 patients with MG. Furthermore, the peripheral blood lymphocyte subsets was examined in 104 cases MG patients. RESULTS: The positive rates of the AchRab and PsMab in MG were very significantly higher than that of control.The positive rate of PsMab in patients with negative AchRab was very significantly higher than that of patients with the positive AchRab.In 202 MG patients, the TNF-? was significantly higher, but the RBC-C3bRR was significantly lower than those of control, while the RBC-ICR has no significant change. CONCLUSION: MG is an autoimmune disease with damaged postsynaptic membrane by the AchRab, and some patients with both the AchRab and PsMab could manifest disfunction in the postsynaptic and presynaptic membrane. While the patients who just has the PsMab maybe damaged only in the presynaptic membrane. Other body fluid immune or/and cell immune mechanism may be involved in MG patients without AchRab and PsMab. [

Thirty cases of pituitary stalk interruption syndrome were divided into normal height (group 1) and short stature (group 2).There was no significant difference in growth hormone or insulin-like growth factor-Ⅰ levels between two groups(P＞0.05).Expected height and body mass index in group 1 were higher than those in group 2,while testosterone,cortisol,and FT4 were lower.Height were positively correlated with age,expected height,birth weight,and body mass index.The height of patients with pituitary stalk interruption syndrome depended on both genetic factors and the hormones from pituitary-target gland.

Objective To study the role of atrial natriuretic peptide (ANP) and renin-angiotensin-aldosterone system (RAAS) in the pathogenesis of salt-sensitive (SS) hypertension and mechanism of the hypotensive effect of benazepril and ANP in patients with SS essential hypertension. Methods Sixty-four patients with essential hypertension were divided into SS (n=30) and non-salt-sensitive (NSS, n=34) groups by modified Sulliran's method. Plasma ANP, angiotensin Ⅱ (AⅡ) and aldosterone (ALD) were determined before and during the period of salt loading test. Thirty healthy subjects as controls were also enrolled. A self-comparative study of benazepril with the placebo was performed in SS group. Before and after the placebo and benazepril therapy, blood pressure (BP) and plasma ANP were determined. Results (1)Basal plasma ANP level in the SS group was significantly lower than that in the NSS group. Basal plasma ANP level in the NSS group was also significantly lower than that in the control group [(110.28±15.40) pmol/L vs NSS (145.52±26.53) pmol/L and control (197.74±26.20) pmol/L]. Plasma ANP in both SS and NSS groups [(133.56±34.03) pmol/L and (169.20±35.91) pmol/L respectively, both P<0.05 vs control]. Percentage of increase of plasma ANP in SS and NSS groups was of no difference (P>0.05) . (2) No significant difference of basal plasma AⅡ and ALD levels were found between SS and NSS groups (P>0.05). There were no significant changes of plasma AⅡ and ALD during salt loading in both SS and NSS groups ( both P>0.05). (3) After the benazepril treatment, plasma ANP was increased significantly [(146.74±31.86) pmol/L , P<0.01]; both systolic and diastolic BP were reduced significantly in SS group. (4) Basal plasma ANP level was negatively correlated with the magnitude of increase of mean arterial pressure (MAP) by salt loading (b=-0.06, P<0.05). Conclusion Deficiency of circulating endogenous ANP may play an important role in the pathogenesis of SS hypertension. Benazepril could reduce BP and increase plasma ANP significantly in patients with SS hypertension.

Objective To investigate the incidence and risk factors of end-stage liver disease (ESLD) in human immunodeficiency virus (HIV) and hepatitis C virus (HCV) co-infected patients after antiretroviral therapy (ART).Methods The demographic and clinical data of HIV-HCV coinfected patients in Zhongnan Hospital,Wuhan University and local Centers for Disease Control and Prevention (CDC) from Jan 2003 to Dec 2010 were analyzed retrospectively. Single factor and multiple factor Logistic regression were used to analyze the correlation between the variables and incidence of ESLD,such as baseline age ≥40,male,previous blood transfusion history,duration of HCV persistent infection,hepatitis B virus (HBV) co-infection (HBsAg positive),HIV RNA≥-1 ×104 copy/mL at last visit,HCV RNA≥ 1.× 105 copy/mL at last visit,CD4 count ＞ 200 / μL at last visit,alanine aminotranferase (ALT) ≥ 2 × upper limit normal (ULN) at last visit,ART containing nevirapine (NVP),follow-up duration,ART duration＞5 years and HCV genotype 1b.The effect of ESLD on the survival of HIV-HCV co-infected patients was analyzed by Kaplan-Meier method.Results Totally 427 HIV-HCV co-infected patients were followed up with average of 3.7 years. Fifty-five patients (12.9％) developed ESLD,and 52 patients (12.2％) died.Factors independently associated with ESLD included baseline age≥40 (OR=2.385,P=0.039),ALT ≥2× ULN (OR=16.374,P=0.000),HBV-coinfection (OR=2.507,P=0.042),duration of ART ＞ 5 years (OR=3.232,P=0.010),and CD4 count ≥200/μL (OR=0.364,P=0.011).The cumulative mortality of HIV-HCV co-infected patients with ESLD was 50.9％,whereas that of HIV-HCV co-infected patients without ESLD was 6.5％ (P=0.000).Conclusion In the ART era,ESLD is common among HIV-HCV co-infected patients in China,which is responsible for reducing the survival time of the patients.

Objective To analyze the incidence,mortality and risk factors of death in human immunodeficiency virus (HIV) and hepatitis C virus (HCV) co-infected patients with combined antiretroviral therapy (cART).Methods A total of 427 HIV/HCV co-infected patients admitted to Zhongnan Hospital of Wuhan University or local disease prevention and control canters from January 2003 to December 2010 were enrolled in the study.The demographic and clinical data of patients were retrospectively studied.Cox progressive regression model was used for data analysis,and Kaplan-Meier method was used to evaluate the effect of end-stage liver diseases on the death.Results of 427 HIV/HCV co-infected patients,53 ( 12.4％ ) died during the follow-up,in which 28 (52.8％) died of liver-related diseases.Male gender ( RR =2.63,P =0.05 ),infection via blood transfusion ( RR =2.15,P =0.04),baseline CD4 + T cells ＜50 cells/μL ( RR =2.83,P =0.02),HIV RNA≥ 104copies/mL at the end of follow-up (RR =2.79,P =0.00 ) and complicated with end-stage liver disease ( RR =7.79,P =0.00) were significantly related to the death.Duration of cART ＞ 5 years is a protective factor for the death ( RR =0.03,P =0.00).Themortality of patients complicated with end-stage liver diseases was 52.7％ ( 29/55 ).Conclusion Liver disease-related death has become the leading cause of death in HIV/HCV co-infected patients,and patients with end-stage liver diseases are of high risk of death.

To investigate the chemical constituents of A. barbadensis, aqueous extract of the plant was subjected to preparative medium pressure liquid chromatography (MPLC). The chemical structures were mainly determined by spectroscopic evidences (UV, IR, HR-MS, 1H NMR, 13C NMR, HSQC, 1H-1H COSY and HMBC) and chemical methods. A new O, O, O-triglucosylated naphthalene derivative, together with two known 6-phenyl-2-pyrone derivatives and four 5-methylchromones, were isolated and identified as 1-((3-((4- O-beta-D-glucopyranosyl)-beta-D-xylopyranosyloxymethyl)-1-hydroxy-8-alpha-L-rhamnopyranosyloxy)naphthalene-2-y])-ethanone (1), 10-O-beta-D-glucopyranosyl aloenin (2), aloenin B (3), aloesin (4), 8-C-glucosyl-(R)-aloesol (5), 8-C-glucosyl-7-O-methyl-(S)-aloesol (6), and isoaloeresin D (7). Compound 1 is a novel naphthalene derivative and named as aloveroside B, compounds 2-3 are isolated from this Aloe species for the first time.

Objective To explore the value of endothelial dysfunction assessment in predicating major adverse cardiac and cerebrovascular events (MACCE) in peritoneal dialysis (PD) patients.Methods A prospective cohort study included 136 end stage renal disease (ESRD) patients from Jan 1,2009 to Dec 31,2011 was conducted.Endothelial function was assessed by flowmediated dilation (FMD) of brachial artery.Kaplan-Meier method was used to estimate survival rate.The survival difference between the two groups was compared by the log-rank test.Multivariate Cox proportional hazards regression was used to determine the independent risk factors of MACCE.Results In the follow-up period,18 patients in low FMD (FMD≤2.7％) group occurred MACCE,and 13 patients in high FMD (FMD ＞ 2.7％) group occurred MACCE.Compared with high FMD group,MACCE-free survival rate in the low FMD group had a significantly decreased (x2=4.190,P=0.041).Multivariate Cox proportional hazards regression analysis showed that higher level of total cholesterol,lower FMD,longer PD time and higher levels of hs-CRP were all independent predictors of MACCE.Conclusion Reduced brachial artery FMD is an independent risk factor of MACCE,and the application of FMD contributes to the risk stratification of cardiac and cerebrovascular disease in PD patients.

Objective To explore a variety of levels of serum marker test applications in the diagnosis of fatty liver .Methods Data were randomly selected from April 2013 to April 2014 for treatment of patients with fatty liver hospital 45 cases ,set the study group ,choose the same period in healthy volunteers to undergo a medical examination in our hospital 45 cases ,it was set to control group ,two groups of subjects were taking a variety of levels of serum markers tested .Comparison and analysis of two groups of subjects to detect a variety of levels of serum markers and positive case detection rate .Results The study group subjects ALT , AST ,TG ,TC index the average level of detection was higher than the control group ,statistically significant differences (P<0 .01);study group subjects ALT ,AST ,TG ,TC index the positive rates were 77 .78% ,93 .33% ,55 .56% ,46 .67% more than 8 .89% in the control group ,4 .44% ,15 .56% ,11 .11% higher ,statistically significant differences (P<0 .05);United biochemical indicator de‐tection of biochemical indicators of detection rate of fatty liver was obviously higher than that of single detection rate ,the difference was statistically significant (P<0 .05) .Conclusion Multiple levels of serum markers of fatty liver diagnostic test in higher detec‐tion rate .