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Objective@#To understand the molecular evolution characteristics of the nucleoprotein (N) genes and epidemiological feature of 118 rabies virus (RABV) strains isolated in Yunnan province, China from 2006 to 2015.@*Methods@#The brain tissue samples from mad dogs, suspicious sick dogs, sick cow, and human brain tissue, saliva and CSF samples from rabies patients were collected in Yunnan province to detect the viral antigen by direct immunofluorescence assay (DFA). The viral RNA from positive samples was extracted. Coding region of N gene was amplified by RT-PCR and sequenced. The phylogenetic tree was constructed by Neighbor-Joining method of MEGA5.0 software.@*Results@#The sequences of N genes of 91 RABV strains in Yunnan from 2012 to 2015 were obtained. With the sequences of N genes of 27 RABV strains in Yunnan from 2006 to 2011 and 29 RABV strains from Southeast Asian Countries, the phylogenetic analysis was performed. RABV strains in Yunnan were divided into clades YN-A (105 strains), YN-B (6 strains), YN-C (7 strains), which belonged to clades China-I, China-VI, China-II respectively. Clade YN-A was epidemic every year from 2006 to 2015, of them, 14 strains from 2006 to 2011 and 91 strains from 2012 to 2015 were distributed in 13 prefectures (cities) of Yunnan. Clades YN-B and YN-C were epidemic only from 2006 to 2010 and from 2008 to 2011 respectively. The regional distribution of clades YN-B and YN-C was limited. The strains of YN-A and YN-C were closely related to the strains of clades China-I and China-II from neighboring Sichuan, Guizhou, Guangxi and Hunan provinces. The strains of YN-B were closely related to the strains from Myanmar, Laos, Vietnam and Cambodia.@*Conclusions@#Three RABV clades with multiple transmission sources were identified in Yunnan. Clade YN-A was widely distributed in rabies endemic area in Yunnan from 2006 to 2015, and it has strong ability to spread as principal clade in Yunnan. Since 2012, clades YN-B and YN-C were not found again in Yunnan.
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Objective·To analyze the clinical characteristics of decompensated cirrhosis patients with intestinal obstruction and related risk factors.Methods·Clinical data of 1 783 decompensated cirrhosis patients treated between March 2010 and March 2016 were collected.Of them,128 (7.18％) patients with intestinal obstruction were screened as the observation group and 128 patients without intestinal obstruction were randomly selected as the control group.Clinical data of two groups were retrospectively investigated,clinical characteristics were compared and analyzed,and related risk factors were analyzed with the Logistic regression analysis.Results·The clinical symptoms of decompensated cirrhosis patients with intestinal obstruction were hidden and misdiagnoses or delayed diagnoses were common.The incidences of abdominal pain,abdominal distension,vomiting,stop exhaust defecate,ascites,electrolyte disorders,fever,and spontaneous peritonitis were significantly higher in the observation group than in the control group (P ＜ 0.05).The results of univariate analysis showed that age,history of abdominal surgery,white blood cell count,serum sodium,serum potassium,neutrophil percentage,and serum albumin were risk factors for decompensated cirrhosis patients with intestinal obstruction.The results of multivariate analysis indicated that age,history of abdominal surgery,white blood cell count,serum sodium,serum potassium,neutrophil percentage,and serum albumin were independent risk factors for decompensated cirrhosis patients with intestinal obstruction.Conclusion·Decompensated cirrhosis patients with age ≥ 50 years old,a history of abdominal surgery,the abdominal cavity infection,low potassium,hyponatremia,and lower serum albumin are likely to develop the intestinal obstruction.
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Objective To evaluate the relationship between serum level of hepatitis C virus (HCV) core protein and antiviral therapy response in patients with hepatitis C. MethodsOne hundred and sixty-two consecutive patients with chronic hepatitis C (CHC) were retrospectively analyzed. Serum level of HCV core protein and HCV RNA were measured. Among them, 35 CHC patients treated with pegylated interferon (PEG-IFN)+ ribavirin (RBV) were followed up before and at week 4, 24, 48 of treatment. The correlations between serum HCV core protein level, HCV RNA and antiviral therapy were evaluated. Comparison between groups was done by t test and comparison of paired data among groups was done by analysis of variance. Results Total 162 patients were divided into six groups according to the HCV virus load: 56 with HCV virus load≤1×103 IU/mL and HCV core protein absorbance (A)=0. 100±0. 029; 9 with 1×103 IU/mL＜ HCV virus load≤1 × 104 IU/mLand A=0. 246±0. 213; 11 with 1 × 104 IU/mL＜ HCV virus load≤1 × 105 IU/mL and A=0. 235±0.179; 28 with 1×105 IU/mL＜ HCVvirusload≤1×106 IU/mL and A=0. 422±0. 319; 51 with 1 × 106 IU/mL＜ HCV virus load≤ 1 × 107 IU/mL and A = 0. 603 ± 0. 330 ; 7 with 1 × 107 IU/mL＜HCV virus load≤ 1 × 108 IU/mL and A = 0. 900± 0. 379. The HCV core protein absorbance was positively correlated with HCV RNA level (r= 0.36, P＜0.05). The HCV core protein absorbance values of 35 CHC patients before therapy, at week 4, 24, and 48 of therapy were 0. 564 ±0. 296,0. 144±0. 062, 0. 091 ±0. 035 and 0. 112±0. 103, respectively. The HCV core protein absorbance values at week 4, 24, 48 all decreased significantly compared with that before therapy (t= 8. 563,9. 195, 9. 250; all P＜0.05), and there was significant difference between those at week 4 and week 24 (t=4. 301, P＜0.05). Conclusion Serum HCV core protein level is closely correlated with HCV viral load during HCV infection process and antiviral treatment.
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Objective To quantitatively investigate the expression levels of the cytosine deaminases,hA3B,hA3F and hA3G in peripheral blood mononuclear cell(PBMC)of human immunodeficiency virus(HIV)infected patients and to analyze the correlation between cytosine deaminases expression and CD4~+ T lymphocyte counts. Methods Peripheral blood samples were collected from 21 HIV-infected subjects who didn't take antiretroviral therapy(ART-),21 HIV-infected subjects receiving ART(ART+),and 10 HIV-uninfected subjects. PBMC were isolated by Ficoll density gradient centrifugation, followed by RNA extraction and cDNA synthesis.hA3B,hA3F and hA3G mRNA levels were determined by real-time fluorescent quantitative polymerase chain reaction(PCR).CD4~+ T lymphocyte counts were determined using flow cytometry. Data were analyzed by t test, t' test or Wilcoxon rank sum test. Results In HIV-infected subjects without or with ART,HIV-uninfected subjects, the levels of hA3B mRNA were 208.4,365.2 and 563.6,hA3F mRNA were 245.5,316.6 and 442.9,hA3G mRNA were 404.6,360.8 and 638.6,respectively.hA3G mRNA level in HIV-infected subjects was lower than that in HIV-uninfected controls(P=0.0131),but there was no statistical difference between ART+ and ART-groups(P=0.7342).There were no correlations between hA3B,hA3F and hA3G mRNA levels and CD4+ T lymphocyte counts in either ART-or ART+HIV-infected subjects(ART-:r=-0.0104,r=-0.0545,r=0.1623,all P＞0.05;ART+:r=0.3220,r=0.2193,r=0.1455,all P＞0.05).In addition,hA3B,hA3F and hA3G mRNA levels were positively correlated with one another in ART-HIV-infected subjects and HIV-uninfected controls(P＜0.05),but not in ART-HIV-infected subjects(P＞0.05).Conclusions hA3B,hA3F and hA3G expression levels do not directly correlate with CD4~+ T lymphocyte counts in HIV-1-infected patients,hA3B,hA3F and hA3G expression levels in PBMCs tend to he decreased after HIV-1 infection, and ART may increase hA3B and hA3F mRNA expression.
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The nucleocapsid protein (N) is a major structural protein of coronaviruses. The N protein of bat SARS-like coronavirus (SL-CoV) has a high similarity with that of SARS-CoV. In this study, the SL-CoV N protein was expressed in Escherichia coli, purified and used as antigen. An Indirect Enzyme-Linked Immunosorbent Assay (indirect ELISA) was developed for detection of SARS- or SL-CoV infections in bat populations. The detection of 573 bat sera with this indirect ELISA demonstrated that SL-CoVs consistently circulate in Rhinilophus species, further supporting the proposal that bats are natural reservoirs of SL-CoVs. This method uses 1-2 μl of serum sample and can be used for preliminary screening of infections by SARS- or SL-CoV with a small amount of serum sample.
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Objectives To analyze the characteristic of HIV-1 viral infectivity factor (Vif) gene variants isolated from Shanghai. To construct the prokaryotic expression vector of HIV-1 vif gene and understand its immunogenieity. Methods HIV-1 vii genes were amplified and sequenced from 23 serum samples of HIV-1 infected patients in Shanghai and then compared with the international standard HIV-1 strain. Subsequently, these amplified Vif fragments were sub cloned into pETS2b(+)expression vector. The recombinant prokaryotie plasmids pETS2b (+)-HIV-1/Vif were then transferred into BL21DS(Star)cells for expressing and purifying HIV-1 Vif protein. HIV-1 Viff rat polyclonal antibody was then preparing by injecting the purified Vif proteins into the mice. ELISA was used to determine the purity of Vif proteins and the immunogenicity of its polyclonal antibodies. Results The nucleotide acid mutation rate of HIV-1 vif gene in Shanghai AIDS patient was (0. 179±0. 006)% compared with the international standard HIV-1 strain. Some similar mutations in vif gene were found in HIV-1 strains isolated from Shanghai while the amino acid sequence between 151 and 240 in Vif protein was conserved. The construction of HIV-1 Vif prokaryotic expression plamids and the preparation of Vif polyclonal antibodies were successfully done in this study. The reaction between recombinant HIV-1 Vif protein and the serum from HIV-1 infected patient was not significantly different from that between the recombinant protein and healthy control serum(P＞0.05). HIV-1 Vif polyclonal antibody reacted differently with recombined HIV-1 Vif protein compared with healthy control serum samples (t=178.61, P＜0.01). Conclusions The Vif gene mutation rate is high in HIV-1 strains isolated from Shanghai compared with international standard HIV-1 strain. The prokaryotic expression plasmids of HIV-1 vif antigen are successfully constructed and Vif polyclonal antibodies are prepared well.