OBJECTIVETo elucidate the variation in characterizations and genetic evolution of the matrix protein 2 or ion channel protein(M2) genes of avian influenza subtype H5N1 viruses in the boundary region of Yunnan province from 2008 to 2012.

METHODSA total of swab samples were collected from foreign poultry such as the junction between Yunnan and Vietnam, Laos,myanmar and wild birds in boundary region of Yunnan province from 2008 to 2012 and screened by H5N1 subtype-specific multiplex RT-PCR. The M genes of H5N1 virus from the positive samples were amplified by RT-PCR and cloned into pMD18-T vectors for sequencing. The alignment and phylogenetic analysis of M2 genes were performed with sequences of the known reference strains.

RESULTSA total of 71 positive samples were found out of 1240 samples and the positive rate was 5.72%. A total of 14 different M2 sequences were obtained from 30 positive samples and were divided into 3 distinct clades or sub-clades(1.2.1, 1.2.2 and 2) by phylogenetic analysis, 5, 7 and 2, respectively. The M2 genes and Hemagglutinin(HA) genes of H5N1 viruses from the boundary region of Yunnan province had showed different relationship of genetic evolution. The substitution or mutation of key amino acids sites had been found among the domains of epitope, adamantane-resistance, and poultry or human original viral strains.

CONCLUSIONThe M2 genes of H5N1 subtype viruses in boundary region of Yunnan province from 2008 to 2012 showed genetic divergence and the virus of clade 1.2.2 had become dominant epidemic strain in this region.

Animals ; Birds ; virology ; Chickens ; virology ; China ; Evolution, Molecular ; Influenza A Virus, H5N1 Subtype ; classification ; genetics ; Influenza in Birds ; virology ; Phylogeny ; Poultry ; virology ; Viral Matrix Proteins ; genetics

OBJECTIVETo elucidate the characteristics of variation and the genetic evolution of non-structural protein (NS1, NS2) genes related to avian influenza subtype H5N1 viruses isolated from the boundary region of Yunnan province.

METHODSSwab samples were collected from foreign poultry and wild birds in the boundary regions of Yunnan province and screened by H5/N1 subtype-specific multiplex RT-PCR. The NS segment of H5N1 virus from the positive samples were amplified by RT-PCR and cloned into pMD18-T vectors for sequencing. The alignment and phylogenetic analysis on those available NS1, NS2 genes were performed with sequences of the known reference strains.

RESULTS71 positive samples were identified from 1240 samples, with the positive rate as 5.72%. Fourteen different NS segment sequences were obtained from 30 representative positive samples and could be divided into 3 distinct clades or sub-clades (I-1, I-2 and II), by phylogenetic analysis. The NS1/NS2 genes and Hemagglutinin (HA) genes of H5N1 viruses from the boundary regions of Yunnan province showed different relationships regarding the characteristics on genetic evolution. The substitution or mutation of key amino acids sites had been noticed in the nuclear location signal domains, effect domain, and other pathogenicity markers.

CONCLUSIONNS genes of H5N1 subtype viruses in boundary region of Yunnan province showed genetic divergence and the virus of clade I-2 and II had become dominant epidemic strains in this region since 2010.

Amino Acid Sequence ; Animals ; Animals, Wild ; Birds ; virology ; China ; epidemiology ; Evolution, Molecular ; Genome, Viral ; Influenza A Virus, H5N1 Subtype ; genetics ; isolation & purification ; Influenza in Birds ; epidemiology ; virology ; Phylogeny

OBJECTIVETo explore the feasibility and the effect of laparoscopic hepatectomy for primary liver cancer(PLC).

METHODSA retrospective study on 61 cases of laparoscopic hepatectomy for PLC was made between November 2002 and June 2007, among which there were 49 male and 12 female, aged from 14 to 71 years. All patients were diagnosed as PLC by type-B ultrasonic, CT or MRI, and APF.

RESULTSFifty-six patients were completed laparoscopically successfully. Five cases underwent conversion to open operation because of hemorrhage. The mean operative time was 60 min (30-150 min). The mean blood loss was 450 ml (100-2000 ml). The mean hepatic portal block time was 20 min (15-30 min). All the patients had excellent recovery without any postoperative surgical complications. The patients were mobilized out of the bed in 24 hours. Oral intake of food started in 1 to 3 days. The average postoperative hospital stay was 6.6 d (5-10 d).

CONCLUSIONLaparoscopic hepatectomy for PLC is safe and feasible by using hepatic portal block instrument.

Adolescent ; Adult ; Aged ; Feasibility Studies ; Female ; Hepatectomy ; methods ; Humans ; Laparoscopy ; Liver Neoplasms ; surgery ; Male ; Middle Aged ; Retrospective Studies ; Treatment Outcome

Objective To elucidate the genetic diversifications of avian influenza subtype H5N1 viruses in the boundary regions of Yunnan province during 2009 to July,2011.Methods Swab samples were collected from foreign poultry and wild birds in boundary regions of Yunnan province during 2009 to July,2011 and tested by H5/N1 subtype-specific multiplex RT-PCR.The HA genes of H5N1 virus from the positive samples were amplified by RT-PCR and cloned into pMD 18-T vectors for sequencing.Both alignment and phylogenetic analysis were performed with sequences of the known reference strains.Results Fifteen different HA sequences were obtained from 36 representative positive samples and could be divided into 2 distinct Clades (2.3.2 and 2.3.4).Through phylogenetic analysis,Clade 2.3.2 and 2.3.4 could then be further divided into 3 ( Ⅱ -1 to Ⅱ -3 ) and 2 smaller clades ( Ⅰ -1 and Ⅰ -2),respectively.The viruses of Clade 2.3.2 Ⅱ -1 and Ⅱ-2 were new variant strains of H5N 1 virus.The cleavage sites of HA from positive samples all possessed molecular characterization of highly pathogenic avian influenza virus.Mutation of key amino acids had been found among receptor binding sites,potential glycosylation sites,neutralizing epitopes and others.Conclusion It seemed evident that the H5N1 subtype viruses showed genetic diversifications and had undergone the evolution progress of multi-clade (2.3.2,2.3.4) to single caide (2.3.2) in the boundary regions of Yunnan province,during 2009 to July,2011.

Objective To evaluate the clinical effects of multiple rewarming interventions in adult hypothermia trauma patients.Methods A systematic search of Cochrane Library,PubMed,EMBASE,Scopus,CINAHL,Chinese Biomedical Literature Database (CBM),Chinese Knowledge Infrastructure (CNKI),VIP and Wan Fang Database was carried out to identify all randomized controlled trials(RCTs) and controlled clinical trials(CCTs) that explored the effects of rewarming interventions in adult hypothermia trauma patients.The quality of the literature was evaluated using JBI 2008 RCT and quasi-experimental study evaluation criteria.Data and network plot were analyzed and drawn by ADDIS 1.16.7 software.Results Totally 6 RCTs and 1 quasi-experimental design were included,involving 10 interventions and 619 patients.There was statistically significant difference in body temperature after rewarming between the warm blankets and the forced-air blankets in all rewarming measures.The results of the top three interventions were carbon-fiber heating blanket(set to 42℃),forced-air blankets,warmed intravenous fluids plus blanket which resulted from the primary outcome indicators.The incidence of chills and cold discomfort decreased with the use of forced-air blankets and chemical heat pad as compared with traditional warm blankets,while the heart rate of the patients who used chemical heating pads and continuous heating of carbon fiber blanket were declined more than those used normal blankets.Conclusion The effects of carbon-fiber heating blanket which set to 42°C was the best method in all rewarming interventions.But this conclusion still requires randomized controlled trials with larger sample size to further verify.

Objective:To obtain the information of alkaloids in Evodia rutaecarpa by HPLC-Q-TOF-MS/MS. Method:Inter Sustain-C₁₈ column (4.6 mm×250 mm,5 μm) was used with 0.2% formic acid water-acetonitrile as the mobile phase for gradient elution. The column temperature was 25℃,the volume flow rate was 1.0 mL·min^-1,and the sample volume is 5 μL. The detection wavelength was 245 nm,and the chromatographic effluent was detected and analyzed by using both positive and negative ions. Result:According to molecular ion peaks and secondary mass spectrometry characteristic fragment ions,as well as the mass spectrometry information of reference substances and relevant literature reports,more than 40 major peaks were analyzed,and 21 alkaloids were identified from the methanol extract of E. rutaecarpa, including 10 kinds of indole alkaloids,10 kinds of quinolone alkaloids,and 1 kind of ephedrine. Main types of alkaloids in E. rutaecarpa were basically clarified. And the research found that the alkaloids have a good response mainly in the positive mode. Conclusion:Based on HPLC-Q-TOF-MS/MS technology, high-performance liquid chromatography (HPLC) separation,mass spectrometry determination of molecular mass,pyrolysis data,literature analysis and retrieval were performed to quickly,accurately and comprehensively identify alkaloids in E. rutaecarpa, so as to provide a scientific basis for the further extraction and separation of the chemical constituents of E. rutaecarpa.

OBJECTIVE: This study demonstrated that dexamethasone (DEX) protects the endothelial glycocalyx from damage induced by the inflammatory stimulus tumor necrosis factor-α (TNF-α) during severe acute pancreatitis (SAP), and improves the renal microcirculation. METHODS: Ninety mice were evenly divided into 3 groups (Sham, SAP, and SAP+DEX). The SAP mice model was established by ligature of pancreatic duct and intraperitoneal injection of cerulein. Renal perfusion and function, and morphological changes of the glycocalyx were evaluated by laser Doppler velocimetry, electron microscopy, and histopathology (hematoxylin and eosin (H&E) staining), respectively. Serum levels of syndecan-1 and TNF-α were assessed by enzyme-linked immunosorbent assay (ELISA). The protective effects of dexamethasone on the glycocalyx and renal microcirculation were evaluated. RESULTS: Significantly high levels of serum TNF-α were detected 3 h after the onset of SAP. These levels might induce degradation of the glycocalyx and kidney hypoperfusion, resulting in kidney microcirculation dysfunction. The application of dexamethasone reduced the degradation of the glycocalyx and improved perfusion of kidney. CONCLUSIONS Dexamethasone protects the endothelial glycocalyx from inflammatory degradation possibly initiated by TNF-α during SAP. This is might be a significant discovery that helps to prevent tissue edema and hypoperfusion in the future.
Acute Disease ; Animals ; Dexamethasone/pharmacology* ; Disease Models, Animal ; Edema/metabolism* ; Endothelium, Vascular/metabolism* ; Enzyme-Linked Immunosorbent Assay ; Glycocalyx/drug effects* ; Kidney/drug effects* ; Male ; Mice ; Mice, Inbred C57BL ; Microcirculation ; Pancreatitis/drug therapy* ; Perfusion ; Protective Agents/pharmacology* ; Tumor Necrosis Factor-alpha/metabolism*