Nucleic acid detection and sequence analysis of arboviruses in Xichang / 中华检验医学杂志

Objective:To understand the prevalence of arboviruses in mosquito samples in Xichang City, Sichuan Province, and enrich the data of arbovirus activity and genetic characteristics in southwestern Sichuan Province.Methods:In June 2018, the nucleic acid was extracted from Culex tritaeniorhynchus mosquitoes collected from different pigsties in three villages and suburbs of Xichang City. The specific primers of Yunnan orbivirus, Banna virus, Tibet orbivirus (S7, S10), Flavivirus and alphavirus were used for quantitative polymerase chain reaction examination, and the positive product was cloned for sequencing analysis. Results:A total of 9 012 mosquitoes were collected, of which Cx. tritaeniorhynchus was the dominant species. A number of 88 batches of these mosquitoes were amplified, and 2 strains of Japanese encephalitis virus (JEV), 7 strains of Banna virus (BAV), 7 strains of Tibet orbivirus (TIBOV) and 1 strain of Yunnan orbivirus virus (YOUV) were detected, respectively. By the results of cluster analysis and evolutionary tree analysis, the 17 newly found virus strains were close to the Yunnan isolates, and 2 JEV strains were located in the GI-b clade. The other 7 strains of BAV were A2 evolutionary clades. Of the 7 TIBOV plants, 6 were located in the same clade. One TOUV was in the same clade as the Yunnan strain. Conclusions:Culex tritaeniorhynchus mosquitoes in Xichang city might carry JEV, BAV, YOUV and TIBOV, among them JEV was GI-b type and BAV was A2 type. The results provide data supporting the detection and analysis of arboviruses in Xichang city.

Epidemiological investigation and phylogenetic analysis of Classical Swine Fever virus in Yunnan province from 2015 to 2021

Background@#Classical swine fever virus (CSFV), the causative agent of classical swine fever (CFS), is a highly contagious disease that poses a serious threat to Chinese pig populations. @*Objectives@#Many provinces of China, such as Shandong, Henan, Hebei, Heilongjiang, and Liaoning provinces, have reported epidemics of CSFV, while the references to the epidemic of CSFV in Yunnan province are rare. This study examined the epidemic characteristics of the CSFV in Yunnan province. @*Methods@#In this study, 326 tissue samples were collected from different regions in Yunnan province from 2015 to 2021. A reverse transcription-polymerase chain reaction (RT-PCR), sequences analysis, and phylogenetic analysis were performed for the pathogenic detection and analysis of these 326 clinical specimens. @*Results@#Approximately 3.37% (11/326) of specimens tested positive for the CSFV by RTPCR, which is lower than that of other regions of China. Sequence analysis of the partial E2 sequences of eleven CSFV strains showed that they shared 89.0–100.0% nucleotide (nt) and 95.0–100.0% amino acid (aa) homology, respectively. Phylogenetic analysis showed that these novel isolates belonged to the subgenotypes 2.1c and 2.1d, with subgenotype 2.1c being predominant. @*Conclusions@#The CSFV was sporadic in China’s Yunnan province from 2015 to 2021. Both 2.1c and 2.1d subgenotypes were found in this region, but 2.1c was dominant.

Isolation and identification of Getah virus from Culicoides in Yunnan / 中华实验和临床病毒学杂志

Objective@#To discuss the taxonomy and molecular characteristics of one virus strain (SZC30) isolated from Culicoides in Yunnan.@*Methods@#Culicoides were collected with light trap method in the Wulong Village of Shizong County of Yunnan Province in July, 2013. BHK-21 and C6/36 cells were used for virus isolation. The positive isolates were inoculated into brain of one-day suckling mice. Alphavirus and Getah virus specific primers were used to amplify the genome of the virus isolation by RT-PCR. The products of RT-PCR were sequenced. Clustal X1.83, DNAStar, Mega5.1 were used for bioinformatics analysis.@*Results@#Totally 3 500 culicoides were collected and divided into 41 batches for virus isolation. One isolate (SZC30) produced cytopathic effect (CPE) on BHK-21 and C6/36 cells; the result of RT-PCR with Alphavirus and Getah virus NS1 specific primer were positive; the sequence analysis of NS1 gene suggested that SZC30 and two Getah virus strains (YN0540, SC1210) from China were in the same evolutionary branching, the nucleotide homology were 97%-100%, and the amino acid was 97%-100%.@*Conclusions@#SZC30 isolated in Yunnan province was identified as Getah virus.

Management strategies of ectopic pregnancy and reproductive outcomes:a report of 648 cases / 中华全科医师杂志

Objective To explore the reproductive outcomes of different management strategies of ectopic pregnancy .Methods A retrospective cohort study was performed for 648 consecutive patients with a diagnosis of ectopic pregnaney between June 2011 and september 2013.The follow-ups were conducted telephone interviews of pregnancy outcomes after various treatments .Results The subsequent intrauterine pregnancy rates after surgical , medical and expectant managements were 89(65.0%), 35(79.6%) and 6 (54.5%) the intrauterine pregnancy was higher for medical treatment than that for surgical treatment (P<0.05).The subsequent infertility rate after surgical , medical and expectant management were 4.5%, 36.4%, 19.0%.The subsequent infertility rate after medical treatment was lower than that for surgical and expectant managament ( P<0.05) .The re-ectopic pregnancy rates of laproscopic versus abdominal operation were [11 (11.7%), 13(30.2%)] with significant difference (P<0.05).And abdominal operation was higher than laproscopic operation .Conclusion For younger ectopic pregnancy patients , medical treatment shold be as possible as we can .If operation is necessary , laproscopic operation is preferred .

Construction and expression analysis of micro-linear vector as a new general gene therapy vector / 生物工程学报

The most difficult field in gene therapy is that vector system should offer both a means of successful transfection and a maximum of safety for the patient. Viral vectors and plasmid vectors are traditional vectors; they may cause unwanted immunological side effects resulting from the expression of nontherapeutic genes. Our aim is to develop a new general gene therapy vector which is suggested to be called as Micro-Linear Vector. The gene expression cassette is capped by our designed cap, including promoter, enhancer, objective gene, and RNA-stabilizing sequence, so it can defend the exnuclease in the eukaryotic cell, at the same time, DNA not encoding the objective gene is reduced to a minimum. The GFP gene is separated from the pEGFP-N3 plasmid, and acts as a reporter gene to construct the Micro-Linear Vector, then both the new vector and the plasmid are transfected to cells, the results are tested by fluorescence microscope and flow cytometry. The results show that the Micro-Linear Vector has a high effective of transfection and safety in 293, 3T3, CNE2 and B95-8 cell lines, at the same time it is less toxicity than the plasmid. We can get the rudiments of conclusion that Micro-Linear Vector has high effection of the transfection and more safety than tradition plasmid in eukaryotic cell.