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Objective:To analyze the prevalence and genotypes of human pegivirus-1 (HPgV-1) among HIV-1-infected men who have sex with men (MSM) in Zhuhai, aiming to elucidate the impact of HPgV-1 on the progression of AIDS.Methods:This study collected 934 serum specimens positive for antibodies against HIV-1 for viral RNA extraction from MSM in Zhuhai from 2012 to 2020. HPgV-1 5′UTR was amplified by nested PCR and then E gene was amplified by nested PCR and sequenced in the 5′UTR-positive specimens. A phylogenetic tree was constructed to analyze genotype distribution. The influence of HPgV-1 infection on the progression of AIDS was evaluated through analyzing HIV-1 viral load and CD4 + cell counts in patients in the early stage of AIDS before antiviral treatment. Results:The positive rate of HPgV-1 in MSM with HIV-1 infection in Zhuhai was 31.05%. A total of 273 valid sequences were obtained after amplification. The main genotype of HPgV-1 was G3 (252, 92.31%), which was highly homologous to the epidemic strains in China and Japan in recent years, followed G2 (21, 7.69%), which was highly homologous to the epidemic strains in France and America. HPgV-1 strains of G1, G4, G5, G6 and G7 genotypes were not detected. There was no significant difference in HIV-1 virus load or CD4 + cell counts between patients with HIV-1 infection alone and those with HIV-1 and HPgV-1 (G3 or G2 genotype) co-infection. Conclusions:According to the data of this study, HPgV-1 infection could not delay the progression of AIDS in MSM in the early stage of AIDS before antiviral therapy. The widespread HPgV-1 of G3 genotype in China did not have a significant impact on the progression of AIDS. Therefore, a systematic in-depth research on various genotypes of HPgV-1 and further study on the pathogenic mechanism of HPgV-1, especially in patients with HPgV-1 and HIV co-infection, were needed to understanding the interaction mechanism between different genotypes of HPgV-1 and HIV-1.
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Objective@#To describe the molecular epidemiological characteristics of norovirus gastroenteritis outbreaks in Zhuhai from 2011 to 2016.@*Methods@#Anal swab specimens were collected from 576 cases with 56 outbreaks of acute norovirus gastroenteritis from 2011 to 2016. Specimens were tested by real-time RT-PCR. Three to four of norovirus positive specimens were selected from every outbreak to amplify the VP1 gene by RT-PCR and one strain was chosen randomly from every outbreaks to determine the genotype by phylogenetic tree analysis.@*Results@#Eight genotypes were identified from 56 outbreaks and all of them belonged to GⅡ genogroup. The genotype of norovirus strain changed with prevalence time. The GⅡ.4/2006b was dominant from 2011 to 2012, and replaced by GⅡ.4/Sydney _2012 during the 2012—2013 norovirus season, and both of them never appeared after Feb. 2013. GⅡ.17 was the only genotype during the 2014—2015 norovirus season. All the 7 outbreaks occurred from 2015 to 2016 were caused by GⅡ.3 norovirus. The GⅡ.17and GⅡ.3 were identified from Apr. to Sep. 2016; GⅡ.p16-GⅡ.2 were the only genotype in 12 outbreaks from Nov. to Dec. 2016. The GⅠ genogrope was never identified from 2011 to 2016 in Zhuhai.@*Conclusions@#The Norovirus GⅡ was the only pathogeny which caused the outbreaks of norovirus gastroenteritis. The recombinant norovirus strain GⅡ.p16-GⅡ.2 emerged and caused large outbreaks in the last two months of 2016 in Zhuhai; several recombinant strains of the GⅡ.p16 RdRp gene were found now, which suggests that attention should be focused on the prevalence and evolution of the recombinant norovirus.
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Objective To understand the epidemiological characteristic of bacterial food-poisoning in Zhuhai City to provide a scientific basis for the judgment ,control and prevention of bacterial food-poisoning .Methods The samples of bacterial food-poison-ing in the Zhuhai Municipal Bacterial Food-Poisoning Laboratory during 2011-2015 were detected according to two version of Mi-crobiological Examination of Food (implementation on 2004-01-01 and 2016-06-01 respectively ) ,and the detection results were sta-tistically analyzed .Results Thirty-seven cases of bacterial food-poisoning(430 samples) occurred in Zhuhai City in the recent five years ,among which 22 cases(124 strains of pathogenic bacteria) were detected ,the cause detection rate of bacterial food-poisoning events was 59 .46% .The pathogenic bacteria causing bacterial food-poisoning were mainly vibrio parahaemolyticus (97 strains , 78 .23% ) and Staphyloccocus aureus (12 strains ,9 .68% ) .The sample detection rate of anal swabs was highest (77 strains ,62 . 10% ) ,followed by feces samples(17 strains ,13 .71% ) .The seasonality distribution was obviously concentrated in the third quarter (99 strains of pathogenic bacteria ,79 .84% ) and second quarter o (16 strains ,12 .90% ) .Conclusion Vibrio parahaemolyticus and Staphyloccocus aureus were the main pathogenic bacteria causing bacterial food-poisoning in Zhuhai City during these recent five years ,and the seasonality distribution was mainly concentrated in the second and third quarter .It is important to improve health awareness of the whole people and strengthen the surveillance and supervision and management work of food-borne pathogenic bac-teria during the food production ,processing and storage process in order to reduce the occurrence of food-poisoning .
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Objective:To analyze the level diversifies of plasma CCL19 and CCL21 in the male drug users infected HCV.Methods: The plasma CCL19 and CCL21,anti-HCV and HCV RNA were detected by ELISA quantitation,ELISA qualitation and Real-time RT-PCR respectively.Compared with 60 healthy man,the level diversifies of plasma CCL19 and CCL21 in 391 male drug users conducted as part of HCV Surveillance Programme in Zhuhai were analyzed.Results: 180 of 391 male drug users were infected HCV and the infection rate was 46.04%.The level of plasma CCL19 and CCL21 in the male drug users[anti-HCV(-)/HCV RNA(-)] were higher than that in the healthy man (P≤0.001).The level of plasma CCL19 and CCL21 in anti-HCV(-)/HCV RNA(+) group was lower than that in the others(P≤0.05) .Compared with that in the healthy man,the level of plasma CCL19 and CCL21 in the drug abuse anti-HCV(+)/HCV RNA(-) group had significant deviation(P<0.05).Conclusion: Drug abuse can heighten the level of plasma CCL19 and CCL21 in man.Increasing the level of plasma CCL19 and CCL21 may conduce to spontaneous HCV clearance.It may prognosticate that HCV infection will be persistent and have a bad consequence when the level of plasma CCL19 and CCL21 were degraded.
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In order to determine whether H1 histone proteins are associated with innate immune antimicrobial response in goldfish, we extracted the total RNA from the hemocytes of goldfish (Carassius auratus), designed 3 pairs of primers based on the previous antimicrobial peptide sequences from fish and performed RT-PCR. Among 3 obtained-PCR products, we identified a novel histone H1 coding sequence of 576 bp which belongs to the histone H1 family and is 78% homologous with the amino acid sequence of histone H1 from Salmon salar that had been found with an important role in salmon defenses against infectious pathogens. The H1 histone of goldfish contained 3 predicting cleavage sites that divided the protein into 4 parts. We successfully cloned and expressed the whole CDs (No ATG) of H1 histone and its N-terminal part (2-38aa) in Pichia pPIC9K expression system. The products of H1 histone and its N-terminal deriving peptide (AEVAPAASAPPAKAPKKKSAAKAKKAGPAVGDLIVKA) show antimicrobial activity. The results suggested that the H1 histone fragment reported in this paper is a novel antimicrobial peptide found in goldfish. H1 histone plays an important role in innate immune responses of goldfish.
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Animals , Amino Acid Sequence , Anti-Bacterial Agents , Pharmacology , Cloning, Molecular , Goldfish , Genetics , Metabolism , Histones , Genetics , Pharmacology , Molecular Sequence Data , Peptide Fragments , GeneticsABSTRACT
Objective] To amplify,clone and express of a gene encoding hexose transporter of Plasmodium falcipuram(PfHT1) from Southern China isolate FCC1/HN for studing the immune of recombinant which protective from malaria parasite infection. [Methods] Cultivation of P.falciparum isolate FCC1/HN in vitro; extraction of genomic DNA from FCC1/HN using the alkali specific cleavage method; PCR amplification of PfHT1 and cloning into eukaryotic expression vector, pEGFPN3. The recombinant was introduced into mammalian cells, HEPG2 by using liposome\|mediated transfection. [Results] The gene encoding PfHT1 was specifically amplified from the genomic DNA of P.falciparum isolate FCC1/HN. The size of amplified fragment was 1 516 base pair. The eukaryotic expression recombinant, pN3\|HT1 , was constructed and expressed steadily in the hepatocarcinoma cell lines, HEPG2. [Conclusion] The gene encoding PfHT1 was successfully amplified and cloned. The pN3\|HT1/HEPG2 cell line was built for expressing fusion protein of GFP\|HT1.