OBJECTIVE: This study was aimed at investigating the carrier rate of, and molecular variation in, α- and β-globin gene mutations in Hunan Province. METHODS: We recruited 25,946 individuals attending premarital screening from 42 districts and counties in all 14 cities of Hunan Province. Hematological screening was performed, and molecular parameters were assessed. RESULTS: The overall carrier rate of thalassemia was 7.1%, including 4.83% for α-thalassemia, 2.15% for β-thalassemia, and 0.12% for both α- and β-thalassemia. The highest carrier rate of thalassemia was in Yongzhou (14.57%). The most abundant genotype of α-thalassemia and β-thalassemia was -α ^3.7/αα (50.23%) and β ^IVS-II-654/β ^N (28.23%), respectively. Four α-globin mutations [CD108 (ACC>AAC), CAP +29 (G>C), Hb Agrinio and Hb Cervantes] and six β-globin mutations [CAP +8 (C>T), IVS-II-848 (C>T), -56 (G>C), beta nt-77 (G>C), codon 20/21 (-TGGA) and Hb Knossos] had not previously been identified in China. Furthermore, this study provides the first report of the carrier rates of abnormal hemoglobin variants and α-globin triplication in Hunan Province, which were 0.49% and 1.99%, respectively. CONCLUSION Our study demonstrates the high complexity and diversity of thalassemia gene mutations in the Hunan population. The results should facilitate genetic counselling and the prevention of severe thalassemia in this region.
Humans ; beta-Thalassemia/genetics* ; alpha-Thalassemia/genetics* ; Hemoglobinopathies/genetics* ; China/epidemiology* ; High-Throughput Nucleotide Sequencing

Child ; Consensus ; Drugs, Chinese Herbal ; Humans ; Medicine, Chinese Traditional ; Respiratory System

Objective: To evaluate the impact of coronavirus disease 2019 ( COVID-19 ) prevention and control measures in Huzhou on influenza epidemic strength and characteristics in 2020, so as to provide reference for formulating influenza prevention measures. Methods: Using the influenza surveillance data of the national influenza sentinel surveillance system from January 2015 to July 2020, the seasonal characteristics of influenza epidemic were analyzed, the proportion of influenza-like illness cases ( ILI% ) and the positive rate of influenza virus in January to July of 2020 were compared with those of the same period in 2015-2019, in order to evaluate the impact of COVID-19 prevention and control measures. Results : The ILI% and the positive rate of influenza virus in Huzhou were 3.90% and 15.32% during 2015-2019, while were 4.41% and 12.63% from January to July of 2020. The trends of ILI% during 2015-2019 fluctuated similar, but continued to drop since January 2020. The positive rate of influenza virus peaked from December to March in 2015-2019, also peaked from December 2019 to January 2020, but decreased to 0 in March. ILI% was positively correlated with the positive rate of influenza virus ( r=0.682, P<0.05). The growth rates of ILI% from January to July 2020 were 4.75%, -11.27%, 0.68%, 19.84% and 0.92%, compared with the same period of 2015-2019, respectively. The growth rates of ILI% in January 2020 were much higher ( >57.00% ) and from April to July were much lower ( <-33.00% ) . The growth rates of influenza virus positive rate from January to July 2020 were -47.96%, -36.53%, -3.44%, -35.92% and -39.37%, compared to the same period of 2015-2019, respectively. The growth rates of influenza virus positive rate in January 2020 were much higher ( >11.00% ) and from February to March were much lower ( <-61.00% ). Conclusion Since COVID-19 prevention and control measures were implemented in January 2020 in Huzhou, the ILI% and the positive rate of influenza virus in sentinel hospitals decreased significantly.

Objective To develop and evaluate a single -tube multiplex RT -real time PCR assay for the simultaneous detection of rotavirus,astrovirus and hepatitis A virus.Methods Gen -bank sequences of rotavirus,astrovirus and hepatitis A virus were included as reference sequences.Primers and probes were designed based on the reference sequences.A multiplex real -time RT -PCR assay was developed,and the reproducibility,specificity and sensitivity of the assay were evaluated.Fecal samples from 1 28 patients with viral diarrhea were detected and verified by gene sequencing.Results There was high reproducibility and specificity of the single -tube multiplex real -time RT -PCR assay for detecting rotavirus,astrovirus and hepatitis A virus.Detection limits of 1 01 copies were achieved in tests of rotavirus,1 02 copies for astrovirus and hepatitis A virus in one reaction.3.1 3% and 1 7.97% of 1 28 clinical specimens were detected positive for astrovirus RNA and rotavirus RNA respectively.And the positive samples were verified by gene sequencing.Conclusion Rotavirus,astrovirus and hepatitis A virus can be detected and identified by the single -tube multiplex RT -real time PCR assay with high specificity and sensitivity.The assay developed in this study can be applied to the clinical diagnosis and epidemiological investigation.

Objective The purpose of this study was to develop RT-PCR assay for Rapidly detect and distinguish between Norovirus genogroup Ⅰ and genogroup Ⅱ with a pair of primers.Methods A pairs of primers specific to capsid prote in ORF2 gene of G Ⅰ and G Ⅱ Norovirus were dsigned according to the published complete genome sequence,with which the RNA of Norovirus was extracted and RT-PCR amplification.The sensitivity,specificity of the RT-PCR assay was estimated and apply it to the detection of Norovirus in clinical specimens.Results The results showed that the assay possessed high specificity for Norovirus detection and without any evident cross-reaction with other viruses,including rotavirus,enteric adenovirus and hepatitis A virus.The detection limit of RT-PCR assay for Norovirus G Ⅰ and G Ⅱ were up to 100 pg/ml and 10 pg/ml respectively.Conclusion The RT-PCR assay provide rapid and sensitive detection of Norovirus G Ⅰ and G Ⅱ and should prove to be useful for Norovirus diagnosis in the outbreaks of acute gastroenteritis.

To study the molecular characteristics of Noroviruses causing outbreaks of acute gastroenteritis in Huzhou. During April 2008 and February 2009, fecal specimens of patients collected from 2 outbreaks of acute gastroenteritis were tested for Norovirus by real-time RT-PCR. Partial sequence of RNA dependent RNA polymerase(RdRp) of the positive samples were amplified by RT-PCR, the PCR products were then purified, sequenced and phylogenetic analysis was conducted. Both genogroup II (GII) and genogroup I (GI) noroviruses were detected in 2 outbreaks. Phylogenetic analysis revealed that two of the GI norovirus strains isolated from 2008 belonged to genotype GI/2 and one of the GI Norovirus strain isolated from 2009 belonged to genotype GI/3. The other GIIú norovirus strains isolated from 2009 had high nucleotide identity with GIIb genotype that had been reported frequently in European countries during 2000 and 2001 and in Asian countries recently. These results suggested that the epidemic strains of norovirus isolated in Huzhou had a high degree of genetic diversity and prevalent genotypes at different times were also different. To our knowledge this is the first report of detecting GIIb variant in outbreaks of acute gastroenteritis in China.
Acute Disease ; Caliciviridae Infections ; epidemiology ; virology ; China ; epidemiology ; Disease Outbreaks ; Feces ; virology ; Gastroenteritis ; epidemiology ; virology ; Genotype ; Humans ; Norovirus ; classification ; genetics ; isolation & purification ; Phylogeny ; RNA, Viral ; genetics ; Sequence Homology, Nucleic Acid

Objective To study the molecular characteristics of Noroviruses causing outbreaks of acute gastroenteritis in Huzhou.Methods From 2008 to 2010,total 119 fecal specimens collected from outbreaks of acute gastroenteritis were tcsted for Norovirus. Partial sequence of RNA dependent RNA polymerase(RdRp) of the positive samples were amplified by RT-PCR,then the PCR production were purified,sequenced and put into phylogenetic analysis. Results 50 of 119 specimens were positive for Norovirus by real-time RT-PCR.Out of those 50 Norovirus positive specimens,9 were Norovirus Genogroup Ⅰ (GI) positive,35 were Norovirus Genogroup Ⅱ (GⅡ)positive,6 was both Norovirus GⅠ and GⅡ positive.12 PCR products for RdRp were selected for further studies on sequencing.Phylogenetic analysis revealed that the 5 GⅠ norovirus isolates were belonged to genotype GⅠ/2 and GⅠ/3.Of the 7 GⅡ norovirus isolates,6 were belonged to genotype GⅡ/4,1 was belonged to genotype GⅡb.Conclusion Norovirus is a major cause of outbreaks of acute gastroenteritis in Huzhou and the epidemic strains of norovirus isolated from Huzhou had a high degree of genetic diversity.

Objective To establish a TaqMan-based multiplex real-time PCR assay for detection of influenza viruses A and B, and the constructed method was primarily applied to clinical samples test.Methods The specific primers and probes were designed in the conserved region of the M and HA gene of influenza viruses A and B, respectively. The reaction conditions were optimized and the sensitivity, specificity and the stab ility of the assay were evaluated. The clinical specimens collected from the patients were detected by this assay. Results For specific detecting the influenza A and B viruses,the detection limits of the assay were 0.1TCID50and 0.01 TCID50. The viral RNA was directly detected from the clinical specimens by this assay. Only three hours were needed for viral RNA extraction and real-time PCR amplification.Conclusion This assay is a rapid, sensitive and specific one for simultaneous detection of influenza viruses A and B.

Objective To detect HBV-LP and HBV DNA of the patients with hepatitis B, and to study the sensitive and specificity of HBV-LP detecting and its evaluate on for clinical application. Methods The ELISA was used for HBV-LP detecting and RT-PCR for HBV DNA detecting. Results The sensitive and specificity of HBV-LP and HBV DNA were 64.89% ,99.68% ,60.63% and 100% respectively(P > 0.05);+LR, -LR were 202.78,60.63, 0.3522 and 0.3937, and there were significance between +LR of the detecting (P<0.005 ). Conclusion The sensitive and specificity of HBV-LP and HBV DNA detecting are considerable, +LRof HBV-LP are higher comparing HBV DNA. The HBV-LP is better serology index for detecting replication of HBV DNA.

OBJECTIVETo investigate the feasibility of transfection of recombinant human endothelial nitric oxide synthase (eNOS) into human hypertrophic scar fibroblasts (HSFbs), and to observe NO secretion and the synthesis of collagen I and III.

METHODSRecombinant human eNOS with karyocyte expressive vector was constructed in vitro, then was transfected into HSFbs which was isolated from hypertrophic scar tissues and cultured in vitro (T group). The HSFbs untransfected (normal culture) or transfected with empty-vector was used as control group and empty-vector group respectively. The mRNA expression of eNOS, collagen I and III was determined by Realtime PCR. The content of NO was determined by NO assay kit.

RESULTSThe expression of eNOS mRNA in T group was 5.92 +/- 0.21, which was obviously higher than that in empty-vector group (0.98 +/- 0.13, P < 0.05). The expression of collagen I mRNA (0.76 +/- 0.15), and collagen III (0.79 +/- 0.08) in T group was significantly lower than those in empty-vector group (0.98 +/- 0.15, 1.02 +/- 0.12, P < 0.05, respectively). The content of NO in T group (36.1 +/- 0.8 micromol/L) was obviously higher than that in empty-vector group (28.4 +/- 1.0 micromol/L, P < 0.01) and control group (27.7 +/- 1.3 micromol/L, P < 0.01).

CONCLUSIONHSFbs can be the target cells for eNOS gene transfection. The transfected cells can express eNOS and produce NO, which inhibit the synthesis of collagen.

Cicatrix, Hypertrophic ; metabolism ; Collagen Type I ; genetics ; metabolism ; Collagen Type III ; metabolism ; Fibroblasts ; metabolism ; Humans ; In Vitro Techniques ; Nitric Oxide ; metabolism ; Nitric Oxide Synthase Type III ; genetics ; RNA, Messenger ; metabolism ; Transfection