Gene Knockdown in Human Rhinovirus 1B Using 2'-OMe-modified siRNAs Results in the Reactivation of the Interferon Response / 生物医学与环境科学（英文）

The aim of this study was to investigate the knockdown efficiency of 2'-O-methylated (2'-OMe)-modified small interfering RNAs (siRNAs) on human rhinovirus 1B (HRV1B) replication and the interferon response. Thus, 24 2'-OMe-modified siRNAs were designed to target HRV1B. The RNA levels of HRV1B, Toll-like receptor 3, melanoma differentiation-associated gene 5, retinoic acid inducible gene-I, and interferons were determined in HRV1B-infected HeLa and BEAS-2B epithelial cells transfected with 2'-OMe-modified siRNAs. The results revealed that all 2'-OMe-modified siRNAs interfered with the replication of HRV1B in a cell-specific and transfection efficiency-dependent manner. Viral activation of Toll-like receptor 3, melanoma differentiation-associated gene 5, retinoic acid inducible gene-I, and the interferon response was detected. In conclusion, the 2'-OMe-modified siRNAs used in this study could interfere with HRV1B replication, possibly leading to the reactivation of the interferon response.

Norovirus Infection and Histo-blood Group Antigens in Children Hospitalized with Diarrhea in Lulong and Chenzhou in China / 生物医学与环境科学（英文）

Norovirus (NoV) is a pathogen that commonly causes viral diarrhea in children. Studies indicate that NoV recognizes human histo-blood group antigens (HBGAs) as cell attachment factors. In order to explore the correlation between of NoV infection and HBGAs, a cross-sectional study was conducted in children less than five years old who were hospitalized with diarrhea in two areas of China between November 2014 and February 2015. Of the paired stool and saliva samples taken from 424 children, NoV was detected in 24 (6%) children, with viral genotypes GII.3 (n=5), GII.4 (n=14), GII.12 (n=1), and GII.17 (n=4). All of the individuals having NoV infection were either secretors (Lea-b+/Lex-y+) or partial secretors (Lea+b+/Lex+y+) except one GII.3 infection of a non-secretor (Lea+b-/Lex+y-). These results suggest that secretor positive is associated with NoV infection, although non-secretors are not absolutely protected from NoV infection.

Whole genome analysis of human group A rotavirus G9p8 strains in Hebei lulong region, 2009-2011 / 病毒学报

Abstract:This study aims to investigate the genetic characteristics of group A rotavirus (GARV) G9P[8] strains from infantile diarrhea samples in Hebei Lulong region from 2009 to 2011. We randomly selected five GARV G9P[8] strains in Hebei Lulong region from 2009 to 2011, amplified the 11 gene fragments of GARVs by RT-PCR, and analyz their full-genome sequences by homology and phylogenetic analysis with DNAStar and MEGA. The nucleotide homology between strains LL11131077 and LL11131083 in 2011 was significantly higher than hat etween them and the other three strains in 2009 and 2010. The G9P[8] GARVs circulating in Hebei Lulong region from 2009 to 2011 elenged to the same genotype as the prevalent G9P[8] GARVs in other parts of the world. However,the two strains in 2011, compared with those in 2009 and 2010, were located in a different sub-branch of the phylogenetic tree and had amino acid mutations at many sites.

A review of detection methods for human bocaviruses / 病毒学报

Human bocavirus (HBoV) 1-4 have been detected both in respiratory and stool samples since the first HBoV was discovered in 2005. HBoV-1 is mostly associated with respiratory infection, while HBoV 2-4 are usually associated with intestinal tract infection. A variety of signs and symptoms have been described in patients with HBoV infection, including cough, wheezing, pneumonia, and diarrhea, but the research on pathogenic mechanism of HBoV is limited because HBoV cannot be cultured in vitro due to the lack of appropriate host cells. Three-dimensional epithelial cell culture, reverse genetics, and viral metagenomics are identified as novel tools that may promote the research on pathogenic mechanism of HBoV and the discovery of new viruses. This review summaries currently available diagnostic approaches such as electron microscopy, cell culture, PCR, and immunoassay in order to provide a method reference for indepth research on HBoV.

Research progress in receptors involved in rotavirus infection / 病毒学报

Rotaviruses, which are recognized as one of the major etiological agents among infants and young children with diarrhea, consist of three concentric layers of protein capsid with the enclosed double-stranded RNA genome. Rotaviruses infect host cells mainly by identifying the specific receptors on cell surfaces and binding to them. Therefore, receptors are important factors for viruses infecting cells. So far, there have been many receptors found to be involved in rotavirus infection, including sialic acid, integrin, Toll-like receptor, and blood group antigen. This article provides an overview of receptors involved in rotavirus infection.

Etiological study of human bocavirus 1-4 in children with acute diarrhea in Lanzhou, China / 病毒学报

This study aimed to study the epidemiological and clinical characteristics of human bocavirus 1-4 (HBoV1-4) in children with acute diarrhea in Lanzhou and to investigate the association between HBoV and acute gastroenteritis. A total of 331 stool samples were collected from children aged under 5 years with acute diarrhea at the Department of Pediatrics, the First Hospital, Lanzhou University, between July 2012 and June 2013. Nested PCR was used to screen for HBoV and a general PCR was employed to screen other common diarrhea viruses. We found human bocavirus 1, 2, 3 and 4 in 26, 15, 7 and 1 cases, respectively. There was no specific seasonal distribution of HBoV, with infections occurring throughout the year. HBoV was mostly found in children aged between 7 and 12 months, with a mean age of 11.04 months (+/- 6.92 months), and 93.88% of affected children were aged under 2 years. Overall, 71.3% of mixed infections were mixed and the majority of other infections were caused by rotavirus. There was no statistical difference in the incidence of fever and vomiting associated with HBoV infection. A rare virus strain, HBoV4 (LZFB086), was identified, which showed highest levels of nucleotide sequence identity (99.0%) with a single Thai HBoV strain (JQ267789). No case of HBoV2B was found. In conclusion, HBoV1 was a major etiological pathogen of HBoV in pediatric cases in Lanzhou. HBoV4 was detected in feces for the first time in China. The rate of mixed infections was high and rotavirus was dominant. The data presented suggests that HBoV is not a major causative agent of gastroenteritis.

Evolutionary relationships of G3 GARV isolated from pigs and humans in Lulong County, Hebei Province, China / 病毒学报

This study aimed to amplify major genome segments (VP7, VP4, VP6, VP2 and NSP2-5) of porcine G3 group A rotavirus (GARV) LLZ212 isolated in our laboratory, determine their genotypes, and explore the evolutionary relationships between G3 GARV strains isolated from humans and pigs in Lulong County, Hebei Province, China. Major genome segments of seven GARV strains were amplified by reverse transcription-polymerase chain reaction (RT-PCR) and the segments were sequenced. The genome segments of seven GARV strains were determined by the online RotaC genotyping tool (RotaC v2.0). The reference sequences of each GARV genome segment were downloaded from GenBank. Homology and phylogenetic evolutionary analyses were conducted using the MEGA 5.0 and DNAStar software packages. LLZ212 isolated from pigs in Lulong had the following genotype: G3-P[8]-I5-C1-N1-T1-E1-H1. All human GARV strains had the following genotype: G3-P[8]-I1-C1-N1-T1-E1-H1. The VP7, VP4, NSP4 and NSP5 genes of the LLZ212 strain had the highest nucleotide identities with the human GARV E885, CMH014/07, Wa and RMC321 strains, respectively, and these clustered together in a sublineage. The VP6, NSP4 and NSP5 genes of the LLZ212 strain shared the highest nucleotide identities with the porcine GARV PRG921 strain, while VP2 associated most closely with porcine GARV OSU strain, and these also clustered in a sublineage. A rare porcine G3-P[8]-I5-C1-N1-T1-E1-H1 GARV strain was identified, which may represent a reassortment between porcine and human viruses. In conclusion, the VP7, VP4, NSP4 and NSP5 genes of LLZ212 share high levels of sequence identity with human GARV, while VP2, VP6, NSP2 and NSP3 cluster with porcine GARV.

The epidemiological characteristics of group C rotavirus in Lulong area and the analysis of diversity of VP6 gene / 中华实验和临床病毒学杂志

<p><b>OBJECTIVE</b>To study the epdimiology characteristics and the diversity of VP6 gene of GCRV in Lulong, and to provide the basis for GCRV in-depth research.</p><p><b>METHODS</b>793 stool specimens from porcine with diarrhea or not from Lulong in 2007 and 2008. GCRV was detected by nested multiple reverse transcription- polymerase chain reaction (nested RT-PCR) , and analyzed the identity and conducted phylogenetic tree by the seqences.</p><p><b>RESULTS</b>The positive rate of GCRV was 16.65%. Porcine GCRV strains of Lulong had significant homology differences. Phylogenetic analysis indicated porcine GCRVs were with significant diversity. Amino acid analysis showed GCRV strains with the same host shared the nearest kinship.</p><p><b>CONCLUSION</b>The infection rate of GCRV was high from 2007 to 2008 in Lulong. Homology and phylogenetic analysis showed that VP6 gene diversity was widespread. The experimental data provided basis for molecular characteristics of porcine GCRVs.</p>

Research progress of real-time quantitative PCR method for group A rotavirus detection / 病毒学报

Group A rotavirus is one of the most significant etiological agents which causes acute gastroenteritis among infants and young children worldwide. So far, several method which includes electron microscopy (EM), enzyme immunoassay (EIA), reverse transcription-polymerase chain reaction (RT-PCR)and Real-time Quantitative PCR has been established for the detection of rotavirus. Compared with other methods, Real-time quantitative PCR have advantages in specificity, sensitivity, genotyping and quantitative accuracy. This article shows a overview of the application of real-time quantitative PCR technique to detecte group A rotavirus.

Etiological and epidemiological study on viral diarrhea among children in Changchun / 中华实验和临床病毒学杂志

<p><b>OBJECTIVE</b>To study the pathogen and characteristics of viral diarrhea in children in Changchun area.</p><p><b>METHODS</b>460 stools specimens were collected from children with acute diarrhea cured in the childrens, hospital of Changchun in 2010. Rotavirus were detected by ELISA, caliceverus and astrovirus were detected by reverse transcription-polymerase chain reactions (RT-PCR), adenovirus were detected by polymerase chain reactions (PCR).</p><p><b>RESULTS</b>A total of 460 specimens were detected. The detection rate of rotavirus, caliceverus, astrovious, adenovious respectively is 35.22%, 20.43%, 9.78%, 3.70%, the detectablerate of mixed infection is 7.61%, children under 2 years old were the major patient. The main genotypes of the virus: rotavirus (G3P[8]), caliceverus (GII-4), astrovious (type I), adenovious (Ad41).</p><p><b>CONCLUSION</b>Rotavirus is the main pathogen in Changchun. Followed by caliceverus, astrovious, adenovious.</p>

Molecular epideiological and clinical feature of human calicivirus and adenovirus among children with diarrhea less than 5 years old from 2010 to 2011 in Lanzhou, Gansu province / 中华实验和临床病毒学杂志

<p><b>OBJECTIVE</b>To investigate the clinical and molecular epidemiology characteristics of calicivirus and adenovirus in children for viral diarrhea in Lanzhou.</p><p><b>METHODS</b>Stool samples were collected from 295 children with diarrhea at the First Hospital of Lanzhou University, Gansu Province,China, between July 2010 and June 2011. Reverse transcription-polymerase chain reaction (RT-PCR) or PCR were used to detected calicivirus and adenovirus. The adenovirus positive samples were typed by nested PCR and multiple PCR.</p><p><b>RESULTS</b>Of the 295 specimens, 13.2% (39/295) were positive for calicivirus, and 5.1% (15/295) were adenovirus. Typing and Phylogenetic analysis revealed that novirus GII-3 and adenovirus 41 were the dominant strains. Both calicivirus and adenovirus predominately affect children under the age of 2. In seasonal distribution, there was no obvious peak.</p><p><b>CONCLUSION</b>Human calicivirus and adenovirus are important pathogens of viral diarrhea,it is important to develop long-term systematic surveillance.</p>

Molecular and epidemiological study on among children under 5 years old in Nanjing / 中华实验和临床病毒学杂志

<p><b>OBJECTIVE</b>To study the infected information, clinical symptom and molecular epidemiological characteristics of HuCV infection among children under 5 years old in Nanjing.</p><p><b>METHODS</b>In Nanjing Children's Hospital of Nanjing Medical University from July 2010 to June 2011, we collected 428 stool specimens from children with diarrhea and 428 asymptomatic controls. Human Calicivirus were tested by using RT-PCR. Then we sequenced the nucleic acid of PCR amplifications and identified the genotype and gene group of prevalent strains.</p><p><b>RESULTS</b>63 (14.72%) out of 428 stool samples were detected as HuCV. 58 were norovirus and 5 were sapovirus, while GII-4 2006b was the predominant strain of NoV. In the 428 control samples, 19 samples were positive for calicivirus, there were 8 NoV and 13 SaV (Including 3 co-infection cases).</p><p><b>CONCLUSION</b>Human caliciviruses with different genotypes circulated among children in Nanjing,and GII. 2006b is the dominant genotype.</p>

Detection and molecular characterization of human parechovirus (HPeV) in children with acute gastroenteritis / 中华实验和临床病毒学杂志

<p><b>OBJECTIVE</b>To study HPeV from stool samples of children with acute gastroenteritis under 5 years old.</p><p><b>METHODS</b>We conducted a real-time PCR to detect HPeV from stool samples and to amply VP1 sequence by nested RT-PCR to identify HPeV type.</p><p><b>RESULTS</b>The results showed that 27 of 306 (8.82%) children with acute gastroenteritis were infected HPeV. 11 strains were typed. 9 strains HPeV1, both HPeV2 and HPeV4 was 1 strain. HPeV was mostly identified in autumn season with a peak in July. HPeV seemed relevant in children >2 years old. The range of nucleotide identity between all isolated strains with reference strains was 79%-92%.</p><p><b>CONCLUSION</b>Epidemiology characteristic of HPeV in Jilin was concordance with that of reports. HPeV3 wasnt detected. It's significant to conduct the large scale and long-term surveillance of HPeV.</p>

Analysis on epidemiologic characteristics of viral diarrhea among infants in Taiyuan, Shanxi province, 2007-2008 / 中华实验和临床病毒学杂志

<p><b>OBJECTIVE</b>To study the epidemiologic characteristics of virus-induced acute diarrhea in children under 5 years old in Taiyuan, Shanxi province.</p><p><b>METHODS</b>Stool specimens and clinical data were collected from 346 inpatients with acute diarrhea from children less than 5 years old. Rotavirus-positive specimens were identified by ELASA kit. Calicivirus and astrovirus were detected by reverse transcription-polymerase chain reaction (RT-PCR). Adenovirus was done by polymerase chain reaction (PCR).</p><p><b>RESULTS</b>Of the 346 specimens, the percentage of samples with Rotavirus, Calicivirus, Astrovirus, and Adenovirus was 40.8%, 7.5%, 6.4% and 3.2%. Among 141 rotavirus positive samples, serotype G1 (42.6%) was the predominant strain. More than 95% of viral diarrhea patients under hospitalization occurred among children younger than 2 years.</p><p><b>CONCLUSION</b>Rotavirus is the major pathogen contributing to the acute diarrhea. The disease generally peaks at autumn/winter. The predominant rotavirus strain circulated was G1P[8].</p>

Detection of Aichi virus in stool samples from children in Lanzhou / 中华实验和临床病毒学杂志

<p><b>OBJECTIVE</b>To identification and analysis Aichi virus from diarrhea and normal children in Lanzhou, and discuss the relationship between Aichi virus and Infant Diarrhea.</p><p><b>METHODS</b>According to the literature published data, Using RT-PCR method to amplified Aichi virus 3CD fragment and the positive products were sequenced and determined, and made the alignment analysis between the nucleotide sequences of the amplified fragment with the known sequence of this virus.</p><p><b>RESULTS</b>There was 1 case detection of Aichi virus in the 46 hospitalized children with diarrhea and 299 children with diarrhea out-patients specifically, Overall detection rate was 0.06%, and there was no Aichi virus was detected in normal control children. 2 viral 3CD gene and the known reference strains of nucleotide sequences were 97%, while phylogenetic analysis showed that genotype of 2 viral belongs to the B.</p><p><b>CONCLUSIONS</b>There existed B Genotype of Aichi virus in China, and more research is needed to clarified the etiology and epidemiology of Aichi virus characteristics.</p>

Sequence the complete sequence of bocavirus I with SISPA-PCR / 中华实验和临床病毒学杂志

<p><b>OBJECTIVE</b>To sequence the complete sequence of bocavirus I with sequence independent single primer amplification (SISPA-PCR).</p><p><b>METHODS</b>To exclude the co-effection samples, all clinical samples of diarrhea cases were screened with special primers of rotavirus, astrovirus, adenovirus, calicivirus and bocavirus I. The virus were enriched through ultracentrifugation. Other nucleic acids, such as human and bacteria genomes, were degradated by DNase I and RNase. DNA of bocavirus was Amplificated with SISPA-PCR, then purificated, cloned and sequenced. The sequences were alighmented in nr with blastn and assembled with DNAstar.</p><p><b>RESULTS</b>A 4834bp sequence of bocavirus I were assembled.</p><p><b>CONCLUSION</b>SISPA-PCR is an economical and efficient technique for sequence a virus complete genome.</p>

Detection of bovine rotavirus G10P11 in a diary farm in Daqing, China / 病毒学报

Group A rotavirus are the most frequently detected viral agent associated with the acute diarrhea in calves. In order to investigate the situation of rotavirus strains circulating in diary farms, a total of 117 fecal specimens were collected from diarrhea calves under 4 weeks-age on Yinluo diary farm in Daqing region in China from 2008 to 2009. Ten specimens were detected to be positive by a Rotavirus Group A Diagnostic Kit, which confirmed that the rotavirus was important viral agent associated with diarrhea in this diary farm. Based on the new classification system, G10P[11] genotype was determined in rotavirus positive samples. Sequence and Phylogenetic analysis indicated DQ-75 strain was introduced into our country with imported bovine.

Study on the epidemiological of rotavirus diarrhea in Lulong in 2008-2009 / 中华实验和临床病毒学杂志

<p><b>OBJECTIVE</b>To analyze the feature of epidemiological of rotavirus diarrhea in Lulong county, Hebei province.</p><p><b>METHODS</b>426 stool specimens were collected from inpatant with acute diarrhea from children less than 5 years old. Rotavirus-positive specimens were identified by ELISA kit. G/P typing assays were confirmed with multiplex seminested RT-PCR.</p><p><b>RESULTS</b>Rotavirus was detected in 202 of 426 (47.42%) specimens. Genotyping of rotavirus showed that G3 was predominant (57.9%), followed by Gmix (16.3%), G9 (14.9% ), G1 (7.9%), G4 (1%), G2 (0.5%), P-genotyping showed that P [8], Pmix, P [4], P [9], type were found in 58.4%, 28.7%, 6.9% and 1% respectively. The most common G/P combination identified was G3P [8].</p><p><b>CONCLUSION</b>Group A rotaviruses was a major pathogen of diarrhea in Children in Lulong. G3P [8] was the predominant type in 2009, Gmix and Pmix abound, and G9 serotypes has become the second predominant after G3 strain in the region.</p>

Pathogens of high incidence of other infectious diarrhea in Guangdong Province, from November 2008 to January 2009 / 病毒学报

From November 2008 to January 2009, a sharp increase of diarrhea in children in Guangdong province appeared, we randomly collected 53 stool specimens from out-patient children with dirrhea in 3 major hospitals (Guangzhou City Children's Hospital, Shenzhen Baoan District Maternal and Child Health Hospital, First Affiliated Hospital of Shantou University). Rotavirus and calicivirus were screened by ELISA and RT-PCR. We found 29 cases of rotavirus infection with diverse serotypes. Only four cases were identified as calicivirus infection. The result indicated that rotavirus was a major pathogen of this high incidence of diarrhea from November 2008 to January 2009 in Guangdong Province.

A method of detect the virus which caused diarrhea / 中华实验和临床病毒学杂志

<p><b>OBJECTIVE</b>Building a method which can examines virus pathogenic in gastroenteritis excrement specimen.</p><p><b>METHODS</b>Choosing six positive specimens which tested in our laboratory, include adenovirus, calicivirus, rotavirus, bocavirus, astrovirus and enterovirus. Through sequence-independent single primer amplification(SISPA) constructs a gene bank. Looks up the viral gene fragment in gene bank.</p><p><b>RESULTS</b>Obtaining corresponding viral acid sequence in six specimens.</p><p><b>CONCLUSION</b>This research can examine enterovirus and the virus which cause diarrhea, It make a foundation for further studies the viral cause of disease which the examination not yet discovered at present.</p>