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Objective:To establish the biology reference interval (RI) of peripheral blood procalcitonin (PCT) for children between 3 days and 6 years old in China.Methods:Totally 3 353 reference individuals with apparent health or no specific diseases were recruited in 18 hospitals throughout the country during October 2020 to May 2021. Reference individuals were divided into four groups: 3-28 days, 29 days - 1 year, 1-3 years and 4-6 years. Vein blood or capillary blood were collected by percutaneous puncture from every reference individual. The PCT level in serum and the capillary whole blood were assayed by Roche Cobas e601 and Norman NRM411-S7 immunoanalyzer. Outliers were deleted and 95th percentiles of every group were provided as RIs. Man-Whitney U test or Kruskal-Wallis test were used performed to assess the difference among different gender, age or method groups. Results:The difference of PCT distribution between male and female is not statistically significant, but the difference between serum and capillary whole blood is statistically significant. The differences between age groups are significant too. For Roche e601, serum PCT RI of 3-28 days group is <0.23 μg/L, 29 days - 6 years are <0.11 μg/L. For NRM411, Serum PCT RI of 3-28 days group is <0.21 μg/L, 29 days - 1 year: <0.09 μg/L, 1 - 6 years: <0.10 μg/L. For whole blood PCT, RI of 3-28 days group is <0.26 μg/L, 29 days - 6 years is <0.15 μg/L.Conclusions:Serum and capillary whole blood PCT have different RIs, however, capillary whole blood PCT testing is valuable in pediatric application. Children in 3-28 days show higher PCT levels than other age group. To establish the RIs and understand the differences among different groups are essential for the interpretation and clinical application of peripheral blood PCT testing results.
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Objective@#To analyze the infection status and recombination of Norovirus in patients with acute gastroenteritis in Ningxia.@*Methods@#The specimens of 10 sentinel hospitals in Ningxia were collected from 2016 to 2017. Real-time quantitative PCR was used for nucleic acid detection. GⅡ-positive samples were amplified by RT-PCR for the RdRp and Capsid regions, then sequenced and genotyped. Evolution analysis was performed using software such as MEGA-X, and recombination analysis was performed using Simplot 3.5.1 and RDP4.@*Results@#The age of the 2 334 cases was 1.42 (0.68, 7.69) years old, 1 133 cases in 2016 and 1 201 cases in 2017, 1 343 and 991 cases for males and females respectively. The positive rate of Norovirus GⅠ genogroup was 0.86% (20/2 334), and GⅡ genogroup was 14.82% (346/2 334). A total of 78 recombinant strains were sequenced and 12 recombinant types were found. GⅡ.Pe/GⅡ.4Sydney_2012 and GⅡ.P12/GⅡ.3 were the main epidemic strains, accounting for 35.90% (28 strains) and 32.05% (25 strain) respectively, followed by GⅡ.P16/GⅡ.2 accounting for 12.82% (10 strains). Among them,GⅡ.P7/GⅡ.6 (2 strains), GⅡ.P12/GⅡ.3 (6 strains), GⅡ.P16/GⅡ.1 (2 strains), GⅡ.P16/GⅡ.2 (5 strains), GⅡ.Pe/GⅡ.4 (7 strains) were detected for the first time in Ningxia. Recombinant strains were all intergenotype recombination, and the recombination breakpionts were all located within ORF1.@*Conclusion@#Norovirus infection in Ningxia area was mainly in GⅡ genogroup from 2016 to 2017, and most of them were recombinant strains. GⅡ.Pe/GⅡ.4Sydney_2012 and GⅡ.P12/GⅡ.3 were the main epidemic strains, followed by GⅡ.P16/GⅡ. 2.
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Objective@#To understand the pathogenic composition of hand, foot and mouth disease (HFMD) in Ningxia Hui Autonomous Region (Ningxia) from 2016 to 2017, and analyze the genetic characteristics of the main pathogens enterovirus (EV)-A71 and coxsackievirus (CV)-A16.@*Methods@#Analysis of the result of nucleic acid testing of HFMD in Ningxia from 2016 to 2017 to determine the pathogenic composition of HFMD. The complete VP1 coding region was amplified by RT-PCR and the gene sequence was determined for the enterovirus strains sent to the National HFMD Network Monitoring Laboratory in Ningxia from 2016 to 2017. BLAST analysis confirmed the serotype of the strain, and the phylogenetic tree was constructed respectively by selecting EV-A71 and CV-A16 isolates.@*Results@#The leading pathogens of HFMD in Ningxia of 2016 and 2017 were other EV (397, 43.72%) and EV-A71 (918, 56.18%) respectively, and the dominant pathogens in different months may differ. The pathogenic composition causing HFMD in the past two years has changed from CV-A16 and other EV to EV-A71 and other EV. The isolated EV-A71 strains were C4a evolutionary branch and the isolated CV-A16 strains were B1b evolutionary branch.@*Conclusions@#Compared to 2016, in 2017 EV-A71, CV-A16 and other EV changed dynamically. Dynamic monitoring of EV-A71 in Ningxia is of great significance to guide the strategy of using EV-A71 vaccine, concentrating medical resources to strengthen the treatment and reduce the mortality rate of severe HFMD cases.
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Background: Disruption of tight junctions between intestinal epithelial cells followed by loss of barrier function is crucial for the pathogenesis and progression of a variety of gastrointestinal disorders.Aims: To investigate the protective effect of ulinastatin on hydrogen peroxide (H2O2)-induced intestinal epithelial barrier disruption.Methods: Model of intestinal epithelial monolayer barrier was established with Caco-2 cells in vitro,and then divided into four groups: blank control group (without any intervention),H2O2 group (500 μmol/L H2O2),low-dose (500 U/mL) and high-dose (3 000 U/mL) ulinastatin groups (ulinastatin + H2O2).Level of malondialdehyde (MDA) and activity of superoxide dismutase (SOD) were detected;transepithelial electrical resistance (TEER) and flux of sodium fluorescein were measured to assess the barrier function;expression and localization of two tight junction proteins,ZO-1 and occludin were evaluated by Western blotting and immunofluorescence;ultrastructure of tight junctions was observed by transmission electron microscopy (TEM).Results: Compared with the blank control group,treatment of Caco-2 cell monolayers with H2O2 resulted in increase in level of MDA,flux of sodium fluorescein and decrease in activity of SOD,TEER and expressions of ZO-1 and occludin (P all ＜0.05).TEM and immunofluorescence showed that the brusher border of Caco-2 cells in H2O2 group was destroyed,the cell-cell junction was vague and the localization of ZO-1 and occludin was discontinuous and the fluorescence intensity was extremely low.While in ulinastatin groups,especially the high-dose group,all the indices above-mentioned were significantly improved (P all ＜0.05).Conclusions: Ulinastatin protects intestinal epithelial monolayer barrier against H2O2-induced disruption at least partially by its antioxidant activity and modulating expression and localization of tight junction proteins.
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Objective To study the genetic characteristics of VP1 region of coxsackievirus A10(Cox A10) strains isolated from hand foot and mouth disease (HFMD) cases in Ningxia Hui Autonomous Region(Ningxia)in 2013. Methods A total of 280 specimens,which were identified as non-enterovirus 71 and non-Cox A16 by real-time PCR,were collected and cultured by using RD cell,and the VP1 genes of isolated strains were amplified by using reverse transcriptase PCR (RT-PCR) with degenerated primers and sequenced. The sequencing results were aligned with the sequences in GenBank with BLAST algorithm to identify the virus genotypes. Homologous comparison and phylogenetic analysis were conducted for all the Cox A10 strains identified. Results Among 36 virus strains isolated from 280 clinical specimens,6 were identified as Cox A10. The homologies of nucleotide and amino acid of the Cox A10 strains isolated in Ningxia were 97.0%-99.8% and 99.0%-99.7% respectively,and the Cox A10 strains isolated in Ningxia shared 76.3%-77.2%,81.6%-83.1%,94.4%-98.9% and 80.0%- 82.3% nucleotide homologies respectively and shared 92.3%-93.0%,94.0%-95.3%,98.0%-99.7% and 90.6%-94.0% amino acid homologies respectively with the representative strains of A,B,C and D genotypes. Phylogenetic tree analysis revealed that Cox A10 strains isolated in Ningxia belonged to genotype C. Conclusion Cox A10 is one of the most common pathogen causing HFMD in Ningxia in 2013. All the Cox A10 stains isolated from HFMD patients in Ningxia belonged to genotype C.
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Objective To study the genetic characteristics of VP1 region of coxsackievirus A10(Cox A10) strains isolated from hand foot and mouth disease (HFMD) cases in Ningxia Hui Autonomous Region(Ningxia)in 2013. Methods A total of 280 specimens,which were identified as non-enterovirus 71 and non-Cox A16 by real-time PCR,were collected and cultured by using RD cell,and the VP1 genes of isolated strains were amplified by using reverse transcriptase PCR (RT-PCR) with degenerated primers and sequenced. The sequencing results were aligned with the sequences in GenBank with BLAST algorithm to identify the virus genotypes. Homologous comparison and phylogenetic analysis were conducted for all the Cox A10 strains identified. Results Among 36 virus strains isolated from 280 clinical specimens,6 were identified as Cox A10. The homologies of nucleotide and amino acid of the Cox A10 strains isolated in Ningxia were 97.0%-99.8% and 99.0%-99.7% respectively,and the Cox A10 strains isolated in Ningxia shared 76.3%-77.2%,81.6%-83.1%,94.4%-98.9% and 80.0%- 82.3% nucleotide homologies respectively and shared 92.3%-93.0%,94.0%-95.3%,98.0%-99.7% and 90.6%-94.0% amino acid homologies respectively with the representative strains of A,B,C and D genotypes. Phylogenetic tree analysis revealed that Cox A10 strains isolated in Ningxia belonged to genotype C. Conclusion Cox A10 is one of the most common pathogen causing HFMD in Ningxia in 2013. All the Cox A10 stains isolated from HFMD patients in Ningxia belonged to genotype C.
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<p><b>OBJECTIVE</b>To study the genetic characteristics of VP1 region of coxsackievirus A10 (Cox A10) strains isolated from hand foot and mouth disease (HFMD) cases in Ningxia Hui Autonomous Region (Ningxia) in 2013.</p><p><b>METHODS</b>A total of 280 specimens, which were identified as non-enterovirus 71 and non-Cox A16 by real-time PCR, were collected and cultured by using RD cell, and the VP1 genes of isolated strains were amplified by using reverse transcriptase PCR (RT-PCR) with degenerated primers and sequenced. The sequencing results were aligned with the sequences in GenBank with BLAST algorithm to identify the virus genotypes. Homologous comparison and phylogenetic analysis were conducted for all the Cox A10 strains identified.</p><p><b>RESULTS</b>Among 36 virus strains isolated from 280 clinical specimens, 6 were identified as Cox A10. The homologies of nucleotide and amino acid of the Cox A10 strains isolated in Ningxia were 97.0%-99.8% and 99.0%-99.7% respectively, and the Cox A10 strains isolated in Ningxia shared 76.3%-77.2%, 81.6%-83.1%, 94.4%-98.9% and 80.0%-82.3% nucleotide homologies respectively and shared 92.3%-93.0%, 94.0%-95.3%, 98.0%-99.7% and 90.6%-94.0% amino acid homologies respectively with the representative strains of A, B, C and D genotypes. Phylogenetic tree analysis revealed that Cox A10 strains isolated in Ningxia belonged to genotype C.</p><p><b>CONCLUSION</b>Cox A10 is one of the most common pathogen causing HFMD in Ningxia in 2013. All the Cox A10 stains isolated from HFMD patients in Ningxia belonged to genotype C.</p>
Subject(s)
Humans , Algorithms , Amino Acids , China , Databases, Nucleic Acid , Enterovirus A, Human , Genetics , Genotype , Hand, Foot and Mouth Disease , Virology , Phylogeny , Real-Time Polymerase Chain ReactionABSTRACT
<p><b>OBJECTIVE</b>To investigate the genetic characteristics of coxsackievirus A10(CV-A10) strains isolated from hand, foot and mouth disease (HFMD) cases in Ningxia province.</p><p><b>METHODS</b>Based on the HFMD laboratory network surveillance system, 2 470 patients clinical specimens including 450 faeces and 2 020 throat swaps were collected from various regions people's hospital in Ningxia Hui Autonomous Region during January, 2013 to December, 2014. All specimens were isolated using rhabdomyosarcoma cells. VP1 regional gene of isolated strains was amplified by RT-PCR using degenerate primers and sequenced. Sequences were compared with the database of GenBank by the Blast algorithm to identify the enterovirus genotypes. All the CV-A10 strains were performed the homology and phylogenetic evolution analysis.</p><p><b>RESULTS</b>450 specimens identified as non-EV-A71, non-CV-A16 enterovirus were collected and 36 CV-A10 strains were isolated, 6 strains were isolated in 2013 and 30 strains were isolated in 2014. The homology of nucleotides and amino acids among 36 CV-A10 strains were 90.6%-100.0% , and 90.2%-100.0%, respectively. Compared 36 strains with genotype A, B, C, D representative strains, it has the highest homology with the genotype C, the nucleotide and amino acids homogeneity were 90.2%-98.9% and 95.7%-99.7%. The phylogenetic tree showed 36 strains and genotype C representative strains located in the same evolutionary branch.</p><p><b>CONCLUSION</b>CV-A10 was one of the most common pathogen of HFMD in Ningxia Hui Autonomous Region. All CV-A10 strains belonged to genotype C and contained wide homology range.</p>
Subject(s)
Humans , China , Enterovirus , Genetics , Genotype , Hand, Foot and Mouth Disease , Virology , Phylogeny , Sequence Homology, Amino Acid , Sequence Homology, Nucleic AcidABSTRACT
Objective To investigate the etiological characteristics of human rotavirus (HRV),human calicivirus (HuCV),human astrovirus (HAstV) and human enteral adenovirus (HAdV)in Ningxia province during 2011. Methods Stool specimen was collected from acute diarrhea case of Ningxia during 2011. HRV was detected by ELISA and serotype/genotype identified on those RT-PCR positive specimens. HuCV,HAstV and HAdV were detected by RT-PCR. Results In this study,a total of 690 specimens were detected,with the infection rates of HRV,HuCV,HAstV and HAdV as 2.17%,21.74%,3.19%and 6.52%,respectively. Co-infections were found in 4.20%of all the samples being tested. Among 15 HRV positive cases,serotypes G1,G3 and P[4]were the most predominant strains. Conclusion Children who were under 2 years of age were the majority among patients infected by diarrhea viruses while HuCV was recognized as the main pathogen responsible for the viral diarrhea casses in Ningxia,2011.
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Objective To investigate the etiological characteristics of human rotavirus (HRV),human calicivirus (HuCV),human astrovirus (HAstV) and human enteral adenovirus (HAdV)in Ningxia province during 2011. Methods Stool specimen was collected from acute diarrhea case of Ningxia during 2011. HRV was detected by ELISA and serotype/genotype identified on those RT-PCR positive specimens. HuCV,HAstV and HAdV were detected by RT-PCR. Results In this study,a total of 690 specimens were detected,with the infection rates of HRV,HuCV,HAstV and HAdV as 2.17%,21.74%,3.19%and 6.52%,respectively. Co-infections were found in 4.20%of all the samples being tested. Among 15 HRV positive cases,serotypes G1,G3 and P[4]were the most predominant strains. Conclusion Children who were under 2 years of age were the majority among patients infected by diarrhea viruses while HuCV was recognized as the main pathogen responsible for the viral diarrhea casses in Ningxia,2011.
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<p><b>OBJECTIVE</b>To investigate the etiological characteristics of human rotavirus (HRV), human calicivirus (HuCV), human astrovirus (HAstV) and human enteral adenovirus (HAdV) in Ningxia province during 2011.</p><p><b>METHODS</b>Stool specimen was collected from acute diarrhea case of Ningxia during 2011. HRV was detected by ELISA and serotype/genotype identified on those RT-PCR positive specimens. HuCV, HAstV and HAdV were detected by RT-PCR.</p><p><b>RESULTS</b>In this study, a total of 690 specimens were detected, with the infection rates of HRV, HuCV, HAstV and HAdV as 2.17%, 21.74%, 3.19% and 6.52%, respectively. Co-infections were found in 4.20% of all the samples being tested. Among 15 HRV positive cases, serotypes G1, G3 and P[4] were the most predominant strains.</p><p><b>CONCLUSION</b>Children who were under 2 years of age were the majority among patients infected by diarrhea viruses while HuCV was recognized as the main pathogen responsible for the viral diarrhea cases in Ningxia, 2011.</p>
Subject(s)
Adolescent , Adult , Aged , Aged, 80 and over , Child , Child, Preschool , Humans , Infant , Infant, Newborn , Middle Aged , Young Adult , Adenoviruses, Human , Caliciviridae , China , Epidemiology , Diarrhea , Virology , Mamastrovirus , RotavirusABSTRACT
Objective To study the genetic characterization of enterovirus 71 (EV71) strains isolated from specimens of patients with hand-foot-mouth disease (HFMD) in Ningxia province in 2008. Methods All the stool, throat swab and vesicle samples that collected from patients with HFMD were cultured. The positive isolates were identified by reverse transcriptase PCR (RT-PCR) with specific primers of EV71. Complete VP1 gene sequences (891 nucleotides) of 29 strains (part of 93 EV71 strains) were determined and compared with A, B and C genotype reference EV71 strains while EV71 China isolates by homogeneity and phylogenetic tree analyses. Results 215 strains of EV were isolated from 439 specimens. Results from RT-PCR indicated that 93 strains belong to EV71. Phylogenetic tree analysis revealed that the selected 29 stains were clustered with reference strains of C4 subgenotype. The nucleotide identity with C4 reference strains was 91.7%-99.4%. The amino acid homogeneity was 96.6%-100.0%. Conclusion The recently identified EV71 strains in Ningxia province belonged to subgenotype C4 which resembled to most of the isolates in China.