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Objective:To understand the pathogen infection and epidemiological characteristics of children with diarrhea in Tianjin.Methods:Stool samples from 1 466 children with diarrhea in Tianjin Children's Hospital from August 2017 to July 2018 were collected, and all samples were tested for five intestinal-related pathogens (norovirus, rotavirus, Clostridium difficile toxin, adenovirus, and astrovirus). Results:Among the 1 466 samples, 627 samples were positive for nucleic acid detection of intestinal pathogens, with a positive rate of 42.8%. The detection rate of norovirus was the highest (26.3%), followed by rotavirus (15.3%), Clostridium difficile toxin (4.6%), adenovirus (4.1%), and astrovirus (1.84%). The infection has obvious seasonality. The positive detection rates of the five pathogens were similar among children of different sexes, and only the positive detection rates of norovirus and rotavirus were statistically significant among different ages ( P<0.05). There were 110 cases of mixed infection, and the mixed infection of norovirus and rotavirus was the most common, with a total of 37 cases. Conclusions:The pathogen spectrum of infant infectious diarrhea in Tianjin is complex and diverse, and the main pathogens are norovirus and rotavirus.
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Objective:To evaluate the prevalence of human rhinovirus (HRV) infection in hospitalized children in Tianjin and investigate the clinical impact of HRV infections.Methods:From July 2017 to December 2019, 2 945 nasopharyngeal secretion specimens were screened for HRV using polymerase chain reaction (PCR). VP4/VP2 sequences of HRV were further characterized. The clinical characteristics of the HRV infection were analyzed. The detection results of HRV for different groups and different months were compared using SPSS 19.0.Results:HRV-positive specimens accounted for 8.15% (240/2 945), of which 74.78% (86/115) were diagnosed with pneumonia, 40.83%(98/240) had co-infections with other common pathogens. HRV infections could be detected throughout the year with peaks in spring (11.00%, 66/660) and autumn (9.29%, 81/872). The positive rate of HRV was 4.14%(29/700) in winter. By VP4/VP2 sequence analysis, HRV-A was the most frequently detected strain(50.00%, 78/156), followed by HRV-C (41.67%, 65/156).46.15% (30/65) of HRV-C infections occurred in October and November. There were several different HRV-A types and HRV-C types. The most commonly detected HRV-A types were A12(11.54%, 9/78), A49(6.41%, 5/78), A22, A101, and A66(5.13%, 4/78), etc. The most common HRV-C types were C2(20.00%, 13/65), C22(9.23%, 6/65), C26, C43, C54 and C53(4.62%,3/65). Patients with HRV-A infections are more likely to show fever symptoms than HRV-C (χ2=5.411, P<0.05). No significant difference in other symptoms were found between the two types. Conclusions:HRV was a commonly detected virus among infants and had a clear seasonal distribution. It′s also possible for the HRV patients to have co-infections with other pathogens.HRV showed high genetic diversity.
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Objective:To analyze the epidemiological characteristics and molecular classification of Human adenovirus (HAdV) and Human bocavirus (HBoV) infection in hospitalized children with acute respiratory infection in Tianjin Children′s Hospital.Methods:A total of 1 171 nasopharyngeal aspirates were collected from children with acute respiratory infection in Tianjin Children′s Hospital from March 2019 to February 2020. The specific primers designed by gene sequence were amplified by polymerase chain reation (PCR), and the positive amplification products were determined by sequencing. The sequences of HAdV and HBoV were compared in GenBank, molecular typed and phylogenetic tree analyzed of HAdV by MEGA7.0.26. The positive rate of HAdV and HBoV in different age groups(<6 months, 6-11 months, 12-23 months, 24-35 months, 36-47 months, ≥48 months) and seasons were compared by SPSS20.0.Results:Thirty HAdV were detected in 1 171 specimens, with a positive rate of 2.56% (30/1 171) and 84 cases with HBoV, with a positive rate of 7.17% (84/1 171).The positive detection rates of HAdV and HBoV in different age groups were 1.02% (4/392)-6.61% (8/121) and 4.09% (7/171)-11.45% (26/227), respectively. There was a significant difference in the positive detection rate of HAdV and HBoV in each age group (χ2=12.862, P=0.025; χ2=14.178, P=0.015).Winter is the peak period of HAdV infection, with a positive rate of 5.54% (15/271). The peak of HBoV infection is autumn and winter with a positive rate of 12.00% (36/300) and 12.5% (34/271), respectively, higher than that of the other two seasons (χ2=43.753, P<0.05). There was a significant difference in different season groups (χ2=13.287, P=0.004; χ2=43.753, P<0.05). The sequences of 29 adenoviruses were HAdV-3, 7 serotypes of HAdV-B subgroup and HAdV-1, 2, 5 serotypes of HAdV-C subgroup. Conclusion:HAdV and HBoV play important roles in children′s respiratory tract infections, and are closely related to factors such as the season and the age of the child. They should attract clinical attention.
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Objective:To investigate the prevalence and clinical characteristics of Mycoplasma pneumoniae( Mp) genotypes and subtypes in children in Tianjin. Methods:Children with pneumonia admitted to Tianjin Children′s Hospital from December 2017 to December 2019 were selected as the research objects. Bronchoalveolar lavage fluid was collected by fiberoptic bronchoscopy. The positive samples were detected by real-time fluorescent quantitative PCR and Mp culture. PCR-restriction fragment length polymorphism(RFLP) and multiple variable number tandem repeats were used for genotyping. Detailed clinical and laboratory data were collected for all cases. Results:The results of RFLP showed that there were 138 cases (78.9%) of typeⅠand 37 cases (21.1%) of type Ⅱ; 37 cases of type M3-5-6-2, including six subtypes B, G, M, S, V and Y; 138 cases of M4-5-7-2 were detected, including seven subtypes of E, J, P, U, X, Z and a. In M3-5-6-2 type, there were 1 case of P1-Ⅰtype (2.7%), 36 cases of P1-Ⅱtype (97.3%), 137 cases of P1-Ⅰ type (99.2%) and 1 case of P1-Ⅱ type (0.7%) in M4-5-7-2 type. There was no significant difference in genotype distribution among different age groups. There were statistical differences in the distribution of four seasons among the 13 genotypes of B, G, M, S, V, Y and E, J, P, U, X, Z, a. All Mp infected children had symptoms of fever and cough. The hospitalization time, fever duration, high fever (>39℃), cough duration, skin changes, digestive system symptoms and liver function injury rate of P1-Ⅰ/M4-5-7-2 pneumonia children were higher than those of P1-Ⅱ/M3-5-6-2 pneumonia children, but the difference was not statistically significant. The WBC count of P1-Ⅱ/M3-5-6-2 types was higher than that of typeⅠand M4-5-7-2; the LDH of P1-Ⅰ/M4-5-7-2 was higher than that of Ⅱ and M3-5-6-2, with statistical difference. There was no significant difference in the incidence of inflammatory consolidation, atelectasis, pleural thickening and pleural effusion among different genotypes. Conclusions:Mp infection in children with pneumonia in Tianjin is mainly P1-Ⅰ/ M4-5-7-2, and P1-Ⅱ is on the rise. P1-Ⅰ and M4-5-7-2 were associated with fever and severe symptoms.
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Objective:To investigate the molecular epidemiological characteristics of norovirus (NoV) in hospitalized children with sporadic acute gastroenteritis in Tianjin in 2019.Methods:Fecal specimens and clinical data were collected from 3 116 hospitalized children with sporadic acute gastroenteritis possibly caused by viral infection in Tianjin Children′ Hospital between January and December, 2019. Real-time quantitative PCR was used to detect NoV. Partial sequences of RNA-dependent RNA polymerase (RdRp) and capsid genes of NoV were amplified by RT-PCR. Sequence alignment and phylogenetic analysis were performed for further analysis.Results:Among the 3 116 specimens, 809 (26.0%) were positive for NoV. There were significant differences in NoV detection rate between different age groups ( P=0.000), and the highest NoV detection rate (31.6%) was observed in the age group of 7-12 months. Moreover, the detection rate of NoV varied with seasons ( P=0.000), and the NoV detection rate was highest in winter (39.0%). Based on the sequence analysis of RdRp and capsid genes, 286 identified NoV strains belonged to six genotypes, which were GⅡ.P12-GⅡ.3, GⅡ.P16-GⅡ.2, GⅡ.P17-GⅡ.17, GⅡ.Pe-GⅡ.2, GⅡ.Pe-GⅡ.3 and GⅡ.Pe-GⅡ.4. The predominant genotype was GⅡ.Pe-GⅡ.4 Sydney 2012 (61.2%), followed by GⅡ.P12-GⅡ.3 (33.6%, 96/286), GⅡ.Pe-GⅡ.3 (2.4%, 7/286), GⅡ.P16-GⅡ.2 (2.1%, 6/286), GⅡ.Pe-GⅡ.2 (0.3%, 1/286) and GⅡ.P17-GⅡ.17 (0.3%, 1/286). Patients carrying the NoV of GⅡ.Pe-GⅡ.4 Sydney 2012 genotype were likely to suffer from vomiting than those positive for NoV of GⅡ.P12-GⅡ.3 genotype. Conclusions:NoV was an important pathogen causing acute gastroenteritis in children. GⅡ.Pe-GⅡ.4 Sydney 2012 and GⅡ.P12-GⅡ.3 were the major genotypes of NoV in hospitalized children with sporadic acute gastroenteritis in Tianjin in 2019.
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Objective:To analyze the molecular epidemiological characteristics of GⅡNorovirus (NoV) infection in children in Tianjin Children′s Hospital in 2018.Methods:Single center study. From January to December 2018, a total of 2 185 stool specimens were collected from the children with acute gastroenteritis suspected caused by virus infection in Tianjin Children′s Hospital. Norovirus was detected by real-time fluorescence reverse transcription polymerase chain reaction (RT-PCR).The capsid protein VP1(VP1) region of positive samples was amplified and sequenced. The phylogenetic tree was constructed by MEGA5.05 for analyzing the results. The detection of NoV in different age groups and different months were compared by SPSS 20.0.Results:Among the 2 185 stool specimens,610 were NoV positive, the positive rate was 27.9% (610/2 185).All of the positive samples were GⅡgenogroup. Seven genotypes were found, with GⅡ.3 subtype accounting for 46.2%(151/327),GⅡ.4 subtype accounting for 40.1%(131/327),GⅡ.2 subtype accounting for 4.6%(15/327), other subtypes accounting for 9.1%(30/327).There was a significant difference in NoV detection rate among different age groups (χ 2=17.050, P=0.002). Among the positive specimens, the detection rate of less than or equal to 3 years old was the highest which is accounting for 89.2%.Also there was a significant difference in NoV detection rate among different months(χ 2=225.153, P<0.001).November and December are the most frequent months. There was significant difference between GⅡ.3 and GⅡ.4 NoV infection in children with granulocytopenia (χ 2=11.270, P=0.001) , and also in children with respiratory symptoms (χ 2=7.257, P=0.007) . Conclusions:GⅡ.3 and GⅡ.4 were the main genotypes of NoV infection in children in Tianjin in 2018. The Multiple genotypes suggests that the monitoring and prevention of NoV infection in children should continue to be strengthened in the future.
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Objective To understand the infection status, epidemiological characteristics and genetic evolution of adenovirus in children with diarrhea in Tianjin. Methods A total of 1609 fecal specimens were collected from hospitalized children with diarrhea from July 2017 to July 2018 in Tianjin Children's Hospital. Viral nucleic acid was extracted and amplified by the hexon gene fragment. Positive specimens were used for nucleic acid sequence determination and sequence alignment and phylogenetic tree analysis with known sequences in GenBank. Rotavirus in the stool specimens was detected by gold standard method, and campylobacter was detected by latex agglutination method. Bocavirus, Norovirus and Clostridium difficile were detected by quantitative real-time polymerase chain reaction (qRT-PCR). Results A total of 69 specimens with adenoviruses were detected in 1609 cases, with a positive rate of 4.3％, in which 82.6％adenovirus-positive children were between 7 and 48 months old. The positive rate of adenovirus peaked in the summer and autumn, and the differences of this value among seasons were statistical significance (χ2=11.467,P=0.009). In these cases, the mixed infection rate of adenovirus was 26.1％(18/69), in which 7 cases were mixed with Norovirus, 7 cases were mixed with rotavirus and 4 cases were mixed with Clostridium difficile. A total of 39 adenovirus-positive PCR products were randomly selected for gene sequence detection, and 7 adenovirus genotypes were detected. Results showed that these viruses were mainly the type 41 enteric adenovirus (46.1％, 18/39), followed by type 31, type 3 and type 7 non-intestinal adenovirus, while types 1, 5 and 6 were relatively rare. Conclusions There are diverse types of adenovirus in children with diarrhea in Tianjin. The adenovirus-infected diarrhea mostly happen in summer and autumn, and is common in children.
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The blood samples of 102 type 1 diabetic children aged under 15 years and 127 normal children were collected and their genomic DNAs were extracted. The single nucleotide polymorphisms rs1990760 and rs35744605 of interferon induced with helicase C domain 1(IFIH1)gene were detected. The results showed that the allele of IFIH1 rs35744605 in diabetes group and control group was the wild type G allele. The frequency of IFIH1 rs1990760 A allele in diabetes group was higher than that in control group(22. 1％ vs 13. 0％ ,P=0. 015), suggesting that IFIH1 rs1990760 A allele is associated with type 1 diabetes in Tianjin area.
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Objective To investigate the correlation between neural tube defects (NTDs) and DACT1 gene, and provide the basic data for disease diagnosis and genetic counseling. Methods Blood samples were obtained from 163 NTDs patients and 480 unrelated healthy individuals. Mutation detection of DACT1 gene and DNA direct sequencing was carried out by PCR amplification. Bioinformatics analysis of these mutated loci was performed. Results Six mutations were found in NTDs patients, including 4 missense mutations (p.R45W, p.D142G, p.N356K and p.V702G). But these mutations were not found in 480 healthy individuals. Three mutated amino acid residues (p.45R, p.142D and p.356N) were highly conservative in evolution, and the mutated carriers were female patients, and suffered from anencephaly. Conclusion DACT1 gene mutation may be a risk factor of NTDs in Han population of northern China.
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Objective To investigate the methylation alteration of genomic DNA (gDNA) and its significance in pedigree neural tube defects (NTDs).Methods Twelve subjects from 3 NTDs pedigrees were enrolled in this study.NTDs patients were served as the case group,and their family members with normal phenotype were served as the control group.Peripheral vein blood was extracted,then gDNA was extracted.The extracted gDNA was treated with sodium bisulfite propagated as DNA segments in the way of whole genome amplification,which was put in I11umina Infinium human methylation 450k bead chip to perform hybridization,elution,extension,and imaging.The chip was scanned by iScan.Genome Studio was used to read the outcome.Illumina methylation analyzer software was used to analyze the methylation data.Results Gene differential methylation analysis showed that differential methylation sites only accounted for 0.2％ of the detected CpG sites and there were 617 differential hypermethylation sites (P ＜ 0.05),and 63 of them represented significant difference(P ＜ 1 × 10-4),including zinc finger E-box binding homebox 2,5,10-methylenetetrahydrofolate dehydrogenase 1 etc;there were 104 differential hypomethylation sites (P ＜ 0.05),and 65 of them represented significant difference (P ＜ 0.01),including Homeobox B7 and runt-related transcription factor 3 etc.Clustering analysis indicated that the tendency of DNA hypermethylation was consistent with NTDs patients,but the tendency of DNA hypomethylation was consistent with the controls.Conclusion In NTDs pedigree,the abnormal DNA methylation alterations may be the risk factor for NTDs occurrence.
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Objective To investigate the methylation alteration of genomic DNA (gDNA) and its significance in pedigree neural tube defects (NTDs).Methods Twelve subjects from 3 NTDs pedigrees were enrolled in this study.NTDs patients were served as the case group,and their family members with normal phenotype were served as the control group.Peripheral vein blood was extracted,then gDNA was extracted.The extracted gDNA was treated with sodium bisulfite propagated as DNA segments in the way of whole genome amplification,which was put in I11umina Infinium human methylation 450k bead chip to perform hybridization,elution,extension,and imaging.The chip was scanned by iScan.Genome Studio was used to read the outcome.Illumina methylation analyzer software was used to analyze the methylation data.Results Gene differential methylation analysis showed that differential methylation sites only accounted for 0.2％ of the detected CpG sites and there were 617 differential hypermethylation sites (P ＜ 0.05),and 63 of them represented significant difference(P ＜ 1 × 10-4),including zinc finger E-box binding homebox 2,5,10-methylenetetrahydrofolate dehydrogenase 1 etc;there were 104 differential hypomethylation sites (P ＜ 0.05),and 65 of them represented significant difference (P ＜ 0.01),including Homeobox B7 and runt-related transcription factor 3 etc.Clustering analysis indicated that the tendency of DNA hypermethylation was consistent with NTDs patients,but the tendency of DNA hypomethylation was consistent with the controls.Conclusion In NTDs pedigree,the abnormal DNA methylation alterations may be the risk factor for NTDs occurrence.
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Objective To explore the association between maternal methylene tetrahydrofolate reductase (MTHFR) C677T polymorphism and neural tube defects (NTDs). Methods CBM, VIP, CNKI, Wanfang, PubMed and Web of Science databases from set up to March, 2014 were electronically searched to identify case-control studies on the relationship between maternal MTHFR C677T polymorphism and NTDs. The data were quantitatively analyzed by RevMan 5.0 software. Results A total of 25 studies were selected including 2 282 cases and 3 420 controls. Overall, the pooled OR (with 95%CI) under co-dominant model and allele contrast were 2.28(1.60-3.24), 1.25(1.02-1.53) and 1.42(1.21-1.67). Subgroup analysis showed significant association between maternal MTHFR C677T polymorphism and NTDs susceptibility in Asian populations. Conclusion The present meta-analysis suggests that MTHFR C677T polymorphism is significantly associated with maternal risk for NTDs, especially in Asian populations.