Objective:To investigate the molecular epidemiological characteristics of coxsackievirus A16 (CVA16) in Fujian province from 2020 to 2023.Methods:The epidemiological characteristics of CVA16 associated hand, food and mouth disease (HFMD) in Fujian province from 2020 to 2023 was analyzed. The complete VP1 gene of CVA16 was amplified by RT-PCR and then sequenced, and genetic evolution was analyzed by MEGA X and other softwares.Results:From 2020 to 2023, there were 13 120 cases of HFMD in Fujian province, and the proportion of HFMD which caused by CVA16 was 16.5% (2 160/13 120). From 2020 to 2023, the proportion of accounted cases was 4.7% (94/2 019), 14.1% (457/3 243), 47.6% (1 521/3 199) and 1.9% (88/4 659) respectively. HFMD caused by CVA16 was mainly concentrated in children aged 1 to 5 years, and most of them were 3 years old. The genetic evolution and genotype analysis of 92 complete VP1 gene sequences obtained from 2020 to 2023 showed that the genetic distance between CVA16 strains in Fujian province and the prototype strain was far away. The CVA16 genotype in Fujian province from 2020 to 2023 has three clusters of B1a, B1b and B1c, among which the composition ratio of B1a and B1b in Fujian province in 2020 was 40% and 60% respectively. In 2021, B1a and B1b accounted for 81.8% and 18.2% respectively. Only B1a in 2022; in 2023, there were B1a, B1b and B1c, which respectively accounted for 44.4%, 7.4% and 48.2%. During the period from January to September, B1a was the main cluster. After October we observed an emergence of B1c cluster, which had never been found in Fujian province and was rare in China, was detected and became the dominant cluster.Conclusions:The evolutionary cluster of CVA16 dominant changed from B1b in 2020 to B1a in 2021-2023. After October 2023, the newly discovered B1c became the dominant cluster in Fujian province.

Objective:To improve the efficiency and success rate of human adenovirus 3 (HAdV-3) whole genome sequencing, a high-throughput sequencing method for the whole genome of HAdV-3 based on multiplex PCR was established.Methods:Multiplex PCR primers suitable for the whole genome amplification of HAdV-3 were designed. The whole genome sequence of HAdV-3 was amplified by multiplex PCR, and the specificity of the amplification product was preliminarily verified by agarose gel electrophoresis. High-throughput sequencing of the multiplex PCR products was performed using Illumina second-generation sequencing. After obtaining the sequence, software such as CLC and IGV were used to analyze the effective data amount, average sequencing depth, and whole genome coverage obtained by high-throughput sequencing, then the sequencing quality was evaluated. Based on the whole genome sequencing result, a phylogenetic tree was constructed to confirm the virus type and analyze homology of the sequences, and then the accuracy of this method was evaluated.Results:A total of 70 (35 pairs) multiplex PCR amplification primers for the whole genome of HAdV-3 were designed, with amplicon size of approximately 1 200 bp. And the expected whole genome coverage could reach 99.8% (with a total genome length of approximately 35 240 bp). Agarose gel electrophoresis analysis showed that the size of the multiplex PCR products was consistent with expectations, and the amplification specificity was high. The high-throughput sequencing result showed that the whole genome sequences obtained by this method were complete and intact, and the genome coverage was consistent with expectations. Sequence quality analysis showed that the high-throughput sequencing method based on multiplex PCR could obtain more effective data and greater sequencing depth, resulting in more uniform whole genome coverage. Phylogenetic analysis showed that the evolutionary typing result of viral DNA sequenced after multiplex PCR amplification were consistent with those of viral DNA sequenced directly and had high homology, indicating that the multiplex PCR method had high amplification fidelity and the results obtained in combination with high-throughput sequencing were accurate.Conclusions:A high-throughput sequencing method for the whole genome of HAdV-3 based on multiplex PCR was established in this study successfully. This method could improve the efficiency and success rate of HAdV-3 whole genome sequencing, aiming to provide better technical support and reference for HAdV-3 pathogen surveillance and epidemic source-tracing based on whole genome sequencing.

Objective This study aimed to examine and propagate the medication experience and group formula of traditional Chinese medicine (TCM) Master XIONG Jibo in diagnosing and treating arthralgia syndrome (AS) through data mining. Methods Data of outpatient cases of Professor XIONG Jibo were collected from January 1, 2014 to December 31, 2018, along with cases recorded in A Real Famous Traditional Chinese Medicine Doctor: XIONG Jibo's Clinical Medical Record 1, which was published in December 2019. The five variables collected from the patients’ data were TCM diagnostic information, TCM and western medicine diagnoses, syndrome, treatment, and prescription. A database was established for the collected data with Excel. Using the Python environment, a customized modified natural language processing (NLP) model for the diagnosis and treatment of AS by Professor XIONG Jibo was established to preprocess the data and to analyze the word cloud. Frequency analysis, association rule analysis, cluster analysis, and visual analysis of AS cases were performed based on the Traditional Chinese Medicine Inheritance Computing Platform (V3.0) and RStudio (V4.0.3). Results A total of 610 medical records of Professor XIONG Jibo were collected from the case database. A total of 103 medical records were included after data screening criteria, which comprised 187 times (45 kinds) of prescriptions and 1 506 times (125 kinds) of Chinese herbs. The main related meridians were the liver, spleen, and kidney meridians. The properties of Chinese herbs used most were mainly warm, flat, and cold, while the flavors of herbs were mainly bitter, pungent, and sweet. The main patterns of AS included the damp heat, phlegm stasis, and neck arthralgia. The most commonly used herbs for AS were Chuanniuxi (Cyathulae Radix), Huangbo (Phellodendri Chinensis Cortex), Cangzhu (Atractylodis Rhizoma), Qinjiao (Gentianae Macrophyllae Radix), Gancao (Glycyrrhizae Radix et Rhizoma), Huangqi (Astragali Radix), and Chuanxiong (Chuanxiong Rhizoma). The most common effect of the herbs was “promoting blood circulation and removing blood stasis”, followed by “supplementing deficiency (Qi supplementing, blood supplementing, and Yang supplementing)”, and “dispelling wind and dampness”. The data were analyzed with the support ≥ 15% and confidence = 100%, and after de-duplication, five second-order association rules, 39 third-order association rules, 39 fourth-order association rules, and two fifth-order association rules were identified. The top-ranking association rules of each were “Cangzhu (Atractylodis Rhizoma) → Huangbo (Phellodendri Chinensis Cortex)” “Cangzhu (Atractylodis Rhizoma) + Chuanniuxi (Cyathulae Radix) → Huangbo (Phellodendri Chinensis Cortex)” “Chuanniuxi (Cyathulae Radix) + Danggui (Angelicae Sinensis Radix) + Gancao (Glycyrrhizae Radix et Rhizoma) → Qinjiao (Gentianae Macrophyllae Radix)” and “Chuanniuxi (Cyathulae Radix) + Danggui (Angelicae Sinensis Radix) +Gancao (Glycyrrhizae Radix et Rhizoma) + Huangbo (Phellodendri Chinensis Cortex) → Qinjiao (Gentianae Macrophyllae Radix)”, respectively. Five clusters were obtained using cluster analysis of the top 30 herbs. The herbs were mainly drying dampness, supplementing Qi, and promoting blood circulation. The main prescriptions of AS were Ermiao San (二妙散), Gegen Jianghuang San (葛根姜黄散), and Huangqi Chongteng Yin (黄芪虫藤饮). The herbs of core prescription included Cangzhu (Atractylodis Rhizoma), Chuanniuxi (Cyathulae Radix), Gancao (Glycyrrhizae Radix et Rhizoma), Huangbo (Phellodendri Chinensis Cortex), Mugua (Chaenomelis Fructus), Qinjiao (Gentianae Macrophyllae Radix), Danggui (Angelicae Sinensis Radix), and Yiyiren (Coicis Semen). Conclusion Clearing heat and dampness, relieving collaterals and pain, and invigorating Qi and blood are the most commonly used therapies for the treatment of AS by Professor XIONG Jibo. Additionally, customized NLP model could improve the efficiency of data mining in TCM.

Objective:To study the epidemiological and molecular characteristics of coxackievirus A2 (CV-A2) associated with hand, foot and mouth disease (HFMD) in Fujian province, 2011-2020.Methods:Descriptive epidemiological method was used to analyze the epidemiological characteristics of CV-A2. Case information was collected in the laboratory of Fujian Provincial Center for Disease Control and Prevention from 2011 to 2020. The VP1 gene obtained by first-generation sequencing and the complete genome obtained by second-generation sequencing were made phylogenetic analysis and recombination analysis by Mega X and Simplot3.5.1.Results:From 2011 to 2020, there were 35 HFMD-associated CV-A2 cases in Fujian province, most of whom aged 1 to 2 years (28 / 35), and the gender ratio of male to female was 2.5∶1 (25 / 10). Phylogenetic tree of VP1 region showed that CV-A2 in Fujian province was mainly distributed in the evolutionary branches of D1 and C1 subgenotype, only one strain (2019FJPT064) belonged to C1 subgenotype and the other 27 strains were grouped under D1 subgenotype. Among them, 4 isolates from 2011 to 2012 belonged to cluster 1 of D1 subgenotype, and the other two isolates from 2011 to 2012 and those after 2012 belonged to cluster 2 of D1 subgenotype. The complete genome length of 6 CV-A2 strains were about 7 393 to 7 402 bp. The homology of the nucleic acid sequences in the VP1 region of 6 CV-A2 strains and the CV-A2 prototype strain were 80.3%-81.7%, while in the other regions were 72.7%-86.2%. The phylogenetic tree of P2 region showed that 6 CV-A2 strains had a close evolutionary distance with the coxackievirus A4 (CV-A4) and the phylogenetic tree of P3 region showed that 2011FJFZ027, 2012FJQZ311 and 2020FJPTN040 had close evolutionary distance with coxackievirus A6 (CV-A6), coxackieviru A14 (CV-A14), coxackievirus A8 (CV-A8), respectively. Recombination analysis showed that 2011FJFZ027, 2012FJQZ311 and 2020FJPTN040 maybe evolved from recombinant CV-A6 strain (GenBank accession number: KR706309), CV-A14 strain (GenBank accession number: KP036482) and CV-A8 strains (GenBank accession number: KM609475 and MT648786) in non-structural protein region.Conclusions:HFMD-associated CV-A2 was sporadic in Fujian province from 2011 to 2020. The D1 subgenotype of CV-A2 was persistently circulating in Fujian province. From 2011 to 2012, the cluster 1 of D1 subgenotype was the dominant type, and then it shifted to the cluster 2 to take its place after 2012. CV-A2 strains in Fujian were frequently recombined in diversity patterns.

Objective:To investigate the gene characteristics and recombination of coxsackievirus A5(CV-A5)in Fujian province.Methods:The VP1 regions of CV-A5 strains from positive samples of hand, foot and mouth disease (HFMD) collected from all cities in Fujian province from 2011 to 2020 were amplified by reverse transcription nested polymerase chain reaction (RT-nPCR), then sequenced and identified the subgenotypes. CV-A5 strains were isolated in rhabdomyosarcoma (RD) cells. The isolated virus strains were sequenced by next generation sequencing to obtain the whole genome sequences. MEGA7.0 was used for phylogenetic analysis. RDP4 and SimPlot 3.5.1 softwares were used to analyze viral gene recombination.Results:Total thirty-one HFMD related CV-A5 cases were detected in Fujian province from 2011 to 2020, mainly in children aged 1 to 2 years (16/31), with a male/female ratio of 2.1∶1 (21/10), including 3 severe cases and 1 deceased case. CV-A5 subgenotyping was successful in 27 cases, all of them belonged to E1 subgenotype. The whole genome sequences of six CV-A5 strains were obtained, and the homology with the prototype strain Swartz was 81.1%-81.6%. Phylogenetic analysis results based on the sequences of whole genome, P1, P2 and P3 regions showed that the virus strain 2017FJFZ239 is a suspected recombinant strain, which was distributed in the same branch as CV-A5 reference strains (GenBank accession number: MW079817 and MN663160) in P2 and P3 regions. Analyzing with RDP4 showed that the virus strain 2017FJFZ239 had the same recombination event with CV-A5 reference strains (GenBank accession number: MW079817 and MN663160) and the potential parental sequences were CV-A2 (GenBank accession number: KX595284) and CV-A5 (GenBank accession number: KP289362). Further analyzing with SimPlot showed that the virus strain 2017FJFZ239 may recombine with CV-A2 strain (GenBank accession number: KX595284) in non-structural protein 2B, 2C and P3 regions.Conclusions:From 2011 to 2020, HFMD related CV-A5 in Fujian province belongs to E1 subgenotype, and there is a recombinant strain.

Objective:To investigate the prevalence of autism spectrum disorders (ASD)among children in kindergartens in Xi′an urban districts.Methods:A stratified cluster sampling method was adopted, and selected all children from 12 kindergartens in 6 urban districts of Xi′an.Primary screening positive children with ASD were identified by filling out the Autism Behavior Checklist (ABC) by their parents and the guardians reports, and then the beha-vioral observations were made to identify suspicious ASD children.Finally these children were diagnosed in the hospital through the autism diagnostic observation schedule, 2 nd edition (ADOS-2). The data were calculated with SPSS 18.0. Results:Totally, 38 cases with ASD were diagnosed among 5 178 children, the prevalence of children ASD in kindergartens in Xi′an urban districts was 7.3‰, and the 95% confidence interval was 4.98‰-9.62‰.The prevalence of ASD in children was statistically significant in different age groups ( χ2=9.914, P<0.05) and gender groups ( χ2=18.812, P<0.05). The accuracy of ASD screening by guardians reports is better than that by ABC. Conclusions:(1)The prevalence of ASD children in kindergartens in Xi′an urban districts is at high level in similar reports in China.If ASD children at the special education institutions and home are considered, the overall prevalence rate is higher, indicating that the prevalence of ASD children in China may be underestimated.(2)With the increase of age, the attendance rate of ASD children in kindergartens has a decreasing trend.

Objective To explore the program of convolutional neural networks for the diagnosis of schizophrenia and evaluate its effects. Methods Using the convolutional neural network,the training model was trained in the lead data of 138 normal people and 183 schizophrenic patients,and the model was valida-ted by 20-fold cross-validation. Results The true positive rate of schizophrenia prediction using the convolu-tional neural network training model was 0. 749, the false positive rate was 0. 275, and the accuracy was 0. 738. Conclusion This model can achieve a strong diagnostic ability for patients with schizophrenia. Therefore,convolutional neural network for the diagnosis of schizophrenia will become an important research direction in the future.

For investigating the epidemiology and genetic characteristics of enterovirus 71 (EV71) in Fujian from 2010 to 2015,we analyzed the surveillance data of EV71 and sequenced VP1 genes of 72 EV71 strains randomly picked from the past 6 years.The overall infection rate was gradually down and one incidence peak (from May to July) was observed each year.Major infectious population were focused on Xiamen,Fuzhou,Nanping and Quanzhou,the ages ranged from one to three years old.Scattered children were the most infected ones.The proportion of EV71 in the severe case was higher than in the HMFD(χ2 =732.064 5,P＜0.000 1).EV71 circulated from 2010 to 2015 in Fujian Province was belonged to subgenotype C4a in consistent with vaccine strain (H07).Compared with the VP1 of vaccine strains,the divergence of complete VP1 nucleotide sequence was gradually expanding as time distance increased,but the sequence of amino acid was not found obvious difference.Variations in 4 key immune epitopes of amino acid had not appeared a regular pattern in year and not consistent with the trend of proportion of EV71 in HMFD.As a result,we considered the epidemiology characteristics of EV71 in Fujian was obvious,72 strains still belonged to C4a subgenotype and had no outstanding antigenic drift or mutation.Extensive epidemiology surveillance and genetic characteristic are needed for the application of EV71 vaccine.

To study the biological characteristics and mutations of influenza A(H1N1)pdm09 virus isolated from one case of pneumonia of unknown etiology (PUE),which would provide references for clinical treatment and disease control,the throat swab specimen from the PUE case was isolated in the Madin-Darby Canine Kidney (MDCK) cells,and then the antigenicity,pathogenicity and drug resistance of influenza A (H1N1) pdm09 virus were analyzed after sequencing.As a result,one influenza virus strain was isolated from the specimen and named as A/FujianGulou/SWL64/2016(H1N1).The similarities of nucleotide sequences and amino acids sequences compared with the vaccine strain A/California/07/2009 (H1N1) were 96.9％-98.9％ and 96.7％-99.5％,respectively.Eighteen amino acids had mutated in the HA and 4 mutations,K163Q,S185T,S203T and D222N,were involved in 3 different epitopes,which indicated that the antigenic drift had occurred in the influenza virus.The D222N mutation associated with receptor binding site made the virus infect lower respiratory tract more easily.The virus was still amantadine-resistance and oseltamivir-sensitive.In conclusion,the influenza A (H1N1) pdm09 virus in this study have occurred antigenic drift and has the molecular characterization of causing severe pneumonia,so further surveillance should be performed to prevent and control the influenza epidemic.

OBJECTIVETo study the molecular epidemiology of hand-foot-mounth disease (HFMD) associated Coxsackievirus A10 (Cox A10) identified in Fujian province.

METHODSA total of 1 525 specimens from non-EV71 non-Cox A16 HFMD patients were collected during 2011-2014. Isolated virus strains were identified and sub-typed. Full-length coding regions for the VP1 gene of the predominant serotype Cox A10 isolates were amplified and sequenced.

RESULTSAmong the 407 non-EV71 non-Cox A16 HFMD cases confirmed by virus isolation and molecular subtyping, 103 (25.3%) were caused by Cox A10, accounting for 11.0%, 6.0%, 18.4% and 9.2% among the HFMD-associated entero-viruses identified in 2011, 2012, 2013 and 2014, respectively, in Fujian province. Compared to the general features observed in the HFMD epidemics, no differences on the Cox A10-specificity rates were observed among factors as geographical origins, gender or age groups, but all with high rates of severity. Data from the nucleotide sequence analyses on VP1 genes showed low homology levels of 76.0%-77.1% among Cox A10 strains from Fujian province, in contrast to the prototype Cox A10 strain, but with high levels of homology in the amino acid sequences (91.9%-93.6%). RESULTS from the Phylogenetic analysis also indicated that Cox A10 isolates from Fujian province were distinct from the prototype strain or other isolates from other countries but was homologous to domestic strains, but the Fujian isolates clustered into multiple branches.

CONCLUSIONSCox A10 remained one of the predominant serotypes of HFMD in Fujian province. Cox A10 isolates identified in Fujian province were co-circulating and co-evolving with other domestic strains.

Benzeneacetamides ; Child ; Child, Preschool ; China ; epidemiology ; Enterovirus A, Human ; classification ; genetics ; isolation & purification ; Epidemics ; Female ; Hand, Foot and Mouth Disease ; epidemiology ; genetics ; virology ; Humans ; Infant ; Male ; Molecular Epidemiology ; Molecular Sequence Data ; Open Reading Frames ; Phylogeny ; Piperidones ; Serogroup