Objective: To investigate the genotypes of enteroviruses causing hand, foot and mouth disease ( HFMD ) in Shaoxing City, so as to provide insights into HFMD control. Methods: The anal and pharyngeal swab specimens were collected from HFMD cases under 14 years of age reported by sentinel hospitals of HFMD in Shaoxing City from 2018 to 2020. The enterovirus genotypes were determined by quantitative real-time PCR (qPCR) assay and sequenced by BLAST, and the constitution and temporal distribution of enterovirus genotypes were descriptively analyzed. Results: A total of 1 267 anal and pharyngeal swab specimens were collected from children with HFMD, and 661 specimens were positive for enteroviruses, with a positive rate of 52.17%. Among the 661 enterovirus-positive specimens, 70 specimens were tested positive for EV71 ( 5.52% ), 208 specimens tested positive for CoxA16 ( 16.42% ), 239 specimens tested positive for CoxA6 ( 18.86% ); 23 specimens tested positive for CoxA10 (1.82%), and 121 specimens tested positive for other 10 genotypes, including CoxA2, CoxA4, and CoxA5. The prevalence rates of EV71 were 14.08%, 0.72% and 1.84% from 2018 to 2020, and the detection of EV71 peaked during the period between April and July, 2018. The prevalence rates of CoxA16 were 10.98%, 26.57% and 11.98 from 2018 to 2020, and the detection of CoxA16 peaked during the period between April and August, and between November and December, 2019. The prevalence rates of CoxA6 were 10.50%, 14.73% and 30.88% from 2018 to 2020, and the detection of CoxA6 peaked during the period between May and December, 2020. Conclusions EV71, CoxA16 and CoxA6 were predominant enterovirus genotypes causing HFMD in Shaoxing City from 2018 to 2020, and the detection of CoxA6 appeared a tendency towards a rise.

Objective : To investigate the clinical and molecular epidemiological characteristics of 10 patients with enterovirus (EV)-D68 infections in the respiratory tract in Shaoxing City, Zhejiang Province, so as to provide insights into formulation of control interventions for EV-D68 infections. Methods: Clinical specimens were sampled from patients with acute respiratory tract infections (ARTIs) admitted to sentinel hospitals in Shaoxing City from 2021 to 2022, and EV-D68 was detected using real-time fluorescent PCR assay and sequencing of the VP1 region of enterovirus. The epidemiological and etiological characteristics of EV-D68 infected cases were descriptively analyzed. Results: A total of 3 009 specimens were sampled from patients with ARTIs from 2021 to 2022, and the detection of EV-D68 was 0.33%. Of all EV-D68-infected patients, there were 6 men and 4 women, and 5 cases under 18 years of age, 2 cases at ages of 18 to 60 years and 3 cases at ages of over 60 years. EV-D68 infection predominantly occurred in summer (5 cases detected between May and July) and autumn (5 cases detected between September and October). The main clinical symptoms included fever (10 cases), sore throat (9 cases) and cough (8 cases), and all 10 cases recovered well, with no deaths reported. Sequencing identified D3 subtype in all 10 specimens positive for EV-D68. Conclusions The ARTIs caused by EV-D68 occurred predominantly among children under 18 years of age in Shaoxing City, and was highly prevalent in summer and autumn. D3 was the predominant enterovirus subtype.

Objective: To investigate the molecular typing and drug resistance of Salmonella in market-sold raw livestock meats in Shaoxing City, Zhejiang Province, so as to provide the basis for the prevention and control of drug-resistant Salmonella. Methods: Salmonella isolates were detected and serotyped from raw livestock meat samples collected from supermarkets, agricultural markets and restaurants in Shaoxing City between March and November 2023. Molecular typing was performed using pulsed-field gel electrophoresis (PFGE) and multilocus sequence typing (MLST). Antibiotic resistance phenotypes and resistance gene analyses were conducted using broth microdilution method and whole genome sequencing. Results: A total of 34 Salmonella isolates were detected from 329 samples and the positive rate was 10.33%. There were 14 serotypes, with S. Rissen (9 isolates, 26.47%) and S. London (8 isolates, 23.53%) being dominant. PFGE typing revealed 31 band patterns and 23 banding pattern clusters of Salmonella, and the similarity ranged from 30.80% to 100.00%, without dominant band patterns. MLST identified 15 sequence typing (ST) types, and S. Rissen were all ST469 and S. London were all ST155. Salmonella strains exhibited high levels of resistance to tetracycline (44.12%) and ampicillin (35.29%). Thirteen resistance patterns were identified, with 8 multidrug resistant patterns (29.41%). There were 8 classes of 14 resistance genes, with aminoglycoside resistance gene aac(6' )-Iaa found in 100.00% of the strains. The highest concordance rate between the resistance phenotype and gene was observed for phenicol antibiotics (97.06%), while there was no concordance between the quinolone resistance phenotype and genes. Conclusions S. Rissen and S. London are the predominant serotypes in market-sold raw livestock meats in Shaoxing City. The PFGE band types of the Salmonella isolates show polymorphism distribution, and MLST are mainly ST469 and ST155. The isolates have high levels of resistance to tetracycline and ampicillin, manifesting multidrug resistance.

Objective: To investigate the epidemiological characteristics of foodborne disease outbreaks in Shaoxing City, Zhejiang Province, from 2012 to 2022, so as to provide the evidence for improving the foodborne disease control strategy. Methods: Foodborne disease outbreaks in Shaoxing City from 2012 to 2022 were collected from National Foodborne Disease Outbreak Monitoring System in China, including populations, places of outbreak, pathogenic factors and suspected foods. The temporal distribution, regional distribution, distribution of outbreak places and pathogenic factors of foodborne disease outbreaks were descriptively analyzed. Results: A total of 89 foodborne disease outbreaks were reported in Shaoxing City from 2012 to 2022, covering totally 699 patients, with an average annual attack rate of 6.35%. The outbreak peaked during the period between June and October (73 outbreaks, 82.02%), and family was the predominant place of outbreak (41 outbreaks, 46.07%). There were 83 outbreaks with known pathogenic factors, including 51 outbreaks caused by microbial factors, with Vibrio parahaemolyticus, Salmonella and norovirus as predominant pathogens, and 29 outbreaks caused by fungi and their toxins, which were all poisonous mushrooms poisoning, resulting in 2 deaths. In addition, there were 3 outbreaks caused by chemical factors. Conclusions The outbreak of foodborne diseases predominantly occurred in summer and autumn in Shaoxing City from 2012 to 2022. Family was the predominant place of outbreak, and toxic mushroom poisoning was the most lethal pathogenic factor.

Objective: To investigate the molecular transmission network characteristics of HIV-1 among men who have sex with men (MSM) in Shaoxing City, Zhejiang Province, so as to provide insights into AIDS prevention and control among MSM. Methods: Newly reported HIV/AIDS cases among MSM in Shaoxing City were selected from January 2021 to September 2023. Plasma samples before the antiviral treatment were collected. The HIV-1 pol gene was amplified using reverse transcription PCR and nested PCR to construct phylogenetic trees for gene subtype analysis. The HIV-TRACE method was used to construct a molecular transmission network with a genetic distance of 1.5% to analyze clustering and the characteristics of cases within molecular clusters. Results: A total of 216 HIV/AIDS cases among MSM were included, and 179 qualified sequences were obtained. The predominant HIV-1 subtypes were CRF07_BC and CRF01_AE, with 95 and 66 cases, respectively. At 1.5% genetic distance, 20 molecular clusters were formed, with 61 nodes and 58 edges. A total of 61 sequences were connected to the transmission network (34.08%). HIV/AIDS cases among MSM from all the counties (cities, districts) in Shaoxing City were included in the network. There was the largest molecular cluster with ≥10 nodes, involving 12 cases from five counties (cities, districts), 3 medium-sized molecular clusters with 4 to 5 nodes, and the 16 small-sized molecular clusters with 2 or 3 nodes. Seven cases with high risk of transmission, each with ≥4 edges, were all CRF07_BC subtypes. Among them, two cases were from the large molecular cluster, and five cases were from the same molecular cluster composed of cases from Shengzhou City and Xinchang County. Conclusions The predominant HIV-1 subtypes among MSM in Shaoxing City were CRF07_BC and CRF01_AE. There was cross-regional HIV transmission, and potential transmission risk might exist in Shengzhou City and Xinchang County.

Objective To predict and identify liner B-cell epitopes in the hemagglutinin ( HA) of human-infected avian-origin H7N9 influenza virus and analyze the specificity of H7 subtype.Methods Three serum samples collected at different times from the same patient who was confirmed to be infected with H7N9 influenza virus were provided by Shaoxing People’s Hospital, and one serum sample from healthy person was collected as the control.The extracellular region of HA protein was predicted by TMHMM Sever v.2.0.The potential B-cell epitopes were predicted by DNAStar Lasergene’ s Protean, BcePred and ABCpred tools, and the immunogenicity of the predicted B cell antigen epitopes was assessed by indirect enzyme-linked immunosordent assay ( ELISA ) .H7 subtype specificity was analyzed by comparing HA protein amino acid sequence with H7N9 and H1-H16 subtype influenza virus from Genbank using Clustal X 2.1 software, and Cn3D 4.3.1 software was used to detect the distribution and 3D structure of predicted epitopes on the HA protein of H7N9.Results The potential B-cell epitopes may be located in 172-183, 363-380, 452-472 and 491-506 of extracellular N-terminus of HA protein.ELISA showed that four predicted eptiopes specifically reacted with positive serums from patient.Multi-sequence alignment demonstrated that peptide 172-183 and 363-380 had higher H7 subtype specificity compared with amino acid sequences of other subtypes.Moreover, the predicted linear B-cell epitopes all located on the surface of HA protein according to the 3D structure analysis.Conclusion Four potential B-cell epitopes were identified, in which peptide 172-183 and 363-380 have higher H7 subtype specificity, and may be used in the design of epitope-based vaccines and diagnostics tests.

Objective:To screen host cell proteins interacting with rotavirus nonstructural protein 2(NSP2) and provide a theoretical basis for the discovery of antiviral targets.Methods:E. coli BL21(DE3) was transformed with a recombinant plasmid NSP2-pGEX-6P-1 and induced by IPTG to express NSP2-GST. GST affinity chromatography was used to purify NSP2-GST and Western blot was performed for verification. NSP2-GST and GST proteins were used as targets to capture interacting proteins in MA104 cells via GST pull-down. After silver staining, differential proteins were screened by in-gel enzymatic hydrolysis and mass spectrometry. Protein pilot platform was used to filter peptides, and the names and the biological functions of the proteins were obtained by Paragon algorithm. The potential connection between the interacting proteins was demonstrated by protein interaction network diagrams and GO functional annotation analysis. Moreover, the molecular docking of the top three proteins with NPS2 was predicted using HDOCK server and verified by docking and confidence scores, so as to investigate the visual docking model between interacting proteins. Results:SDS-PAGE and Western blot showed the successful purification of the recombinant protein NSP2. Ten host proteins including PKM2, which might interact with NSP2, were identified by GST pull-down and protein profiling. GO analysis and interaction diagrams revealed that RPS4X, EZR, SUPT16H and EIF2S3 mediated molecular expression; PKM2, LDHA and ATP5A1 participated in energy metabolism; HSP90, ACTB and ANXA2 were involved in biological movement. Besides, there were functional connections and interaction networks among them. Molecular docking further verified PKM2, HSP90 and RPS4X did interact with NSP2, and the interaction force were strong enough to form a stable structure.Conclusions:This study successfully discovered several host proteins including PKM2, HSP90 and RPS4X that could interact with NSP2, providing reference for investigating the process of rotavirus infection and making related prevention and control strategies.

OBJECTIVETo investigate the clinical results of one stage surgical treatment in congenital scoliosis (CS) patients associated with split cord malformation (SCM).

METHODSBetween January 2007 and December 2010, 50 patients underwent one stage surgical treatment for CS associated with SCM. Among of them, 38 patients (13 male and 25 female) with an average age of (15 ± 6) years, who were followed up in the clinic at least 2 years longer, were include in the study. There were 12 patients with Type I SCM and 26 patients with Type II SCM Pre-operative, post-operative and the follow-up imaging data were collected and compared by paired t-test, while imaging data between Type I SCM group and Type II SCM group were compared by group t-test. Bony spur was first resected to the Type I SCM while nothing was done to the Type II SCM. Then, all patients were followed by posterior corrective procedure in one stage. Meanwhile, duraplasty were only applied in 5 patients whose dural cleft were more than 1 cm longer.

RESULTSThe average follow-up was 41 ± 13 months (range, 26-68 months). The average operation time was 491 ± 152 minutes (range, 105-780 minutes) and the average blood loss was (1 933 ± 1 516) ml (range, 1 000-8 000 ml). The mean major coronal curve was corrected from 70° ± 26° preoperatively to 312° ± 16° postoperatively with a correction rate of 57% ± 18%, and 33° ± 17° at the final follow-up with a correction rate of 54% ± 20%. The mean major sagittal curve was corrected from 43° ± 31° to 26° ± 16°, and 27° ± 15° at the final follow-up. The postoperative complication occurred in 2 patients (5.3%) with Type I SCM, including neurological deterioration in 1 patient (2.6%) and cerebrospinal fluid leakage in 1 patient (2.6%). There were no paralysis and other serious complications. The patients who suffered from neurological deterioration recovered to the preoperative neurological status at 30 months postoperatively and no further improvement at the final follow-up.

CONCLUSIONIt is safe and efficient to treat the CS associated with SCM by one stage surgery without increasing the risk of neurological complications postoperatively.

Adolescent ; Child ; Female ; Follow-Up Studies ; Humans ; Male ; Neural Tube Defects ; complications ; surgery ; Retrospective Studies ; Scoliosis ; complications ; congenital ; surgery ; Treatment Outcome ; Young Adult