Objective To understand the epidemiological characteristics of hand-foot-mouth disease(HFMD) in China, and provide evidence for the prevention and control of HFMD and vaccine clinical research design.Methods Using PubMed,China National Knowledge Infrastructure(CNKI) and Wanfang databases as retrieval sources, the Chinese and English articles on epidemiological characteristics of HFMD in China published from 2012 to 2021 were systematically retrieved, and relevant information was extracted. Chi-square test was used for the statistical analysis of the enterovirus detection rate, type composition, the distribution of gender, age, year, season, region and symptoms.Results A total of 327 articles were included in this study, of which 295 were included in the overall detection rate of HFMD enterovirus. The total number of people involved was 4 786 389, the number of enterovirus nucleic acid positive cases was 709 086, and the laboratory-diagnosed cases accounted for about 14. 81% of the total number. The four types with the highest detection rates were Coxsackievirus A6(CA6), enterovirus 71(EV71), CA16 and CA10. A total of 327 articles with 61 serotypes were included in the type proportion, and the four types more than 1% were EV71, CA16, CA6 and CA10, and other types such as CA4, CA2 and CA5accounted for a lower proportion. Among the laboratory-diagnosed cases, there were more men than women, with a sex ratio of about 1. 72∶1; children under 5 years of age were at high risk of HFMD, with 1 year of age being the most and 0 year of age being the least; the composition ratio of enteroviruses varied with the year and season, the proportion of EV71 showed an overall downward trend from 2008 to 2019, and the peak season for HFMD was from April to June. The distribution of HFMD exhibited geographical differences, and the top 5 provinces were: Guangdong, Jiangsu, Hebei, Hunan and Zhejiang; HFMD was mainly common type/mild type, and the proportion of EV71 in severe cases and deaths was significantly higher than that of other types.Conclusion The type composition of HFMD enteroviruses in China was mainly EV71, CA16, CA6 and CA10,while other types such as CA4, CA2 and CA5 accounted for a relatively low proportion. Type distribution varied in sex, age,year, season, region and symptoms.

Objective:To analyze the relationship between the expression level of calumenin(CALU)and the prognosis of hepatocellular carcinoma(HCC)patients by bioinformatics tools and establish the prognostic prediction nomogram,and to clarify its possible mechanism.Methods:The data of 374 HCC tissue samples were downloaded from The Cancer Genome Atlas(TCGA)database and the data of 160 normal tissue samples were down loaded from Genotype-Tissue Expression(GTEx)database.Paired sample t-test was used to analyze the difference in CALU expression between the HCC tissue samples and the paired adjacent normal tissue samples.Human Protein Atlas(HPA)database was used to verify the results.DESeq2 package was used to screen the differentially expressed genes(DEGs)between CALU low expression group and CALU high expression group in the HCC tissue samples.R package pROC was used to analyze the receiver operating characteristic(ROC)curve.Univariate and multivariate Cox regression analyses were used to confirm the prognosis value of CALU in the HCC patients with different clinicopathological characteristics,and ggplot2 package was used to construct the forest plot.R packages rms and survival were used to construct the nomogram and its calibration curve,and the diagnostic value of CALU in distinguishing HCC tissue from normal tissue was analyzed.The data from Kaplan-Meier Plotter database were used to further verify the relationship between CALU and the prognosis of HCC patients.The gene transcriptional expression data of 216 HCC samples obtained from GSE14520 dataset were used to verify the prediction accuracy of the nomogram.Gene Ontology(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG)enrichment analyses were used to determine the function and enrichment pathways of the DEGs,and Gene Set Enrichment Analysis(GSEA)was used to obtain the significantly enriched gene sets of the DEGs.Single-cell sequencing data of 10 HCC tissue samples and 8 adjacent normal tissue samples obtained from GSE149614 dataset were used to verify the relationship between CALU and the prognosis of HCC patients and its mechanism.Results:Compared with normal tissue,the expression level of CALU mRNA in HCC tissue was significantly increased(P＜0.001),and the expression level of CALU protein in HCC samples was also increased.A total of 928 DEGs were identified between CALU low expression group and CALU high expression group in the HCC samples,including 784 upregulated DEGs and 144 downregulated DEGs.The ROC analysis results indicated that CALU showed high diagnostic value in distinguishing cancer tissue from adjacent non-cancer tissue with an area under curve(AUC)of 0.839.Kaplan-Meier survival analysis showed that the survival rate of HCC patients in CALU high expression group was significantly lower than that in CALU group low expression(P＜0.001).Univariate and multivariate Cox regression analyses results demonstrated that high expression of CALU was an independent risk factor of the prognosis in HCC patients,and a prognosis prediction nomogram was constructed.The applicability of nomogram on the prognosis of HCC was verified by GSE14520 dataset.The GO enrichment analysis results showed that DEGs were mainly enriched in pathways related to the oxidative stress,ferroptosis and cuproptosis(P＜0.05).The KEGG enrichment analysis results showed that DEGs were mainly enriched in the pathways related to extracellular matrix(ECM)receptor interaction,linoleic acid metabolism and neuroactive ligand receptor interaction(P＜0.05).The GSEA results showed that high expression of CALU may promote the G1-S phase transition of the cell cycle,ubiquitination protein polymerization and HCC progression,while low expression of CALU may activate oxidative stress,ferroptosis and cuproptosis in HCC cells.Single-cell sequencing analysis results showed that the expression level of CALU mRNA was significantly increased in HCC cells with advanced tumor stages.HCC_CALU_High cell subset was mainly related to ubiquitination,p53 and cell cycle(P＜0.01),and HCC_CALU_Low cell subset was mainly related to oxidative stress,ferroptosis,and histone binding(P＜0.01).Conclusion:The high expression of CALU may be related to the poor prognosis of HCC patients.The constructed nomogram of HCC prognosis shows favourable effect in predicting the survival rate of the HCC patients.The up-regulation of CALU may promote HCC progression by regulating the G1-S phase of the cell cycle and ubiquitination of protein,while down-regulation of CALU may inhibit HCC progression by inducing oxidative stress,ferroptosis and cuproptosis in cells.

Objective To evaluate the efficacy and safety of ozone injection therapy for lumbar disc hemiation(LDH).Methods A computerized retrieval of academic papers concerning the randomized controlled trial(RCT)on ozone injection therapy for LDH from the databases of Embase,PubMed,Cochrane library and Web of science was conducted.The retrieval time period was from the establishment of the database to February 2023.The literature retrieval,screening,and data extraction were independently performed by two researchers.Cochrane bias risk assessment tool was used to assess the quality of the included literature.Stata 17.0 software was used to make meta-analysis.Results A total of 9 RCTs including 702 patients were finally included in this study.The results of meta-analysis showed that compared with radiofrequency thermocoagulation,percutaneous rotation and other treatments for LDH,the combination use of ozone injection could signifiicantly improve the effective rate based on Macnab efficacy evaluation criteria(RR=1.097,95％CI:1.038～1.159,P=0.001)and the excellent rate(RR=1.185～95％CI:1.074～1.309,P=0.001),and decrease the visual analog scale(VAS)pain score(WMD=-0.810～95％CI:-1.205～-0.414,P=0.000),and the differences in the above indexes were statistically significant.Conclusion Compared with the simple use of radiofrequency thermocoagulation,percutaneous rotation,and other treatment for LDH,the combination use of ozone injection therapy can significantly improve the effective rate and excellent rate based on Macnab efficacy evaluation criteria,decrease VAS score,with a high clinical safety.Limited by the quantity and quality of the original studies included in this study,the above conclusions need to be further verified by multi-center,large-sample and high-quality studies.

Objective:To establish a TAL gel-clot bacterial endotoxin test for aluminum hydroxide adjuvant.Methods:According to the bacterial endotoxin test in general chapter 1143 of the Chinese Pharmacopoeia 2020,pre-interference test was performed using 3 types of buffer solutions and Ca-Mg additive by orthogonal design.The interference test and sample preparation validation were carried out using different batches of aluminum hydroxide adjuvant from 2 TAL manufacturers.Results:No interference was obtained through pre-interference test.Under the validated conditions,aluminum hydroxide adjuvant combined with phosphate buffered saline and Ca-Mg additive was employed to remove interference in bacteria endotoxin test.Conclusion:The established TAL gel-clot method is applicable to bacterial endotoxin test for aluminum hydroxide adjuvant.

Objective:To preliminarily investigate the applicability of a poliovirus pseudovirus-based neutralization assay in evaluating the immunogenicity of recombinant poliovirus vaccines and their in vivo potency in rats. Methods:Serum samples from rats immunized with recombinant poliovirus vaccines were tested using both the pseudovirus neutralization assay and the live-virus neutralization assay with Sabin strain. The consistency and correlation of the two methods were analyzed using the Kappa test and Spearman′s rank correlation.Results:For the neutralizing antibodies against typeⅠ, Ⅱ, and Ⅲ polioviruses, the Kappa values for consistency analysis of the two methods were 0.914, 1.000, and 0.751, respectively ( P<0.001), and the correlation coefficients ( R values) were 0.833, 0.927, and 0.859, respectively ( P<0.001). Conclusions:The test results of the two methods are consistent and show a good correlation, indicating that the pseudovirus neutralization assay can be applied to evaluating the immunogenicity of poliovirus vaccines and also can be used in rat potency tests.

Objective:To systematically evaluate the prevalence of norovirus causing sporadic acute gastroenteritis in China.Methods:Relevant articles on acute gastroenteritis caused by norovirus in China published between January 2010 and October 2023 were retrieved from Wanfang, CNKI and PubMed database. The articles met inclusion and exclusion criteria were enrolled in this study. Excel software and SPSS20.0 software were used for statistical analysis. The epidemiological features of sporadic cases of acute gastroenteritis caused by norovirus in China were summarized using descriptive statistical analysis.Results:A total of 500 articles were included in this study, involving 784 486 cases of acute gastroenteritis and 670 292 samples in 32 provinces and regions. Norovirus GⅡ was the predominant genogroup causing acute gastroenteritis in China in recent years, but there were significant differences in the distribution of genotypes and epidemic strains at different times. GⅡ.4 was the predominant genotype in each year, and GⅡ.4/2006b and GⅡ.4 /Sydney_2012 were the main epidemic strains. Norovirus-related diarrhea occurred throughout the year, especially between the months of October and December. The incidence of norovirus infection was high in children under five years old and varied in different regions.Conclusions:Norovirus GⅡ was the predominant genogroup causing norovirus-related sporadic acute gastroenteritis in China, but there was an obvious genetic evolutionary trend in the epidemic strains. Factors such as epidemic strains, season and geographical region should be considered when making strategies for the prevention and control of norovirus-related diarrhea and developing vaccines.

Objective: To evaluate the caries preventive effect and economic outcome of an oral health promotion model in rural primary schools in Guangxi Zhuang Autonomous Region. Methods: In November 2015, a baseline survey was conducted on the caries prevalence of rural first graders in program areas, and then a program intervention was carried out on the first graders of target primary schools. The intervention methods included the distribution of oral health care products, oral health education, teacher training, etc. In 2020, the method of cluster random sampling was adopted to include the sixth graders who received the program intervention as first graders and were still available in 2020 as the intervention group. During this period, the sixth graders transferred from foreign schools who failed to receive the program intervention as first graders were included in the non-intervention group. The caries preventive effect was evaluated by analyzing the caries epidemiological data of 2 652 children aged 11-13 years, comparing the intervention and non-intervention group. The cost of the program was estimated by the structural analysis method. A decision tree model was established by TreeAge pro2019 and incorporated into the effect parameters and cost parameters for cost-effectiveness analysis and sensitivity analysis. Results : The prevalence of caries in the intervention group was 54.8%, and the mean DMFT was 1.36 ± 1.64, both of which were lower than those in the non-intervention group. The difference in prevalence and mean DMFT between the two groups was statistically significant (P < 0.05). It costs CNY 319.83 per child to reduce suffering from caries. The number of patients with caries in the intervention group was the most sensitive indicator of an economic effect. The probability of a cost-effectiveness advantage for the program was 92.2%. Conclusion This oral health promotion model used in rural schools demonstrated a caries preventive effect that was very likely economically advantageous. It is of practical significance to improve and promote this model in the Guangxi Zhuang Autonomous Region.

Lactic acid bacteria (LAB) are a representative probiotic. As the dominant flora in the human intestinal tract, LAB can regulate the balance of human intestinal flora and improve host health. The purpose of this study was to isolate and screen LAB that are well suited to the intestinal characteristics of the Chinese population, with excellent probiotics and high antibacterial activity. After 16S ribosomal RNA (rRNA) homology and phylogenetic tree analysis, potential probiotics were tested for their antibacterial activity, resistance to artificial gastrointestinal fluid and drugs, surface hydrophobicity, and safety. Three strains of LAB with acid resistance, bile salt resistance, epithelial cell adhesion, and no multidrug resistance were selected:

Objective:To retrospectively analyze the prevalence and genotype distribution of human papillomavirus (HPV) among women in mainland China from the introduction of cervical cancer vaccine to its marketing in China, so as to provide data for comparing HPV prevalence after the application and gradual promotion of HPV vaccine.Methods:Related articles published during 2006 to 2016 were retrieved from PubMed, China National Knowledge Infrastructure (CNKI) and Wanfang databases. Data about HPV infection in women receiving physical examination or routine cervical cancer screening and female outpatients were collected and analyzed.Result:A total of 270 articles were retrieved, in which the sampled women were aged from 13 to 90 years old. Seventy-two articles were about HPV infection in women screened for cervical cancer with a sample size of 580 308 people. The overall results showed that during the 11 years, the five most common high-risk HPV genotypes were HPV52, 16, 58, 68 and 18, and the common low-risk HPV genotypes were HPV81, 43 and 6. The distribution of HPV genotypes varied in different regions, but HPV16, 52 and 58 were always the predominant genotypes. Among the 270 collected articles, 200 analyzed HPV infection in female outpatients with a total of 675 035 cases. The results showed that the top five high-risk HPV genotypes with high prevalence were HPV16, 52, 58, 18 and 33, and the common low-risk types were HPV6, 11 and 81. The analysis of HPV infection rate in seven geographical regions indicated that, although the distribution of the commonly circulating high-risk HPV genotypes varied in different regions, HPV16, 52 and 58 remained the predominant genotypes.Conclusions:The present study showed that the prevalence and genotype distribution of HPV infection varied by region, but HPV16, 52 and 58 were always the most common high-risk genotypes and HPV6 was the predominant low-risk genotype in both women screened for cervical cancer and female outpatients. Besides that, the infections of other high-risk genotypes, such as HPV53 and 68, should also be of concern.

Objective To investigate the physicochemical properties and immunogenicity of virus like particles(VLPs) in two different conformations assembled from the essential capsid protein VP1 of GⅡ.4 norovirus(NoV) in Hansenula polymorpha. Methods NoV GⅡ.4 VLPs in two different conforma-tions were prepared from high-density fermentation of recombinant engineered strains and VLPs purification. Physicochemical properties of the two forms of VLPs were identified by Western blot,size-exclusion high per-formance liquid chromatography (SEC-HPLC), dynamic light scattering(DLS) and transmission electron microscopy. Serum VLPs binding activities and blocking activities against VLPs binding to histo-blood group antigen(HBGA-VLPs) were evaluated after immunization of BALB/c mice with the two forms of VLPs. Re-sults VLPs of two different diameters with high homogeneity were obtained after purification. DLS results showed that particle sizes of two VLPs were 53.98 nm and 45.18 nm,respectively. The two VLPs were sim-ilar in binding abilities to HBGA receptors. Serum VLPs binding activities and blocking activities against HBGA-VLPs were found higher in NoV-VLP-L than NoV-VLP-S,but the difference was not statistically sig-nificant (P>0.05). Conclusion VLPs in two different conformations were obtained by expressing NoV GⅡ.4 VP1 proteins in Hansenula polymorpha. Though they were similar in physicochemical properties and immunogenicity,the NoV-VLP-L might be potential antigen candidates for the development of recombinant human norovirus vaccine.