Objective: To evaluate the immunogenicity and safety of co-administration with bivalent human papillomavirus (HPV) vaccine and hepatitis E virus (HEV) vaccine, so as to provide reference for optimizing the vaccination schedule. Methods: Females aged 18 to 25 years were recruited from September to October 2021 in Hengdian College of Film & Television in Zhejiang Province and randomly divided into the HPV+HEV group, the HPV group, and the HEV group. The vaccination procedures were one dose each at 0, 1, and 6 months. Immunogenicity was evaluated by detecting the geometric mean titers (GMT) of HPV16 IgG, HPV18 IgG, and/or HEV IgG antibodies before the first vaccination and one month after the full course of immunization, and comparing the difference in seroconversion, and the GMT ratio. The non-inferiority margin was set at a seroconversion difference of ≤5%, and the lower limit of the 95%CI of the GMT ratio was >0.5. Safety was evaluated by collecting conjunctive local reactions/events and systemic reactions/events within 7 days after each dose, non-conjunctive adverse events within 30 days after each dose, and serious adverse events throughout the observation period (0 to 7 months). Results: A total of 240 females were included, among whom 236 completed the full vaccination program, including 79 in the HPV+HEV group, 77 in the HPV group, and 80 in the HEV group. One month after the full course of immunization, the seroconversion rates of HPV16 IgG and HPV18 IgG antibodies in both the HPV+HEV group and the HPV group were 100%, and the differences in seroconversion rates were 0 (95%CI: -3.39%-+∞). The seroconversion rates of HEV IgG antibodies in both the HPV+HEV group and the HEV group were 100%, and the difference in seroconversion rates was 0 (95%CI: -3.27%-+∞). The GMT of HPV16 IgG and HPV18 IgG antibodies in the HPV+HEV group was 393.88 and 284.86 IU/mL respectively, which was not inferior to 489.39 and 341.24 IU/mL in the HPV group, and the GMT ratios were 0.80 (95%CI: 0.66-+∞) and 0.83 (95%CI: 0.68-+∞), respectively. The GMT of HEV IgG in the HPV+HEV group was 13.55 U/mL, which was not inferior to 12.72 U/mL in the HEV group, and the GMT ratio was 1.07 (95%CI: 0.92-+∞). The incidences of pain, pruritus, and induration in the HPV+HEV group were 54.43%, 21.52% and 40.51% respectively, which were significantly higher than 10.39%, 0, and 0 in the HPV group (all P<0.05). The incidences of redness/swelling, muscle pain/general weakness in the HPV+HEV group were 2.53% and 0, respectively, which were significantly lower than 12.50% and 16.25% in the HEV group (both P<0.05). Conclusion The co-administration of the bivalent HPV vaccine and HEV vaccine is not inferior to individual vaccination in terms of immunogenicity and safety, and the vaccination plan can be optimized through co-administration.

Objectives:To investigate the prognostic value of the CHA2DS2-VASc and R2CHA2DS2-VASc scores in patients with atrial fibrillation(AF)and heart failure(HF).Methods:Patients with AF and HF from hospitals diagnosed by the Heart Failure Center in Guangdong Province between January 2017 and December 2021 were selected.Major adverse cardiovascular events(MACE)were used as the follow-up endpoint.Statistical methods such as the area under the receiver operating characteristic(ROC)curve(AUC),net reclassification index(NRI),and integrated discrimination improvement(IDI)were applied to evaluate the predictive value of the CHA2DS2-VASc and R2CHA2DS2-VASc scores in patients with AF and HF.Results:A total of 1 839 patients were enrolled in this study,comprising 703 patients in the MACE group and 1 136 patients in the non-MACE group.Compared with the non-MACE group,the MACE group exhibited significantly advanced age,higher prevalence of New York Heart Association class Ⅳ and coronary artery disease,lower diastolic blood pressure and estimated glomerular filtration rate levels,and elevated serum N-terminal pro-B-type natriuretic peptide concentrations(all P<0.05).Additionally,significantly lower proportions of patients in the MACE group received angiotensin-converting enzyme inhibitors/angiotensin receptor blockers/angiotensin receptor-neprilysin inhibitors,beta-blockers,mineralocorticoid receptor antagonists,or anticoagulant therapy(all P<0.05).Multivariable logistic regression analysis revealed that each 1-point increment in both CHA2DS2-VASc and R2CHA2DS2-VASc scores was associated with approximately 10%increased risk of MACE.ROC curve analysis demonstrated that the AUC values for predicting MACE in AF patients with HF were 0.555(95%CI:0.528-0.582,P<0.001)for CHA2DS2-VASc and 0.576(95%CI:0.549-0.608,P<0.001)for R2CHA2DS2-VASc,indicating marginally superior discriminatory capacity of the R2CHA2DS2-VASc score.Delong's test confirmed statistically significant differences between the two scoring systems(P=0.001).The R2CHA2DS2-VASc score demonstrated a NRI of 0.259(95%CI:0.166-0.352,P<0.001)and an IDI of 0.007(95%CI:0.005-0.010,P<0.001)compared with the conventional CHA2DS2-VASc score.Although the R2CHA2DS2-VASc score exhibited slightly better predictive accuracy and outcome discrimination capacity than the original scoring system,both scores demonstrated suboptimal clinical predictive performance.Conclusions:Both the R2CHA2DS2-VASc and CHA2DS2-VASc scores show suboptimal performance for predicting the risk of MACE in patients with AF and HF,and the predicting performance of R2CHA2DS2-VASc score is marginally superior to CHA2DS2-VASc score in this patient cohort.

Objective:To investigate the distribution of CGG repeat numbers in the FMR1 gene among reproductive-age women in China, providing data reference for carrier screening and genetic counseling of Fragile X syndrome. Methods:This cross-sectional study recruited 16 610 reproductive-age women from 12 medical institutions between July 2022 and October 2023. Peripheral venous blood samples (3 mL) were collected, and genomic DNA was extracted. The number of CGG repeats in the FMR1 gene was determined using the triplet-primed polymerase chain reaction (TP-PCR) combined with capillary electrophoresis technology. Statistical analyses were performed to assess the prevalence and distribution of CGG repeat expansions. Results:Among 16 610 women of childbearing age, 5 684 (34.220%) women had the same number of CGG repeats in the two alleles of FMR1 gene, and 10 926 (65.780%) women had different numbers of repeats in the two alleles. Among the 33 220 FMR1 alleles in 16 610 women of reproductive age, the most common CGG repeat numbers were 29 [48.645% (16 160/33 220)] and 30 [26.276% (8 729/33 220)], while the most frequent CGG genotype was CGG 29/29 [24.726% (4 107/16 610)]. The CGG repeat numbers of FMR1 gene were normal in 16 498 women (99.326%). Among the 112 women (0.674%) with CGG repeat abnormities, 96 (0.578%) women were classified as intermediate carriers, 15 (0.090%) as premutation carriers, and 1 (0.006%) as a full mutation carrier, whose CGG genotype was (36, >200). Conclusion:In the general reproductive-age female population in China, the normal CGG repeat numbers of the FMR1 gene account for 99.326%, while the intermediate carrier rate is 0.578%, and the combined carrier rate of the premutation and full mutation types is 0.096%.

Objective To explore the potential mechanism of Gegen qinlian decoction(GGQLD)containing serum in ameliorating nonalcoholic fatty liver disease(NAFLD)in human hepatocellular carcinoma HepG2 cells based on transcriptomics.Methods An in vitro model of NAFLD was constructed by free fatty acid(FFA)-induced fat accumulation in HepG2 cells,and cells were treated with different proportions of GGQLD and pioglitazone-containing serum.The lipid deposition in each group was detected by oil red O staining,and the lipid content in each group was evaluated by triglyceride level.Transcriptome technology was used to detect the differentially expressed genes between the intervention groups,and GO annotation analysis,KEGG enrichment analysis and protein interaction(PPI)network analysis were performed to verify the differentially expressed genes by RT-PCR.Results Compared with normal control group,the number of red lipid droplets in the model control group increased,and the triglyceride content increased significantly(P＜0.01).Compared with model control group,the content of red lipid droplets in the GGQLD medium dose group showed a decreasing trend,and the intracellular triglyceride content decreased significantly(P＜0.05).A total of 608 differentially expressed genes were identified by transcriptome analysis,of which 163 differentially expressed genes were up-regulated and 445 differentially expressed genes were down-regulated.GO enrichment analysis showed that the differentially expressed genes were mainly involved in the regulation of MAP kinase phosphatase activity.KEGG analysis showed that the differentially expressed genes were mainly involved in MAPK signaling pathway.RT-PCR results showed that GGQLD up-regulated the expression level of MAP2K6 mRNA and down-regulated the expression levels of FOSL1,CTSL,DUSP5,DUSP1,JUN,HSPA6,IL1A,IL11 and RELB mRNA,which may be mainly involved in MAPK signaling pathway.Conclusion GGQLD has the effect of improving NAFLD,which may be related to MAPK signaling pathway.

Objective:To investigate the distribution of CGG repeat numbers in the FMR1 gene among reproductive-age women in China, providing data reference for carrier screening and genetic counseling of Fragile X syndrome. Methods:This cross-sectional study recruited 16 610 reproductive-age women from 12 medical institutions between July 2022 and October 2023. Peripheral venous blood samples (3 mL) were collected, and genomic DNA was extracted. The number of CGG repeats in the FMR1 gene was determined using the triplet-primed polymerase chain reaction (TP-PCR) combined with capillary electrophoresis technology. Statistical analyses were performed to assess the prevalence and distribution of CGG repeat expansions. Results:Among 16 610 women of childbearing age, 5 684 (34.220%) women had the same number of CGG repeats in the two alleles of FMR1 gene, and 10 926 (65.780%) women had different numbers of repeats in the two alleles. Among the 33 220 FMR1 alleles in 16 610 women of reproductive age, the most common CGG repeat numbers were 29 [48.645% (16 160/33 220)] and 30 [26.276% (8 729/33 220)], while the most frequent CGG genotype was CGG 29/29 [24.726% (4 107/16 610)]. The CGG repeat numbers of FMR1 gene were normal in 16 498 women (99.326%). Among the 112 women (0.674%) with CGG repeat abnormities, 96 (0.578%) women were classified as intermediate carriers, 15 (0.090%) as premutation carriers, and 1 (0.006%) as a full mutation carrier, whose CGG genotype was (36, >200). Conclusion:In the general reproductive-age female population in China, the normal CGG repeat numbers of the FMR1 gene account for 99.326%, while the intermediate carrier rate is 0.578%, and the combined carrier rate of the premutation and full mutation types is 0.096%.

Objectives:To investigate the prognostic value of the CHA2DS2-VASc and R2CHA2DS2-VASc scores in patients with atrial fibrillation(AF)and heart failure(HF).Methods:Patients with AF and HF from hospitals diagnosed by the Heart Failure Center in Guangdong Province between January 2017 and December 2021 were selected.Major adverse cardiovascular events(MACE)were used as the follow-up endpoint.Statistical methods such as the area under the receiver operating characteristic(ROC)curve(AUC),net reclassification index(NRI),and integrated discrimination improvement(IDI)were applied to evaluate the predictive value of the CHA2DS2-VASc and R2CHA2DS2-VASc scores in patients with AF and HF.Results:A total of 1 839 patients were enrolled in this study,comprising 703 patients in the MACE group and 1 136 patients in the non-MACE group.Compared with the non-MACE group,the MACE group exhibited significantly advanced age,higher prevalence of New York Heart Association class Ⅳ and coronary artery disease,lower diastolic blood pressure and estimated glomerular filtration rate levels,and elevated serum N-terminal pro-B-type natriuretic peptide concentrations(all P<0.05).Additionally,significantly lower proportions of patients in the MACE group received angiotensin-converting enzyme inhibitors/angiotensin receptor blockers/angiotensin receptor-neprilysin inhibitors,beta-blockers,mineralocorticoid receptor antagonists,or anticoagulant therapy(all P<0.05).Multivariable logistic regression analysis revealed that each 1-point increment in both CHA2DS2-VASc and R2CHA2DS2-VASc scores was associated with approximately 10%increased risk of MACE.ROC curve analysis demonstrated that the AUC values for predicting MACE in AF patients with HF were 0.555(95%CI:0.528-0.582,P<0.001)for CHA2DS2-VASc and 0.576(95%CI:0.549-0.608,P<0.001)for R2CHA2DS2-VASc,indicating marginally superior discriminatory capacity of the R2CHA2DS2-VASc score.Delong's test confirmed statistically significant differences between the two scoring systems(P=0.001).The R2CHA2DS2-VASc score demonstrated a NRI of 0.259(95%CI:0.166-0.352,P<0.001)and an IDI of 0.007(95%CI:0.005-0.010,P<0.001)compared with the conventional CHA2DS2-VASc score.Although the R2CHA2DS2-VASc score exhibited slightly better predictive accuracy and outcome discrimination capacity than the original scoring system,both scores demonstrated suboptimal clinical predictive performance.Conclusions:Both the R2CHA2DS2-VASc and CHA2DS2-VASc scores show suboptimal performance for predicting the risk of MACE in patients with AF and HF,and the predicting performance of R2CHA2DS2-VASc score is marginally superior to CHA2DS2-VASc score in this patient cohort.

Objective To explore the potential mechanism of Gegen qinlian decoction(GGQLD)containing serum in ameliorating nonalcoholic fatty liver disease(NAFLD)in human hepatocellular carcinoma HepG2 cells based on transcriptomics.Methods An in vitro model of NAFLD was constructed by free fatty acid(FFA)-induced fat accumulation in HepG2 cells,and cells were treated with different proportions of GGQLD and pioglitazone-containing serum.The lipid deposition in each group was detected by oil red O staining,and the lipid content in each group was evaluated by triglyceride level.Transcriptome technology was used to detect the differentially expressed genes between the intervention groups,and GO annotation analysis,KEGG enrichment analysis and protein interaction(PPI)network analysis were performed to verify the differentially expressed genes by RT-PCR.Results Compared with normal control group,the number of red lipid droplets in the model control group increased,and the triglyceride content increased significantly(P＜0.01).Compared with model control group,the content of red lipid droplets in the GGQLD medium dose group showed a decreasing trend,and the intracellular triglyceride content decreased significantly(P＜0.05).A total of 608 differentially expressed genes were identified by transcriptome analysis,of which 163 differentially expressed genes were up-regulated and 445 differentially expressed genes were down-regulated.GO enrichment analysis showed that the differentially expressed genes were mainly involved in the regulation of MAP kinase phosphatase activity.KEGG analysis showed that the differentially expressed genes were mainly involved in MAPK signaling pathway.RT-PCR results showed that GGQLD up-regulated the expression level of MAP2K6 mRNA and down-regulated the expression levels of FOSL1,CTSL,DUSP5,DUSP1,JUN,HSPA6,IL1A,IL11 and RELB mRNA,which may be mainly involved in MAPK signaling pathway.Conclusion GGQLD has the effect of improving NAFLD,which may be related to MAPK signaling pathway.

Abstract: Objective　To evaluate the application of variable number tandem repeats (VNTR) and whole genome sequencing (WGS) in tracing the transmission of Mycobacterium tuberculosis in a tuberculosis outbreak in school, and explore the roles of these genetic methodologies in addressing tuberculosis transmission in school environments. Methods Identification of mycobacterial strains, spacer oligonucleotide typing (Spoligotyping), VNTR, and WGS were performed on six Mycobacterium tuberculosis strains obtained from TB cases in the first multidrug-resistant tuberculosis outbreak happened in a school in Beijing. Genotyping characteristics were analyzed to identify the transmission chain. Results The 6 culture-positive strains involved 6 students from 4 classes across 2 grades. One of them was the first case, three were close contacts of the first case, and the other two were general contacts. All 6 strains were identified as Mycobacterium tuberculosis. Spoligotyping results indicated that 5 of the strains were of the Beijing genotype; the other one had no result. VNTR genotyping divided the 6 strains into 3 clusters with a clustering rate of 66.7%. The largest one of the 3 clusters contained 4 strains with the same genotype, indicating a significant level of recent transmission. The remaining two strains, differing by 1.0-1.6 copies at 1-2 loci from the other four strains, were identified as individual strains. The results of WGS showed that the genomic SNP differences among the 6 strains were greater than 12 SNPs. According to the molecular biology identification criteria of this study, the strains exhibited significant heterogeneity with no homology. Conclusions WGS offers higher accuracy and advantages over VNTR genotyping in evaluating the recent transmission of tuberculosis. WGS can more accurately characterize recent TB transmission in the case of TB outbreaks in schools, especially when there are drug-resistant TB cases, and should be used as a supplement to traditional epidemiology.

Objective:To report the clinical and genetic characteristics of autosomal recessive spastic ataxia of Charlevoix-Saguenay (ARSACS) induced by a new homozygous mutation in the SACS gene, and to improve the clinicians′ recognition of the disease. Methods:Detailed nervous system physical examination was performed on the patient and his parents from a consanguineous family admitted to the Genetics and Metabolism Clinic of the Department of Neurology, the First Affiliated Hospital of Kunming Medical University in March 2022. The peripheral blood DNA of the patient and his parents was extracted, and whole exon sequencing (WES) was performed on the patient and his parents using second-generation sequencing technology. The mutation sites were verified by Sanger sequencing, and the mutation sites were analyzed by software.Results:The 18-year-old Han ethnic male patient developed a progressive stiffness of his bilateral lower limbs and gait unsteadiness since the age of 3. He had pyramidal tract sign in his bilateral lower limbs, cerebellar ataxia, pes cavus and hammer toes. Brain magnetic resonance imaging (MRI) showed symmetrical low signal of bilateral pons, cerebellar atrophy and thinning of corpus callosum in T 2WI and T 2 fluid attenuated inversion recovery (FLAIR) sequences. Neuroelectrophysiological examination showed sensory motor peripheral neuropathy. Ophthalmic examination revealed concomitant exotropia and ametropia in both eyes. WES revealed a homozygous variant of c.6958T>C (p.Tyr2320His) in exon 10 of the SACS gene of the patient, and his parents were heterozygous variant carriers confirmed by Sanger sequencing. The variant was classified as possibly pathogenic (PM1+PM2+PP3+PP4) according to the American Society for Medical Genetics and Genomics. The patient was clearly diagnosed as ARSACS caused by homozygous mutation of c.6958T>C in the SACS gene. Conclusions:A novel pathogenic variant (c.6958T>C) in the SACS gene identified in this study leads to the manifestation of ARSACS. The primary clinical manifestations include cerebellar ataxia, pyramidal tract signs, and sensorimotor peripheral neuropathy. Head MRI examination of T 2WI and T 2FLAIR sequences with symmetrical low signal on both sides of the pons helps to narrow down the scope of differential diagnosis.

Objective To investigate the epidemiological characteristics and mutation spectrum of monogenic diseases in Chinese population through a large-scale,multicenter carrier screening.Methods This study was conducted among a total of 33 104 participants(16 610 females)from 12 clinical centers across China.Carrier status for 223 genes was analyzed using high-throughput sequencing and different PCR methods.Results The overall combined carrier frequency was 55.58%for 197 autosomal genes and 1.84%for 26 X-linked genes in these participants.Among the 16 669 families,874 at-risk couples(5.24%)were identified.Specifically,584 couples(3.50%)were at risk for autosomal genes,306(1.84%)for X-linked genes,and 16 for both autosomal and X-linked genes.The most frequently detected autosomal at-risk genes included GJB2(autosomal recessive deafness type 1A,393 couples),HBA1/HBA2(α-thalassemia,36 couples),PAH(phenylketonuria,14 couples),and SMN1(spinal muscular atrophy,14 couples).The most frequently detected X-linked at-risk genes were G6PD(G6PD deficiency,236 couples),DMD(Duchenne muscular dystrophy,23 couples),and FMR1(fragile X syndrome,17 couples).After excluding GJB2 c.109G>A,the detection rate of at-risk couples was 3.91%(651/16 669),which was lowered to 1.72%(287/16 669)after further excluding G6PD.The theoretical incidence rate of severe monogenic birth defects was approximately 4.35‰(72.5/16 669).Screening for a battery of the top 22 most frequent genes in the at-risk couples could detect over 95%of at-risk couples,while screening for the top 54 genes further increased the detection rate to over 99%.Conclusion This study reveals the carrier frequencies of 223 monogenic genetic disorders in the Chinese population and provides evidence for carrier screening strategy development and panel design tailored to the Chinese population.In carrier testing,genetic counseling for specific genes or gene variants can be challenging,and the couples need to be informed of these difficulties before testing and provided with options for not screening these genes or gene variants.