Disease-Associated Streptococcus pneumoniae Genetic Variation.

Streptococcus pneumoniae is an opportunistic pathogen that causes substantial illness and death among children worldwide. The genetic backgrounds of pneumococci that cause infection versus asymptomatic carriage vary substantially. To determine the evolutionary mechanisms of opportunistic pathogenicity, we conducted a genomic surveillance study in China. We collected 783 S. pneumoniae isolates from infected and asymptomatic children. By using a 2-stage genomewide association study process, we compared genomic differences between infection and carriage isolates to address genomic variation associated with pathogenicity. We identified 8 consensus k-mers associated with adherence, antimicrobial resistance, and immune modulation, which were unevenly distributed in the infection isolates. Classification accuracy of the best k-mer predictor for S. pneumoniae infection was good, giving a simple target for predicting pathogenic isolates. Our findings suggest that S. pneumoniae pathogenicity is complex and multifactorial, and we provide genetic evidence for precise targeted interventions.

Focused ultrasound: a novel therapy for improving vaginal microecology in patients with high-risk HPV infection.

OBJECTIVES: To investigate changes in vaginal microecology in women with high-risk human papillomavirus (HR-HPV) infection after focused ultrasound (FU) treatment. MATERIALS AND METHODS: We collected vaginal secretions at the time of admission and 3 months after FU treatment from 169 women who received FU treatment for cervical HR-HPV infection between July 2020 and September 2022. Among them, there were 101 patients with cute vaginitis, we also collected their vaginal secretions after one week of drug treatment. These samples were evaluated for vaginal microecology and HPV-DNA examination. RESULTS: Of the 169 patients, 101 (59.7%) suffered from acute vaginitis at the time of admission. After one week of targeted antibiotics drug treatment, there were no pathogens or pus cells on the field of microscopic vision, but there was no significant difference(p > 0.05) in the diversity and density of vaginal flora, the proportion and function of Lactobacillus (H2O2 negative rate) between one week after treatment and at the time of admission. At the time of admission of the 169 patients, the normal flora rate was 40.3%, which increased to 93.5% three months after FU treatment. The differences in vaginal secretion parameters at the time of admission and 3 months after FU treatment were as follows: H2O2 negative rate (37.3% vs. 3.6%), leukocyte esterase positive rate (54.4% vs. 5.9%), sialidase positive rate (38.5% vs. 4.1%), bacterial vaginitis positive rate (55% vs. 4.7%), fungal vaginitis positive rate (44.4% vs. 5.9%), and trichomonal vaginitis positive rate (7.1% vs. 0). The difference was statistically significant (p < 0.01). The pH value and Nagent score at the time of admission were significantly higher than those three months after FU. Three months after FU, the positive rate of HPV was 5.8% in the group of patients with normal vaginal microecology at the time of admission and post-FU; it was 6.7% in the group of patients with abnormal vaginal microecology at the time of admission and normal vaginal microecology post-FU; and it was 100% in the group patients with abnormal vaginal microecology at the time of admission and post-FU. A significant difference was observed among the three groups (p < 0.01). CONCLUSION: FU is an effective treatment for patients with cervical HR-HPV infection. FU does not interfere with the vaginal microecology of HR-HPV positive patients with normal vaginal microecology.FU followed by antibiotic drug therapy for pathogens is beneficial to restore the function of Lactobacillus vaginalis in HR-HPV positive patients with acute vaginitis,so as to improve the vaginal microecology of HR-HPV positive patients with abnormal vaginal microecology.

[Screening of pathogenic molecular markers of Staphylococcus aureus in children based on whole genome sequencing technology]. / åŸºäºŽå ¨åŸºå› ç»„æµ‹åºæŠ€æœ¯ç­›é€‰å„¿ç«¥é‡‘é»„è‰²è‘¡è„çƒèŒçš„è‡´ç— ç›¸å ³åˆ†å­æ ‡å¿—ç‰©.

OBJECTIVES: To explore the molecular characteristics of Staphylococcus aureus (S. aureus) in children, and to compare the molecular characteristics of different types of strains (infection and colonization strains) so as to reveal pathogenic molecular markers of S. aureus. METHODS: A cross-sectional study design was used to conduct nasopharyngeal swab sampling from healthy children in the community and clinical samples from infected children in the hospital. Whole genome sequencing was used to detect antibiotic resistance genes and virulence genes. A random forest method to used to screen pathogenic markers. RESULTS: A total of 512 S. aureus strains were detected, including 272 infection strains and 240 colonization strains. For virulence genes, the carrying rates of enterotoxin genes (seb and sep), extracellular enzyme coding genes (splA, splB, splE and edinC), leukocytotoxin genes (lukD, lukE, lukF-PV and lukS-PV) and epidermal exfoliating genes (eta and etb) in infection strains were higher than those in colonization strains. But the carrying rates of enterotoxin genes (sec, sec3, seg, seh, sei, sel, sem, sen, seo and seu) were lower in infection strains than in colonization strains (P<0.05). For antibiotic resistance genes, the carrying rates of lnuA, lnuG, aadD, tetK and dfrG were significantly higher in infection strains than in colonization strains (P<0.05). The accuracy of cross-validation of the random forest model for screening pathogenic markers of S. aureus before and after screening was 69% and 68%, respectively, and the area under the curve was 0.75 and 0.70, respectively. The random forest model finally screened out 16 pathogenic markers (sem, etb, splE, sep, ser, mecA, lnuA, sea, blaZ, cat(pC233), blaTEm-1A, aph(3')-III, ermB, ermA, ant(9)-Ia and ant(6)-Ia). The top five variables in the variable importance ranking were sem (OR=0.40), etb (OR=3.95), splE (OR=1.68), sep (OR=3.97), and ser (OR=1.68). CONCLUSIONS: The random forest model can screen out pathogenic markers of S. aureus and exhibits a superior predictive performance, providing genetic evidence for tracing highly pathogenic S. aureus and conducting precise targeted interventions.

Molecular epidemiology of Streptococcus pneumoniae isolated from pediatric community-acquired pneumonia in pre-conjugate vaccine era in Western China.

BACKGROUND: Streptococcus pneumoniae (S. pneumoniae) is one of the most common pathogens which can cause morbidity and mortality in pediatric infections worldwide. This study aimed to describe the phenotypic and molecular characteristics of community-acquired pneumonia (CAP)-causing S. pneumoniae recovered from children in Western China. METHODS: We retrospectively enrolled pediatric patients younger than 5 years diagnosed with CAP. All 419 S. pneumoniae isolates were tested for antibiotic susceptibility, serotypes, virulence genes, resistance genes, and sequence types. The potential relationships between molecular characteristics were tested by correspondence analysis. RESULTS: Most of S. pneumoniae isolates were resistant to erythromycin, tetracycline, clindamycin and trimethoprim-sulfamethoxazole, with 93.8% isolates classified as multidrug resistant. The dominant STs were ST271 (30.8%) and ST320 (12.2%), while the prevailing serotypes were 19F (46.8%), 6B (11.5%), 23F (9.5%) and 19A (9.3%). The coverage rates of PCV-7 and PCV-13 were 73.03% and 86.16%, while the coverage rates of PCV13 among children aged < 1 year and 1-2 years were high in 93.18% and 93.62%. We also observed that CC271 expressed more of mef (A/E), lytA, rlrA and sipA than non-CC271 isolates. Moreover, there were strong corresponding relationships between molecular characteristics. CONCLUSIONS: The high coverage rate of PCV13 suggests the necessity of introducing the PCV13 vaccine in Western China. Our findings underscore the value of monitoring multiple molecular characteristics to provide new guidance for developing future pneumococcal vaccines.

Comparison of Three Immunoassay Systems for Determining Serum Anti-Mullerian Hormone.

BACKGROUND: Serum anti-Mullerian hormone (AMH) is a dimeric glycoprotein of the transforming growth factor-ß (TGF-ß) family of growth and serves as a key biomarker of folliculogenesis as well as steroidogenesis within ovaries. Cobas E601, UniCel DxI 800 and iFlash3000 are automated immunoassay systems which are widely used to measure serum AMH levels in the blood. However, the correlations of serum AMH measured by the three automated immunoassay systems are not well recognized, the aim of this study is to compare the performance of the three automated immunoassay systems and record the correlation of serum AMH concentrations. METHODS: Serum AMH concentrations were measured in 100 serum samples using the UniCel DxI 800, Cobas E601, and iFlash 3000 automated immunoassay systems on the same day. Passing-Bablok regression analysis and Bland-Altman plot analysis were used to compare the system methods. The concordance correlation coefficient was analyzed to explain interrater agreement between the three immunoassay systems. RESULTS: Bland-Altman plot showed that the concentrations of AMH measured by UniCel DxI 800 were about 0.15 times higher than that measured using Cobas E601, the concentrations of AMH measured by UniCel DxI 800 were about 0.05 times higher than that measured using iFlash 3000, serum AMH concentrations measured by the iFlash3000 were about 0.19 times higher than that measured using Cobas E601. The concordance correlation coefficient ρc was 0.921, 0.909 and 0.978 for the UniCel DxI 800 versus the Cobas E601, the iFlash 3000 versus the UniCel DxI 800, and the iFlash3000 versus the Cobas E601, respectively. CONCLUSIONS: The three measurement systems have good correlation with each other for determining serum AMH.

Diagnostic Value of Serum DNASE1L3 in Hepatitis B Virus-Related Hepatocellular Carcinoma.

BACKGROUND: Deoxyribonuclease 1-like 3 (DNASE1L3) is an endonuclease associated with many autoimmune diseases and tumors. However, the serum DNASE1L3 level in hepatitis B virus (HBV)-related hepatocellular carcinoma (HCC) remains unreported. Thus, this study compared the diagnostic value of DNASE1L3 and alpha-feto-protein (AFP) individually and in combination in HBV-related HCC. METHODS: The study population consisted of 88 patients with HBV-related HCC, 80 patients with HBV-related liver cirrhosis (LC) and 88 control subjects. The serum DNASE1L3 levels were measured using an enzyme-linked immunosorbent assay. The serum AFP was also assayed. RESULTS: Our data showed that the serum DNASE1L3 levels were significantly higher in patients with HBV-related HCC than in the healthy controls and patients with LC. When the two biomarkers were analyzed individually, the receiver operating characteristic curve analysis showed that the areas under the curve of DNASE1L3 and AFP were 0.898 and 0.866, respectively. When DNASE1L3 and AFP were combined, the area under the curve was 0.951. The sensitivities of DNASE1L3 and AFP were 72.73% and 74.81%, respectively, and the specificities were 93.18% and 92.05%, respectively, in the diagnosis of HBV-related HCC. The sensitivity of the two combined could be improved to 89.77%. However, no correlation was found between serum DNASE1L3 and AFP in HBV-related HCC patients (r = 0.005, p = 0.734). CONCLUSIONS: Serum DNASE1L3 has high sensitivity and specificity in the diagnosis of HCC. DNASE1L3 combined with AFP has higher sensitivity and can improve the diagnostic efficiency of HBV-related HCC.

[Clustering and influencing factors of Streptococcus pneumoniae carriage among kindergarten children].

OBJECTIVE: To study the clustering and influencing factors of Streptococcus pneumoniae carriage among kindergarten children, and to provide a scientific basis for the prevention of pneumococcal diseases. METHODS: The multi-stage stratified cluster sampling method was used to collect nasal swabs via the nasal vestibule at both sides from 1 702 kindergarten children in Liuzhou, China. A questionnaire survey was performed on their parents. The chi-square test and the random effects logistic regression analysis were used to analyze data. RESULTS: The carriage rate of Streptococcus pneumoniae was 13.16% (224/1 702) among kindergarten children. The clustering analysis showed that the class-level random effect of Streptococcus pneumoniae carriage was statistically significant (Z=2.07, P=0.038), with an intraclass correlation coefficient of 5.9%. The random effects logistic regression analysis showed that the children aged 5-7 years had a lower risk of Streptococcus pneumoniae carriage than those aged 2-< 5 years (OR=0.55, 95%CI:0.40-0.76, P=0.001), and the children with ≥ 5 family members living together had a higher risk of Streptococcus pneumoniae carriage than those with < 5 family members (OR=1.34, 95%CI:1.01-1.79, P=0.043). CONCLUSIONS: Streptococcus pneumoniae carriage shows a class-level clustering effect, and age and the number of cohabitants are important influencing factors for Streptococcus pneumoniae carriage in children.

Etiology of acute otitis media and phenotypic-molecular characterization of Streptococcus pneumoniae isolated from children in Liuzhou, China.

BACKGROUND: The etiology and epidemiology of acute otitis media (AOM) are poorly understood in China. This study aimed to describe the etiology of AOM and the phenotypic and molecular characteristics of AOM-causing Streptococcus pneumoniae (S.pneumoniae) recovered from Chinese children. METHODS: A retrospective study was conducted to enrol patients younger than 18 years diagnosed as AOM. Middle ear fluid specimens were collected then cultured for bacterial pathogens. All S.pneumoniae isolates were tested for antibiotic susceptibility, serotypes, virulence genes, antibiotic resistant determinants and sequence types. RESULTS: The dominant otopathogen among AOM children was S.pneumoniae (54.4%). Among S.pneumoniae isolates, there were 97.3, 97.3 and 75.7% isolates resistant to erythromycin, tetracycline and trimethoprim-sulfamethoxazole, respectively. There was 72.8% S.pneumoniae with multidrug resistance. The dominant sequence types (STs) were ST271 and ST320, whereas the prevailing serotypes were 19F and 19A. The 7-valent and 13-valent pneumococcal conjugate vaccine (PCV) coverage among AOM children were 73.0 and 94.6%, respectively. Additionally, we found that CC271 expressed more of mef(A/E) (P < 0.001), pspA (P = 0.022) and sipA (P < 0.001) than non-CC271 isolates. CONCLUSION: The high prevalence of international multidrug-resistant clone (Taiwan19F-14) in China necessitates continued dedication to expand PCV13 immunization and better control of antibiotic use in China.

Interactions between genetic variants involved in the folate metabolic pathway and serum lipid, homocysteine levels on the risk of recurrent spontaneous abortion.

BACKGROUND: The interaction between folate pathway gene polymorphisms and homocysteine, serum lipid leverls are poorly understood in patients with recurrent spontaneous abortion (RSA). The aim of this study is to explore the effects of folate pathway gene polymorphisms (the 5-10-methylenetetrahydrofolate reductase, MTHTR C677T, MTHFR A1298C and the methionine synthase reductase, MTRR A66G) and their interactions with homocysteine on serum lipid levels in patients with RSA. METHODS: A total of 403 RSA women and 342 healthy women were randomly selected. Genotyping of the MTHFR C677T, A1298C and MTRR A66G were performed by TaqMan-MGB technique. Serum homocysteine, folate, fasting glucose, fasting insulin, Interleukin 6, Tumor necrosis factorα (TNFα) and lipid profiles were measured according to the kits. Continuous variables were analyzed using 2-sample t-tests. Categorical variables were analyzed and compared by χ2 or Fisher's exact tests. Unconditional logistic regression model was applied to test the interactions of gene polymorphisms on RSA. RESULTS: The distribution of genotype of CC, CT TT and T allele of MTHFR C677T, genotype of AA and C allele of MTHFR A1298C, and genotype of AA, AG and G allele of MTRR A66G were different between cases and controls (all p were < 0.05). There were significant interactions between MTHFR C677T-A1298C and MTHFR A1298C-MTRR A66G in RSA group and control group, with ORs of 1.62 (95%CI: 1.28-2.04, p < 0.001) and 1.55 (95%CI: 1.27-1.88, p < 0.001), respectively. Serum TNFα level and insulin resistant status (HOMR-IR) were higher in RSA group than in control group (p = 0.038, 0.001, respectively). All the three gene SNPs except MTRR 66AG gene variant had detrimental effects on HOMA-IR (all p were < 0.05). RSA group who carried the MTHFR 677CT, TT, CT/TT genotypes and MTRR 66AG, AG/GG genotypes had detrimental effects on serum homocysteine levels, the MTHFR 677CT, CT/TT genotype carriers had favorable effects on serum folate levels, the MTHFR 677TT, CT/TT, 1298 AC, AC/CC genotype carriers had detrimental effects on serum low-density lipoprotein cholesterol (LDL-C) levels, and the MTRR 66AG genotype carriers had lower high-density lipoprotein cholesterol (HDL-C) levels than the AA genotype carriers (all p were < 0.05). CONCLUSIONS: Interaction between the MTHFR C677T, A1298C and MTHFR A1298C, MTRR A66G are observed in our RSA group. Besides, all the three gene SNPs except MTRR 66AG gene variant had detrimental effects on HOMA-IR. MTHFR C677T and MTRR A66G gene variants had detrimental effects on serum homocysteine levels and insulin resistance status, while MTHFR C677T, A1298C and MTRR A66G gene variants had detrimental effects on certain serum lipid profiles.

Serotype distribution and antimicrobial resistance of Streptococcus pneumoniae causing invasive diseases in China: a meta-analysis.

BACKGROUND: To summarize information about invasive pneumococcal disease (IPD) among children in mainland China. METHODS: Sixteen eligible studies were included in this systematic review and the random effect model was used to estimate the pool prevalence of IPD. RESULTS: The most predominant serotypes circulating in children were 19F (27.7, 95% confidence interval (95% CI): 17.7-37.6%), 19A (21.2%, 16.4-26.1%), 14 (16.5%, 12.8-20.1%), 6B (8.6%, 5.2-10.8%) and 23F (7.3%, 5.2-9.5%). The serotype coverage of the available pneumococcal conjugate vaccines PCV7, PCV10, and PCV13 was 60.8% (52.5-69.4%), 65.1% (57.7-72.4%), and 90.0% (87.1-92.8%), respectively. The pooled antibiotic resistance rates of Streptococcus pneumoniae revealed a resistance to penicillin prevalence rate of 32.0% (12.1-51.9%). Approximately 94.4% (90.7-98.1%) and 92.3% (87.4-97.3%) of isolates were resistant to erythromycin and clindamycin. eBURST analysis revealed great diversity among isolates, with 102 sequence types (STs) for 365 isolates. The major predominant clonal complexes (CCs) were CC271 (43.6%, 159/365), CC876 (13.4%, 49/365), CC81 (5.2%, 19/365), and CC90 (4.1%, 15/365). Long-term and regional surveillance of S. pneumoniae is necessary. CONCLUSIONS: Based on our pooled results showing that PCV13 coverage of the reported serotypes was 90% and that most serotypes contributed to the distribution of antibiotic-resistant isolates, implementation of PCV13 into the Chinese Expanded Program on Immunizations (EPI) would achieve health benefits in Chinese children.