To evaluate the performance of simultaneous amplification and testing assay for group B Streptococcus detection: comparison with real-time PCR and ddPCR assays.

BACKGROUND: To evaluate the performance of simultaneous amplification and testing (SAT) assay for the detection of group B Streptococcus (GBS) in maternal vaginal and perianal swabs compared with real-time polymerase chain reaction (RT-PCR). METHODS: We obtained vaginal and perianal swabs from 1474 pregnant women at the Obstetrics and Gynecology Hospital of Fudan University (Shanghai, China) between April 2023 and June 2023. Vaginal and perianal swabs were collected at 35-37 weeks of gestation. Swabs were tested for GBS simultaneously by using the SAT assay and RT-PCR, and a comparative analysis (kappa coefficient) was performed. Furthermore, we conducted additional droplet digital PCR (ddPCR) tests to confirm the results when there were controversial results between SAT and RT-PCR. In addition, we compared the limit of detection, technical specificity, repeatability and reproducibility of SAT-GBS with those of routine RT-PCR assays. RESULTS: In our study, the detection rate of clinical GBS according to the SAT assay was 11.5% (169/1471). The SAT assay showed a sensitivity of 91.8%, a specificity of 99.9%, a diagnostic accuracy of 98.9%, a positive predictive value (PPV) of 99.4% and a negative predictive value (NPV) of 98.8%. The kappa value between RT-PCR and SAT was 0.917. CONCLUSIONS: This SAT assay for the detection of group B Streptococcus is not only easy to perform but can also detect GBS sensitively and specifically and may be used in the regular molecular diagnosis of GBS infection among pregnancies.

Clinical application of expanded noninvasive prenatal testing for fetal chromosome abnormalities in a cohort of 39,580 pregnancies.

The aim of this study was to determine the predictive value of expanded noninvasive prenatal testing (NIPT-plus) for fetal chromosome abnormalities in the second trimester (12-26 weeks). We conducted a retrospective cohort study of 39,580 pregnancies with NIPT-plus. Screening positive cases were diagnosed with karyotyping and single-nucleotide polymorphism array analysis (SNP array)/copy number variation sequencing (CNV-seq) with follow-up. The positive predictive values (PPVs) of trisomy 21, 18, and 13 (T21, T18, and T13), sex chromosome aneuploidies (SCAs), and microdeletion and microduplication syndromes (MMS) by NIPT-plus were recorded. We assessed the predictive value of NIPT-plus based on maternal age and conventional indications. Of 39,580 pregnancies with NIPT-plus, 511 (1.3%) had prenatal screening positive results of fetal chromosome abnormality, of which 87.7% (448/511) had invasive prenatal diagnosis. NIPT-plus performed better in predicting fetal SCAs and chromosome aneuploidies for pregnancies with advanced maternal age (AMA) than young maternal age (YMA). Besides, the PPVs of T21, T13, and chromosome aneuploidies showed an upward trend when comparison was based on maternal age in 5-year subintervals. The termination rates of 45,X, 47,XXX, 47,XXY, and 47,XYY were 100% (11/11), 20.0% (3/15), 91.7% (22/24), and 7.1% (1/14) with postnatal follow-up. Last but not least, the PPV for MMS is 41.7% (30/72), which may have a positive correlation between the size of CNVs. Pregnant women with screen-positive results for common trisomies (T13, T18, and T21) were more willing to conduct invasive prenatal diagnosis compared to those with positive results for SCAs or MMS. However, the current study demonstrated SCAs and MMS had the lowest PPV. This highlights the importance of confirmatory prenatal diagnosis in those patients and the potential impact on genetic counseling and informative decision-making.

Antifungal Susceptibility Profiles and Drug Resistance Mechanisms of Clinical Lomentospora prolificans Isolates.

Lomentospora prolificans is an opportunistic fungal pathogen with low susceptibility to current antifungal drugs. Here, we tested the in vitro susceptibility of 8 drugs against 42 clinical L. prolificans isolates. All isolates showed high MICs to voriconazole (MIC90>16 µg/ml), itraconazole (MIC90>16 µg/ml), posaconazole (MIC90>16 µg/ml), isavuconazole (MIC90>16 µg/ml), amphotericin B (MIC90>16 µg/ml), and terbinafine (MIC90>64 µg/ml) and high minimum effective concentrations (MECs) to micafungin (MEC90>8 µg/ml), with the exception of miltefosine showing an MIC90 value of 4 µg/ml. We examined six different in vitro drug combinations and found that the combination of voriconazole and terbinafine achieved the most synergistic effort against L. prolificans We then annotated the L. prolificans whole genome and located its Cyp51 and Fks1 genes. We completely sequenced the two genes to determine if any mutation would be related to azole and echinocandin resistance in L. prolificans We found no amino acid changes in Cyp51 protein and no tandem repeats in the 5' upstream region of the Cyp51 gene. However, we identified three intrinsic amino acid residues (G138S, M220I, and T289A) in the Cyp51 protein that were linked to azole resistance. Likewise, two intrinsic amino acid residues (F639Y, W695F) that have reported to confer echinocandin resistance were found in Fks1 hot spot regions. In addition, three new amino acid alterations (D440A, S634R, and H1245R) were found outside Fks1 hot spot regions, and their contributions to echinocandin resistance need future investigation. Overall, our findings support the notion that L. prolificans is intrinsically resistant to azoles and echinocandins.

Molecular epidemiology and risk factors of Stenotrophomonas maltophilia infections in a Chinese teaching hospital.

BACKGROUND: Stenotrophomonas maltophilia (S. maltophilia) is an important opportunistic pathogen that can be isolated in hospitals. With the abuse of broad spectrum antibiotics and invasive surgical devices, the rate of S. maltophilia infection is increasing every year. This study was an epidemiological analysis of the clinical and molecular characteristics of S. maltophilia infection in a Chinese teaching hospital. The goal was to obtain a comprehensive understanding of the status of S. maltophilia infection to provide strong epidemiological data for the prevention and treatment of S. maltophilia infection. RESULTS: A total of 93 isolates from Renji Hospital affiliated with the Shanghai Jiaotong University School of Medicine were included, in which 62 isolates were from male patients. In addition, 81 isolates were isolated from sputum samples. A total of 86 patients had underlying diseases. All patients received antibiotics. Multilocus sequence typing (MLST) analysis indicated that 61 different sequence types (STs) were found (including 45 novel STs), and MLST did not show significantly dominant STs. Pulsed field gel electrophoresis (PFGE) results showed that 93 isolates could be divided into 73 clusters, and they also showed weak genetic linkages between isolates. The resistant rates to trimethoprim/sulfamethoxazole (TMP/SMX) and levofloxacin were 9.7 and 4.3%, respectively, and all isolates were susceptible to minocycline. Four virulence gene's loci Stmpr1, Stmpr2, Smf-1, and Smlt3773 were positive in 79.6, 91.4, 94.6, and 52.7% of the isolates, respectively. Three biofilm genes rmlA, spgM, and rpfF were positive in 82.8, 92.5, and 64.5% of the isolates, respectively. Mean biofilm forming level of OD492 was 0.54 ± 0.49. We did not find any significant difference between different genders and different age-groups. We retrospectively analyzed data from patients in the intensive care unit (ICU) and the control group. The independent risk factors of those who were infected in the ICU included immunosuppression and the increased antibiotic usage. CONCLUSIONS: Most of the patients had prior medical usage histories and baseline diseases. The positive rate of virulence genes was high, the drug resistance rate of S. maltophilia was low, and the biofilm formation ability was strong. The increased use of antibiotics was an independent risk factor for S. maltophilia infection, which should receive more attention. No obvious clonal transmissions were found in the same departments.

Characterization of vaginal microbiota in Chinese women with cervical squamous intra-epithelial neoplasia.

OBJECTIVES: Although persistent human papillomavirus (HPV) infection is a major cause of cervical squamous intra-epithelial neoplasia, the relationship between vaginal microbiota and different grades of squamous intra-epithelial neoplasia is not well established. We explored the possible relationship between the vaginal microbiota and the progression of cervical squamous intra-epithelial neoplasia. METHODS: We evaluated 69 women who attended the Obstetrics and Gynecology Hospital of Fudan University. The vaginal bacterial composition of three groups of women was characterized by deep sequencing of bar-coded 16S rRNA gene fragments (V3-4) using Illumina MiSeq. Exclusion criteria were any previous hysterectomy, history of cervical or other lower genital cancer, and/or destructive therapy of the cervix. Women who had autoimmune disorders, who were HIV positive, who received antibiotics within 15 days of sampling, or who had engaged in sexual intercourse or douching within 48 hours prior to sampling were also excluded. P values for age and proportions of organisms were calculated using one-way ANOVA and p values for HPV status and community state types (CSTs) were calculated using a χ2 test. RESULTS: The vaginal bacterial composition of three groups of women, those without an intra-epithelial lesion or malignancy (n=31), those with a low-grade squamous intra-epithelial lesion (LSIL) (n=22), and those with a high-grade squamous intra-epithelial lesion (HSIL) (n=16) were analyzed. Lactobacillus was the most dominant genus overall. Prevotella and Streptococcus were increased in the HSIL group. Cervical disease progression was associated with the prevalence of high-risk HPV infection. Squamous intra-epithelial neoplasia converted the vaginal bacterial community structure from CSTs IV to II. Microbiota diversity was more pronounced in CST types II and IV (p<0.001), especially in type II. We found a significant enrichment in the Peptostreptococcaceae family, Pseudomonadales order, and other types of bacteria in the group of women without intra-epithelial lesions or malignancy compared with women with squamous intra-epithelial neoplasia. We found enrichment in Delftia in the LSIL and HSIL groups compared with the group without an intra-epithelial lesion or malignancy. CONCLUSIONS: Our results show that the vaginal microbiota is directly or indirectly related to the progression of squamous intra-epithelial neoplasia, and Delftia might be a microbiological hallmark of cervical pre-cancerous lesions.

ERG11 couples oxidative stress adaptation, hyphal elongation and virulence in Candida albicans.

Candida albicans is a major fungal opportunistic pathogen for humans. In the treatment of C. albicans, azole drugs target the sterol 14α-demethylase (CYP51) encoded by ERG11 gene. Most studies have focused on the fact that the ERG11 mutant results in drug resistance, but its mechanism of action as a drug target has not been described yet. Our results showed that deletion of ERG11 reduced filamentous and invasive growth, and impaired hyphal elongation in sensing serum. Lack of ERG11 increased susceptibility to H2O2 and was defective in clearing reactive oxygen species. ERG11 may affect oxidative stress adaptation by specifically downregulating CAT1 expression. In addition, C. albicans cells lacking ERG11 were more efficiently killed by macrophages and became avirulent in vivo. This study is the first to indicate that ERG11 plays an essential role in hyphal elongation, oxidative stress adaptation and virulence in C. albicans. We speculated that azole drugs not only inhibit the growth of C. albicans, but also assist the host immune system in clearing the fungal organism. The new understanding of mechanisms of action of antifungal drugs should facilitate the development of treatment strategies for resistant fungal infections.

Clonal spread and azole-resistant mechanisms of non-susceptible Candida albicans isolates from vulvovaginal candidiasis patients in three Shanghai maternity hospitals.

In our multicenter study, 43 fluconazole non-susceptible and 45 fluconazole-susceptible isolates were collected from vulvovaginal candidiasis (VVC) patients from three Shanghai maternity hospitals to analyze their molecular epidemiological features and fluconazole resistant mechanisms. Cross-resistance to fluconazole, itraconazole and voriconazole was observed in 53.5% of the nonsusceptible isolates. Though we acquired 12 clonal complexes (CCs) of diploid sequence types (DSTs) in clinical isolates by a multilocus sequence typing method, fluconazole nonsusceptible isolates all belonged to CC69 with a predominant genotype of DST 79. Increased expressions of efflux pump genes (CDR1, CDR2, and MDR1) were observed only in minor fluconazole non-susceptible isolates by real-time quantitative polymerase chain reaction (PCR). However, ERG11 genes of fluconazole SDD and resistant isolates had significantly higher expression levels than fluconazole-susceptible isolates. Moreover, 13 distinct amino acid substitutions in Erg11p were found in clinical isolates. Three of the substitutions were novel amino acid substitutions (T123I, P98S, and Y286D), which were not in the susceptible isolates. Only two heterozygous amino acid substitutions (A18P/A and R365G/R) in Erg3p were found in two isolates with cross-resistance to fluconazole, itraconazole, and voriconazole. Taken together, we observed the clonal spread of CC69 in fluconazole non-susceptible isolates of Candida albicans from VVC patients with the dominant genotype DST79. ERG11 gene mutations and overexpression predominantly contributed to fluconazole resistance instead of the more common increased expressions of efflux pump genes (CDR1, CDR2, and MDR1).

Epidemiological analysis of maternal lipid levels during the second trimester in pregnancy and the risk of adverse pregnancy outcome adjusted by pregnancy BMI.

BACKGROUND: Pregnancy is accompanied by profound changes in lipid metabolism. We aimed to assess whether effects of second trimester body mass index and maternal lipid concentrations are associated with an increased risk of adverse pregnancy outcomes. METHODS: We investigated the serum levels of maternal lipids during the second trimester in pregnancy, and analyzed associations between the lipid levels and the risk of adverse pregnancy outcome. Seven hundred and seventy-four pregnant women were enrolled in this study between February 2016 and June 2016. Multivariate logistic regression analysis was conducted to estimate the relative risk between maternal lipids and adverse pregnancy outcome. RESULTS: Compared with the control group, during the second trimester of pregnancy, BMI, TG, and Lp(a) were risk factors for gestational diabetes mellitus; middle trimester pregnancy BMI, Lp(a), and APO-B were risk factors for pre-eclampsia; second trimester BMI and TG/HDL-C were risk factors for macrosomia; age and Lp(a) were uterine atony postpartum hemorrhage risk factors, while APO-AI was a protective factor of uterine inertia and postpartum hemorrhage; second trimester BMI, TCH, Lp(a), and TG/HDL-C were risk factors for fetal distress, while parity was a protective factor against fetal distress. CONCLUSION: Abnormal blood lipid levels in pregnancy are significantly associated with GDM, pre-eclampsia, and other adverse pregnancy outcomes.

Serum levels of human epididymis protein 4 are more stable than cancer antigen 125 in early and mid-term pregnancy.

AIM: Previous studies have shown serum human epididymis protein 4 (HE4) and cancer antigen 125 (CA125) levels could serve as predictive markers in the diagnosis of ovarian cancer. However, HE4 levels have been less well studied during pregnancy, especially in threatened abortion, while CA125 levels were fluctuated. Our study aimed to assess the stability of HE4 and CA125 levels through trimesters and in cases of threatened abortion. METHODS: Forty-six nonpregnant women (control), 167 healthy pregnant women and 46 pregnant threatened abortion (TA; 8-12 weeks pregnancy) women who visited the Obstetrics and Gynecology Hospital of Fudan University from September to December 2017 were recruited. The healthy pregnant women group included 57 women in the first trimester (T1), 55 in the second (T2) and 55 in the third (T3). Serum levels of HE4 and CA125 were measured by electrochemiluminescent immunoassay. RESULTS: Both HE4 and CA125 levels in the T3 group were significantly higher than in the control group (P < 0.001). There was no difference in HE4 between the control, T1 and T2 groups, while levels of CA125 in T1 group were elevated (P < 0.001). There was no difference in HE4 levels between the TA, control and T1 groups. However, the levels of CA125 in the TA group were much higher (P < 0.05). CONCLUSION: The level of HE4 was found to be more stable than that of CA125 in early and mid-pregnancy and in cases of threatened abortion.

A newly identified amino acid substitution T123I in the 14α-demethylase (Erg11p) of Candida albicans confers azole resistance.

The increasing prevalence of azole resistance in Candida albicans poses a growing problem for clinical treatment. Amino acid substitution of the 14α-demethylase (Erg11p) encoded by the ERG11 gene is one of the most common mechanisms involved in azole resistance. Although amino acid substitutions of Erg11p have been observed in many clinical isolates, only a few amino acid substitutions have been confirmed to be related to azole resistance. In this study, by amplifying and sequencing the open reading frame of the ERG11 gene from 55 clinical isolates, we identified 27 fluconazole-resistant isolates that harbor a novel amino acid substitution, T123I, in Erg11p, in addition to the previously described homozygous substitution Y132H. We investigated both the contribution of this novel substitution T123I and its synergistic effect with substitution Y132H to azole resistance by heterogeneously expressing the C. albicans Erg11p with different substitution forms in Saccharomyces cerevisiae. Results showed that S. cerevisiae cells harboring the substitution T123I displayed higher (4-fold) minimum inhibitory concentration values to both fluconazole and voriconazole than the cells expressing the wild-type version of C. albicans Erg11p, but this was not true for itraconazolele. More importantly, a synergistic effect of substitutions T123I and Y132H was observed in an assay of voriconazole resistance. These results indicate that amino acid substitutions of Erg11p are prevalent among azole-resistant isolates and that the substitution T123I confers resistance to both fluconazole and voriconazole.

Antifungal susceptibility and molecular typing of 115 Candida albicans isolates obtained from vulvovaginal candidiasis patients in 3 Shanghai maternity hospitals.

In our multicenter study, we studied the distribution of Candida species in vulvovaginal candidiasis patients and investigated antifungal susceptibility profile and genotype of Candida albicans in vaginal swab. A total of 115 Candida albicans strains were detected in 135 clinical isolates. Minimum inhibitory concentration determinations showed that 83% and 81% of the 115 Candida albicans strains were susceptible to fluconazole and voriconazole. Randomly amplified polymorphic DNA analysis (RAPD) was applied to identify clonally related isolates from different patients at the local level. All tested strains were classified into genotype A (77.4%), genotype B (18.3%), and genotype C (4.3%). Genotype A was further classified into five subtypes and genotype B into two subtypes.Candida albicans was the dominant pathogen of vulvovaginal candidiasis, the majority belonging to genotype A in this study. Exposure to azoles is a risk factor for the emergence of azole resistance among Candida albicans isolated from VVC patients.

[Analysis of serum lipids levels and the establishment of reference intervals for serum lipids in middle and late pregnancy].

OBJECTIVE: To explore the serum lipids levels in healthy pregnant women, and to establish the reference intervals of serum lipids in middle and late pregnancy. METHODS: Triglyceride (TG), total cholesterol (TCH), high density lipoprotein (HDL), low density lipoprotein (LDL), apo-lipoprotein-A(APO-A) and apo-lipoprotein-B (APO-B) were measured in 3 200 pregnant women and 3 200 healthy women of childbearing age(the control group) from January 2014 to Febuary 2015 in Obstetrics and Gynecology Hospital of Fudan University. In the healthy pregnant women, serum lipids were measured at 14-20, 24-28 and 37-40 gestational weeks, respectively. All the parameters were detected by Hitachi 7180 automatic biochemical analyzer. The test results were calculated and determined by the C28-A3 standard of the National Clinical and Laboratory Standards Institute. And the normal reference intervals of serum lipids in middle and late pregnancy were defined as 2.5%-97.5%. RESULTS: (1) The levels of TG, TCH, HDL, LDL, APO-A and APO-B in the control group were 0.8, 4.2, 1.0, 2.7 mmol/L and 1.1, 0.8 g/L, respectively. The levels of TG, TCH, HDL, LDL, APO-A and APO-B in middle and late pregnancy were significantly higher than those in the control group (P<0.05). (2) The serum lipids levels at 14-20, 24-28 and 37-40 gestational weeks in healthy pregnant women were compared with the control group as following. The TG levels were 1.9, 3.8 and 4.4 folds of the control group; the TCH levels were 1.1, 1.5 and 1.5 folds of the control group; the HDL levels were 1.2, 1.6 and 1.5 folds of the control group; the LDL levels were 1.1, 1.4 and 1.4 folds of the control group; the APO-A levels were 1.3, 1.4 and 1.5 folds of the control group; and the APO-B levels were 1.1, 1.5 and 1.5 fold of the control group respectively. The TG level was the most increased, and it increased gradually with gestational age (P<0.01). (3) The median of LDL to HDL cholesterol ratio in the healthy pregnancy group at 14-20, 24-28 and 37-40 gestational weeks were 2.7, 2.5, 2.6, respectively, which were significantly lower than that of the control group (2.8; P<0.05). (4) Reference intervals of serum lipids at 14-20, 24-28 and 37-40 gestational weeks in healthy pregnant women were as following. The TG levels were 0.7-3.9, 1.7-6.3 and 1.6-8.1 mmol/L, respectively; the TCH were 3.3-6.9, 4.3-8.3, 4.3-8.7 mmol/L, respectively; the HDL were 0.8-1.8, 1.0-2.1 and 1.0-2.1 mmol/L, respectively; the LDL were 2.1-4.5, 2.7-5.1 and 2.6-5.2 mmol/L, respectively; the APO-A were 1.1-1.8, 1.2-1.9 and 1.1-2.4 g/L, respectively; and the APO-B were 0.6-1.4, 0.9-1.8 and 0.8-2.1 g/L, respectively. The LDL/HDL ratios were 2.3-3.1, 2.2-2.9 and 2.1-3.0, respectively. CONCLUSIONS: Serum lipids increased physiologically with gestational age in middle and late pregnancy. The establishment of reference intervals for serum lipids in pregnancy will help to distinguish abnormal serum lipid levels in middle and late pregnancy.

Association of serum inhibin a with pre-eclampsia: An observational cohort study with propensity score matching.

OBJECTIVE: To assess whether serum inhibin A at 14-20 weeks of gestation is associated with the occurrence of pre-eclampsia. METHODS: A retrospective cohort study using propensity score matching was conducted on 11 682 singleton pregnant women with established deliveries at the Obstetrics and Gynecology Hospital of Fudan University between January 2017 and July 2019. We investigated serum inhibin A levels at 14-20 weeks of gestation and calculated the relative risk between inhibin A and pre-eclampsia by multifactorial logistic regression analysis. Smoothed, fitted curves were used to observe the effect of inhibin A in relation to the occurrence of pre-eclampsia. RESULTS: The risk of pre-eclampsia occurrence increased with elevated serum inhibin A. After full adjustment for confounders, the risk ratio for pre-eclampsia in the group of pregnant women with high inhibin A was 2.92 (95% confidence interval [CI] 2.08-4.11) compared with those with normal inhibin A. The results of sensitivity analysis suggested a consistent effect of inhibin A on the risk of pre-eclampsia in different populations. CONCLUSION: Elevated serum inhibin A at 14-20 weeks of gestation is associated with pre-eclampsia and may provide an early warning signal for pregnancy outcomes associated with pre-eclampsia.

Diagnostic value of dual-fluorescence staining in bacterial vaginosis.

OBJECTIVE: The aim of this study was to investigate the epidemiology of bacterial vaginosis (BV) in Shanghai, China, and to explore the value of a dual-fluorescence staining method in the diagnosis of BV. METHODS: Specimens were collected from women with vaginitis at the Obstetrics and Gynecology Hospital of Fudan University from January 2020 to December 2021, and the proportions of various vaginitis types (such as Candida vaginitis, Trichomonas, and bacterial vaginitis) were analyzed statistically. To explore the diagnostic value of dual-fluorescence staining for BV, we first executed a dual-fluorescence staining method to analyze the vaginal secretions of 265 patients, then confirmed our diagnoses by consulting clinical physicians and by using Nugent scoring of Gram staining. RESULTS: There were 16,905 patients who were diagnosed with vaginitis over the previous 2 years, with a median age of 32 (minimum age of 9 years and maximum of 84 years). Of these patients, we noted 10,887 cases (64.40%) of BV. Our staining results revealed that the dual-fluorescence method was consistent with Gram staining in the diagnosis of BV, with a P value of less than .001 using a χ 2 test and a consistency kappa value of 0.896. Compared with Gram staining, the dual-fluorescence staining method required an acceptable time (2.2 min vs 2.5 min, respectively) and exhibited different visual effects (green and yellow vs purple and red, respectively). CONCLUSION: Dual-fluorescence staining for the detection of bacterial diseases of the vagina exhibited acceptable consistency with Gram staining and performed well with respect to dyeing time, stability, and the interpretation of results. We argue that this method should be used in outpatient services.

CPY27B1-related vitamin D deficiency contributes to the impaired function of CD4+T cells in Recurrent spontaneous miscarriage.

The study aimed to explore the relationship between the expression of cytochrome P450 family 27 subfamily B member 1 (CYP27B1), vitamin D, and impaired T cell subsets in recurrent spontaneous miscarriage (RSM). A Total of 779 healthy women of childbearing age and 1031 women with a history of RSM were involved in this study. The results of flow cytometry showed that the proportion of Tregs was higher in healthy women than in the women with RSM. For cytokines, the levels of interleukin-17 (IL-17) and interferon-gamma (IFN-γ) were significantly higher in RSM patients than in healthy women, while IL-10 was notably lower in RSM patients. Furthermore, compared to healthy individuals, RSM patients had lower levels of serum 25(OH)D detected by chemiluminescence. The frequency of Tregs was negatively correlated with 25(OH)D. Specifically, for every 10 ng/ml increase in 25(OH)D, the percentage of Tregs increased by 0.58 as calculated. IL-17 and IFN-γ were inversely correlated with 25(OH)D, while the serum interleukin-10 (IL-10) level was positively correlated with 25(OH)D. CYP27B1 was found to be expressed in both cytotrophoblast and extracellular villi trophoblast cells. However, reduced expression of CYP27B1 was observed in the placenta with RSM. Notably, the level of 25(OH)D increased in the supernatant of CYP27B1 knockdown BeWo compared to normal cells, while human chorionic gonadotropin (hCG) was significantly reduced. The hCG secretion of CYP27B1 KO BeWo cells was partially restored after 1,25(OH)2D3 supplementation. In addition, 1,25(OH)2D3 treatment could induce more CD4+ T cells to convert to Foxp3+iTreg, which in turn inhibited the secretion of IL-17, IFN-γ. In summary, this research unveiled a connection between reduced CYP27B1 and vitamin D deficiency in RSM. Our study underscores the potential benefits of vitamin D treatment supplementation in the context of RSM. However, it is important to note that further research is imperative to validate these observations.

Serum CYFRA21-1 and SCC-Ag levels in women during pregnancy and their diagnostic value for cervical cancer.

OBJECTIVES: The incidence of cervical cancer increases every year during pregnancy. Cervical cytology in pregnant women has a unique morphology and liquid-based cytology methods are prone to cause false positives. The aim of this study was to investigate the serum cytokeratin 19 fragment antigen 21-1 (CYFRA21-1) and squamous cell carcinoma associated antigen (SCC-Ag) concentrations in healthy pregnant women during pregnancy and to assess their diagnostic value for cervical cancer in pregnancy. METHODS: In this prospective study, 165 healthy non-pregnant women, 441 healthy pregnant women and 22 patients with cervical cancer in pregnancy were recruited. The healthy pregnant women group included 143 women in the first trimester (T1), 147 in the second (T2) and 151 in the third (T3). RESULTS: Both SCC-Ag and CYFRA21-1 levels were significantly different in the healthy pregnant women group compared to the control group. The CYFRA21-1 and SCC-Ag were higher in the T1 and T3 than in the control groups. However, there was no statistically significant difference in serum CYFRA21-1 and SCC-Ag levels in the T2 group compared to the control group. The AUCs of CYFRA21-1, SCC-Ag and CYFRA21-1 combined with SCC-Ag were 0.674, 0.792, and 0.805, respectively. The cut-off values of CYFRA21-1 and SCC-Ag were 6.64 ng/mL and 1.75 ng/mL, respectively. CONCLUSIONS: Serum CYFRA21-1 and SCC-Ag levels were higher in pregnant women during early and late pregnancy compared to non-pregnant individuals, while they were not statistically different from non-pregnant women during mid-trimester. CYFRA21-1 and SCC-Ag have diagnostic value for cervical cancer in pregnancy.

Prospective prenatal cell-free DNA screening for genetic conditions of heterogenous etiologies.

Prenatal cell-free DNA (cfDNA) screening uses extracellular fetal DNA circulating in the peripheral blood of pregnant women to detect prevalent fetal chromosomal anomalies. However, numerous severe conditions with underlying single-gene defects are not included in current prenatal cfDNA screening. In this prospective, multicenter and observational study, pregnant women at elevated risk for fetal genetic conditions were enrolled for a cfDNA screening test based on coordinative allele-aware target enrichment sequencing. This test encompasses the following three of the most frequent pathogenic genetic variations: aneuploidies, microdeletions and monogenic variants. The cfDNA screening results were compared to invasive prenatal or postnatal diagnostic test results for 1,090 qualified participants. The comprehensive cfDNA screening detected a genetic alteration in 135 pregnancies with 98.5% sensitivity and 99.3% specificity relative to standard diagnostics. Of 876 fetuses with suspected structural anomalies on ultrasound examination, comprehensive cfDNA screening identified 55 (56.1%) aneuploidies, 6 (6.1%) microdeletions and 37 (37.8%) single-gene pathogenic variants. The inclusion of targeted monogenic conditions alongside chromosomal aberrations led to a 60.7% increase (from 61 to 98) in the detection rate. Overall, these data provide preliminary evidence that a comprehensive cfDNA screening test can accurately identify fetal pathogenic variants at both the chromosome and single-gene levels in high-risk pregnancies through a noninvasive approach, which has the potential to improve prenatal evaluation of fetal risks for severe genetic conditions arising from heterogenous molecular etiologies. ClinicalTrials.gov registration: ChiCTR2100045739 .

Copy number variations of the F8 gene are associated with venous thromboembolism.

BACKGROUND: Venous thromboembolism (VTE) is a complex disease and several inherited and acquired factors are relevant to its occurrence. Among these, an elevated level of plasma coagulation factor VIII (FVIII) is an established risk factor for VTE; copy number variations (CNVs) have also been discovered to be associated with many diseases. OBJECTIVE: To explore the proposed association between CNV of the F8 gene and the risk of VTE. METHODS: A case-control study including 179 VTE patients and 176 healthy individuals were enrolled in this study. Activity of plasma factor VIII was measured. Genomic DNA was extracted for subsequent quantitative real-time PCR analysis of CNVs of the F8 gene. RESULTS: Plasma factor VIII levels were significantly higher in VTE patients than in healthy controls (251% vs. 99%, p<0.01). Copy number of the F8 gene in VTE patients was significantly higher than in healthy controls. (male: p=6.1×10(-14), OR=12, 95%CI: 6.0-25; female: p=4.3×10(-10), OR=9.5, 95%CI: 4.5-20). Plasma factor VIII levels in the samples with high copies of the F8 gene were higher than in those individuals with normal copy number (male: p=0.023; female: p=0.036). CONCLUSIONS: Amplification of the F8 gene copy number seems to enhance factor VIII activity and was associated with VTE.

Elevated Serum Uric Acid Is Associated With Gestational Diabetes Mellitus: An Observational Cohort Study.

CONTEXT: Elevated serum uric acid may be closely related to the occurrence of gestational diabetes mellitus (GDM). OBJECTIVE: We aimed to elucidate the relationship between changes in serum uric acid before 24 weeks of gestation and the risk of GDM and associated adverse pregnancy outcomes and provide clinical epidemiological evidence for the involvement of uric acid in the etiology of GDM. METHODS: We conducted a retrospective cohort study of 23 843 singleton pregnant women between February 2018 and June 2022. The exposure factor was serum uric acid before 24 weeks of gestation, primary outcome was gestational diabetes diagnosed at 24 to 28 weeks of gestation, and secondary outcomes were GDM A2 (GDM requiring pharmacotherapy), GDM combined with pre-eclampsia, preterm delivery, and large for gestational age infants. Adjusted risk ratios (RRs) were calculated using multivariate predictive marginal proportions from logistic regression models. RESULTS: Among 23 843 singleton pregnant women, 3204 (13.44%) were diagnosed with GDM at 24 to 28 weeks of gestation, and elevated uric acid before 24 weeks of gestation was strongly associated with the risk of GDM. Compared with uric acid <240 µmol/L, the RR for GDM was 1.43 (95% CI 1.29-1.56) when uric acid was between 240 and 300 µmol/L; when uric acid was >300 µmol/L, the RR for GDM was 1.82 (95% CI 1.55-2.15). In secondary outcomes uric acid had a similar relationship with GDM A2, preterm birth, and GDM combined with pre-eclampsia. CONCLUSION: Elevated uric acid levels before 24 weeks of gestation are associated with subsequent GDM; the best time to test for uric acid is before 18 weeks of gestation. Pregnant women with low and intermediate risk for GDM development may benefit more from serum uric acid measurements before 18 weeks of gestation.