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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.

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 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.

To elucidate whether uric acid changes in early pregnancy are associated with the development of preeclampsia and their association with preeclampsia-related adverse pregnancy outcomes. We conducted a retrospective cohort study of 4725 singleton pregnant women between January 2017 and July 2019 using propensity score matching. The primary outcome of the cohort was preeclampsia, and the secondary outcomes were preterm delivery, preterm preeclampsia and low birth weight infants. Multivariable predicted marginal proportions from logistic regression models were used to compute adjusted risk ratios. The quantitative-effect relationship between serum uric acid and preeclampsia development was observed by a dose‒response graph, and the effect of serum uric acid on the week of gestation at delivery was assessed using the Kaplan‒Meier method and the log-rank test. The risk of preeclampsia development increased with higher serum uric acid levels. After adjusting for confounders, the risk ratio for the development of preeclampsia with uric acid levels ≥240 µmol/l was 1.25 (95% CI: 0.96-1.65) compared with the group with uric acid levels240 µmol/l. In the subgroup analysis of KM (Kaplan-Meier) curves, the gestational week at delivery was earlier when uric acid levels ≥240 µmol/l occurred at 8-12 weeks of gestation. Elevated serum uric acid levels before 20 weeks of gestation are associated with the development of preeclampsia, especially in the first 8-12 weeks of gestation, and the effect is attenuated with increasing gestational weeks, which suggests that elevated uric acid levels in early pregnancy may be a causative factor in preeclampsia. Elevated serum uric acid levels before 20 weeks of gestation are associated with the development of preeclampsia, especially in the early 8-12 weeks of gestation, and the effect attenuates with increasing gestational weeks, which suggest that elevated uric acid in early pregnancy may be a causative factor in preeclampsia.

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 is an epidemiological analysis of the clinical and molecular characteristics of S. maltophilia infection in a Chinese teaching hospital. The goal is to obtain a comprehensive understanding of the status of S. maltophilia infection in order to provide strong epidemiological data for the prevention and treatment of S. maltophilia infection.Results: A total of 93 isolates from Renji Hospital affiliated to Shanghai Jiaotong University School of Medicine were included, in which 62 isolates were from male patients and 81 isolates were isolated from sputum samples. A total of 86 patients had underlying diseases. All of the patients had received antibiotic therapeutics. The multilocus sequence typing (MLST) analysis indicated that 61 different sequence types (STs) were found (including 45 novel STs), and the MLST did not show significantly dominant STs. Pulsed field gel electrophoresis (PFGE) results showed the 93 isolates could be divided into 73 clusters, and they also showed weak genetic linkage between isolates. The resistant rates to Trimethoprim/sulfamethoxazole (TMP/SMX) and levofloxacin were 9.7% and 4.3%, respectively, and all of the isolates were susceptible to minocycline. The positive rates of the four virulence genes Stmpr1, Stmpr2, Smf-1, and Smlt3773 locus were 79.6%, 91.4%, 94.6%, and 52.7%, respectively. The positive rates of the three biofilm genes rmlA, spgM, and rpfF were 82.8%, 92.5%, and 64.5%, respectively. The mean biofilm forming level of OD492 was 0.54 ± 0.49, and there was no significant difference between different genders and different age groups. Data from patients in the intensive care unit (ICU) and the control group were analyzed retrospectively, and the independent risk factors of those who were infected in the ICU included immunosuppression and the increased of the antibiotics 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 transmission was found in the same department.

Aortic aneurysms are characterized by vascular inflammation, neovascularization, and extracellular matrix destruction of the aortic wall. Although experimental studies indicate a potential role of CD248 in microvessel remodeling, the functions of CD248 in human vascular pathologies remain unexplored. Here we aimed to study how CD248 interferes with pathological vascular remodeling of human aortic aneurysms. Immunofluorescent staining showed that CD248 expression was mainly localized in the CD8+ T cells infiltrating in the adventitia and media of aortic walls of patients with ascending thoracic aortic aneurysms. qPCR and immunofluorescent staining analyses revealed increased aortic CD248 expression and infiltrating CD248+CD8+ T cells in aortic aneurysms than in nonaneurysmal aortas. Flow cytometry analysis of human peripheral blood further identified a fraction of circulating CD248+ cells which was confined in the CD8+ T-cell compartment. The increased infiltrating of CD248+CD8+ T cells was coincident with reduced circulating CD248+CD8+ T cells in patients with ascending TAA when compared with patients with coronary artery diseases and healthy donors. The CD248+CD8+ T cells were characterized by upregulated IL-10 and downregulated IL-1β/INF-γ expression when compared with CD248-CD8+ T cells. Moreover, when co-cultured with human aortic endothelial cells, the CD248+CD8+ T cells not only downregulated endothelial expression of ICAM1/VCAM1 and MMP2/3 but also suppressed endothelial migration. This study shows that CD248 reduces pathological vascular remodeling via anti-inflammatory CD248+CD8+ T cells, revealing a CD248-mediated cellular mechanism against human aortic aneurysms.

Objective: Our aim was to assess the relationship between serum lactate levels at intensive care unit (ICU) admission and all-cause mortality in the pediatric ICU. Methods: We used the pediatric intensive care (PIC) database (a large pediatric intensive care database in China from 2010 to 2018) to conduct a retrospective analysis to evaluate the serum lactate levels at ICU admission of 12,213 critically ill children admitted to the ICU. We analyzed the association between serum lactate and all-cause mortality. Adjusted smoothing spline plots, subgroup analysis, and segmented multivariate logistic regression analysis were conducted to estimate the relative risk between proportional risk between serum lactate and all-cause mortality. Results: Of the 12,213 children, 755 (6.18%) died. After fully adjusting for confounding factors, serum lactate was an independent risk factor for all-cause mortality in pediatric ICU (adjusted OR=1.14, 95% CI: 1.12, 1.17). The results of sensitivity analysis showed that in different stratified analyses, the effect of serum lactate on all-cause mortality remained stable. Conclusions: Admission serum lactate is a risk factor, which is independent of the presence of acid-base disorders, inflammation, malnutrition, and renal or hepatic dysfunction, for all-cause mortality in the pediatric intensive care unit.

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.

Aim. To evaluate the diagnosis value of serum human epididymis protein 4 (HE4), cancer antigen 125 (CA125), the Risk of Ovarian Malignancy Algorithm (ROMA), and Copenhagen Index (CPH-I) at early stages for differentiating borderline ovarian tumors from epithelial ovarian cancer. Methods. We recruited 144 borderline ovarian tumors in FIGO stages I and II (BOT I+II), 108 epithelial ovarian cancers in FIGO stages I and II (EOC I+II), and 238 benign ovarian tumor patients with surgical treatment in the retrospective study. The concentration of HE4 and CA125 and the values of CPH-I and ROMA were assessed separately. Results. The HE4 level and ROMA and CPH-I values of EOC I+II were all higher than that of BOT I+II and benign groups whether in all, pre-, or postmenopausal groups (P<0.01). When distinguishing BOT I+II from EOC I+II, the AUC-ROC of CPH-I and HE4 were bigger than CA125 (P<0.001), while the CPH-I has the highest sensitivities in all and postmenopausal groups (78.7%, 85.1%), and HE4 has the highest specificity and PPV (90.91%, 88.64%) in postmenopausal groups. Under pathological stratification, HE4, ROMA, and CPH-I of the serous EOC I+II were higher than that of BOT I+II (P<0.001) and the AUC of the three indices were significantly bigger than CA125 (P<0.001). However, the concentration of HE4 and CA125 and the values of CPH-I and ROMA have no significant difference between the two endometrioid subgroups. The index with the highest sensitivity and NPV among the four indices of different pathological subtype groups was CPH-I, and the index with the highest specificities and PPV was HE4. Conclusion. CPH-I was more valuable than CA125 for differentiating BOT I+II from EOC I+II regardless of menopausal status, while HE4 might be better than CA125 for postmenopausal subgroups. HE4 and CPH-I were more favorable than CA125 for differentiating BOT I+II from EOC I+II in the case of unknown pathology or in serous type.

Objective: Low birth weight (LBW) is a major cause of fetal mortality and morbidity. This study aims to assess the relationship between maternal serum folate levels and LBW in early pregnancy in eastern China. Methods: We conducted a retrospective study including 124 newborns with LBW (born≥1500g to Results: After controlling the covariates, maternal underweight before pregnancy (AOR = 5.15: 95% CI 2.40, 11.05), folate deficiency in early pregnancy (AOR = 2.06: 95% CI 1.03, 4.09), preeclampsia during pregnancy (AOR = 6.02: 95% CI 2.83, 12.81), and premature delivery (AOR =23.81: 95% CI 12.90, 43.95) were risk factors for LBW. However, multipara (AOR = 0.53:95% CI 0.31, 0.91) was a protective factor for LBW. Conclusions: This study revealed different factors associated with LBW in eastern China. Sufficient folate before 18 weeks of pregnancy is a protective factor for LBW. Serum folate > 32.5 nmol/L can reduce the risk of LBW. Therefore, maternal screening of folate statue in the first trimester of pregnancy could be helpful.

Background: Long-chain per- and polyfluoroalkyl substances (PFASs) and their short-chain alternatives have been produced and used extensively in China. However, it is unclear whether these compounds contribute to the risk of gestational diabetes mellitus (GDM) in women residing in contaminated areas. Objective: The study was performed to explore the association between PFASs varying in chain length and the risk of developing GDM. Method: A nested case-control study was conducted in a prospective cohort of 2,460 pregnant women between July 1, 2017, and January 31, 2019 in Shanghai, China. Twelve PFASs of interest were measured using ultra-performance liquid chromatography/quadrupole time-of-flight mass spectrometry (UPLC-Q/TOF MS) in the sera of pregnant women at 16–20 weeks. GDM was diagnosed by an oral glucose tolerance test administered over 24–28 gestational weeks. The cases and controls were matched by maternal age. The relationship between maternal serum PFAS level and GDM risk was determined by conditional logistic and linear regression analyses. Results: A total of 165 GDM cases and 330 controls were enrolled in the study cohort. The frequencies of detection of PFHpA, PFDS, and PFOSA were all ≤80%. Hence, they were excluded from any further risk analysis. Perfluorooctane sulfonate (PFOS) and perfluorooctanoic acid (PFOA) were detected at relatively high serum levels (medians 6.57 ng/mL and 8.07 ng/mL, respectively). The serum levels of perfluorobutanesulfonic acid (PFBS) and perfluorododecanoic acid (PFDoA) were significantly higher in the GDM group than they were in the control group (P = 0.02 and P

Candida krusei attracts attention from medical professionals mainly for its intrinsic resistance to fluconazole and the limited number of drugs available to treat C. krusei vulvovaginal candidiasis. Miltefosine was demonstrated to have good antifungal activity both in vitro and in vivo. Here, we determined the susceptibility profiles of 57 clinical C. krusei isolates from vulvovaginal candidiasis patients and assessed the antifungal activity of miltefosine against C. krusei. All isolates were susceptible to voriconazole and itraconazole, whereas 1.8% of the isolates were of non-wild-type phenotype to amphotericin B. In contrast, miltefosine showed low MICs against all C. krusei isolates with fungicidal activity. The checkerboard assay showed that the synergistic effect of miltefosine in combination with amphotericin B was observed in 25% of the tested planktonic C. krusei isolates and 18.8% of the tested preformed biofilms, whereas miltefosine in combination with fluconazole showed indifferent interaction for all tested planktonic isolates. The presence of sorbitol in the broth microdilution assay did not influence the MIC values of miltefosine against C. krusei, but the presence of ergosterol increased the MIC values. Visible changes in cell content in cells treated with miltefosine were observed. We found that cells treated with miltefosine showed decreased cell viability and chromatin condensation under PI staining, which indicates that miltefosine may induce apoptosis-like cell death in C. krusei. In conclusion, we found miltefosine has a good activity against C. krusei isolates and exerts its fungicidal effect by binding to ergosterol in the cell membrane and inducing apoptosis.

ObjectivesAlthough 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.MethodsWe 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.ResultsThe 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 (pDelftia in the LSIL and HSIL groups compared with the group without an intra-epithelial lesion or malignancy.ConclusionsOur 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.

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 (MIC(90)>16 μg/ml), itraconazole (MIC(90)>16 μg/ml), posaconazole (MIC(90)>16 μg/ml), isavuconazole (MIC(90)>16 μg/ml), amphotericin B (MIC(90)>16 μg/ml), and terbinafine (MIC(90)>64 μg/ml) and high minimum effective concentrations (MECs) to micafungin (MEC(90)>8 μg/ml), with the exception of miltefosine showing an MIC(90) 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.

Candida auris, a globally emerging human fungal pathogen, has arisen as a public health concern worldwide because of its ability to cause nosocomial outbreaks and its resistance to multiple antifungal drugs ([1][1], [2][2]). C. auris is resistant to fluconazole in up to 90% of isolates and exhibits

Background With the gradual severe bacterial resistance and slow development of antibiotics, drug-resistant bacterial strains are widely distributed and have become a serious public health problem. Group B Streptococcus (GBS) which cause Group B strep-related disease is the major cause of severe infection in newborns. However, Clindamycin resistance of GBS induced by Erythromycin is emerging and become important clinical concerns today. Methods A retrospective study was conducted on the drug resistance analysis of GBS strains isolated from Obstetrics and Gynecology Hospital from Jan 2016 to Dec 2017. The clinical and microbiological data including patient demographics, antimicrobial susceptibility testing, relative distribution drug resistance-associated genes mefA & ermB to Erythromycin, and multilocus sequence typing (MLST typing) were collected and analyzed. The Kirby-Bauer and VITEK2-compact were used to perform the susceptibility testing. The double disk diffusion method (D-test) was used for the detection of inducible clindamycin resistance. MLST was employed to identify sequence types of these strains. Polymerase Chain Reaction (PCR) was conducted to detect the drug resistance genes mefA & ermB to Erythromycin. Results A total of 1021 strains were cultured and isolated from 31894 specimens. Erythromycin and clindamycin resistance was 53.6%（547/1021）and 50.1 % (512/1021), respectively, in which 74.4%（407/547）had harbored constitutive macrolide, lincosamide and streptogramin B resistance (cMLS B ), 45.0%（63/140）were inducible MLS B (iMLS B ). Additionally, MLST identified 12 different ST types including a new ST type ST1072 in 63 iMLS B GBS strains and the dominant STs were ST12 (30.1%) and ST19 (25.4%). The resistance ratio of ST19 to Levofloxacin (75.0%) was higher than that of other ST types. The relevance resistance ratio of mefA and ermB was respectively 27.0% and 41.3% among 63 GBS isolates. Conclusion Our study not only demonstrated a genetic diversity in iMLS B GBS in Shanghai through the analysis of MLST typing and resistance genes, but also found that there exist different distribution patterns of resistance and related resistance genes between different ST types. These findings would provide theoretical support for clinical prevention and treatment of resistant iMLS B GBS infection.

Additional file 2: Table S1. Primers for MLST, biofilm, and virulence genes of S. maltophilia.

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

The precise pathophysiological mechanisms of preeclampsia (PE) and preventative strategies remain unknown. Laboratory markers which can help in identifying PE patients from pregnant women and assessing the severity of PE during pregnancy are worthy to be explored. In this study, a retrospective case-control study was designed to assess whether the serum levels of albumin (ALB), total protein (TP), prealbumin (PA), alkaline phosphatase (ALP), lactic dehydrogenase (LDH), D-dimer, fibrinogen (Fbg), platelet (PLT) count, mean platelet volume (MPV), and platelet distribution width (PDW) can help in assessing PE and evaluate its severity. 256 pregnant women were enrolled and classified into 3 groups: mild preeclampsia (mPE,n=85), severe preeclampsia (sPE,n=78), and healthy normotensive controls (control,n=93). Our result showed that the serum levels of ALP, LDH, and D-dimer were significantly higher in mild or severe PE patients compared with the healthy controls (66 (52.5-76.5) vs. 168 (141.5-201.25) vs. 182.5 (120-191.5), 152 (139.75-166.25) vs. 183.5 (163.25-307) vs. 282 (215.25-306), 1.05 (0.65-1.57) vs. 3.05 (2.25-4.08) vs. 5.65 (2.29-7.71)), while ALB, TP, and PA are lower (38 (37-42) vs. 31.5 (25.5-34.5) vs. 28.5 (24-33), 65 (63-68.25) vs. 56.5 (52-61) vs. 51.5 (49-58),219.14±68.25vs.167.88±52.21vs.143.22±50.46). On the other hand, compared with the mPE group, the sPE group showed significantly lower PLT count but higher level of LDH, D-dimer, and Fbg. No significant differences in MPV or PDW were found between any of the two groups. In conclusion, the above markers except for the MPV and PDW may be correlated with PE severity in this patient cohort, indicating possible values of these potential biomarkers in auxiliary diagnosis and severity assessment of PE.

The objective of this study is to evaluate the predictive value of sperm DNA fragmentation Index (DFI) in unexplained recurrent spontaneous abortion (RSA) and to investigate its correlation with conventional sperm parameters. Besides, we aimed to reveal the necessity of establishing a DFI clinical threshold of each laboratory for the prognostic diagnosis of RSA and establish our own DFI threshold. Semen samples were collected from male partners of RSA patients (n = 139) and healthy recent fathers (control, n = 200). DFI was tested using SCSA and conventional semen analysis was performed using an automatic semen analyzer. The DFI value and distribution were compared between the two groups using corresponding statistical software. The diagnostic threshold value was established by ROC curve. The correlation between DFI and the conventional semen parameters of the 139 cases was further analyzed using Student's t test and Mann-Whitney U test. Our result showed that DFI was significantly higher in RSA patients compared with normal donor controls. We established our own DFI threshold at 13.59%. There was only a weak partial correlation between DFI values and conventional sperm analysis parameters. Our present study suggested that DFI might be used as a valuable predictor for RSA independent of conventional sperm parameters. Additionally, we recommend that each laboratory should establish its own clinical DFI threshold for more precise prediction of RSA and we recommend that sperm DNA fragmentation test should be included in complete sperm quality assessment in addition to conventional semen analysis for RSA male partners.

Autophagy is an essential metabolic pathway mediated by lysosomal degradation, which is involved in scavenging and recycling senescent or damaged organelles and biological macromolecules in eukaryotic cells. The present study explored the association between the autophagic activity and chemotherapy resistance of leukaemia cells, and the possibility of using autophagy inhibitors to combat leukemic drug resistance. It was found that the levels of basic autophagy in multidrug‑resistant leukaemia cells (K562/ADM) were significantly higher compared with sensitive cells (K562), and that Adriamycin (ADM) was capable of inducing autophagic activity in K562 and K562/ADM cells. K562 and K562/ADM cells were treated with a series of hydroxychloroquine (HCQ) concentrations to inhibit cellular autophagy and detect cell sensitivity to ADM. The results demonstrated that the sensitivity of K562 cells to ADM was mildly enhanced by HCQ, and that the sensitivity of K562/ADM cells to ADM was markedly strengthened by HCQ. In addition, more typical morphological changes associated with apoptosis emerged, and the ratio of Bax/Bcl‑2 and activity of caspase‑3 were markedly increased in K562/ADM cells treated with HCQ. Notably, the expression of mdr1 mRNA and P‑glycoprotein (P‑gp) in drug‑resistant K562/ADM cells was upregulated along with increasing autophagic activity induced by ADM. Furthermore, HCQ significantly reduced the increase in P‑gp expression by inhibiting autophagic activity. Collectively, these findings indicated that the inhibition of autophagy significantly promoted the sensitivity of K562/ADM cells to ADM by facilitating apoptosis. Furthermore, inhibition of autophagy attenuated the expression of P‑gp; therefore, P‑gp may be involved in autophagic regulation in drug‑resistant cells.

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.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.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).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.

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.

Background The dynamic methylation of human papillomavirus (HPV) 16 DNA is thought to be associated with the progression of cervical lesions. Previous studies that did not consider the physical status of HPV 16 may have incorrectly mapped HPV 16 methylomes. In order to identify reliable biomarkers for squamous cervical cancer (SCC), we comprehensively evaluated the methylation of HPV 16 depending on the integration incidence of each sample. Methods Based on the integration status of 115 HPV 16-infected patients (50 SCC, 30 high-grade squamous intraepithelial lesion [HSIL], and 35 low-grade squamous intraepithelial lesion [LSIL]) and HPV 16-infected Caski cell lines by PCR detection of integrated papillomavirus sequences, we designed a series of primers that would not be influenced by breakpoints for a high-resolution melting (HRM) PCR method to detect the genome methylation. Results A few regions with recurrent interruptions were identified in E1, E2/E4, L1, and L2 despite scattering of breakpoints throughout all eight genes of HPV 16. Frequent integration sites often occurred concomitantly with methylated CpG sites. The HRM PCR method showed 100% agreement with pyrosequencing when 3% was set as the cutoff value. A panel of CpG sites such as nt5606, nt5609, nt5615, and nt5378 can be combined in reweighing calculations to distinguish SCC from HSIL and LSIL patients which have high sensitivity and specificity (88% and 92.31%, respectively). Conclusions Our research shows that combination of CpG sites nt5606, nt5609, nt5615, and nt5378 can be used as potential diagnosis biomarkers for SCC, and the HRM PCR method is suitable for clinical methylation analysis. Electronic supplementary material The online version of this article (10.1186/s13148-018-0445-8) contains supplementary material, which is available to authorized users.

The objective of this work was to investigate correlations between Acinetobacter baumannii isolates from neurosurgical intensive care unit patients and its environment. This is a prospective, observational study. The minimal inhibitory concentrations of antimicrobial agents against 27 clinical and 28 environmental isolates were determined by the agar dilution method. Molecular genotyping was performed by enterobacterial repetitive intergenic consensus PCR (ERIC-PCR), pulsed-field gel electrophoresis (PFGE) and multi-locus sequence typing (MLST). The presence of carbapenemase and metallo-β-lactamase genes were analyzed by specific PCRs and DNA sequencing. From the clinical A. baumannii isolates, 25.9% were found resistant to minocycline, 51.9% to cefoperazone-sulbactam, 59.3% to imipenem and 70% resistant to other antimicrobial agents. Environmental isolates were more sensitive compared with clinical isolates (P

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).

The rapid expansion of carbapenem-resistant Acinetobacter baumannii (CRAB) clinical isolates is a big issue. We investigated the antibiotic susceptibility, molecular epidemiology and resistance gene of A. baumannii collected at two hospitals in Shanghai, China. Besides, the A. baumannii PCR-based replicon typing method (AB-PBRT) was conducted to categorize the plasmids into homogeneous groups on the basis of replicase genes. Most CRAB isolates showed high-level resistance to almost all antibiotics but retain susceptibility to colistin and tigecycline. A total of 101 isolates carried blaOXA-51-like gene. Sequencing identified the presence of blaOXA-66 for CRAB isolates. blaOXA–23 gene were discovered in all CRAB isolates. Each CRAB isolate contained 1–3 of 19 different plasmid replicase (rep) gene homology groups (GRs) and the GR6 (repAci6) was ubiquitous. Genotyping by Multilocus Sequence Typing (MLST) showed seven defined MLST patterns and three novel STs were found. eBURST analysis indicated they were all grouped in CC92 (GCII) with the most frequent ST208 (50%). Two blaOXA–23-bearing transposons were found: Tn2006 and Tn2008. Tn2008 were detected in 54 (96.4%) isolates and Tn2006 in two remaining isolates. The blaOXA–23 carbapenem gene was vitally associated with repAci6 plasmid belong to CC92 clonal group. Our survey revealed severe drug resistance in A. baumannii isolates. Tn2008-containing CC92 A. baumannii were endemic, which may facilitate the blaoxa23 dissemination.

In order to investigate the prevalence of fosfomycin-resistance (fos) determinants in Enterococcus faecium, clinical strains were collected from hospitals throughout China between January 2008 and December 2009. Antimicrobial susceptibility testing was performed, after which the fos genes in all isolates and van genes in vancomycin-resistant isolates were characterized by PCR and sequencing. Conjugation experiments were carried out with fosB-positive E. faecium, DNA fragments flanking the fosB3 gene were sequenced and the genetic environment of fosB3 was analysed. Fosfomycin-resistant E. faecium (FREF) strains were characterized further by multilocus sequence typing (MLST) and PFGE. Among 145 E. faecium clinical isolates, 10 were resistant to fosfomycin with MICs >1024 mg l−1 including six vancomycin-resistant strains of which five were vanA-positive and the sixth vanM-positive. All ten FREF strains harboured the fosB3 gene. Fosfomycin resistance and fosB3 could be transferred by conjugation from nine isolates. The fosB3 and tnpA genes were located in a circular DNA intermediate in all FREF strains and reversely inserted into vanA transposon Tn1546 in four vanA-positive FREF isolates. Ten different PFGE types and seven MLST types were found among the ten fosB3-positive isolates, while all strains belonged to the common clonal complex CC17. In conclusion, the transferable fosfomycin-resistance determinant fosB3 plays an important role in E. faecium resistance to fosfomycin in China.

To explore the serum lipids levels in healthy pregnant women, and to establish the reference intervals of serum lipids in middle and late pregnancy.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%.(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 (P0.05). (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 (P0.01). (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; P0.05). (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.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.