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Objectives Physiological changes during pregnancy can affect the results of renal function tests (RFTs). In this population-based cohort study, we aimed to establish trimester-specific reference intervals (RIs) of RFTs in singleton and twin pregnancies and systematically investigate the relationship between RFTs and adverse pregnancy outcomes. Methods The laboratory results of the first- and third-trimester RFTs, including blood urea nitrogen (BUN), serum uric acid (UA), creatinine (Crea) and cystatin C (Cys C), and the relevant medical records, were retrieved from 29,328 singleton and 840 twin pregnant women who underwent antenatal examinations from November 20, 2017 to January 31, 2021. The trimester-specific RIs of RFTs were estimated with both of the direct observational and the indirect Hoffmann methods. The associations between RTFs and pregnancy complications as well as perinatal outcomes were assessed by logistic regression analysis. Results Maternal RFTs showed no significant difference between the direct RIs established with healthy pregnant women and the calculated RIs derived from the Hoffmann method. In addition, elevated levels of RFTs were associated with increased risks of developing various pregnancy complications and adverse perinatal outcomes. Notably, elevated third-trimester RFTs posed strong risks of preterm birth (PTB) and fetal growth restriction (FGR). Conclusions We established the trimester-specific RIs of RFTs in both singleton and twin pregnancies. Our risk analysis findings underscored the importance of RFTs in identifying women at high risks of developing adverse complications or outcomes during pregnancy.

Objective The purpose of this study was to evaluate the diagnostic performance of the following hemostatic markers in hypertensive disorder of pregnancy (HDP): tissue-type plasminogen activator and inhibitor-1 complex (tPAI-C), thrombomodulin, thrombin-antithrombin complex, plasmin inhibitor-plasmin complex, D-dimer, and fibrinogen degradation products. Methods A total of 311 individuals diagnosed with HDP and 187 healthy controls (HC) of matched gestational age were admitted, including 175 subjects with gestational hypertension, 94 with mild preeclampsia, and 42 with severe preeclampsia. Results Compared with those of the HC group, the plasma concentrations of all the hemostatic markers continuously increased with the clinical severity of the hypertensive disorder, regardless of their statistical significance. In the receiver operating characteristic analysis, tPAI-C displayed the best discrimination performance. Conclusion The tPAI-C level was consistently and significantly elevated across the different HDP groups when compared with the HC group, suggesting aggravated fibrinolysis disorder increasing with the severity of the HDP.

Background Disturbances in maternal lipid metabolism may increase the risk of developing pregnancy complications and adverse perinatal outcomes. However, there is no consensus as to what constitutes normal serum lipid ranges during pregnancy. Our study was aimed to establish trimester-specific serum lipid reference intervals (RIs) and investigate the associations between maternal dyslipidaemia and adverse outcomes in a population-based study. Methods The first- and third-trimester lipid profiles were derived from 16,489 singlet pregnant women for regular antenatal check-ups between 2017 and 2019. The serum samples were assayed for total cholesterol (TC), triglycerides (TG), high-density lipoprotein-cholesterol (HDL-C), and low-density lipoprotein-cholesterol (LDL-C) in the institutional clinical laboratory. The trimester-specific lipid RIs were estimated with both of the direct observational and the indirect Hoffmann methods. The associations between maternal lipid profiling and pregnancy complications and perinatal outcomes were assessed statistically. Results Serum levels of TC, TG, LDL-C and HDL-C were all increased significantly in the third trimester of pregnancy. There was no significant difference between the observed RIs established with healthy pregnant women and the calculated RIs derived from the Hoffmann method. A trend towards increased risks of gestational complications and adverse perinatal outcomes was observed in the subjects with elevated levels of TC, TG, and LDL-C or decreased level of HDL-C. Conclusions In pregnancy, increased serum levels of TC, TG and LDL-C, and a decreased level of HDL-C posed higher risks of developing pregnancy complications and adverse perinatal outcomes.Key messagesIt is necessary to establish trimester-specific reference intervals for serum lipids including TC, TG, LDL-C and HDL-C that were found significantly increased as the gestational age went up. More importantly, around the upper reference limits of TC, TG and LDL-C (or the lower reference limit of HDL-C), the higher the serum lipid levels were (or the lower the HDL-C level was), the higher risks of developing pregnancy complications and adverse perinatal outcomes were observed.

Objective. The incidence of cervical cancer is increasing year by year, which seriously threatens the health of female patients. This study is aimed at investigating the association of sentinel lymph node biopsy (SLNB) with clinicopathological features in cervical cancer patients. Methods. Patients diagnosed with cervical cancer in our hospital from February 1, 2019, to June 30, 2021, were selected as the research subjects. Statistical analysis was performed on the SLN examination of patients with cervical cancer with different pathological characteristics and the correlation between the positive rate of SLN detection and the pathological characteristics of cervical cancer. Results. A total of 59 patients with cervical cancer were included in this study, the SLNB detection rate was 94.92%, 15 patients had lymph node metastasis, and the metastasis rate was 25.42% confirmed by histopathology. Thirteen of them had SLN metastases, and the other 2 had non-SLN metastases. The sensitivity of SLNB was 86.67%, and the false negative rate was 13.33%. Statistical analysis results showed that there was no significant difference in the positive rate of SLN among cervical cancer patients with different FIGO stages, pathological types, degree of differentiation, depth of invasion, and tumor size. In addition, the results of Pearson’s correlation analysis showed that the positive rate of SLN was not significantly correlated with the FIGO stage, pathological type, degree of differentiation, depth of invasion, and tumor size of cervical cancer. Conclusion. SLNB has a high sensitivity, safety, and feasibility in the diagnosis and evaluation of lymph node metastasis in cervical cancer. There is no significant correlation between SLNB and the clinicopathological features of cervical cancer.

Background Lysophosphatidylcholine (LPC) plays pivotal roles in several physiological processes and their disturbances are closely associated with various disorders. In this study, we described the development and validation of a reliable and simple flow injection analysis–tandem mass spectrometry (FIA‐MS/MS)‐based method using dried blood spots (DBS) for quantification of four individual LPC (C20:0, C22:0, C24:0, and C26:0). Methods Lysophosphatidylcholines were extracted from 3.2 mm DBS with 85% methanol containing 60 ng/ml internal standard using a rapid (30 min) and simple procedure. The analytes and the internal standard were directly measured by triple quadrupole tandem mass spectrometry in multiple reactions monitoring mode via positive electrospray ionization. Results Method validation results showed good linearity ranging from 50 to 2000 ng/ml for each LPC. Intra‐ and inter‐day precision and accuracy were within the acceptable limits at four quality control levels. Recovery was from 70.5% to 107.0%, and all analytes in DBS were stable under assay conditions (24 h at room temperature and 72 h in autosampler). The validated method was successfully applied to assessment of C20:0‐C26:0LPCs in 1900 Chinese neonates. C26:0‐LPC levels in this study were consistent with previously published values. Conclusion We propose a simple FIA‐MS/MS method for analyzing C20:0‐C26:0LPCs in DBS, which can be used for first‐tier screening., We proposed a reliable and simple flow injection analysis–tandem mass spectrometry (FIA‐MS/MS)‐based method using dried blood spots (DBS) for quantification of four individual LPC (C20:0, C22:0, C24:0, and C26:0). The elaborated method was successfully applied to assessment of C20:0‐C26:0LPCs in 1900 Chinese neonates.

Background Preeclampsia (PE) prediction has been shown to improve the maternal and fetal outcomes in pregnancy. We aimed to evaluate the PE prediction values of a series of serum biomarkers. Methods The singleton pregnant women (20–36 gestational weeks) with PE‐related clinical and/or laboratory presentations were recruited and had the blood drawn at their first visits. The following markers were tested with the collected serum samples: soluble fms‐like tyrosine kinase 1 (sFlt‐1), placental growth factor (PlGF), thrombomodulin (TM), tissue plasminogen activator inhibitor complex (tPAI‐C), complement factors C1q, B, H, glycosylated fibronectin (GlyFn), pregnancy‐associated plasma protein‐A2 (PAPP‐A2), blood urea nitrogen (BUN), creatinine (Cre), uric acid (UA), and cystatin C (Cysc). Results Of the 196 recruited subjects, 25% (n = 49) developed preeclampsia before delivery, and 75% remained preeclampsia negative (n = 147). The serum levels of sFlt‐1, BUN, Cre, UA, Cysc, and PAPP‐A2 were significantly elevated, and the PlGF level was significantly decreased in the preeclampsia‐positive patients. In the receiver operating characteristics (ROC) analyses, the area under the curves were listed in the order of decreasing values: 0.73 (UA), 0.67 (sFlt‐1/PlGF), 0.66 (Cysc), 0.65 (GlyFn/PlGF), 0.64 (PAPP‐A2/PlGF), 0.63 (BUN), 0.63 (Cre), and 0.60 (PAPP‐A2). The positive predictive values of these serum markers were between 33.1% and 58.5%, and the negative predictive values were between 80.9% and 89.5%. Conclusions The serum markers investigated in current study showed better performance in ruling out than ruling in PE. Absence of pre‐defined latency period between blood draw and the onset of PE limits the clinical utility of these markers., In present study, we evaluated the predictive values of the following serum biomarkers in a prospective study with the women suspected to develop preeclampsia (PE): soluble fms‐like tyrosine kinase 1 (sFlt‐1), placental growth factor (PlGF), pregnancy‐associated plasma protein‐A2 (PAPP‐A2), glycosylated fibronectin (GlyFn), thrombomodulin (TM), tissue plasminogen activator inhibitor complex (tPAI‐C), uric acid (UA), blood urea nitrogen (BUN), creatinine (Cre), and cystatin C (Cysc). The graphical image showed the ROC analyses of the serum markers for PE prediction in the prospective cohort with PE‐related clinical or laboratory presentations. A. ROC analyses for sFlt‐1 and sFlt‐1/PlGF; B. BUN, Cre, UA, and Cysc; C. PAPP‐A2, GlyFn/PlGF and PAPP‐A2/PlGF; D. PlGF.

Breast cancer is the most commonly diagnosed cancer in women and the leading cause of cancer death. About one million new cases are diagnosed each year in China [1]. According to the latest data released by the World Health Organization in 2017, the number of breast cancer deaths in China reached 49,011, accounting for 0.52% of the total deaths. The age-adjusted mortality rate is 5.76 deaths per 100,000 people, ranking China 179 globally. Worldwide, there will be an estimated 2.1 million newly diagnosed breast cancer cases for women in 2018, accounting for a quarter of all cancers in women. And there will be approximately 0.63 million breast cancer death in 2018 [2]. Recently, the incidence of this cancer is increasing in developing countries. Breast cancer is the most frequently diagnosed cancer in most countries (154 of 185 countries) and is also the leading cause of cancer deaths in more than 100 countries; the main exceptions are Australia/New Zealand, Northern Europe, North America (where lung cancer is the top one), and many countries in sub-Saharan Africa (due to the increased incidence of cervical cancer [2]). The incidence of breast cancer is the highest in countries such as Australia/New Zealand, Northern Europe (e.g., UK, Sweden, Finland, and Denmark), Western Europe (Belgium, Netherlands, and France), Southern Europe (Italy), and North America [2].

Background To study the prevalence of the exposure of pregnant women to antimicrobials, a sensitive and reliable liquid chromatography‐tandem mass spectrometry (LC‐MS/MS) method was developed and validated to determine nine antimicrobials, namely sulfadimidine, sulfapyridine, sulfadiazine, sulfathiazole, ofloxacin, ciprofloxacin, norfloxacin, tetracycline, and lincomycin, in human serum. Methods The sample preparation procedure included protein precipitation followed by a cleanup step with solid phase extraction (SPE). Separation was carried out using a CORTECS T3 column (100 × 2.1 mm, 2.7 µm) by gradient elution with a runtime of 8.0 min. Detection was performed on a triple quadruple tandem mass spectrometer with scheduled multiple reaction monitoring (sMRM) in positive ion scan mode. Results The calibration curves were linear over the concentration range of 0.5–50 ng/ml, and the limit of quantitation was between 0.01 and 0.2 ng/ml. For each level of quality control samples, the inter‐ and intra‐assay precision values were less than 12.0%, and the accuracy ranged from 86.1% to 109.0%. No significant matrix effect or carryover was observed. The antimicrobials of interest were stable under all investigated conditions. The validated method was applied to analyze clinical samples from pregnant women in China, and 10 out of 500 samples showed the presence of antimicrobial residues. Moreover, compared with the time‐resolved fluoro‐immunoassay (TRFIA) method, the developed method showed greater sensitivity and specificity. Conclusion This study provides a simple and rapid LC‐MS/MS method for the simultaneous measurement of nine antimicrobials in serum samples, which could be a useful tool in clinical utilization., We proposed a simple SPE LC‐MS/MS method for the analysis of nine antimicrobials in serum. The elaborated method was applied to investigate the exposure of antimicrobials in Chinese pregnant women, and the results have been compared with those obtained from TRFIA method.

Background Phe-restricted diets have been the basis of therapy for phenylketonurics; however, little is known how this treatment effects homeostasis of other important amino acids. This study aimed to describe blood amino acid patterns in neonates with phenylketonuria (PKU) and identify any effects of Phe restriction on these patterns in young children.Methods Neonate group (age 0-4 weeks): 45 PKU patients, 45 age-/sex-matched controls without PKU; 1-4 year-old group: 27 diet-treated PKU patients, 27 age-/sex-matched children without PKU. Concentrations of 11 amino acids were measured using liquid chromatography-tandem mass spectrometry (LC-MS/MS) performed on dried blood spots.Results Elevated blood phenylalanine (Phe), arginine (Arg), citrulline (Cit), valine (Val) and methionine (Met) concentrations were observed in PKU neonates relative to controls (Phe, Arg, Cit, Val: P < 0.001; Met: P < 0.05), of which Phe, Arg, and Met levels could be either partially or completely restored with dietary intervention. Diet had no effect on elevated Cit and Val. Decreased blood tyrosine (Tyr) and proline (Pro) concentrations were observed in PKU neonates compared to controls (P < 0.001). Both amino acids could be near completely restored to normal with dietary treatment. No significant differences in alanine (Ala), leucine (Leu), ornithine (Orn) and glycine (Gly) concentrations were found in the PKU neonates and 1-4 year-old groups (P > 0.05).Conclusions Blood amino acid homeostasis is disrupted in neonates and young children with PKU. Although dietary intervention adjusts amino acid homeostasis in the direction of a healthy equilibrium, complete restoration is not achieved. This persistent disruption may represent a clinically significant barrier to achieving the best possible therapy for those with PKU. Use of laboratory technologies such as LC-MS/MS enable characterization of persistent blood amino acid disequilibrium in the treated phenylketonuric. Testing of this kind presents opportunity for customized treatment feedback that may allow even greater optimization of therapy for neonates and children with PKU.

Background Thromboelastography (TEG) provides global assessment of hemostatic function and has been recommended to monitor potential coagulopathies during pregnancy in which hypercoagulable state is favored. In present study, we established the reference intervals (RIs) of the TEG parameters (R, K, MA, and α‐angle) with Chinese pregnant women of third trimester. In addition, we examined the diagnostic efficacies of the TEG parameters in the patients diagnosed of gestational hypertension (GH), gestational diabetes mellitus (GDM), or preeclampsia (PE). Methods With specified including and excluding criteria, non‐pregnant controls, healthy pregnant women, and pregnant women with GH, GDM, or PE had their venous blood drawn at Beijing Obstetrics and Gynecology Hospital, followed by TEG tests performed in the clinical laboratory. Results The RIs determined with the healthy pregnant women (in third trimester) for R, K, MA, and α‐angle were 4.0‐7.7, 1.2‐3.2, 51.9‐70.1, and 41.4‐74.4, respectively. When compared with the healthy pregnancy group, the K value was significantly decreased in GH patients but increased in PE patients; MA was significantly lower in the PE group. In the receiver operating characteristic curve (ROC) analyses, K value was able to efficiently distinguish normal pregnancy from the GH patients, with an AUC of 0.86 which is far better than those of R (AUC = 0.57) and MA (AUC = 0.56). For the PE patients, the AUC of MA (0.69) was significantly greater than that of R (0.50). Conclusions Thromboelastography may provide more accurate experimental basis for monitoring coagulation functions especially in pregnant women with complications of GH and PE., Thrombelastography (TEG) provides global assessment of hemostatic function and has been recommended to monitor potential coagulopathies during pregnancy in which hypercoagulable state is favored. Here we established the reference intervals (RIs) of the TEG parameters (R, K, MA and α angle) with Chinese pregnant women of third trimester. In addition, we examined the diagnostic efficacies of the TEG parameters in the patients diagnosed of gestational hypertension (GH), gestational diabetes mellitus (GDM) or preeclampsia (PE). The figure presents the boxplots of the TEG parameters R (A), K (B), MA (C) and α angle (D) in non‐pregnant (NP), healthy pregnancy (HP), gestational hypertension (GH), gestational diabetes mellitus (GDM) and preeclampsia (PE) groups. Student’s t test was performed to calculate the p values (* P

Background Due to the low concentration of androgens in women and the limitation of immunoassays, it remains a challenge to accurately determine the levels of serum androgens in polycystic ovary syndrome (PCOS) patients for clinical laboratories. In this report, a liquid chromatography‐tandem mass spectrometry (LC‐MS/MS) method was developed and validated for simultaneous quantitation of testosterone (T), androstenedione (A4), dehydroepiandrosterone sulfate (DHEAS), dihydrotestosterone (DHT), and 17‐hydroxyprogesterone (17‐OHP) that are associated with PCOS. Methods The serum samples were processed by protein precipitation and solid phase extraction before analysis with the in‐house developed LC‐MS/MS. The chromatographic separation was achieved with a C18 column, using a linear gradient elution with two mobile phases: 0.02% formic acid in water (phase A) and 0.1% formic acid in methanol (phase B). The separated analytes were detected by positive or negative electrospray ionization mode under multiple reaction monitoring (MRM). Results The assay for all the five analytes was linear, stable, with imprecision less than 9% and recoveries within ±10%. The lower limits of quantification were 0.05, 0.05, 5, 0.025, and 0.025 ng/mL for T, A4, DHEAS, DHT, and 17‐OHP, respectively. In the receiver operating characteristic curve (ROC) analyses with the PCOS (n = 63) and healthy (n = 161) subjects, the AUC of the four‐androgen combined was greater than that of any single androgen tested in PCOS diagnosis. Conclusions The LC‐MS/MS method for the four androgens and 17‐OHP showed good performance for clinical implementation. More importantly, simultaneous quantitation of the four androgens provided better diagnostic power for PCOS., An in‐house LC‐MS/MS method was developed and validated for simultaneous quantitation of testosterone (T), androstenedione (A4), dehydroepiandrosterone sulfate (DHEAS), dihydrotestosterone (DHT), and 17‐hydroxyprogesterone (17‐OHP) that are associated with polycystic ovary syndrome (PCOS). With PCOS patients and healthy controls, the combinational detection of the four androgens (T, A4, DHEAS, DHT) was superior to any of the single androgen in PCOS diagnosis. (A: The complete chromatograms of the five analytes resolved by the established LC‐MS/MS method: DHEAS (1), A4 (2), T (3), 17‐OHP (4), and DHT (5). B: The ROC analyses were performed with the 63 PCOS patients and 161 healthy controls. The combination detection referred to the simultaneous measurements of the four androgens of T, A4, DHEAS, DHT).

Parkinson’s disease (PD) is the second most common progressive neurodegenerative disease, which is characterized by the loss of dopaminergic neurons in the substantia nigra pars compacta (SNpc). Iron deposit was found in the SNpc of PD patients and animal models, however, the mechanisms involved in disturbed iron metabolism remain unknown. Identifying the relationship between iron metabolism and PD is important for finding new therapeutic strategies. In this study, we found that transgenic overexpression (OE) of Drosophila mitoferrin (dmfrn) or knockdown of Fer3HCH significantly mitigated the reduced mitochondrial aconitase activity, abnormal wing posture, flight deficits and mitochondrial morphology defects associated with PINK1 loss-of-function (LOF). Further work demonstrated that dmfrn OE or Fer3HCH knockdown significantly rescued the impaired mitochondrial respiration in PINK1 LOF flies, indicating that dmfrn or Fer3HCH may rescue PINK1 LOF phenotypes through elevating mitochondrial bioavailable iron levels to promote mitochondrial respiration.

Background: Preeclampsia (PE) is one of the main causes of maternal and fetal morbidity and mortality worldwide. This study was aimed to explore the potential metabolic alterations in women diagnosed with PE and reveal the underlying pathogenesis of disease.Methods: Healthy pregnant women and patients diagnosed with PE were recruited from August 2017 to February 2018. The metabolomic analysis of serum (n=90) and placenta (n=9) samples collected from the two groups were performed with the high performance liquid chromatography coupled with quadrupole-time-of light mass spectrometry (HPLC-QTOF-MS).Results: In serum, 16 metabolites that were present in different concentrations between the two groups were identified, of which pyroglutamic acid (pGlu), methionine, glutamine and taurocholic acid may be used as potential PE diagnosis biomarkers with the area under ROC curve of 0.901, 0.909, 0.892 and 0.873 respectively. Furthermore, the metabolic pathways analysis with differential metabolites in serum and placenta samples showed that linoleic acid and alpha- linolenic acid metabolism, phenylalanine, tyrosine and tryptophan biosynthesis, D-glutamine/D-glutamate metabolism, phenylalanine metabolism, glutathione (GSH) metabolism and tryptophan metabolism were significantly altered and might be involved in PE pathogenesis.Conclusions: These results showed the altered metabolic pathways could contribute to the pathophysiologic mechanisms of PE.

Background Early preeclampsia (PE) prediction has been shown to improve the maternal and fetal outcomes in pregnancy. We aimed to evaluate the PE prediction values of a series of serum biomarkers. Methods The singleton pregnant women with PE-related clinical and/or laboratory presentations were recruited and had the blood drawn at their first visits. The prospective cohort was further divided into the PE-positive and PE-negative groups based on the follow-up results. The following markers were tested with the collected serum samples: sFlt-1, PlGF, M, tPAI-C, compliment factors C1q, B, H, BUN, GlyFn, PAPP-A2, BUN, Cre, UA and Cysc. Results Totally 196 women suspected for PE were recruited with follow-up medical records. Twenty-five percent of the recruited subjects developed PE before delivery and 75% remained PE-negative. The serum levels of sFlt-1, BUN, Cre, UA, Cysc and PAPP-A2 were significantly elevated and the PlGF was significantly decreased in the PE-positive patients. The AUCs were listed in the order of decreasing values: UA (AUC = 0.73), sFlt-1/PlGF (AUC = 0.67), Cysc (AUC = 0.66), GlyFn/PlGF (AUC = 0.65), PlGF (AUC = 0.64), PAPP-A2/PlGF (AUC = 0.64), sFlt-1 (AUC = 0.63), BUN (AUC = 0.63), Cre (AUC = 0.63), and PAPP-A2 (AUC = 0.60) in the ROC analyses. The Logistic regression analysis showed that UA and PAPP-A2 were independent risk factors for PE development with the odds ratios of 3.3 and 2.2 respectively. Moreover, the PPVs of UA and PAPP-A2 were 48.9%, and 40.4%; the NPVs of UA and PAPP-A2 were 82.1% and 81.9%. Conclusions Further studies are warranted to confirm the clinical utilities of the serum markers in PE prediction.

Background Macroprolactin mostly composed of an immunoglobulin G (IgG) and a monomeric prolactin (PRL) represents the major circulating PRL form in the patients with macroprolactinemia that are usually asymptomatic and may not require treatment. In this study, we aimed to evaluate the prevalence of antithyroid and antinuclear antibodies, as well as the IgG subclass distributions in the patients suspected for macroprolactinemia. Methods From January to July in 2018, totally 317 patients with elevated PRL were subjected to the polyethylene glycol (PEG) precipitation assay. The patients with recovery rates of ≤60% were subjected for IgG subclass determination and autoantibody testing including thyroid peroxidase antibody (aTPO), antithyroglobulin antibody (aTG), and antinuclear antibodies (ANA). Results The higher the post‐PEG PRL recovery rates, the less typical hyperprolactinemia symptoms and the higher prevalence of autoantibodies were observed. The IgG1 and IgG3 were the predominant subclasses in the PRL‐IgG complexes according to the immunoprecipitation experiments. Conclusion The patients with post‐PEG PRL recovery rates of, To evaluate the prevalence of associated autoantibodies, as well as the IgG subclass distributions in the patients suspected for macroprolactinemia, totally 317 patients with elevated prolactin were recruited for the initial screening with the polyethylene glycol precipitation assay. The patients with recovery rates of ≤60% were further subjected to IgG subclass determination and autoantibody testing including thyroid peroxidase antibody, antithyroglobulin antibody, and antinuclear antibodies, followed by proper statistical analysis.

Profound physiological changes during pregnancy may affect the requirement of retinol and tocopherol, which are essential micronutrients for the maintenance of maternal health and foetal development. However, the current reference intervals (RIs) of retinol and tocopherol are based on non‐pregnant population. In the present study, a liquid chromatography–tandem mass spectrometry quantitation method for serum retinol and α‐tocopherol was established and validated. In addition, we established trimester‐specific RIs of retinol and α‐tocopherol using the data from paired screening test for 31,301 outpatients who participated in the prenatal vitamins A/E evaluation program at our hospital using the Hoffmann method, which is a simple indirect RI estimation method that does not require the recruitment of healthy subjects. Further, to explore the associations between the levels of retinol and α‐tocopherol and the parameters of complete blood count (CBC), the results of retinol, α‐tocopherol, and CBC of 1,977 pregnant outpatients in the third trimester were analysed. The testing interval between the levels of vitamins and CBC was no more than 7 days. Although no significant changes were noticed in the levels of retinol, the α‐tocopherol levels continuously increased with normal physiological changes throughout pregnancy. Lower retinol levels were associated with the higher incidence of anaemia, whereas higher levels of retinol and lower levels of α‐tocopherol were associated with higher platelet count.

Background Sialic acids are widely distributed in animal tissues, and aberrantly expressed in a variety of cancer types. High expression of sialic acid contributes to tumor aggressiveness by promoting cell proliferation, migration, angiogenesis, and metastasis. Sialidases are responsible for removal of sialic acids from glycoproteins and glycolipids. Methods N-glycomics of bladder cancer cells were detected by MALDI-TOF mass spectrometry. Sialic acid modification in bladder cancer tissue was determined by lectin blot. The down-regulation of NEU1 in bladder cancer cells was determined by high resolution liquid chromatography mass spectrometry (HR LC-MS). The effects of sialidase NEU1 expression on proliferation and apoptosis of human bladder cancer cells were examined by western blot, RT-PCR, confocal imaging and flow cytometry. Moreover, the function of sialic acids on fibronectin-integrin α5β1 interaction were assayed by immunoprecipitation and ELISA. The importance of NEU1 in tumor formation in vivo was performed using BALB/c-nu mice. Expression of NEU1 in primary human bladder cancer tissue samples was estimated using bladder cancer tissue microarray. Results (1) Downregulation of NEU1 was primarily responsible for aberrant expression of sialic acids in bladder cancer cells. (2) Decreased NEU1 expression was correlated with bladder cancer progression. (3) NEU1 overexpression enhanced apoptosis and reduced proliferation of bladder cancer cells. (4) NEU1 disrupted FN-integrin α5β1 interaction and deactivated the Akt signaling pathway. (5) NEU1 significantly suppressed in vivo tumor formation in BALB/c-nu mice. Conclusions Our data showed that NEU1 inhibited cancer cell proliferation, induced apoptosis, and suppressed tumor formation both in vitro and in vivo, by disrupting interaction of FN and integrin β1 and inhibiting the Akt signaling pathway. Our observations indicate that NEU1 is an important modulator of the malignant properties of bladder cancer cells, and is a potential therapeutic target for prognosis and treatment of bladder cancer. Graphical abstract

Purpose. Candida pathogens are commonly found in women and can cause vulvovaginal candidiasis (VVC), whose infection rate is further increased during pregnancy. We aimed to study the Candida prevalence and strain distribution in pregnant Chinese women with a molecular beacon assay. Methodology. From March 2016 to February 2017, a total of 993 pregnant women attending routine antenatal visits at the Beijing Obstetrics and Gynecology Hospital were enrolled. For Candida detection and identification, a unique molecular beacon assay was presented and compared with a traditional phenotypic method. Antifungal susceptibility was tested with the following agents: 5-flucytosine, amphotericin B, fluconazole, itraconazole and voriconazole. Results. The prevalence of Candida was found to be 21.8 % when using the molecular method and 15.0 % when using the phenotypic method. The distribution of the Candida spp. was listed in order of decreasing prevalence: Candida albicans (79.8 %), Candida glabrata (13.5 %), Candida parapsilosis (3.7 %), Candida krusei (2.2 %) and Candida tropicalis (1.1 %). We found that 90.7 % of the Candida detection results were consistent between the molecular and the phenotypic methods. In the cases where the sequencing analyses for the Candida isolates resulted in inconsistent identification, the molecular method showed higher sensitivity than the phenotypic method (96.0 vs 64.6 %). C. albicans, C. glabrata and C. parapsilosis were essentially susceptible to all five antifungal agents tested, whereas C. tropicalis and C. krusei were susceptible to voriconazole and amphotericin B. Conclusion. By exhibiting good sensitivity and specificity, the molecular assay may offer a fast and accurate Candida screening platform for pregnant women.

A 31-year-old Chinese woman with bilateral upper extremity swelling presented to the emergency department at the 20th week of gestation. According to the patient, “normal fetal movement” had stopped 3 days before admission. No abdominal pain or obvious vaginal bleeding was noted. Her body temperature was 36.5 °C and blood pressure was 110/70 mmHg with a heart rate of 80 beats/min. However, there was no fetal heart rate or movement that could be detected by ultrasonography examination, suggesting spontaneous abortion. Although there was no significant family history, the patient had 1 stillbirth 3 years ago at 27 weeks of gestation, accompanied by eclampsia before her labor induction. Because her fetus showed no signs of life with ultrasonography examination, the patient agreed to have ethacridine (Rivanol)-induced labor on day 3 of her hospitalization. On the day of her induction, her blood pressure went up to 158/82 mmHg with proteinuria (urine chemistry: protein +4) and low platelet count (90 × 109/L, normal range 100–300 × 109/L), consistent with a diagnosis of preeclampsia. Magnesium sulfate was administrated during and after the delivery for seizure prophylaxis. After delivery, her blood pressure decreased to 133/81 mmHg and other preeclampsia symptoms resolved in the next few days before discharge. According to the patient's medical record, she had unremarkable endocrine status and negative screening results for infections [toxoplasmosis, rubella cytomegalovirus, herpes simplex, HIV, syphilis, HBV (hepatitis B virus), and HCV (hepatitis C virus)]. The patient had a triple test (α-fetoprotein, human chorionic gonadotropin, and estriol) in her second trimester that showed low risk for Down syndrome. The fetus was sent for autopsy and developmental anomalies were excluded. Relevant laboratory values are shown in Table 1. View this table: Table 1. APS testing and autoimmune antibody testing results. ### QUESTIONS TO CONSIDER 1. How is recurrent pregnancy loss (RPL)4 defined and what …

Additional file 1. Supplementary information/Methods. (Cell lines and cell culture; Antibodies and reagents; Derivatization and separation of N-glycans; Derivatization and separation of N-glycans; Stable isotope labeling by amino acids in cell culture (SILAC); Semi-quantitative and quantitative real-time RT-PCR analysis; Cell transfection; NEU1 gene silencing (siRNA); Proliferation (MTT) assay; Western blotting; Lectin blotting; Immunofluorescence staining; Immunohistochemistry; Wound assay for motility; Cell adhesion assay; Determination of apoptosis by flow cytometry; Tissue microarray (TMA) analysis; Co-immunoprecipitation (co-IP); Extraction of detergent-soluble microdomain (DSM) and detergent-insoluble microdomain (DIM) fractions; TUNEL assay; Cell cycle assay). Table S1. Annotation and quantitative analysis by MALDI-TOF/TOF-MS of N-linked glycan. Table S2. Patient information for sialic acid lectin blot. Table S3. Patient information for analysis of NEU1 expression in tumor and adjacent tissues. Table S4. Patient information for survival analysis. Fig. S1. The intensity of NEU1 protein in LC-MS/MS analysis. Fig. S2.NEU1 mRNA expression in five bladder cancer or epithelial cell lines. Fig. S3. Cell motility during EMT. Fig. S4. Sialidase activity and sialic acid expression in NEU1-overexpressing cells. Fig. S5. Adhesion capacity of YTS-1/Ctrl and YTS-1/NEU1 cells. Fig. S6. EMT marker proteins in YTS-1/Ctrl and YTS-1/NEU1 cells. Fig. S7. NEU1 mRNA level in bladder cancer tissue. Fig. S8. Ki67 and TUNEL staining of mice tumor tissue.

Background The pulmonary surfactant especially lipids in amniotic fluid can reflect the development stage of fetal lung maturity (FLM). However, the conventional lecithin/sphingomyelin (L/S) ratio method by thin layer chromatography (TLC) is insufficient and inconvenient for FLM prediction in clinical practice. Methods The amniotic fluid samples were collected from the pregnant women in labor or undergoing amniocentesis and analyzed for its lipid contents with the liquid chromatography coupled with high‐resolution mass spectrometry (LC‐HRMS) method and the lamellar body count (LBC) method. To reveal the lipidomic profiling of different FLM stages, three groups of amniotic fluid samples including 8 from premature group (gestational week (GW) 38) were compared with the control group (n = 6) of 18 GWs separately. Results In the FLM prediction study, the sensitivity of the LC‐HRMS method and LBC method was 91% and 73%, respectively; the specificity was 100% and 95%, respectively. The most significant metabolic pathway was linoleic acid metabolism between the premature group and the control group. Both glycerophospholipid metabolism and glycosylphosphatidylinositol‐anchor biosynthesis were enriched in the mature groups. In search of potential FLM prediction markers in amniotic fluid, 8 phosphatidylcholines, 1 sphingomyelin, and 1 phosphatidylethanolamine were significantly increased in the mature groups compared with the premature group. Conclusion An efficient LC‐HRMS method for L/S ratio in predicting FLM was established. The linoleic acid metabolism may play an important role in the fetal lung development.

Background Many efforts have been focused on the alternative glycemic marker glycated albumin (GlyA) and its application in pregnancy during which profound physiological changes take place. Our objective was to determine the reference intervals (RIs) of GlyA in healthy Chinese pregnant women and to assess the predictive value of serum GlyA in adverse pregnancy outcomes. Methods Totally 421 healthy subjects including 137 in the first trimester, 152 in the second trimester, and 132 in the third trimester were enrolled from March to July 2019, for the purpose of establishing the trimester-specific RIs of GlyA. In addition, 67 pregnant women diagnosed with GDM were enrolled at 24–28 weeks of gestation. The diagnostic value of GlyA for GDM patients was evaluated and compared with that of fasting plasma glucose (FPG) at 24–28 weeks of gestation. The association between GlyA in the late pregnancy and the adverse pregnancy outcomes was analyzed with the data collected from January to June 2018 at our hospital. Results The estimated RIs of GlyA in present study were 11.26–15.10%, 10.04–13.50%, and 9.76–13.09% in the first, second, and third trimesters respectively. The areas under receiver operating characteristic (ROC) curves were 0.503 for GlyA and 0.705 for FPG. More importantly, the GlyA level in the third trimester was not more elevated in the patients with adverse pregnancy outcomes including large for gestational age (LGA), preterm delivery, hypertension and preeclampsia (PE). The exception was made with the GDM patients who suffered from postpartum hemorrhage and had significantly higher GlyA levels than the control group. Conclusions Our results showed that the GlyA was continuously decreased as the gestational age went up. The GlyA testing has limited value in diagnosing GDM and predicting adverse pregnancy outcomes.

Background As one of the most common pregnant complications, the gestational diabetes mellitus (GDM) is associated with significant adverse pregnant outcomes and it is crucial to accurately monitor the glycemic states of GDM patients. The HbA1c which is a traditional long-term glycemic marker used in diabetic patients, is not recommended in GDM patients during pregnancy. Recently, many efforts have been focused on the alternative marker glycated albumin (GA) and its application in pregnancy during which profound physiological changes take place. Our objective was to determine the reference intervals (RIs) of GA in healthy Chinese pregnant women and to assess the predictive value of serum GA in adverse pregnant outcomes. Methods Totally 479 healthy subjects including 153 in the first trimester, 174 in the second trimester, and 152 in the third trimester were enrolled from March to July 2019, for the purpose of establishing the trimester-specific RIs of GA. The diagnostic value of GA for GDM patients was evaluated and compared with that of fasting plasma glucose (FPG) at 24-28 weeks of gestation. The association between GA in the late pregnancy and the adverse pregnant outcomes was analyzed retrospectively with the data collected from January to June 2018 at our hospital. Results The estimated RIs of GA in present study were 10.87-15.09 %, 10.04-13.50 %, and 9.78-13.03 % in the first, second, and third trimesters respectively. The areas under receiver operating characteristic (ROC) curves were 0.503 for GA and 0.705 for FPG. More importantly, the GA levels of the third trimester did not show significant changes in women with large-for-date birth weight, preterm delivery, postpartum hemorrhage or hypertension when compared in women with normal pregnancy outcomes. The exception was that the GDM patients who developed preeclampsia did have a lower GA level in their late pregnancy. Conclusions Our results show that the GA was continuously decreased as the gestational age went up. It has limited value in diagnosing GDM and predicting adverse pregnancy outcomes.

Background Some of the non‐criteria autoantibodies, especially non‐conventional antiphospholipid (aPL) antibodies, were present with high prevalence in sporadic miscarriages and recurrent pregnant loss. However, whether these autoantibodies are associated with miscarriage patients without apparent causes remain unclear. Methods The subjects were recruited from the female patients visiting the Infertility Center at the Beijing Obstetrics and Gynecology Hospital from January 2017 to March 2018. The women who experienced one sporadic miscarriage (n = 89) or recurrent pregnancy loss (n = 125) were enrolled. The control participants (n = 59) were those women with normal pregnancy history and with no miscarriage or thrombosis experience. The collected serum specimens from above patients and controls were subjected to the 13 non‐criteria autoantibody examinations, targeting non‐conventional phospholipids, thyroid, sperm, endometrial, and anti‐nuclear antigens. Results When compared with the controls, the following non‐criteria antibodies stood out in present study with significantly increased frequency and were listed in the order of decreasing positive rates: aPE IgM (40.0%), ANA (15.2%), aEM IgG (13.6%), aPE IgG (12.8%), and aPT IgM (10.4%). Except for ANA, the presence of aPE IgM, aEM IgG, aPE IgG, and aPT IgM was not associated with positivity of LA tests. In receiver operating characteristic analyses, the combined aPE IgG and aEM IgG biomarker panel had the best discriminating power between miscarriage patients and healthy controls. Conclusion Our findings suggested that the non‐criteria could be included as part of the pregnancy loss evaluation when apparent causes are absent, and the conventional aPLs tests failed to provide interpretations.

Background TORCH including the pathogens of Toxoplasma gondii (TOX), rubella virus (RV), cytomegalovirus (CMV), and herpes simplex virus (HSV) causes intrauterine infections and poses a worldwide threat to women especially in pregnancy. In this study, we described the seasonal difference in TORCH infection and analyzed the anti‐TORCH IgM multipositive serum samples by the indirect immunofluorescence assays (IFA). Methods To observe the seasonal influence of the anti‐TORCH IgG and IgM antibodies, a retrospective study was conducted with 10 669 women (20–40 y old) before pregnancy from August 2016 to July 2017. Totally 199 ELISA anti‐TORCH IgM multipositive serum samples were further tested by IFAs for false‐positive analysis. Results The prevalence of positive HSV1‐IgM, RV‐IgM, HSV2‐IgM, CMV‐IgM, and TOX‐IgM in the present population was 6.30%, 2.55%, 1.94%, 1.24%, and 0.67%, respectively. Additionally, the prevalence of positive RV‐IgM, CMV‐IgM, and HSV1‐IgM was statistically different among four seasons, with the highest positive rates of RV‐IgM (4.12%) in autumn, CMV‐IgM (1.75%) in summer, and HSV1‐IgM (7.53%) in winter. The confirmatory IFAs showed that the positive rates of RUV‐IgM, CMV‐IgM, and HSV2‐IgM were significantly different from those in ELISA screening experiments. Interestingly, only 32.7% (65/199) of the TORCH IgM multipositive results were consistent with those by the IFA, indicating that cross‐reaction caused false positives were common in ELISA IgM antibody screening. Conclusion The TORCH infection displayed different prevalence among four seasons in our 12‐month retrospective study. The IgM multipositives by ELISA screening may need further confirmation analysis due to its relatively high cross‐reaction rate.