Exposure to volatile organic compounds induces cardiovascular toxicity that may involve DNA methylation.

Volatile organic compounds (VOCs) are common air pollutants and water contaminants. We previously found maternal exposure to VOCs was associated with offspring congenital heart disease (CHD). However, little information is available about the effects of VOCs on cardiovascular development at embryonic stage and the underlying mechanism remains unclear. In this study, we aimed to investigate the effects of a mixture of six VOCs on cardiovascular development in zebrafish embryos. Embryos were exposed to different concentrations of VOCs mixture (32 mg/L, 64 mg/L and 128 mg/L) for 96 h, cardiovascular abnormalities including elongated heart shape, increased distance between sinus venosus and bulbus arteriosus, slowed circulation and altered heart rate were observed in a dose- and time-dependent manner. Meanwhile, VOCs exposure increased global DNA methylation levels in embryos. Analysis identified hundreds of differentially methylated sites and the enrichment of differentially methylated sites on cardiovascular development. Two differentially methylated-associated genes involved in MAPK pathway, hgfa and ntrk1, were identified to be the potential genes mediating the effects of VOCs. By enzyme-linked immunosorbent assay, altered human serum hgf and ntrk1 levels were detected in abnormal pregnancies exposed to higher VOCs levels with fetal CHD. For the first time, our study revealed exposure to VOCs induced severe cardiovascular abnormalities in zebrafish embryos. The toxicity might result from alterations in DNA methylation and corresponding expression levels of genes involved in MAPK pathway. Our study provides important information for the risk of VOCs exposure on embryonic cardiovascular development.

Prognosis and outcome of intrauterine treatment of fetuses with critical congenital heart disease.

BACKGROUND: Intrauterine valvuloplasty is an innovative therapy, which promotes ventricular growth and function in some congenital heart diseases (CHDs). The technique remains challenging and can only be performed in a few centers. This study aimed to assess the feasibility and mid-term outcomes of fetal cardiac intervention (FCI) in fetuses with critical CHD in an experienced tertiary center. METHODS: Five fetal aortic valvuloplasty (FAV) or fetal pulmonary valvuloplasty (FPV) procedures were performed in our fetal heart center between August 2018 and May 2022. Technical success was defined as crossing the aortic or pulmonary valve and balloon inflation, followed by evidence of increased blood flow across the valve and/or new regurgitation. Follow-up clinical records and echocardiography were obtained during the prenatal and postnatal periods. RESULTS: Five fetuses received FAV or FPV, including critical aortic stenosis (n = 2) and pulmonary atresia with intact ventricular septum (n = 3). The mean maternal age was 33.0 ± 2.6 years. The median gestational age (GA) at diagnosis was 24 weeks (range, 22-26 weeks). The median GA at intervention was 29 weeks (range, 28-32 weeks). All five cases underwent successful or partially successful procedures. One patient had pulmonary valve perforation without balloon dilation. No procedure-related deaths or significant complications occurred. However, one neonatal death occurred due to heart and renal failure. The median follow-up period was 29.5 months (range, 8.0-48.0 months). The four surviving patients had achieved biventricular circulation, exhibited improved valve, and ventricular development at the last follow-up visit. CONCLUSION: Intrauterine FCI could be performed safely with good prognosis in critical CHD.

Characteristics and outcomes of fetal ventricular aneurysm and diverticulum: combining the use of a new technique, fetal HQ.

Objectives: Congenital ventricular aneurysms or diverticulum (VA/VD) are rare cardiac anomalies with lack prenatal evaluation data. The present study aimed to provide the prenatal characteristics and outcomes from a tertiary center and the use of new techniques to evaluate the shape and contractility of these fetuses. Methods: Ten fetuses were diagnosed with VA or VD, and 30 control fetuses were enrolled. Fetal echocardiography was performed to make the diagnosis. The prenatal echo characteristics and follow-up data were carefully reviewed. The shape and contractility measurements of the four-chamber view (4CV) and both ventricles were measured and computed using fetal fetal heart quantification (HQ). Results: A total of 10 fetuses were enrolled, including 4 cases of left ventricular diverticulum, 5 cases of left ventricular aneurysm, and 1 case of right ventricular aneurysm (RVA). Four cases chose to terminate the pregnancy. The RVA was associated with a perimembranous ventricular septal defect. Two cases had fetal arrhythmia, and one case had pericardial effusion. After birth, one case underwent surgical resection at five years old. The 4CV global sphericity index (SI) of free-wall located ventricular outpouching (VO) was significantly lower than the apical ones and the control group (p < 0.01). Four of five apical left VOs had significant higher (>95th centile) SI in base segments, and three of four left VOs in the free-wall had significant lower (< 5th centile) SI in the majority of 24 segments. Compared to the control group, the left ventricle (LV) global longitudinal strain, ejection fraction, and fractional area change were significantly decreased (p < 0.01), while the LV cardiac output of the cases was in the normal range. The transverse fraction shortening of the affected segments of ventricles was significantly lower than the other ventricle segments (p < 0.01). Conclusions: Fetal HQ is a promising technique to evaluate the shape and contractility of congenital ventricular aneurysm and diverticulum.

Relationship of the levels of reactive oxygen species in the fertilization medium with the outcome of in vitro fertilization following brief incubation.

Embryo selection in in vitro fertilization-embryo transfer (IVF-ET) mostly relies on morphological assessment using a conventional microscope or the time-lapse monitoring system, which is not comprehensive. Inappropriate levels of reactive oxygen species (ROS) in the fertilization medium may cause damage to gametes, eventually leading to adverse IVF outcomes. The present study aimed to identify the optimal oxidation-reduction level in the fertilization medium for IVF outcomes by measuring the static oxidation-reduction potential (sORP) using a highly accurate and sensitive MiOXSYS system. A total of 136 patients undergoing IVF following brief incubation were divided equally into 4 groups in this prospective cohort study. The sORP value in the fertilization medium was detected using the MiOXSYS system, and its relationship with IVF outcomes was analyzed. The primary outcome was pregnancy outcomes, including live birth rate (LBR), clinical pregnancy rate (CPR), biochemical pregnancy rate (BPR), and implantation rate (IR). The secondary outcome was embryo quality, including fertilization rate (FR), cleavage rate (CR), available embryo rate (AER), and good-quality embryo rate (GQER). Group II (sORP: 228.7-235.3 mV) showed a higher LBR, CPR, BPR, and IR compared with Group III (sORP: 235.4-242.7 mV), presented as follows: LBR (32.0% for Group II vs 3.6% for Group III, P = 0.033), CPR (32.0% for Group II vs 3.6% for Group III, P = 0.033), BPR (36.0% for Group II vs 3.6% for Group III, P = 0.019), and IR (31.3% for Group II vs 2.7% for Group III, P = 0.003). The FR in Groups I and II had lower significant differences compared with that in Groups III and IV (71.7% and 70.3% for Groups I and II vs 83.5% and 80.4% for Groups III and IV, P = 0.000). The GQER in Group I to Group IV was 32.7%, 37.4%, 26.5%, and 33.3%, respectively (P = 0.056). This study indicated that the sORP value in the fertilization medium might be a potential indicator of embryo quality and pregnancy outcome.

Maternal and neonatal blood vitamin D status and neurodevelopment at 24 months of age: a prospective birth cohort study.

BACKGROUND: This study aimed to explore the relationship of 25-hydroxyvitamin D [25(OH)D] in three trimesters and at birth with neurodevelopment at 24 months of age. METHODS: From 2013 to 2016, pregnant women from the Shanghai Birth Cohort in China were recruited for the study. Altogether, 649 mother-infant pairs were included. Serum 25(OH)D was measured with mass spectrometry in three trimesters, and cord blood was divided into deficiency (< 20 and < 12 ng/mL, respectively), insufficiency (20-30 and 12-20 ng/mL, respectively), and sufficiency (≥ 30 and ≥ 20 ng/mL, respectively). Bayley-III scale was used to assess cognitive, language, motor, social-emotional, and adaptive behavior development at 24 months of age. The Bayley-III scores were grouped into quartiles, and scores within the lowest quartile were defined as suboptimal development. RESULTS: After adjusting for confounding factors, cord blood 25(OH)D in the sufficient group was positively correlated with cognitive [ß = 11.43, 95% confidence interval (CI) = 5.65-17.22], language (ß = 6.01, 95% CI = 1.67-10.3), and motor scores (ß = 6.43, 95% CI = 1.73-11.1); cord blood 25(OH)D in the insufficient group was also positively correlated with cognitive scores (ß = 9.42, 95% CI = 3.74-15.11). Additionally, sufficient vitamin D status in the four periods and persistent 25(OH)D ≥ 30 ng/mL throughout pregnancy were associated with a lower risk of suboptimal cognitive development in adjusted models, although the effects were attenuated after applying the false discovery rate adjustment. CONCLUSIONS: Cord blood 25(OH)D ≥ 12 ng/mL has a significant positive association with cognitive, language, and motor development at 24 months of age. Sufficient vitamin D status in pregnancy might be a protective factor for suboptimal neurocognition development at 24 months of age.

Association between Suboptimal 25-Hydroxyvitamin D Status and Overweight/Obesity in Infants: A Prospective Cohort Study in China.

This study aimed to investigate whether 25-hydroxyvitamin D (25(OH)D) concentrations are correlated to overweight/obesity in infants and to explore a threshold of 25(OH)D. A total of 1205 six-month-old infants from two community hospitals in Shanghai were randomly recruited, and 925 of them were followed up at 12 months. Concentration of 25(OH)D, weight, and length were measured at two time points. Overweight/obesity was defined as a weight-for-length Z-score >97th percentile. The prevalence of overweight/obesity at 6 and 12 months was 6.88% and 5.26%, respectively. The occurrence of vitamin D (VitD) deficiency (<20 ng/mL) at 6 and 12 months was 6.56% and 2.05%, respectively. Concentration of 25(OH)D at the corresponding age was negatively associated with weight-for-length percentile (WLP) at both 6 (adjusted ß: −0.14; 95% CI: −0.27, −0.02; p = 0.02) and 12 months (adjusted ß: −0.22; 95% CI: −0.41, −0.02; p = 0.03), while the relationship between 25(OH)D at 6 months and WLP at 12 months was nonlinear, where 35 ng/mL was identified as an inflection point. Those with a concentration of 25(OH)D <35 ng/mL at 6 months had a higher risk of overweight/obesity (adjusted OR: 1.42; 95% CI: 1.06, 1.91; p = 0.02) compared to the group with a concentration of 25(OH)D ≥35 ng/mL. Moreover, a concentration of 25(OH)D <35 ng/mL at two time points significantly increased the risk of overweight/obesity at 12 months compared to the group with 25(OH)D concentration ≥35 ng/mL at two time points (adjusted OR: 2.91; 95% CI: 1.13, 7.46; p = 0.03). A suboptimal 25(OH)D concentration <35 ng/mL significantly increases the risk of overweight/obesity in infants.

Congenital aortocaval fistula combined with patent ductus arteriosus in an infant: a case report.

Congenital aortocaval fistula (ACF) is a rare cardiac malformation. While it can occur in combination with patent ductus arteriosus (PDA), this has not been reported. In this case, a 1-year-old infant had a heart murmur found in a routine physical examination, and PDA was revealed by transthoracic echocardiography and abdominal ACF was detected by three-dimensional coronary artery computed tomography. Percutaneous interventional therapy, used for ACF and PDA, was performed to occlude the malformation. The patient presented good health without any discomfort at a 1-year follow-up. The percutaneous closure of ACF and PDA with an Amplatzer vascular device can be considered an appropriate option.

A new perspective on semen quality of aged male: The characteristics of metabolomics and proteomics.

Background: Semen quality is negatively correlated with male age and is mainly quantified by a routine semen analysis, which is descriptive and inconclusive. Sperm proteins or semen metabolites are used as the intermediate or end-products, reflecting changes in semen quality, and hold much promise as a new biomarker to predict fertility in advanced-aged males. Objectives: In this study, we sought to assess whether the semen metabolome and proteome of aged males can affect semen quality and serve as biomarkers for predicting semen quality. Materials and methods: We retrospectively analyzed 12825 males that underwent semen routine analysis to understand the age-dependent changes in sperm quality. To identify the difference between aged and young adults, metabolomics (n=60) analyses of semen and proteomics (n=12) analyses of sperm were conducted. Finally, integrated machine learning of metabolomics was conducted to screen biomarkers to identify aging semen. Results: We discovered that male age was positively correlated with sperm concentration as well as DNA fragmentation index(DFI), and negatively with progressive motile sperm count, total sperm count, sperm volume and progressive sperm motility. The differential metabolites were significantly enriched in various metabolic pathways, and four of these differential metabolites (Pipamperone, 2,2-Bis(hydroxymethyl)-2,2',2''-nitrilotriethanol, Arg-Pro and Triethyl phosphate) were utilized to establish a biomarker panel to identify aging semen. Proteomic analysis showed that differential proteins were significantly enriched in protein digestion and absorption and some energy-related pathways. An integrated analysis of the metabolome and proteome identified differential energy metabolism and oxidative stress-related proteins, which could explain the decreased motility and the increased DFI of aging sperm. Discussion and conclusion: We provide compelling evidence that the changes in semen metabolome and sperm proteome are related to the decline of semen quality in aged males. Moreover, a biomarker panel based on four metabolites was established to identify aging semen.

Associations between maternal vitamin D status during three trimesters and cord blood 25(OH)D concentrations in newborns: a prospective Shanghai birth cohort study.

PURPOSE: Prenatal vitamin D (VitD) deficiency influences children's health in later life. We aimed to test the associations between maternal VitD status in each of the three trimesters of pregnancy and cord blood 25(OH)D concentrations in newborns. METHODS: Participants were pregnant women recruited from the Shanghai Birth Cohort (SBC) (n = 1100). Of all the participants, 946 completed the collection of venous blood at early (< 16 weeks, T1), mid- (24-28 weeks, T2), and late (32-34 weeks, T3) pregnancy as well as the corresponding cord blood in the newborns. Maternal serum 25(OH)D concentrations were measured by LC-MS/MS, and the information on confounding factors was obtained through a standardized questionnaire. RESULTS: The mean 25(OH)D concentrations at time points T1, T2, T3 in maternal blood and cord blood of the newborns were 26.31 ng/mL, 31.92 ng/mL, 35.62 ng/mL, and 19.77 ng/mL, respectively. Neonatal 25(OH)D level in cord blood was positively correlated with maternal serum 25(OH)D levels at each trimester, and the strongest correlation was found at time point T3. CONCLUSION: Maternal 25(OH)D concentrations at each trimester were positively associated with neonatal VitD status in cord blood, and the strongest correlation was found in the late stage of pregnancy, which could be considered as a sensitive time window. Attention should be paid to the nutritional status of VitD during pregnancy to better prevent the VitD deficiency in neonates.

Urine concentrations of perfluoroalkyl acids in children and contributions of dietary factors: a cross-sectional study from Shanghai, China.

The production and emission of short-chain perfluoroalkyl acids (PFAAs) has increased over the years to replace long-chain PFAAs, leading to frequent detection in the environment and raising global concerns about the potential impacts on human health. In this study, the specific urine levels of 10 PFAAs were obtained from 189 children (age 8-12 years) from two primary schools located in urban and suburban areas of Shanghai in 2019, and the contributions of dietary factors were investigated. Perfluorohexanoic acid (PFHxA), perfluoroheptanoic acid (PFHpA), and perfluorobutane sulfonate (PFBS) were detected in 100%, 99.5%, and 87.3% of the samples, with median concentrations of 20.20 ng/L, 46.50 ng/L, and 20.95 ng/L, respectively. The most abundant PFAA was perfluorooctanoic acid (PFOA), with a median concentration of 78.90 ng/L. The concentration of ∑PFAAs ranged from 61.10 to 4108.93 ng/L, with a median concentration of 253.12 ng/L. Children aged 8-9 years had higher median levels of PFBS, perfluorohexane sulfonate (PFHxS), and perfluorooctane sulfonate (PFOS) than children aged 10-12 years. Obese/overweight children had lower levels of PFHpA, PFBS, and PFOS. The intake of red meats, tubers, sugared beverages, fish and seafood, and eggs contributed to higher concentrations of PFAAs, while frequent intake of poultry and soy milk was associated with lower PFAA concentrations.

Pre-pregnancy exposure to fine particulate matter (PM2.5) increases reactive oxygen species production in oocytes and decrease litter size and weight in mice.

Exposure of females to fine particulate matter ≤2.5 µm in diameter (PM2.5) prior to pregnancy could produce adverse impact on fertility and enhances susceptibility of the offspring to a variety of diseases. In the current study, female C57BL/6 mice (6 weeks of age) were exposed to either concentrated PM2.5 or filtered air (average PM2.5 concentration: 115.60 ± 7.77 vs. 14.07 ± 0.38 µg/m-3) using a whole-body exposure device for 12 weeks. Briefly, PM2.5 exposure decreased anti-Müllerian hormone level (613.40 ± 17.36 vs 759.30 ± 21.90 pg mL-1, P＜0.01) and increased reactive oxygen species (ROS) level (45.39 ± 0.82 vs 24.20 ± 0.85 arbitrary unit in fluorescence assay, P＜0.01) in oocytes. The exposure increased oocyte degeneration rate (21.5% vs 5.1%, respectively (P＜0.01) and decreased the 2-cell formation rate (71.9% vs 86.0%, P < 0.01). Transcriptome profiling using RNA sequencing showed wide spectrum of abnormal expression of genes, particularly those involved in regulating the mitochondrial respiratory complex in oocytes and metabolic processes in blastocysts. The exposure decreased litter size (6 ± 0.37 vs 7 ± 0.26, P＜0.05) and weight (1.18 ± 0.02 vs 1.27 ± 0.02 g, P＜0.01). In summary, PM2.5 exposure decreased female fertility, possibly through increased ROS production in oocytes and metabolic disturbances in developing embryos. The cause-effect relationship, however, requires further investigation.

Effects of circulating vitamin D concentrations on emotion, behavior and attention: A cross-sectional study in preschool children with follow-up behavior experiments in juvenile mice.

BACKGROUND: Although few studies show that vitamin D (VitD) deficiency has a negative effect on children's emotion and behavior, the effects of the excessive VitD and the appropriate 25(OH)D concentration have never been reported. We investigated the effect of the deficient and excessive VitD on emotion, behavior and attention. METHODS: 351 preschool children in a multicenter study in Shanghai, China that had serum 25(OH)D measurements and emotion, behavior and attention measures were included in the analyses. In animal experiments, C57 mice were randomly assigned to three groups (n = 8): control (C) group, VitD deficiency (VDD) group, and VitD overdose (VDO) group. The emotion, behavior and attention of juvenile mice were evaluated through the behavioral experiments. RESULTS: There was an "U" relationship between serum 25(OH)D concentration and emotion, behavior and attention. Compared with 20-40 ng/mL group, the odds ratios (ORs) were 1.5 (1.0, 4.8) for emotional problem, 3.8 (1.2, 12.1) for conduct problem and 1.8 (1.1, 5.7) for inattention in <20 ng/mL group. Meanwhile, compared with 20-40 ng/mL group, ORs were 9.5 (2.9, 31.4) for impulsive hyperactivity, and 3.9 (1.2, 12.9) for conduct problem in >40 ng/mL group. Consistent with the results in children, animal experiments showed that the attention level decreased in VDD group, while the anxiety level, hyperactive level and aggressiveness in VDD group and VDO group were significantly increased, respectively. LIMITATIONS: 25(OH)D measurements were only available in one season. CONCLUSION: The deficient and excessive VitD status both adversely affected children's emotion, behavior and attention.

Development of a sensitive HPLC-MS/MS method for 25-hydroxyvitamin D2 and D3 measurement in capillary blood.

BACKGROUND: Measurement of 25-hydroxyvitamin D [25(OH)D)] levels is important. The current method requires a relatively large volume of serum. To minimize the amount of serum needed, we established a high-performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS) method to measure 25(OH)D in capillary serum. METHODS: Venous blood and fingertip blood were collected from 90 participants. Volumes of 100 µL of venous serum and 20 µL of capillary serum were collected. The serum samples were pretreated by protein removal, extraction and concentration, and an HPLC-MS/MS method based on chromatographic separation and multi reactive ion monitoring was conducted. The intra- and inter-batch variation coefficients were less than 10% for both 25-hydroxyvitamin D3 [25(OH)D3 ] and 25-hydroxyvitamin D2 [25(OH)D2 )]. For venous specimens, the accuracies were 3.87% and 4.91%, respectively. For capillary specimens, the accuracies were 1.65% and 5.32%, respectively. RESULTS: The limit of detection (LOD) of 25(OH)D3 was 0.01 ng/mL, and the LOD of 25(OH)D2 was 0.05 ng/mL. The results showed that the mean concentration of 25(OH)D in venous blood was 22.56 ± 9.50 ng/mL, while the mean concentration of 25(OH)D in capillary blood was 18.14 ± 7.86 ng/mL. Furthermore, the adjusted capillary blood 25(OH)D level was 22.99 ± 10.24 ng/mL by the correction formula in our study. Similarly, the mean concentration of 25(OH)D3 in capillary blood was 17.98 ± 7.98 ng/mL. The adjusted capillary blood 25(OH)D3 level was 22.85 ± 10.42 ng/mL. No difference in the content of 25(OH)D or 25(OH)D3 was found between venous serum and corrected capillary serum. The correlation coefficients between venous and corrected capillary concentrations of 25(OH)D and 25(OH)D3 were 0.7941 and 0.8103, respectively, and the areas under the receiver operating characteristic curve were 0.9367 and 0.9565, respectively. CONCLUSIONS: This capillary blood method requires minimal sample preparation and is suitable for routine use in the 25(OH)D detection.

Neonatal outcome of cases with isolated prenatal ventricular disproportion with a dominant right ventricle.

OBJECTIVES: Isolated prenatal ventricular disproportion with a dominant right ventricle represents a challenge in decision-making for both physicians and pregnant women. In the current study, we sought to delineate the postnatal outcomes of these cases. METHODS: This retrospective analysis included consecutive cases of isolated ventricular disproportion identified using complete fetal echocardiography at the Fetal Heart Center of Xinhua Hospital from January 2014 to October 2017. Postnatal cardiac outcome was examined using transthoracic echocardiography within the first 6 months after birth. RESULTS: A total of 90 fetuses were included in the final analysis. The median gestational age (GA) at diagnosis was 29 weeks (range 24 to 36). At postnatal examination, cardiac malformations were detected in 39 cases (43.3%), including 25 (27.8%) cases of congenital cardiac septal defects, eight (8.9%) of persistent left superior vena cava, four (4.4%) of left-sided obstructive diseases, and one (1.1%) case of coronary fistula. Nineteen cases (21.1%) with fetal cardiac malformations had significant lower GA at diagnosis (P = .01) and greater right to left ventricle ratio (1.38 vs 1.30, P = .02). Neonatal surgical intervention was not required in any of the cases. CONCLUSIONS: Isolated prenatal ventricular disproportion with a dominant right ventricle comprises minor postnatal cardiac malformations and doesn't require neonatal intervention.

Vitamin D deficiency during pregnancy affects the function of Th1/Th2 cells and methylation of IFN-γ gene in offspring rats.

The effects of maternal vitamin D status on offspring's Th1/Th2 cell function and the related mechanisms have not been reported. In this study, we established the rat model of vitamin D deficiency during pregnancy. 48 female Sprague-Dawley rats (8 weeks old) were randomly assigned to three groups (n = 16/group): control group (fed with standard AIN-93 G diet until parturition), vitamin D deficiency group (VDD group, fed with vitamin D deficient diet until parturition) and vitamin D supplementation group (VDS group, fed with vitamin D deficient diet prior to mating and with standard AIN-93 G diet during pregnancy). At 4 weeks of age, the ratio of T helper type 1/ T helper type 2 (Th1/Th2) cells and the levels of Th1/Th2 cytokines (IFN-γ, IL-4, IL-5, IL-6, IL-10 and IL-13) in offspring rats were determined by Flow Cytometry and Meso Scale Discovery, respectively. Furthermore, DNA methyltransferase (DNMT) activity as well as the methylation levels of IFN-γ and IL-4 genes were measured. As a result, rats in the VDD group showed a significant decrease in Th1/Th2 ratio and IFN-γ level and an increase in IL-4 level. Additionally, up-regulated DNMT activity and increased methylation rate of IFN-γ gene was shown in VDD offspring rats. Supplementation with vitamin D during pregnancy reversed the above abnormalities. In conclusion, maternal vitamin D deficiency affected the function of Th1/Th2 cells and methylation of IFN-γ gene in offspring rats. Meanwhile, maternal vitamin D deficiency had the potential to regulate DNMT activity, which may determine the status of methylation.

Rare Copy Number Variants Identify Novel Genes in Sporadic Total Anomalous Pulmonary Vein Connection.

Total anomalous pulmonary venous connection (TAPVC) is a rare congenital heart anomaly. Several genes have been associated TAPVC but the mechanisms remain elusive. To search novel CNVs and candidate genes, we screened a cohort of 78 TAPVC cases and 100 healthy controls for rare copy number variants (CNVs) using whole exome sequencing (WES). Then we identified pathogenic CNVs by statistical comparisons between case and control groups. After that, we identified altogether eight pathogenic CNVs of seven candidate genes (PCSK7, RRP7A, SERHL, TARP, TTN, SERHL2, and NBPF3). All these seven genes have not been described previously to be related to TAPVC. After network analysis of these candidate genes and 27 known pathogenic genes derived from the literature and publicly database, PCSK7 and TTN were the most important genes for TAPVC than other genes. Our study provides novel candidate genes potentially related to this rare congenital birth defect (CHD) which should be further fundamentally researched and discloses the possible molecular pathogenesis of TAPVC.

Variant Alleles of the ESR1, PPARG, HMGA2, and MTHFR Genes Are Associated With Polycystic Ovary Syndrome Risk in a Chinese Population: A Case-Control Study.

Polycystic ovary syndrome (PCOS) is the most common endocrinopathy in women of reproductive age, with a prevalence of 6-8%. Although the etiology of PCOS has been investigated extensively, the association between genetic predisposition and PCOS risk is largely unknown. In this study, we genotyped 63 SNPs in 10 genes among 361 PCOS patients and 331 healthy controls in a Chinese Han population. The following variant alleles were significantly associated with decreased PCOS risk: ESR1 rs9340799 (P = 0.000), PPARG rs709154 (P = 0.013), and rs1151996 (P = 0.013), HMGA2 rs2272046 (P = 0.000), MTHFR rs1801133 (P = 0.000). Accordingly, the following genotypes at various loci were associated with reduced PCOS risk: GA genotype at rs9340799 (P < 0.0001) in ESR1, TA genotype at rs709154(P < 0.0001) in PPARG and CA genotype at rs2272046 (P < 0.0001) in HMGA2. Moreover, GA genotype at rs1999805 (P = 0.013) in ESR1 and TT genotype at rs1801133 in MTHFR (P < 0.0001) correlated with elevated PCOS risk. Furthermore, haplotype analysis revealed significant differences in haplotype distributions of CYP11A1, ESR2 and PPARG gene between cases and controls. In addition to confirming that ESR1 rs9340799, HMGA2 rs2272046 and MTHFR rs1801133 are related to the risk of PCOS, these findings also provide the first evidence that PPARG rs709154 and ESR1 rs1999805 are significantly associated with PCOS risk in a Chinese population. Further functional studies are warranted to elucidate the underlying biological mechanisms.

Vitamin D deficiency and high serum IL-6 concentration as risk factors for tubal factor infertility in Chinese women.

OBJECTIVES: The aim of this study was to investigate the relationship between 25-hydroxyvitamin-D [25(OH)D] and female infertility and to further explore the role of inflammatory cytokines. METHOD: We recruited 356 infertile women diagnosed with tubal factor infertility (TFI) or polycystic ovary syndrome (PCOS) or endometriosis, as well as 180 fertile women. Serum concentrations of 25(OH)D, interleukin (IL)-6, IL-1 ß, and interferon-α were measured. RESULTS: The 25(OH)D concentration in TFI women was the lowest (16.9 ng/mL) and was significantly different from that in the fertile women (19.4 ng/mL; P <0.05)]; whereas women with TFI had higher IL-6 concentrations. After adjusting for confounders, 25(OH)D deficiency presented a risk factor for TFI (odds ratio [OR], 4.2; 95% confidence interval [CI], 1.5-11.3). There was a dose-effect relation between IL-6 tertiles and TFI: the higher the IL-6, the higher the risk for TFI (middle versus low: OR, 3.7; 95% CI, 1.5-9.5; high versus low: OR, 13.2; 95% CI, 4.8-36.4). IL-6 showed a negative correlation with 25(OH)D (r = -0.19). Women with both high IL-6 and low 25(OH)D had the highest risk for TFI (OR, 10.6; 95% CI, 4.2-26.3). CONCLUSIONS: Both vitamin D deficiency and high serum IL-6 concentration are risk factors for TFI. Serum 25(OH)D concentration was significantly and negatively correlated with serum IL-6. There was an interaction between IL-6 and 25(OH)D for the risk for TFI-related infertility. We hypothesized that vitamin D might reduce the risk for TFI through suppressing the production of IL-6.

Metal element excretion in 24-h urine in patients with Wilson disease under treatment of D-penicillamine.

Wilson disease is an inherited autosomal recessive disorder causing copper accumulation and consequent toxicity. D-Penicillamine, a potent metal chelator, is an important therapy for Wilson disease. To investigate the changes of metal elements under the treatment of D-penicillamine, we determined the levels of Cu, Zn, Mg, Ca, Fe, Se, Mn, Pb, Hg, Cd, As, Tl, and Al by ICP-MS in 24-h urine of 115 Wilson disease patients who had received treatment with D: -penicillamine for 1 month to 22 years at maintenance doses, as well as 115 age-matched, healthy controls. The levels of Cu, Mg, Ca, Zn, Hg, Pb, Tl, Cd, and Mn in the 24-h urine of the cases were significantly higher than those of the controls (P < 0.05), and the observed increases in the levels of Mg, Ca, and Zn were directly correlated with the treatment duration with Pearson Correlation Coefficient (R) of 0.356 (Mg), 0.329 (Ca), and 0.313 (Zn), respectively (P < 0.05). On the other hand, the levels of Al and As in the 24-h urine were lower than those of the controls (P < 0.05) and were negatively correlated with the treatment time with R of -0.337 (Al) and -0.398 (As), respectively, (P < 0.05). Thus, this study indicates that the levels of metal elements may be altered in patients with Wilson disease under the treatment of D-penicillamine.

Brain-derived neurotrophic factor inhibits phenylalanine-induced neuronal apoptosis by preventing RhoA pathway activation.

Phenylketonuria (PKU) is neuropathologically characterized by neuronal cell loss, white matter abnormalities, dendritic simplification, and synaptic density reduction. The neuropathological effect may be due to the 'toxicity' of the high concentration of phenylalanine, while little is known about the related treatments to block this effect. In this study, we reported that brain-derived growth factor (BDNF) protected neurons from phenylalanine-induced apoptosis and inhibition of Trk receptor by K252a or downregulation of TrkB abrogated the effect of BDNF. We further demonstrated that phenylalanine-induced RhoA activation and myosin light chain phosphorylation were inhibited by pretreatment with BDNF, while phenylalanine activates the mitochondria-mediated apoptosis through the RhoA/Rho-associated kinase pathway. Thus our studies indicate that the protective effect of BDNF against phenylalanine-induced neuronal apoptosis is probably mediated by suppression of RhoA signaling pathway via TrkB receptor. Taken together, these findings suggest a potential neuroprotective action of BDNF in prevention and treatment of PKU brain injury.