Genetic testing for fetal loss of heterozygosity using single nucleotide polymorphism array and whole-exome sequencing.

The study explored the clinical significance of fetal loss of heterozygosity (LOH) identified by single-nucleotide polymorphism array (SNP array). We retrospectively reviewed data from pregnant women who underwent invasive diagnostic procedures at prenatal diagnosis centers in southeastern China from December 2016 to December 2021. SNP array was performed by the Affymetrix CytoScan 750 K array platform. Fetuses with LOH were further identified by parental verification, MS-MLPA, and/or trio whole-exome sequencing (trio-WES). The genetic results, fetal clinical manifestations, and perinatal outcome were analyzed. Of 11,062 fetuses, 106 (0.96%) had LOH exhibiting a neutral copy number, 88 (83.0%) had LOH in a single chromosome, whereas 18 (17.0%) had multiple LOHs on different chromosomes. Sixty-six fetuses had ultrasound anomalies (UAs), most frequently fetal growth restriction (18/66 (27.3%)). Parental SNP array verification was performed in 21 cases and trio-WES in 21 cases. Twelve cases had clinically relevant uniparental disomy, five had pathogenic variants, four had likely pathogenic variants, six had variants of unknown significance, and eight had identity by descent. The rate of adverse pregnancy outcomes in fetuses with LOH and UAs (24/66 (36.4%)) was higher than in those without UAs (6/40 (15.0%)) (p < 0.05). LOH is not uncommon. Molecular genetic testing techniques, including parental SNP array verification, trio-WES, methylation-specific multiplex ligation-dependent probe amplification, regular and systematic ultrasonic monitoring, and placental study, can accurately assess the prognosis and guide the management of the affected pregnancy.

Genetic analysis of chorionic villus tissues in early missed abortions.

Chromosomal abnormalities are the most common etiology of early spontaneous miscarriage. However, traditional karyotyping of chorionic villus samples (CVSs) is limited by cell culture and its low resolution. The objective of our study was to investigate the efficiency of molecular karyotyping technology for genetic diagnosis of early missed abortion tissues. Chromosome analysis of 1191 abortion CVSs in early pregnancy was conducted from August 2016 to June 2021; 463 cases were conducted via copy-number variations sequencing (CNV-seq)/quantitative fluorescent-polymerase chain reaction (QF-PCR) and 728 cases were conducted using SNP array. Clinically significant CNVs of CVSs were identified to clarify the cause of miscarriage and to guide the couples' subsequent pregnancies. Among these, 31 cases with significant maternal cell contamination were removed from the study. Among the remaining 1160 samples, 751 cases (64.7%) with genetic abnormalities were identified, of which, 531 (45.8%) were single aneuploidies, 31 (2.7%) were multiple aneuploidies, 50 (4.3%) were polyploidies, 54 (4.7%) were partial aneuploidies, 77 (6.6%) had submicroscopic CNVs (including 25 with clinically significant CNVs and 52 had variants of uncertain significance), and 8 cases (0.7%) were uniparental disomies. Our study suggests that both SNP array and CNV-seq/QF-PCR are reliable, robust, and high-resolution technologies for genetic diagnosis of miscarriage.

[Investigation on the current situation of the development of intensive care units in Inner Mongolia Autonomous Region in 2022].

OBJECTIVE: To investigate the development present situation of the department of critical care medicine in Inner Mongolia Autonomous Region (hereinafter referred to as Inner Mongolia), in order to promote the standardized and homogeneous development of critical care medicine in Inner Mongolia, and also provide a reference for discipline construction and resource allocation. METHODS: A survey study was conducted in comprehensive intensive care unit (ICU) of tertiary and secondary hospitals in Inner Mongolia by online questionnaire survey and telephone data verification. The questionnaire was based on the Guidelines for the Construction and Management of Intensive Care Units (Trial) (hereinafter referred to as the Guidelines) issued by the National Health Commission in 2009 and the development trend of the discipline. The questionnaire covered six aspects, including hospital basic information, ICU basic information, personnel allocation, medical quality management, technical skill and equipment configuration. The questionnaire was distributed in September 2022, and it was filled out by the discipline leaders or department heads of each hospital. RESULTS: As of October 24, 2022, a total of 101 questionnaires had been distributed, 85 questionnaires had been recovered, and the questionnaire recovery rate had reached 84.16%, of which 71 valid questionnaires had been collected in a total of 71 comprehensive ICU. (1) There were noticeable regional differences in the distribution of comprehensive ICU in Inner Mongolia, with a relatively weak distribution in the east and west, and the overall distribution was uneven. The development of critical care medicine in Inner Mongolia was still lacking. (2) Basic information of hospitals: the population and economy restricted the development of ICU. The average number of comprehensive ICU beds in the western region was only half of that in the central region (beds: 39.0 vs. 86.0), and the average number of ICU beds in the eastern region was in the middle (83.6 beds), which was relatively uneven. (3) Basic information of ICU: among the 71 comprehensive ICU surveyed, there were 44 tertiary hospitals and 27 secondary hospitals. The ratio of ICU beds to total beds in tertiary hospitals was significantly lower than that in secondary hospitals [(1.59±0.81)% vs. (2.11±1.07)%, P < 0.05], which were significantly lower than the requirements of the Guidelines of 2%-8%. The utilization rate of ICU in tertiary and secondary hospitals [(63.63±22.40)% and (44.65±20.66)%, P < 0.01] were both lower than the bed utilization rate required by the Guidelines (75% should be appropriate). (4) Staffing of ICU: there were 376 doctors and 1 117 nurses in tertiary hospitals, while secondary hospitals had 122 doctors and 331 nurses. There were significant differences in the composition ratio of the titles of doctors, the degree of doctors, and the titles of nurses between tertiary and secondary hospitals (all P < 0.05). Most of the doctors in tertiary hospitals had intermediate titles (attending physicians accounted for 41.49%), while most of the doctors in secondary hospitals had junior titles (resident physicians accounted for 43.44%). The education level of doctors in tertiary hospitals was generally higher than that in secondary hospitals (doctors: 2.13% vs. 0, masters: 37.24% vs. 8.20%). The proportion of nurses in tertiary hospitals was significantly lower than that in secondary hospitals (17.01% vs. 24.47%). The ratio of ICU doctors/ICU beds [(0.64±0.27)%, (0.59±0.34)%] and ICU nurses/ICU beds [(1.76±0.56)%, (1.51±0.48)%] in tertiary and secondary hospitals all failed to meet the requirements above 0.8 : 1 and 3 : 1 of the Guidelines. (5) Medical quality management of ICU: compared with secondary hospitals, the proportion of one-to-one drug-resistant bacteria care in tertiary hospitals (65.91% vs. 40.74%), multimodal analgesia and sedation (90.91% vs. 66.67%), and personal digital assistant (PDA) barcode scanning (43.18% vs. 14.81%) were significantly higher (all P < 0.05). (6) Technical skills of ICU: in terms of technical skills, the proportion of bronchoscopy, blood purification, jejunal nutrition tube placement and bedside ultrasound projects carried out in tertiary hospitals were higher than those in secondary hospitals (84.09% vs. 48.15%, 88.64% vs. 48.15%, 61.36% vs. 55.56%, 88.64% vs. 70.37%, all P < 0.05). Among them, the placement of jejunal nutrition tube, bedside ultrasound and extracorporeal membrane oxygenation were mainly completed independently in tertiary hospitals, while those in secondary hospitals tended to be completed in cooperation. (7) Equipment configuration of ICU: in terms of basic equipment, the ratio of the total number of ventilators/ICU beds in tertiary and secondary hospitals [0.77% (0.53%, 1.07%), 0.88% (0.63%, 1.38%)], and the ratio of injection pump/ICU beds [1.70% (1.00%, 2.56%), 1.25% (0.75%, 1.88%)] didn't meet the requirements of the Guidelines. The equipment ratio was insuffcient, which means that the basic needs of development had not been met yet. CONCLUSIONS: The development of comprehensive ICU in Inner Mongolia has tended to mature, but there is still a certain gap in the development scale, personnel ratio and instruments and equipment compared with the Guidelines. Moreover, the comprehensive ICU appears the characteristics of relatively weak eastern and western regions, and the overall distribution is uneven. Therefore, it is necessary to increase efforts to invest in the construction of the department of critical care medicine.

High dietary copper intake induces perturbations in the gut microbiota and affects host ovarian follicle development.

Increasing evidence has shown that gut microbes play an important role in the reproductive endocrine system and the development of polycystic ovary syndrome (PCOS). However, whether environmental factors are involved in these gut microbiota alterations has seldom been studied. In this study, we aimed to explore the crucial role of an imbalanced gut microbiota on abnormal ovarian follicle development induced by Cu. A 1:1 matched case-control study with 181 PCOS patients and 181 controls was conducted using a propensity score matching protocol. Information regarding dietary Cu intake was obtained from a face-to-face dietary intake interview. Alterations in the gut microbiota were detected by high-throughput 16 S rDNA sequencing. The results showed that dietary Cu intake was positively correlated with the risk of PCOS, and the risk threshold was approximately 1.992 mg/d. Compared with those with dietary Cu intakes lower than 1.992 mg/d, those who had a higher dietary Cu intake had a 1.813-fold increased risk of PCOS (OR=1.813, 95% CI: 1.150-2.857). PCOS patients had a lower relative abundance of Bacteroides than controls (P = 0.003), and Bacteroides played a partial mediating role between dietary Cu exposure and PCOS (Pindirect effect=0.026, 95% CI: 0.002-0.072). In addition, an animal model of Cu exposure through the diet showed that Cu can induce gut microbiota disorder; increase serum levels of LPS, MDA, and IL-6; and alter host ovarian steroidogenesis to affect ovarian follicle development. Staphylococcus played a partial mediating role between Cu exposure and CYP17A1 (Pg_Staphylococcus=0.083, 95% CI: 0.001-0.228). Overall, this study shows that long-term exposure to high dietary Cu levels can affect the composition of the gut microbiota, cause inflammation and oxidative stress, and then interfere with hormone signaling, ultimately affecting ovarian follicle development.

Effectiveness of drug-coated balloon in acute myocardial infarction: A protocol for systematic review and meta-analysis.

BACKGROUND: The management of acute myocardial infarction (AMI) has improved dramatically over the past 3 decades and is evolving. Percutaneous coronary intervention is an alternative means of achieving coronary revascularization. Previous studies comparing the published literature on drug-coated balloon (DCB) and drug-eluting stents have drawn divergent conclusions. We perform a protocol for systematic review and meta-analysis to compare the efficacy and safety of DCB and drug-eluting stent in the management of AMI. METHODS: This systematic review was registered in the PROSPERO network (registration number: CRD42023397266). We will follow the Preferred Reporting Items for Systematic Reviews and Meta-analysis Protocol to accomplish the systematic review protocol. A systematic search will be conducted in PubMed, Embase, Cochrane Library, Web of Science, Chinese National Knowledge Infrastructure, Wanfang Database, and Weipu Database without any language restrictions from their inception to February 2022. The risk of bias will be assessed independently by 2 authors using parameters defined in the Cochrane Handbook for Systematic Reviews of Interventions criteria. Statistical analysis will be performed using the STATA13.0 software (IBM, USA). RESULTS: The results of this systematic review and meta-analysis will be publicly available and published in a peer-reviewed journal. CONCLUSION: The results of the study will provide the evidence for the application of DCB in the treatment of AMI.

Prenatal genetic analysis of fetal aberrant right subclavian artery with or without additional ultrasound anomalies in a third level referral center.

To evaluate the correlation between chromosomal abnormalities and fetal aberrant right subclavian artery (ARSA) with or without additional ultrasound anomalies (UAs). A total of 340 fetuses diagnosed with ARSA by ultrasound between December, 2015, and July, 2021, were included. All cases were subdivided into three groups: (A) 121 (35.6%) cases with isolated ARSA, (B) 91 (26.8%) cases with soft markers, and (C) 128 (37.6%) cases complicated with other UAs. Invasive testing was performed via amniotic fluid or cord blood karyotyping and chromosomal microarray analysis (CMA) in parallel, and pregnancy outcomes were followed. Karyotype abnormalities were identified in 18/340 (5.3%) patients. Karyotype abnormalities in Groups A, B, and C were 0/121 (0.0%), 7/91 (7.7%), and 11/128 (8.6%), respectively. CMA abnormalities with clinically significant variants were detected in 37/340 (10.9%) cases, of which 22q11.2 deletion syndrome and trisomy 21 accounted for 48.6% (18/37). The overall abnormal CMA with clinically significant variant detection rates in Groups A, B, and C were 3/121(2.5%), 13/91 (14.3%), and 21/128 (16.4%), respectively. There were significant difference in clinically significant CMA anomalies detection rate between Groups A and C (p < 0.05), as well as Groups A and B (p < 0.05). Comparing CMA to karyotyping showed a clinically significant incremental yield in Group C (7.8%, 10/128) compared to Groups A (2.5%, 3/121) and B (6.6%, 6/91) (p > 0.05). Fetal ARSA with additional UAs, concurred with cardiac and extra-cardiac anomalies, constitutes a high-risk factor for chromosomal aberrations, especially for pathogenic or likely pathogenic copy number variants.

Efficiency of expanded noninvasive prenatal testing in the detection of fetal subchromosomal microdeletion and microduplication in a cohort of 31,256 single pregnancies.

Noninvasive prenatal testing (NIPT) is widely used to screen for common fetal chromosomal aneuploidies. However, the ability of NIPT-Plus to detect copy number variation (CNV) is debatable. Accordingly, we assessed the efficiency of NIPT-Plus to detect clinically significant fetal CNV. We performed a prospective analysis of 31,260 singleton pregnancies, included from June 2017 to December 2020. Cell-free fetal DNA was directly sequenced using the semiconductor sequencing platform for women with high-risk CNV with clinically significant results. Fetal karyotyping and chromosomal microarray analysis (or next-generation sequencing) are recommended for invasive diagnostic procedures. Women at low risk with no other abnormal results continued their pregnancies. We analyzed the expanded NIPT results, diagnostic test results, and follow-up information to evaluate its performance in detecting fetal CNV. Of the 31,260 pregnant women who received NIPT-Plus, 31,256 cases were tested successfully, a high risk of clinically significant CNV was detected in 221 cases (0.71%); 18 women refused further diagnosis; 203 women underwent invasive prenatal diagnosis; and 78 true positive cases and 125 false positive cases, with an overall positive predictive value (PPV) of 38.42% and a false positive rate of 0.40%. For known microdeletion/microduplication syndromes (n = 27), the PPVs were 75% DiGeorge syndrome (DGS), 80% 22q11.22 microduplication, 50% Prader-Willi syndrome, and 50% cri-du-chat. For the remaining clinically significant fetal CNVs (n = 175), the combined PPVs were 46.5% (CNVs > 10 Mb) and 28.57% (CNVs ≤ 10 Mb). NIPT-Plus screening for CNV has certain clinical value. NIPT-Plus yielded relatively high PPVs for 22q11.2 microduplication syndrome and DGS, and low to moderate PPVs for other CNVs.

The Impact of Malignancy on Assisted Reproductive Outcomes for Cancer Survivors: A Retrospective Case-Control Study.

Background: Related studies have shown that it is safe for cancer patients to undergo assisted reproduction. However, studies on whether a history of cancer affects long-term reproductive outcomes in women who undergo assisted reproductive technology (ART) are scarce. In this study, we evaluated the long-term reproductive outcomes of patients with malignant tumors undergoing ART treatment and explored the impact of malignancy history on ART outcomes. Methods: This retrospective study analyzed the clinical outcomes of patients with malignant tumors undergoing their first in vitro fertilization/intracytoplasmic sperm injection (IVF/ICSI) cycles compared with those of age-matched healthy infertile women at Fujian Maternity and Child Health Hospital between January 2003 and October 2020. We evaluated ovarian stimulation outcome, the pregnancy rate, the live birth rate, the risk of adverse obstetric outcomes and birth outcomes. Results: This study included 59 patients in the cancer group for data analysis who had a history of malignancy. By matching, a total of 118 healthy infertile women were included in the control group. No statistically significant association was found in terms of age, duration of infertility, BMI, or insemination type between the two groups of patients. Thyroid cancer(45.8%) and gynecologic malignancies (44.07%) were the major cancer types in this study. There were statistically significant differences in the antral follicle count (AFC) (12.00 ± 7.86 vs. 14.90 ± 8.71, P=0.033), length of ovarian stimulation (9.98 ± 2.68 vs. 11.42 ± 2.43, P=0.033) and endometrial thickness on the trigger day (10.16 ± 3.11 vs. 10.84 ± 2.17, P<0.001) between the two groups. The total gonadotropin dose, number of oocytes retrieved, fertilization rate, cleavage rate, high-quality embryo rate, blastocyst rate and first-time embryo-transfer (ET) implantation rate were nonsignificantly lower in the cancer group than in the control group (P>0.05). There were no significant differences in the clinical pregnancy rate per ET cycle (32% vs. 40.39%, P=0.156), live birth rate per ET cycle (27% vs. 35.96%, P=0.119), miscarriage rate per ET cycle (5% vs. 4.43%, P=0.779), or preterm delivery rate per ET cycle (11.11% vs. 17.80%, P=0.547) between the two groups. Additionally, regression analysis showed that a history of malignancy was not a risk factor for reproductive outcomes. Conclusions: Overall, it is feasible for women with a history of cancer to conceive using ART is feasible and their long-term reproductive outcomes are similar to these of healthy infertile women. A history of cancer does not decrease the number of retrieved oocytes, increase the risk of adverse obstetric outcomes or affect birth outcomes.

Arsenic exposure diminishes ovarian follicular reserve and induces abnormal steroidogenesis by DNA methylation.

Arsenic contamination is a worldwide public health problem, and the effect of arsenic on male reproduction has been extensively studied; however, data on the biotoxicity of arsenic in terms of female reproduction are more scarce. In this study, a human-cell-animal translational strategy was applied to explore the effect of arsenic exposure on ovarian steroidogenesis and its potential mechanism. We conducted a 1:1 propensity score matched case-control study involving 127 diminished ovarian reserve (DOR) cases and 127 healthy controls. The ovarian follicular fluid levels of 21 metal elements, including arsenic, were measured. The results showed that there were significant differences in follicular fluid metal profiles between DOR patients and controls and that arsenic, molybdenum, and strontium played important roles in DOR progression [OR (95 % CI): 2.203 (1.385, 3.503), 2.308 (1.490, 3.575) and 2.922 (1.864, 4.580), respectively]. In the primary ovarian granulosa cell culture model, we found that treatment with 8 µM arsenic for 24 and 48 h induced a decrease in human granulosa cell viability. The estradiol (E2) level was significantly decreased after arsenic exposure (P < 0.05), which was dependent on significant alterations (P < 0.05) in key enzymes in steroidogenesis. In addition, a model for sodium arsenite exposure through water in rats from weaning to sexual maturity was established. We evaluated ovarian development by monitoring the estrous cycle, observing ovarian pathology, and calculating the follicular proportion. RT-qPCR, Western blotting, and bisulfite-sequencing PCR were used to investigate the effect of arsenic exposure on ovarian steroidogenesis and its possible mechanism. The results indicated that steroidogenic factor-1 (SF-1) was an important target of the steroidogenesis disorder induced by arsenic exposure. Arsenic significantly increased the DNA methylation level (P < 0.05) in the promoter region of SF-1 to reduce its expression, subsequently decreasing the levels of steroidogenic acute regulatory protein (StAR), P450 cholesterol side-chain cleavage enzyme (CYP11A1), and aromatase (CYP19A1) (P < 0.05), leading to premature depletion of ovarian follicles.

Renal Doppler Ultrasound in the Evaluation of Renal Function in Patients with Sepsis.

This study was aimed to explore the renal Doppler ultrasound in the evaluation of renal function in patients with sepsis. Fifty patients with sepsis or septic shock were classified into the acute kidney injury (AKI) group (n =25) and the non-AKI group (n =25) according to whether they had AKI. The measurements of renal resistance index (RRI) and power Doppler ultrasound (PDU) were performed on all patients within 7 days of admission to the intensive care unit (ICU). The patient's renal function was assessed. The results showed that the RRI of the two groups showed a slight upward trend over time, and the RRI of the AKI group was higher than that of the non-AKI group. After 7 days in AKI group, the areas under the receiver operating characteristic (ROC) curves of RRI were 0.745, 0.683, 0.729, 0.856, 0.793, 0.819, and 0.836 (P <0.05). There were no statistically considerable differences in areas under ROC curves between the two groups (P >0.05). The grouping of AKI and the time were both fixed effects, and the individual patients were randomized effects. Besides, the linear models were statistically analyzed. The results showed that the differences between the two groups were statistically insignificant (P >0.05). There was no significant difference in the PDU scores measured at different times within 7 days after ICU admission between the two groups (P >0.05). In conclusion, renal Doppler ultrasound had a good adoption effect in the evaluation of the renal function of patients with severe sepsis, which is worth promoting in clinical practice.

Neonatal exposure of bisphenol A led to increased lipolysis of visceral adipose tissue in adult rats with DNA hypomethylation of Atgl being one of the possible underlying mechanisms.

Accumulating evidence suggests the role of developmental exposure of bisphenol A (BPA) in metabolic disorders. However, the underlying mechanism remains unclear. Using a rat model, we investigated the neonatal exposure of BPA on lipid metabolism in adult and the underlying mechanisms. From postnatal day1(PND1) to PND10, male rats were exposed to BPA via daily subcutaneous injection with 10 µg/100 µL BPA (1.24-0.5 mg/kg body weight/day, a dose below the US-EPA LOAEL). After fasting for 8 h, adult rats aged 80 days showed elevated levels of serum free fatty acid (FFA), glycerol and glucose, and increased levels of FFA and glycerol in visceral adipose tissue. The expression levels of key enzymes of lipolysis, adipose triglyceride lipase (Atgl) and hormone-sensitive lipase (Hsl), were increased in visceral adipose tissue from BPA-exposed rats after fasting. On the other hand, transcription levels of lipogenic genes remained unchanged. Differentiation of visceral adipocyte in rats takes place neonatally. In our study, neonatal BPA exposure induced DNA hypomethylation of Atgl in visceral adipose tissue. In 3T3-L1 cell, administration of 10-7 mol/L BPA throughout the differentiation stage led to DNA hypomethylation and increased expression of Atgl. Our results suggest that neonatal exposure of BPA led to increased lipolysis of visceral adipose tissue in young adults, which will predispose individuals to multiple metabolic disorders. The DNA hypomethylation of Atgl might be one of the mechanisms underneath the long-lasting effect of neonatal BPA exposure.

Detection of copy number variation associated with ventriculomegaly in fetuses using single nucleotide polymorphism arrays.

Etiopathogenesis of fetal ventriculomegaly is poorly understood. Associations between fetal isolated ventriculomegaly and copy number variations (CNVs) have been previously described. We investigated the correlations between fetal ventriculomegaly-with or without other ultrasound anomalies-and chromosome abnormalities. 222 fetuses were divided into four groups: (I) 103 (46.4%) cases with isolated ventriculomegaly, (II) 41 (18.5%) cases accompanied by soft markers, (III) 33 (14.9%) cases complicated with central nervous system (CNS) anomalies, and (IV) 45 (20.3%) cases with accompanying anomalies. Karyotyping and single nucleotide polymorphism (SNP) array were used in parallel. Karyotype abnormalities were identified in 15/222 (6.8%) cases. Karyotype abnormalities in group I, II, III, and IV were 4/103 (3.9%), 2/41 (4.9%), 4/33 (12.1%), and 5/45 (11.1%), respectively. Concerning the SNP array analysis results, 31/222 (14.0%) were CNVs, CNVs in groups I, II, III, and IV were 11/103 (10.7%), 6/41 (14.6%), 9/33 (27.3%), and 5/45 fetuses (11.1%), respectively. Detections of clinical significant CNVs were higher in non-isolated ventriculomegaly than in isolated ventriculomegaly (16.81% vs 10.7%, P = 0.19). SNP arrays can effectively identify CNVs in fetuses with ventriculomegaly and increase the abnormal chromosomal detection rate by approximately 7.2%, especially ventriculomegaly accompanied by CNS anomalies.

Prenatal diagnosis of PLP1 duplication by single nucleotide polymorphism array in a family with Pelizaeus-Merzbacher disease.

A family with a history of Pelizaeus-Merzbacher disease (PMD) received prenatal diagnosis of PLP1 gene duplication in a fetus using a single nucleotide polymorphism (SNP) array. A 27-year-old pregnant woman was referred for genetic counseling due to her four-year-old son being diagnosed with a suspected classic type of PMD. Amniocentesis was performed at 18 and 3/7 weeks of gestation, and the SNP array was carried out on DNA from the mother, her affected son, and fetus, then further confirmed by multiplex ligation-dependent probe amplification (MLPA). Cytogenetic analysis of the fetus showed 46,XY. SNP array analysis revealed that the male fetus did not carry PLP1 gene duplication but the affected boy did, and the mother was a carrier for the duplication of the PLP1 gene. All SNP array results were further confirmed by MLPA. SNP array and MLPA analyses of peripheral blood verified the nonduplication of the PLP1 gene in the infant after birth. At present, the child (without PLP1 duplication) is developing normally. This study preliminarily suggests that SNP array is a sensitive and accurate technology for identifying PLP1 duplication and is feasible for reliable diagnosis, including for the prenatal diagnosis of PMD resulting from PLP1 duplication.

Prenatal diagnosis and molecular cytogenetic identification of small supernumerary marker chromosomes: analysis of three prenatal cases using chromosome microarray analysis.

Small supernumerary marker chromosomes cannot be accurately identified by G-banding, and the related phenotypes vary greatly. It is essential to specify the origin, size, and gene content of marker chromosomes using molecular cytogenetic techniques. Herein, three fetuses with de novo marker chromosomes were initially identified by G-banding. Single nucleotide polymorphism array and fluorescence in situ hybridization were performed to characterize the origins of the marker chromosomes. The karyotypes of the three fetuses were 47,XY,+mar, 46,X,+mar[32]/45,X[68], and 45,X[62]/46,X,+mar[9]. In case 1, the karyotype was confirmed as 47,XY,+ idic(22)(q11.2). Therefore, the sSMC originated from chromosome 22 and was associated with cat eye syndrome. In case 2, the marker chromosome derived from ring chromosome X, and the karyotype was interpreted as 45,X[68]/46,X,+r(X)(p11.1q21.31)[32]. Meanwhile, the karyotype of case 3 was defined as 45,X[62]/46,X,idic(Y)(q11.2) and the marker chromosome originated from chromosome Y. Case 1 continued the pregnancy, whereas the other two pregnancies underwent elective termination. The detailed characterization of marker chromosomes can facilitate informed decision making, prevent uncertainty, and provide proper prognostic assessments. Our findings emphasize the importance for combining cytogenetic and molecular genetic techniques in marker chromosome characterization.

Transcriptome and metabolite analysis reveal the drought tolerance of foxtail millet significantly correlated with phenylpropanoids-related pathways during germination process under PEG stress.

BACKGROUND: Foxtail millet [Setaria italica (L.) P. Beauv.] is an excellent crop known for its superior level of drought tolerance across the world. Especially, less water is needed during its germination period than the other cereal crops. However, the knowledge of the mechanisms underlying the abiotic stress effects on seed germination of foxtail millet is largely unknown. RESULTS: The water uptake pattern of foxtail millet seeds was ploted during germination period, according to which the germination time course of millet was separated into three phases. We sequenced the transcriptome of foxtail millet seeds, which were treated by PEG during different germination phases after sowing. The transcriptional studies revealed that more DEGs were identified during the further increase in water uptake period (phase III) than during the rapid initial uptake period (phase I) and the plateau period (phase II) under PEG stress. The pathway analysis of DEGs showed that the highly enriched categories were related to phenylpropanoid biosynthesis, plant hormone signal transduction and phenylalanine metabolism during phase III. The 20 phenylpropanoids-related genes of germinating foxtail millet were found to be down-regulated during the further increase in water uptake period under PEG stress. Further expression analysis identified 4 genes of phenylalanine ammonia-lyase, 4-coumarate-CoA ligase 3, cinnamoyl-CoA reductase 1, cationic peroxidase SPC4 in phenylpropanoids-related pathway, which played important roles in foxtail millet in response to PEG stress during different germination periods. The studies of metabolites in phenylpropanoid biosynthesis pathway revealed that higher amount of cinnamic acid was accumulated in germinating seeds under PEG stress, while the contents of p-coumaric acid, caffeic acid, ferulic acid and sinapic acid were decreased. And the effects of five phenolic compounds on germination and growth of foxtail millet showed that 1 mM concentration of cinnamic acid inhibited shoot and root growth, especially root development. Ferulic acid, caffeic acid, sinapic acid and p-coumaric acid could increase the root length and root/sprout in lower concentration. CONCLUSIONS: These findings suggest that key genes and metabolites of foxtail millet related with phenylpropanoids pathway may play prominent roles in the regulation of resistance to drought during germination. Foxtail millet can probably avoid drought by regulating the levels of endogenous allelochemicals.

Foxtail millet (Setaria italica (L.) P. Beauv) CIPKs are responsive to ABA and abiotic stresses.

CBL-interacting protein kinases (CIPKs) have been shown to regulate a variety of environmental stress-related signalling pathways in plants. Foxtail millet (Setaria italica (L.) P. Beauv) is known worldwide as a relatively stress-tolerant C4 crop species. Although the foxtail millet genome sequence has been released, little is known about the functions of CIPKs in foxtail millet. Therefore, a systematic genome-wide analysis of CIPK genes in foxtail millet was performed. In total, 35 CIPK members were identified in foxtail millet and divided into four subgroups (I to IV) on the basis of their phylogenetic relationships. Phylogenetic and gene structure analyses clearly divided all SiCIPKs into intron-poor and intron-rich clades. Cis-element analysis subsequently indicated that these SiCIPKs may be involved in responses to abiotic stimuli, hormones, and light signalling during plant growth and development, and stress-induced expression profile analysis revealed that all the SiCIPKs are involved in various stress signalling pathways. These results suggest that the CIPK genes in foxtail millet exhibit the basic characteristics of CIPK family members and play important roles in response to abiotic stresses. The results of this study will contribute to future functional characterization of abiotic stress responses mediated by CIPKs in foxtail millet.

[Impact of GOLPH3 expression in cumulus granulosa cells on outcomes of intracytoplasmic sperm injection].

OBJECTIVE: To evaluate the impact of GOLPH3 expression in cumulus granulosa cells on the outcomes of intracytoplasmic sperm injection (ICSI). METHODS: A total of 119 women receiving ICSI due to male infertility at our center between April, 2012 and June, 2014 were enrolled in the study. Cumulus granulosa cells were collected from the women for detection of GOLPH3 expressions using immunocytochemistry, Western blotting, and real-time PCR. GOLPH3 expression rate was compared between women with and without clinical pregnancy following ICSI, and the associations of GOLPH3 expression with the laboratory indicators of ICSI outcomes were assessed. RESULTS: Immunocytochemistry showed that GOLPH3 expression was located mainly in in the plasma of the cumulus granulosa cells. The rate and intensity of GOLPH3 expression in the cumulus granulosa cells differed significantly between women with and without clinical pregnancy following ICSI (P<0.05). GOLPH3 expression was found to positively correlate with the numbers of punctured follicles, grade III oocyte cumulus complex, ICSI oocytes, fertilized oocytes, cleavage, high quality embryos, blastocysts, high quality blastocysts, and frozen embryos (all P<0.01). The results of RTPCR and Western blotting revealed significant differences in GOLPH3 expressions at both the mRNA and protein levels in the cumulus granulosa cells between the pregnant and non-pregnant groups after ICSI (t=14.560, P=0.000). Western blot analysis revealed significant difference of GOLPH3 protein expression in cumulus granulosa cells between women with and without clinical pregnancy following ICSI. CONCLUSION: GOLPH3 expression in the cumulus granulosa cells plays an important role in the development of oocytes and promotion of conception to affect the outcomes of ICSI.

Effect of GOLPH3 on cumulus granulosa cell apoptosis and ICSI pregnancy outcomes.

Previous studies have shown that GOLPH3 mediates cell growth, proliferation and differentiation and inhibits cell apoptosis; however, the role of GOLPH3 in cumulus granulosa cells and the value of GOLPH3 in predicting ICSI pregnancy outcomes remain unknown until now. Our findings showed higher positive expression rate, score of staining intensity, and immunohistochemical score of GOLPH3 in the cumulus granulosa cells of the pregnant women relative to non-pregnant women, and a higher apoptotic rate of cumulus granulosa cells was detected in non-pregnant women than in pregnant women. Pearson correlation analyses revealed that pregnancy correlated negatively with GOLPH3 expression and apoptosis of cumulus granulosa cells, and positively with the number of follicles punctured, number of grade III oocytes, number of eggs retrieved for ICSI, number of zygotes, number of cleavage-stage embryos, number of top-quality embryos, number of blastocysts, number of top-quality blastocysts, and number of frozen embryos. GOLPH3 may be involved in the apoptosis of cumulus granulosa cells, which may correlate with oocyte maturation and egg development. GOLPH3 expression in cumulus granulosa cells may facilitate the selection of top-quality eggs and embryos, the prediction of the clinical pregnancy outcomes of ICSI, and the increase of the pregnancy rate.

Potential Dermal Exposure to Flonicamid and Risk Assessment of Applicators During Treatment in Apple Orchards.

Exposure and risk assessments of flonicamid for applicators were performed in apple orchards in Korea. Fifteen experiments were done with two experienced applicators under typical field conditions using a speed sprayer. In this study, cotton gloves, socks, masks, and dermal patches were used to monitor potential dermal exposure to flonicamid, and personal air samplers with XAD-2 resin and glass fiber filter were used to monitor potential inhalation exposure. The analytical methods were validated for the limit of detection, limit of quantitation, reproducibility, linearity of the calibration curve, and recovery of flonicamid from various exposure matrices. The results were encouraging and acceptable for an exposure study. The applicability of XAD-2 resin was evaluated via a trapping efficiency and breakthrough test. During the mixing/loading, the average total dermal exposure was 22.6 µg of flonicamid, corresponding to 4.5×10(-5)% of the prepared amount. For the spraying, the potential dermal exposure was 9.32 mg, and the ratio to applied amount was 1.9 × 10(-2%). The primary exposed body parts were the thigh (2.90 mg), upper arm (1.75 mg), and lower leg (1.66 mg). By comparison, absorbable quantity of exposure was small, only 1.62 µg (3.2×10(-6)%). The margin of safety (MOS) were calculated for risk assessment, in all sets of trials, MOS > 1, indicating the exposure level of flonicamid was considered to be safe in apple orchards. Although this was a limited study, it provided a good estimate of flonicamid exposure for orchard applicators.

Sodium 1-naphthalenesulfonate-functionalized reduced graphene oxide stabilizes silver nanoparticles with lower cytotoxicity and long-term antibacterial activity.

Silver nanoparticles (AgNPs) are increasingly used in daily life for their antibacterial properties, but their low stability and high cytotoxicity hamper practical applications. In this work, sodium 1-naphthalenesulfonate-functionalized reduced graphene oxide (NA-rGO) was used as a substrate for AgNPs to produce a AgNP-NA-rGO hybrid. This hybrid showed substantially higher antibacterial activity than polyvinyl pyrrolidone (PVP)-stabilized AgNPs, and the AgNPs on NA-rGO were more stable than the AgNPs on PVP, resulting in long-term antibacterial effects. More importantly, this hybrid showed excellent water solubility and low cytotoxicity, suggesting the great potential application as sprayable reduced graphene oxide based antibacterial solutions.