[Prenatal genetic diagnosis of the fetuses with isolated corpus callosum abnormality].

Objective: To explore the application value of chromosome karyotype analysis, chromosomal microarray analysis (CMA) and whole exome sequencing (WES) in prenatal diagnosis of isolated corpus callosum abnormality (CCA) fetus. Methods: Fetuses diagnosed with isolated CCA by ultrasound and MRI and receiving invasive prenatal diagnosis in Guangzhou Women and Children's Medical Center and Qingyuan People's Hospital from January 2010 to April 2021 were selected. Karyotype analysis and/or CMA [or copy number variation sequencing (CNV-seq)] were performed on all fetal samples, and WES was performed on fetal samples and their parents whose karyotype analysis and/or CMA (or CNV-seq) results were not abnormal. Results: Among 65 fetuses with isolated CCA, 38 cases underwent karyotype analysis, and 3 cases were detected with abnormal karyotypes, with a detection rate of 8% (3/38). A total of 49 fetuses with isolated CCA underwent CMA (or CNV-seq) detection, and 6 cases of pathogenic CNV were detected, the detection rate was 12% (6/49). Among them, the karyotype analysis results were abnormal, and the detection rate of further CMA detection was 1/1. The karyotype results were normal, and the detection rate of further CMA (or CNV-seq) detection was 14% (3/21). The detection rate of CMA as the first-line detection technique was 7% (2/27). A total of 25 fetuses with isolated CCA with negative results of karyotyping and/or CMA were tested by WES, and 9 cases (36%, 9/25) were detected with pathogenic genes. The gradient genetic diagnosis of chromosomal karyotyping, CMA and WES resulted in a definite genetic diagnosis of 26% (17/65) of isolated CCA fetuses. Conclusions: Prenatal genetic diagnosis of isolated CCA fetuses is of great clinical significance. The detection rate of CMA is higher than that of traditional karyotyping. CMA detection could be used as a first-line detection technique for fetuses with isolated CCA. WES could increase the pathogenicity detection rate of fetuses with isolated CCA when karyotype analysis and/or CMA test results are negative.

Hypoxia Modulates Melanoma Cells Proliferation and Apoptosis via miRNA-210/ISCU/ROS Signaling.

In this study, luciferase reporter assay was used to establish the relationship between miR-210 and ISCU. This research was performed on both cell lines (A2058, G361, and 293T) and tissue samples. We found that miR-210 was upregulated in hypoxia and was elevated in melanoma in comparison with adjacent normal tissues. Expression of ISCU protein was decreased in melanoma tissues, and ISCU gene is a direct target of miR-210. ISCU knockdown with miR-210 enhanced ROS production. The results of our study showed that miR-210/ISCU/ROS axis can serve as a novel therapeutic target in melanoma.

[Analysis of families with fetal congenital abnormalities but negative prenatal diagnosis by whole exome sequencing].

Objective: To evaluate the value of whole exome sequencing (WES) in prenatal clinical application. Methods: A total of 1 152 cases of congenital abnormal [including structural malformation, nuchal translucency (NT) thickening and intrauterine growth restriction] with traditional prenatal diagnosis [including G-band karyotype analysis and chromosome microarray analysis (CMA)] negative were analyzed. The congenital abnormal fetuses were divided into retrospective group and prospective group according to the time of WES detection, that is whether the pregnancy termination or not. According to the specific location of fetal malformation and their family history, the cohort was divided into subgroups. The clinical prognosis of all fetuses were followed up, and the effect of WES test results on pregnancy decision-making and clinical intervention were analyzed. According to the follow-up results, the data of fetuses with new phenotypes in the third trimester or after birth were re-analyzed. Results: Among 1 152 families who received WES, 5 families were excluded because of nonbiological parents. Among the remaining 1 147 families, 152 fetuses obtained positive diagnosis (13.3%,152/1 147), including 74 fetuses in the retrospective group (16.1%,74/460) and 78 fetuses in the prospective group (11.4%,78/687). In fetuses with negative CMA and G-band karyotype analysis results but new phenotypes in the third trimester or after birth, the positive rate by WES data re-analysis was 4.9% (8/163). A total of 34 (21.3%, 34/160) fetuses were directly affected by the corresponding positive molecular diagnosis. Among 68 cases of live births with diagnostic variation grade 4, 29 cases (42.7%, 29/68) received appropriate medical intervention through rapid review of WES results. Conclusions: WES could increase the detection rate of abnormal fetuses with negative G-banding karyotype analysis and CMA by 13.3%. Prenatal WES could guide pregnancy decision-making and early clinical intervention. It might be an effective strategy to pay attention to the special follow-up of the third trimester and postnatal fetus and to re-analyze the WES data.

Whole exome sequencing as a diagnostic adjunct to clinical testing in fetuses with structural abnormalities.

OBJECTIVES: To evaluate the diagnostic yield of prenatal whole exome sequencing (WES) for monogenic disorders in fetuses with structural malformations and normal results on cytogenetic testing, and to describe information on pathogenic variants that is provided by WES. METHODS: Karyotyping, chromosomal microarray analysis (CMA) and WES were performed sequentially on stored samples from a cohort of 3949 pregnancies with fetal structural abnormalities detected on ultrasound and/or magnetic resonance imaging, referred between January 2011 and December 2015. Diagnostic rates of the three techniques were investigated overall, for phenotypic subgroups and for proband-only vs fetus-mother-father samples. Information on pathogenic variants was identified by WES. RESULTS: Overall, 18.2% (720/3949) of fetuses had an abnormal karyotype. Pathogenic copy number variants were detected on CMA in 8.2% (138/1680) of fetuses that had a normal karyotype result. WES performed on a subgroup of 196 fetuses with normal CMA and karyotype results revealed the putative genetic variants responsible for the abnormal phenotypes in 47 cases (24%). The molecular diagnosis rates for fetus-mother-father and proband-only samples were 26.5% (13/49) and 23.1% (34/147), respectively. Variants of uncertain significance were detected in 12.8% (25/196) of fetuses, of which 22 were identified in the fetal proband-only group (15%; 22/147) and three in the fetus-mother-father group (6.1%; 3/49). The incidental finding rate was 6.1% (12/196). CONCLUSIONS: WES is a promising method for the identification of genetic variants that cause structural abnormalities in fetuses with normal results on karyotyping and CMA. This enhanced diagnostic yield has the potential to improve the clinical management of pregnancies and to inform better the reproductive decisions of affected families. Copyright © 2017 ISUOG. Published by John Wiley & Sons Ltd.

Effect of feeding Rumen-protected capsule containing niacin, K2SO4, vitamin C, and gamma-aminobutyric acid on heat stress and performance of dairy cows.

This study was conducted to evaluate the effects of supplemental rumen-protected capsule (RPC) on animal performance, serological indicators, and serum heat shock protein 70 (HSP70) of lactating Holstein cows under heat stress (HS). During summer months, 30 healthy multiparous lactating Holstein cows with a parity number of 3.1 ± 0.44, 70 ± 15 d in milk, an average body weight of 622 ± 62kg, and an average milk yield of 32.28 ± 0.96kg/d, were used. The cows were randomly allocated to two groups: a control group and an RPC-supplemented group (0.13373kg K2SO4, 0.02488kg vitamin C, 0.021148kg niacin, and 0.044784kggamma-aminobutyric acid per cow). During the 42-d experiment, ambient air temperature and relative humidity inside and outside the barn were recorded hourly every day for the determination of temperature-humidity index (THI). Milk and blood samples were collected every week, and body weight and body condition scoring were measured on day 0. Based on the THI values, the animals had moderate HS. On day 42, the RPC group had lower HSP70, adrenocorticotropic hormone (P = 0.0001), lactate dehydrogenase (P = 0.0338), and IL-6 (P = 0.0724) levels than the control group, with no significant differences in creatine kinase, glucocorticoid, or IL-2 levels. Milk yield, energy-corrected milk, and dry matter intake were higher in RPC than in the control group (P = 0.0196). There were no significant differences in milk fat or daily protein levels between the two groups; however, daily protein and milk fat levels were higher in the RPC group than in the control group (P = 0.0114 and P = 0.0665, respectively). Somatic cell counts were no different between the two groups. In conclusion, RPC may alleviate HS and improve dairy cow performance.

[Application of chromosomal microarray analysis for fetuses with talipes equinovarus].

OBJECTIVE: To investigate the application of fetuses with talipes equinovarus(TE)using chromosomal microarray analysis(CMA)technology. METHODS: From May 2012 to June 2015, 54 fetuses were found with TE and with or without other structural anomalies by prenatal ultrasound. Karyotyping was taking for them all, and the fetuses with normal karyotypes took another CMA test. The data were analyzed with CHAS software. Finally all the cases were followed up to know about their pregnancy outcomes. RESULTS: One of the 54 cases was detected with abnormal karyotype which was trisomy 18(2%, 1/54). CMA was undertaken to the remaining fetuses, they were divided into 2 groups, including isolated TE group(n= 38)and complex TE group(n=15). The detection rate of clinical significant copy number variations(CNV)by CMA was 11%(6/53), while isolated and complex TE group were 5%(2/38)and 4/15, respectively(P= 0.047). Of the 53 cases, 51 cases were successfully followed up. Eleven cases were found without TE after birth, and the false positive rate(FPR)of TE was 22%(11/51). CONCLUSIONS: Whole-genome high-resolution CMA increased the detection rate by 11% in fetuses with TE. With the FPR and the detection rate of the clinical significant CNV of 2 groups, whole-genome CMA could be recommended to the fetuses with complex TE group but normal karyotypes. A series of ultrasonic tests should be suggested to the isolate TE group, while with the abnormal ultrasound, fetuses would be suggested to have CMA test for decreasing the rates of invasive prenatal diagnosis and FPR.

A novel missense mutation nt737T>G of JK gene with Jk(a-b-) phenotype in Chinese blood donors.

OBJECTIVES: The aim of this study was to investigate the molecular mechanism of the JK-null phenotype in the Chinese population. BACKGROUND: The Jk(a-b-) phenotype is vanishingly rare and the molecular basis differs between ethnic groups. The information regarding the molecular basis of JK-null alleles in the Chinese population is limited. MATERIALS AND METHODS: Three unrelated Jk(a-b-) phenotype donors were selected from 52 260 randomly blood samples through the urea lysis test and serological analysis. The JK gene-coding regions were amplified by the polymerase chain reaction and the products were sequenced directly. RESULTS: Sequencing results revealed that one sample of JK(*) B alleles carried the well-known Polynesian Jk(a-b-) mutation IVS5-1g>a. Another null allele, also on the JK(*) B background, presented with two heterozygous missense mutation, including nt222C>A(Asn74Lys) in exon 5 and nt896G>A(Gly299Glu) in exon 9. The third null allele carried two heterozygous missense mutations, nt222C>A and a novel allele nt737T>G(Leu246Arg) in exon 8. The family investigation revealed that the proband was JK(*) A(737T>G)/JK(*) B(222C>A). CONCLUSION: The Jk(a-b-) phenotype in the Chinese population shows several different molecular mechanisms. A novel missense mutation nt737T>G of JK gene was found as associated with Jk(a-b-) phenotype.

A 1st-trimester combined screening test in pregnant women of advanced maternal age in a Chinese population.

The purpose of this study was to evaluate the performance of the 1st-trimester combined screening test for trisomy 21 in different maternal age groups in a Chinese population. In this retrospective study, data on the 1st-trimester combined screening test (maternal age, fetal nuchal translucency, free ß-human chorionic gonadotrophin, pregnancy-associated plasma protein-A) were analysed. The study population of 17,556 pregnant women was subdivided into three groups according to maternal age: 16,113 were < 35 years of age; 1,228 were 35-39 years of age; and 215 were ≥ 40 years of age. The detection and false-positive rates of the 1st-trimester screening test for trisomy 21 or trisomy 18 in the three groups of women were 89.5 and 1.7%; 90.9 and 6.8%; and 100 and 22.3%, respectively. With increasing maternal age, the odds of being affected given a positive result (OAPR) were increased. The balance between the detection rate and false-positive rate of the 1st-trimester combined screening test is more favourable in women < 36 years with comparable OAPR. Although the false-positive rate increases with increasing maternal age, the performance of the 1st-trimester combined screening test in women ≥ 35 years is more effective than screening based on maternal age alone.

Increased first-trimester nuchal translucency associated with thanatophoric dysplasia type 1.

Thanatophoric dysplasia (TD) is the most frequent form of lethal skeletal dysplasia. Prenatal diagnosis is commonly accomplished in the second-trimester scan, but occasionally TD is found to be associated with increased nuchal translucency (NT) at first-trimester screening for aneuploidies. TD may not be clearly distinguished from the other skeletal dysplasias. A definite diagnosis can be established by molecular genetic analysis to find out the abnormal mutations in the fibroblast growth factor receptor 3 (FGFR3) gene. We reported a case of TD type 1 detected by first-trimester NT measurement, and confirmed by molecular analysis of FGFR3 gene using high-resolution melting analysis.

The inhibitory role of miR-214 in cervical cancer cells through directly targeting mitochondrial transcription factor A (TFAM).

Mitochondrial transcription factor A (TFAM) is a high-mobility group (HMG) protein and acts as a key regulator in mitochondrial DNA (mtDNA) replication, transcription, and inheritance. Accumulating evidence has demonstrated that TFAM plays an important role in tumorigenesis; however, the regulatory mechanism of TFAM in cervical cancer has not been revealed. In the current study, the au- thors found that with malignancy of cervical cancer, the protein expression of TFAM was gradually increased, while the expression of miRNA-214 was gradually downregulated. They further identified that TFAM is a target of miR-214. Forced overexpression of miRNA-214 significantly suppressed cell proliferation, cell cycle progression, colony-formation, and migration of cervical cancer Hela and Caski cells; however, upregulation of TFAM notably promoted cell proliferation, cell cycle progression, colony-formation, and migration of Hela and Caski cells. The authors further showed that miR-214 enhanced the susceptibility of Hela and Caski cells to the chemotherapy drug cisplatin. In conclusion, the current study provides a new sight for the regulatory pattern of miRNA-214 and TFAM in cervical cancer in vitro, indicating that miRNA-214 and MTFA may become important candidates for developing promising therapeutic strategies for the treatment of cervical cancer.

Chorionic villus sampling for early prenatal diagnosis: Experience at a mainland Chinese hospital.

The aim of this study was to describe the experience of transabdominal chorionic villus sampling (CVS) at a mainland Chinese hospital. During a 7-year period, 1,172 pregnant women chose to have CVS for prenatal diagnosis. Details and outcome of all of these cases were reviewed. The median maternal age was 29 years (range 19-45). The median gestational age was 12 weeks (range 10-14). Fetal karyotyping and thalassaemia couples were the main indications (97.2%). Overall, 112 (9.7%) chromosomal abnormalities were identified. There were 91 (7.8%) major chromosomal abnormalities, including autosomal trisomy in 70 patients, sex chromosomal abnormalities in 17, triploidy in two and unbalanced chromosomal rearrangement abnormality in two. Additionally, 137 fetuses with severe thalassaemia syndrome were found, including 86 homozygous ß-thalassaemia, and 51 homozygous α-thalassaemia or non-deletional haemoglobin H disease. The procedure failed to obtain an adequate sample in four (0.3%) patients. There were 229 pregnancies terminated for medical indications after CVS. There were three (0.3%) potentially procedure-related fetal losses. CVS is a safe and reliable prenatal diagnostic technique. It should be one of the options available to pregnant women who require prenatal diagnosis.

Effect of integrative medicines on 28-day mortality from sepsis: a systematic review and network meta-analysis.

OBJECTIVE: The aim is to perform a network meta-analysis to evaluate the effect of different Chinese medicines combined with Western medicine treatment (WMT) on the 28-day mortality of patients with sepsis. MATERIALS AND METHODS: We searched multiple databases for randomized controlled trials (RCTs), using keywords such as sepsis, Shenfu, Shenmai, Shengmai, Dahuang Fuzi, Xuebijing, and mortality. The databases we searched included the China Knowledge Network (CNKI), Wanfang database, the Chinese scientific journal database (VIP), the Chinese biomedical literature database (CBM), PubMed, Embase and Cochrane. The collection time was from inception through September 22, 2020. Two researchers independently screened the articles, extracted data, and assessed the quality of the evidence. Stata15 was used for network meta-analysis, and Excel was used to summarize the list of adverse reactions. RESULTS: Twenty-nine studies with a total of 3201 patients were included in this study. Combined with WMT, Shenfu injection [OR = 0.63, 95% CI = (0.47, 0.83)] and Xuebijing injection [OR = 0.71, 95% CI = (0.52, 0.96)] can significantly reduce 28-day mortality among patients with sepsis, while Shenmai injection [OR = 0.40, 95% CI = (0.16, 1.03)] and Dahuang Fuzi decoction [OR = 1.00, 95% CI = (0.48, 2.10)] cannot significantly improve on the 28-day mortality. The efficacy ranking is as follows: Shenmai group (level of evidence: low) > Shenfu group (level of evidence: low) > Xuebijing group (level of evidence: very low) > WMT group > Dahuangfuzi group (level of evidence: very low). CONCLUSIONS: Combined with WMT, Shenfu injection or Xuebijing injection can reduce the 28-day mortality among patients with sepsis. The Shenfu group had the best effect on outcomes, and its level of evidence was higher than that of the Xuebijing group.

Mussel-Inspired Hydrogels for Fluoride Delivery and Caries Prevention.

Fluoride agents hold promise for the repair and prevention of caries lesions, but their interaction with enamel is often hampered and diminished because of the dynamic wet environment in the oral cavity, which affects the efficacy of fluoride delivery and limits treatment success. We herein developed a mussel-inspired wet adhesive fluoride system (denoted TS@NaF) fabricated by the self-assembly of tannic acid (TA), silk fibroin (SF), and sodium fluoride (NaF). TS@NaF demonstrated remarkable biological stability and biocompatibility, showed reliable wet adhesion, released fluoride ions (F-) topically, and induced significant deposition of calcium fluoride (CaF2) onto enamel in vitro. Furthermore, TS@NaF provided an anticaries effect in vitro and induced a detectable increase in enamel mineral density. Advanced fluoride-releasing bioadhesives are therefore promising candidates for caries prevention and highlight the great potential of mussel-inspired dental materials in clinical applications.

Acoustic analysis of a rectangular cavity with general impedance boundary conditions.

A Fourier series method is proposed for the acoustic analysis of a rectangular cavity with impedance boundary conditions arbitrarily specified on any of the walls. The sound pressure is expressed as the combination of a three-dimensional Fourier cosine series and six supplementary two-dimensional expansions introduced to ensure (accelerate) the uniform and absolute convergence (rate) of the series representation in the cavity including the boundary surfaces. The expansion coefficients are determined using the Rayleigh-Ritz method. Since the pressure field is constructed adequately smooth throughout the entire solution domain, the Rayleigh-Ritz solution is mathematically equivalent to what is obtained from a strong formulation based on directly solving the governing equations and the boundary conditions. To unify the treatments of arbitrary nonuniform impedance boundary conditions, the impedance distribution function on each specified surface is invariantly expressed as a double Fourier series expansion so that all the relevant integrals can be calculated analytically. The modal parameters for the acoustic cavity can be simultaneously obtained from solving a standard matrix eigenvalue problem instead of iteratively solving a nonlinear transcendental equation as in the existing methods. Several numerical examples are presented to demonstrate the effectiveness and reliability of the current method for various impedance boundary conditions, including nonuniform impedance distributions.

Single-incision laparoscopic appendectomy performed above the pubic symphysis - a new scarless approach.

The three-port method is commonly used for laparoscopic appendectomy. To obtain a better cosmetic result, we have selected the single-incision laparoscopic appendectomy to be performed above the pubic symphysis. We performed six single-incision laparoscopic appendectomies above the pubic symphysis. During each operation, a 2 cm transverse incision was made in the pubic hair area 3-4 cm above the pubic symphysis. A 5 mm trocar was then placed as an observation port with another two 5 mm trocars as main- and side-operating ports. The laparoscopic appendectomy was performed using this single-incision method. Six patients (five male and one female; body mass indexes of 18.07, 19.27, 21.67, 18.34, 26.83 and 22.46 kg/m(2), respectively) underwent successful single-incision laparoscopic appendectomy above the pubic symphysis. Operating times were recorded at 55, 58, 47, 51, 42 and 33 minutes, the corresponding post-operative anal ventilation times were 25, 24, 22, 18, 7 and 10 hours, while post-operative hospitalization periods stand at 3, 5, 2, 1, 3 and 2 days, respectively. No complications occurred during or post-operation. Our initial work shows that a single-incision laparoscopic appendectomy performed above the pubic symphysis is feasible and safe and yields excellent post-operative cosmetic results.

[Management strategy for the resumption of regular diagnosis and treatment in gastrointestinal surgery department during the outbreak of coronavirus disease 2019 (COVID-19)].

Acute abdomen, abdominal trauma, gastrointestinal bleeding and gastrointestinal tumors are the main conditions that are routinely treated in gastrointestinal surgery department with high incidence and critical condition. These conditions need emergency or selective operations. During the outbreak of the coronavirus disease 2019 (COVID-19), it's a great challenge for us to meet the patients' requirement under the situation. As the COVID-19 was brought under control in China, the Department of General Surgery in Nanfang Hospital resumed regular medical services gradually. Based on our clinical practice, the four major measures of strengthening pre-hospital screening, perioperative prevention and control, medical staff protection, and ward management were adopted. These main measures include the strict implementation of the appointment system and triage system before admission; the conduction of epidemiological and preliminary screening of viral nucleic acids; the chest CT examination during the perioperative period to re-screen COVID-19; the reduction of the risk of droplets and aerosol transmission; the minimally invasive surgery combined with enhanced recovery program in order to reduce patient's susceptibility and shorten the length of postoperative hospital stay; the reinforcement of specific infection control training for medical staff; the strict implementation of hierarchical protection; the establishment of gastrointestinal surgery prevention and control system; the rehearsal of emergency exercise; the installation of quarantine wards; the screening and management of family care-givers; the strict disinfection of environment and materials. Our preliminary practice shows that following the work guidelines issued by the Guangdong Province COVID-19 Prevention and Control Office and adopting precise management strategies in combination with the specific clinical features of gastrointestinal surgery, it is possible to safely resume regular care for the patients and comply to epidemic control at the same time.