Trimethylamine N-Oxide increases soluble fms-like tyrosine Kinase-1 in human placenta via NADPH oxidase dependent ROS accumulation.

BACKGROUNDS: Preeclampsia (PE) is characterized as placental vascular disturbance and excessive secretion of soluble fms-like tyrosine kinase 1 (sFlt-1) into the maternal circulation. Trimethylamine N-oxide (TMAO, a gut microbe-derived metabolite) is strongly associated with various cardiovascular and cerebrovascular diseases. Recently, we observe that higher maternal circulating TMAO and sFlt-1 in patients with PE. The aims of the present study are to explore the effects of TMAO on placental sFlt-1 production and the underlying mechanism in human placenta. METHODS: Human placental explants, human placental primary trophoblasts and the extravillous trophoblasts (EVT) cell line (HRT-8/SVneo) were exposured to various concentrations of TMAO (100, 150, 300, and 600 µM). The mRNA expression and protein secretion of sFlt-1 in placental explants, primary trophoblasts and HRT-8/SVneo cells were determined with qPCR and ELISA, respectively. The levels of intracellular reactive oxygen species (ROS) production in primary trophoblasts and HRT-8/SVneo cells were measured by peroxide-sensitive fluorescent probe dichlorofluorescein diacetate. RESULTS: Exposure of placental explants, primary trophoblasts and HRT-8/SVneo cells to TMAO significantly enhanced sFlt-1 at both mRNA and protein levels in a dose dependent manner. Moreover, inhibition of NADPH oxidase with apocynin significantly attenuated TMAO-induced ROS production in primary trophoblasts and HRT-8/SVneo, and suppressed sFlt-1 secretion in placental explants, primary trophoblasts and HRT-8/SVneo. CONCLUSIONS: Our findings indicated the NADPH oxidase dependent ROS pathway played a critical role in mediating TMAO-induced sFlt-1 generation in human placenta. TMAO may become a potential novel target for pharmacological or dietary interventions to reduce the risk of developing PE.

Cloning, bioinformatics, expression mode analysis of crocetin glycosyltransferase UGTCs4 / 中草药

Objective: With the aim to obtain crocetin glycosyltransferase UGTCs4 in cultured saffron suspension cell, and carry out its bioinformatics and expression mode analysis. Methods: A homologous cloning strategy and 5’ RACE methods were adopted, the full-length cDNA sequence of a crocetin glycosyltransferase, designated UGTCs4 (GeneBank number: KX398932), was obtained. The characteristics of physiochemical properties, structure and function of the deduced UGTCs4 protein were determined using a series of bioinformatics tools. Semi-quantitative PCR was used for gene expression analysis. Results: The results showed that the full length cDNA of UGTCs4 was 1 380 bp in length and encoding a 459 amino acid; UGTCs4 had high identities (83.2%) with UGTCs2 protein from saffron; UTGTCs4 had the same evolutionary tree as UGTCs2. UGTCs4 transcripts were constitutively expressed in the leaves, stems, and roots. UGTCs4 gene could respond to multiple treatments of indoleacetic acid (IAA), abscisic acid (ABA), gibberellin (GA), hydrogen peroxide (H2O2), and methyl jasmonate (MJA), which promoted its transcription. Conclusion: cDNA of crocetin glycosyltransferase was cloned from suspension cells for the first time and the response of UGTCs4 to different inducers was confirmed. Molecular characterization of UGTCs4 will be useful for further functional determination of the gene involving in the crocin biosynthesis and expression regulation.

Levels and correlates of 24-hour movement behaviors in Chinese children aged 6-13 years / 中国学校卫生

Objective@#To investigate the different patterns of 24-h movement behaviors, and their associations with sociodemographic factors in a nationally representative sample of Chinese children aged 6-13 years.@*Methods@#This study was based on a national multi-centered cluster intervention study involving 31 362 children aged 6-13 years from 7 provinces in China. Questionnaires were used to collect moderate to vigorous physical activity (MVPA), screen time (ST) and sleep duration, as well as sociodemographic variables including age, gender, area of residence, parents’ education level and family income. Generalized linear mixed model (GLMM) analyses were conducted for the 24-hour movement behaviors according to sociodemographic variables.@*Results@#The proportions of individuals meeting the MVPA, ST, and sleep guidelines were 32.2%, 78.5%, 30.1%, respectively. The proportion that meet 0, 1, 2 and 3 recommended items was 9.6%, 47.7%, 35.0% and 7.7%, respectively. Age, gender, parents’ education levels and family income showed associations with PA, ST and sleep. Compared with low parents’ education group, the risk for unhealthy behavioral patterns was lower in those with high parents’ education level(P<0.05).@*Conclusion@#The current status of 24-hour physical activity for children aged 6-13 in China is not ideal, and social demographics should be considered when designing targeted interventions to promote children’s health.

Association between hypertensive disorders of pregnancy and risk of autism in offspring: a systematic review and meta-analysis of observational studies.

BACKGROUND: Autism spectrum disorder (ASD) is a common severe pervasive neurodevelopmental disorder of undetermined etiology. Environmental exposures, especially pregnancy complications, have been increasingly recognized as a potential risk factor for ASD. Our aim was to (1) systematically evaluate the association between hypertensive disorders of pregnancy (HDP) and the risk of ASD in offspring, (2) specifically draw a subgroup analysis of disease severity in patients with HDP to achieve more sufficient evidence on this issue. RESULTS: A total of 21 studies were identified with more than 6.5 million participants, including 31,027 ASD probands. A comparative meta-analysis established that offspring born premature to HDP were significantly associated with ASD than matched controls (OR = 1.42, 95% CI: 1.34-1.50). Subgroup analysis of clinical classification include: (1) gestational hypertension, (2) pre-eclampsia, (3) chronic hypertension complicating pregnancy (CHP). The offspring of mothers with pre-eclampsia and CHP have slightly higher risk (OR = 1.43; OR = 1.48, respectively) of ASD than those of mothers with gestational hypertension (OR = 1.37). In consistence with most previous researches, higher ASD prevalence was observed in male than female (OR = 1.38), indicating a potential role for gender in the pathophysiology of ASD. MATERIALS AND METHODS: We conducted a systematic literature search on PubMed, EMBASE, Web of Science, PsycINFO database and China National Knowledge Infrastructure up to Jun. 2017. Statistical analysis was performed using Stata 10.0. CONCLUSIONS: This meta-analysis implies a possible link between HDP and the risk of ASD in offspring. However, further investigation should be conducted to confirm this conclusion, and intensive prenatal surveillance and early prediction for ASD is needed.

[Enhanced recovery after preserving the left colonic artery during laparoscopic anterior resection for rectal cancer].

OBJECTIVE: To evaluate the postoperative outcomes of preserving the left colonic artery during laparoscopic anterior resection for rectal cancer. METHODS: The clinicopathologic data of 91 rectal cancer patients (pathologic Stage II) undergoing laparoscopic anterior resection was retrospectively analyzed. During the surgeries, the left colonic artery was preserved in 40 patients (preserved group) and ligated in 51 patients (unpreserved group). The operating time, intraoperative blood loss, time to first flatus and defecation, duration of postoperative abdominal distension and pain, number of retrieved lymph nodes, ileum fistulation and anatomical leakage rate were compared between the two groups. RESULTS: The surgeries were completed in all the 91 patients laparoscopically without conversion. There was no intraoperative complications including rectal perforation, injury to vessel or ureter in either group. The operating time, blood loss and number of retrieved lymph nodes were similar between the groups (P>0.05). Three patients in preserved group and 5 in ligation group received preventive ileum fistulation due to low rectal cancer. Anatomical leakage occurred in three patients of unpreserved group. The average duration of postoperative abdominal distension and pain was 2.14∓0.35 days in preserved group and 3.15∓0.42 days in ligation group. The time to first flatus and defecation was 37.15∓12.62 h and 3.16∓0.52 days in preserved group and 62.25∓11.75 h and 4.25∓0.75 days in ligation group. Postoperative hospital stay was 4.54∓0.42 days in preserved group and 6.23∓0.51 days in ligation group. Total hospitalization cost in the two groups was 34 525.32∓1206.36 Yuan and 41 215∓1051.32 Yuan, respectively. Significant differences were found the in duration of postoperative abdominal distension and pain, postoperative hospital stay, and total cost between the two groups (P<0.05). CONCLUSION: During laparoscopic anterior resection for rectal cancer, preserving the left colonic artery effectively ensures the blood supply to the anastomosis and the remaining descending colon to promote the recovery of the patients after surgery.

Heavy Metals Induce Decline of Derivatives of 5-Methycytosine in Both DNA and RNA of Stem Cells.

Toxic heavy metals have been considered to be harmful environmental contaminations. The molecular mechanisms of heavy-metals-induced cytotoxicity and carcinogenicity are still not well elucidated. Previous reports showed exposures to toxic heavy metals can cause a change of DNA cytosine methylation (5-methylcytosine, 5-mC). However, it is still not clear whether heavy metals have effects on the recently identified new epigenetic marks in both DNA and RNA, i.e., 5-hydroxymethylcytosine (5-hmC), 5-formylcytosine (5-foC), and 5-carboxylcytosine (5-caC). Here, we established a chemical labeling strategy in combination with liquid chromatography-electrospray ionization-mass spectrometry (LC-ESI-MS/MS) analysis for highly sensitive detection of eight modified cytidines in DNA and RNA. The developed method allowed simultaneous detection of all eight modified cytidines with improved detection sensitivities of 128-443-fold. Using this method, we demonstrated that the levels of 5-hmC, 5-foC, and 5-caC significantly decreased in both the DNA and RNA of mouse embryonic stem (ES) cells while exposed to arsenic (As), cadmium (Cd), chromium (Cr), and antimony (Sb). In addition, we found that treatments by heavy metals induced a decrease of the activities of 10-11 translocation (Tet) proteins. Furthermore, we revealed that a content change of metabolites occurring in the tricarboxylic acid cycle may be responsible for the decline of the derivatives of 5-mC. Our study shed light on the epigenetic effects of heavy metals, especially for the induced decline of the derivatives of 5-mC in both DNA and RNA.

The Significance of Abnormal Muscle Response Monitoring During Microvascular Decompression for Hemifacial Spasm.

BACKGROUND: Despite the wide adoption of the abnormal muscle response (AMR) to electrical stimulation of the facial nerve during microvascular decompression (MVD) surgery, the value of AMR in the prognosis of the postoperative outcome is still controversial. In order to better use this intraoperative electrophysiology, it is necessary to further address the relationship between AMR and postoperative results. METHODS: Three hundred and thirty-two patients with hemifacial spasm (HFS) in whom MVD surgery was performed and in whom AMR was available were finally enrolled in this study. The intraoperative AMR changes were classified as amplitude ≥ 50 %, <50 %, and disappearance. These changes were retrospectively analyzed in association with intraoperative findings and postoperative outcomes. The follow-up period ranged from 11 to 62 months, with an average of 34.1 months. RESULTS: Among the 332 patients with a typical AMR wave recorded at the beginning of the operation, the AMR disappeared in 305, and amplitude was <50 % in 11 and ≥50 % in 16. Of those with AMR disappearance plus those with amplitude < 50 %, 98.4 % achieved relief on the first postoperative day and at the latest follow-up, while of those with amplitude ≥50 %, 18.8 % and 25 %, respectively, achieved relief on the first postoperative day and at the latest follow-up (P < 0.01). Accordingly, a more than 50 % decrease of AMR amplitude may predict a good prognosis. The accuracy, sensitivity, and specificity of AMR monitoring were 97.5 %, 99 %, and 72.2 %, respectively. CONCLUSIONS: AMR could be a good tool for successful MVD in patients with HFS when a rational analysis is conducted in association with the intraoperative findings. Persistence of AMR may imply that the real offending vessel was missed. If the entire facial nerve root is cleared of any vessel, a remaining AMR amplitude of less than 50 % might be acceptable. Otherwise, neurocombing is suggested before finishing the operation.

The operation experience of endoscopic thyroidectomy by areola and axilla approach.

To explore the feasibility of endoscopic thyroidectomy via breast areola and axilla approach. The clinical data of 36 cases that underwent endoscopic thyroidectomy via breast areola and axilla approach from February 2012 to December 2013 were reviewed. All cases were completed, the mean operation time was 136.3 min (95-183 min), intraoperative blood loss was 15.8 ml (5-60 ml). The average hospitalization time was 5 days (4-6 days). There were no conversions to open surgery, no permanent nerve injuries, and no cases of hypoparathyroidism. Three patients had postoperative subcutaneous ecchymosis who were cured spontaneously after 1 month. Endoscopic thyroidectomy is safe and feasible for patients with thyroid diseases with good cosmetic results, and is worthy of being widely applied for patients who have cosmetic demand.

[Identification of C(2)M interacting proteins by yeast two-hybrid screening].

The synaptonemal complex (SC) is a huge structure which assembles between the homologous chromosomes during meiotic prophase I. Drosophila germ cell-specific nucleoprotein C(2)M clustering at chromosomes can induce SC formation. To further study the molecular function and mechanism of C(2)M in meiosis, we constructed a bait vector for C(2)M and used the yeast two-hybrid system to identify C(2)M interacting proteins. Forty interacting proteins were obtained, including many DNA and histone binding proteins, ATP synthases and transcription factors. Gene silencing assays in Drosophila showed that two genes, wech and Psf1, may delay the disappearance of SC. These results indicate that Wech and Psf1 may form a complex with C(2)M to participate in the formation or stabilization of the SC complex.

The effects of microcystin-LR on the mRNA expression levels of base excision repair genes and genes related to apoptosis / 中华劳动卫生职业病杂志

<p><b>OBJECTIVE</b>To explore the effects of microcystin-LR (MCLR) on the expression of base excision repair genes and genes related to apoptosis.</p><p><b>METHODS</b>The BRL-3A cells were exposed to different concentrations of MCLR for various periods of time and the cell viability was measured by MTT. The mRNA expression was determined with the quantitative real-time polymerase chain reaction (QRT-PCR).</p><p><b>RESULTS</b>The viability of BRL-3A cells significantly reduced in a concentration- and time-dependent manner. In 30 µg/ml group, the mRNA expression level (1.327 ± 0.028) of p53 increased significantly at 24 h after exposure, as compared with the other groups (1.005 ± 0.117, 0.862 ± 0.154, 1.028 ± 0.056 and 1.015 ± 0.091) (P < 0.05). The mRNA expression levels (5.080 ± 0.729, 5.820 ± 0.373, 6.018 ± 0.359 and 6.183 ± 0.515) of Bax in all exposure groups were significantly higher than that (1.024 ± 0.277) in control group at 24 h after exposure. However, the Bax mRNA expression level (0.604 ± 0.146) in the 30 µg/ml group at 72 h after exposure was significantly lower than those (1.004 ± 0.107, 0.811 ± 0.142, 0.855 ± 0.101 and 0.814 ± 0.056) in other groups (P < 0.05). When compared with control group (1.006 ± 0.132) and 1 µg/ml group (1.034 ± 0.241), the mRNA expression level (0.488 ± 0.147) of PARP1 in 30 µg/ml group at 48 h after exposure decreased significantly (P < 0.05). Furthermore, the mRNA expression levels (0.594 ± 0.180, 0.491 ± 0.015 and 0.305 ± 0.091) of JWA, XRCC1 and PARP1 in 30 µg/ml group at 72 h after exposure decreased significantly, as compared with the other groups (P < 0.05).</p><p><b>CONCLUSION</b>The induction of gene expression is a transient phenomenon that occurred at different times of exposure for different genes. Inhibition of MCLR on the base excision repair gene expression may play important role in the course of MCLR promoting liver tumor.</p>

[Characterization of the beta-actin gene of the rice field eel and its phylogeny in fish].

Beta-actin is a member of the actin family of genes,which play important roles in maintaining cytoskeletal structure, cell motility, cell division, intracellular movements and contractile processes. We report here the identification of a beta-actin cDNA of the rice field eel,a teleost fish with a characteristic of natural sex reversal. The cDNA sequence of this gene was 1860 bp in length,encoding a 375 amino acid protein. Amino acid identities of the beta-actin between the rice field eel and other vertebrates including human, chicken, and other fishes, were more than 98%. RT-PCR showed expression of the rice field eel beta-actin in testis, ovotestis, ovary, heart, liver, spleen and brain,suggesting a ubiquitous expression pattern. Phylogenetic tree including all beta-actin cds of teleost fishes was constructed,which suggested consistently that teleost beta-actin can be classified into four types,but no fish has been found contains all the four types of beta-actin gene in its genome. The results that suggest lineage-specific beta-actin loss might happen in the radical evolution of teleost fishes.

[Ribosomal protein genes highly expressed in swamp eel gonads].

8 cDNA clones have been isolated from a cDNA library prepared from swamp eel testies by macroarray. DNA sequence analysis and database search showed that they encode 8 proteins which are highly homologous to 40S ribosomal proteins S4,S9,S16,S17,S20 and 60S riobosomal proteins L7, L18a,L29. Phylogenetic trees (ML) based on ribosomal protein genes from swamp eel and other organisms has been reconstructed, which showed that ribosomal protein genes were highly conserved during evolution. These results suggested that ribosomal protein genes as house keeping genes may play roles in developmental regulation such as sexual differentiation and can also be used as markers for the study of molecular evolution.

[Cloning of four members of giant panda Dmrt genes].

Sex determining genes Mab-3 of C. elegans and Doublesex of Drosophila contain a common DNA binding motif called DM (Doublesex and Mab-3) domain, both of which regulate similar aspects of sexual development. Human Doublesex-related gene DMRT1 has been identified, which also contains the conserved DM-related DNA-binding domain and plays an essential role in gonadal differentiation. We amplified genomic DNA of the giant panda using the DM degenerate primers and detected two bands, approximately 140 bp and 250 bp. After cloned into T-easy vector and sequenced, four sequences showed high homology with the DM domain. Amino acid sequence of the first clone is 100% identical with the Dmrt1 of human, mouse and pig, hence we named it as pDmrt1. The second clone is 96% identical with human DMRTB1, and the third one 100% with the Dmrt3 of mouse and medaka, which were named as pDmrtb1 and pDmrt3 respectively. The last sequence contains an intron of 116 bp within the DM domain, which encodes an amino acid sequence 100% identical with human DMRTC2, accordingly we named it as pDmrtc2. Based on similarities of amino acid sequences of the DM domain, Dmrt protein sequences from human, mouse and giant panda were included in a phylogenetic tree. They revealed seven distinct subgroups: Dmrt1, Dmrt2, Dmrt3, Dmrt4 (DMRTA1), Dmrt5 (DMRTA2), Dmrt6 (DMRTB1) and Dmrt7 (DMRTC2). Our results further reveal the unexpected complexity and the evolutionary conservation of the DM domain gene family in both invertebrates and vertebrates.