COVID-19: Variant screening, an important step towards precision epidemiology

Precision epidemiology using genomic technologies allows for a more targeted approach to COVID-19 control and treatment at individual and population level, and is the urgent need of the day. It enables identification of patients who may be at higher risk than others to COVID-19-related mortality, due to their genetic architecture, or who might respond better to a COVID-19 treatment. The COVID-19 virus, similar to SARS-CoV, uses the ACE2 receptor for cell entry and employs the cellular serine protease TMPRSS2 for viral S protein priming. This study aspires to present a multi-omics view of how variations in the ACE2 and TMPRSS2 genes affect COVID-19 infection and disease progression in affected individuals. It reports, for both genes, several variant and gene expression analysis findings, through (i) comparison analysis over single nucleotide polymorphisms (SNPs), that may account for the difference of COVID-19 manifestations among global sub-populations; (ii) calculating prevalence of structural variations (copy number variations (CNVs) / insertions), amongst populations; and (iii) studying expression patterns stratified by gender and age, over all human tissues. This work is a good first step to be followed by additional studies and functional assays towards informed treatment decisions and improved control of the infection rate.

Banxia Xiexin Decoction () Treats Diabetic Gastroparesis through PLC-IP / 中国结合医学杂志

OBJECTIVE@#To test the effect of Banxia Xiexin Decoction (, BXD) on the contraction and relaxation of gastric smooth muscle (SM) in diabetic gastroparesis (DGP) model rats, and to explore the mechanism of BXD in the prevention and treatment of DGP through experiments of signal pathway both in vivo and in vitro.@*METHODS@#Sixty Sprague-Dawley rats were divided into 6 groups according to a random number table: control group, model group, high-, medium- and low-dose BXD groups (9.2, 4.6 and 1.8 g/(kg·d), respectively), and domperidone group (10 mg/(kg·d)), 10 rats per group. DGP model was established initially by a single intraperitoneal injection of streptozotocin (STZ), and was confirmed by recording gastric emptying, intestinal transport velocity and gastric myoelectric activity of rats after 2 months. Each group was treated with a corresponding drug for 4 weeks. The mRNA and protein expressions of phospholipase C (PLC), inositol triphosphate (IP@*RESULTS@#Compared with the model group, high- and medium-dose BXD and domperidone significantly increased the expressions of PLC, IP@*CONCLUSIONS@#Treatment with high- and medium-dose BXD significantly attenuated STZ-induced experimental DGP in rats. The therapeutic effect of BXD on DGP rats might be associated with the PLC-IP

Expression profiles of PI3K, NF-κB, and STAT1 in peripheral blood mononuclear cells in children with bronchial asthma / 中国当代儿科杂志

<p><b>OBJECTIVE</b>To study the expression profiles of PI3K, NF-κB, and STAT1 in peripheral blood mononuclear cells (PBMCs) in children with bronchial asthma, as well as their roles in the pathogenesis of asthma.</p><p><b>METHODS</b>Thirty children with acute exacerbation of bronchial asthma were enrolled as the asthma group, and 20 healthy children were enrolled as the control group. RT-PCR and Western blot were used to measure the mRNA and protein expression levels of PI3K, NF-κB, and STAT1 in PBMCs. A spirometer was used to compare the pulmonary function between the two groups. The correlations between the mRNA expression of PI3K, NF-κB, and STAT1 and pulmonary function in children with bronchial asthma were analyzed.</p><p><b>RESULTS</b>The asthma group had significantly higher mRNA and protein expression levels of PI3K, NF-κB, and STAT1 than the control group (P<0.05). Compared with the control group, the asthma group showed significant reductions in pulmonary function indices such as FEV1%, FEV1/FVC, and PEF% (P<0.05). In children with bronchial asthma, the mRNA expression levels of PI3K, NF-κB, and STAT1 were negatively correlated with FEV1%, FEV1/FVC, and PEF% (P<0.05).</p><p><b>CONCLUSIONS</b>The expression levels of PI3K, NF-κB, and STAT1 increase in children with asthma, and are negatively correlated with pulmonary function indices, suggesting that PI3K, NF-κB and STAT1 are involved in the development and progression of bronchial asthma in children.</p>

Structure design of and experimental research on a two-stage laval foam breaker for foam fluid recycling.

Environmental, economical and efficient antifoaming technology is the basis for achievement of foam drilling fluid recycling. The present study designed a novel two-stage laval mechanical foam breaker that primarily uses vacuum generated by Coanda effect and Laval principle to break foam. Numerical simulation results showed that the value and distribution of negative pressure of two-stage laval foam breaker were larger than that of the normal foam breaker. Experimental results showed that foam-breaking efficiency of two-stage laval foam breaker was higher than that of normal foam breaker, when gas-to-liquid ratio and liquid flow rate changed. The foam-breaking efficiency of normal foam breaker decreased rapidly with increasing foam stability, whereas the two-stage laval foam breaker remained unchanged. Foam base fluid would be recycled using two-stage laval foam breaker, which would reduce the foam drilling cost sharply and waste disposals that adverse by affect the environment.

Biological effect of micro ribonucleic acid-21 on human tongue squamous cell carcinoma lines / 华西口腔医学杂志

<p><b>OBJECTIVE</b>To detect the biological influence to human tongue squamous cell carcinoma (TSCC) cells of micro ribonucleic acid-21 (miR-21).</p><p><b>METHODS</b>Referring to mature miR-21 sequence, the sense and antisense oligonucleotide (sense-miR-21 and AS-miR-21) modified by 2'O-Me were designed to transfect into TSCC cells (Tca8113 and high metastasis cells) by liposome transfection technology, in order to establish an in vitro TSCC cell model. The expression changes of miR-21 in the transfected cells were detected with real-time fluorescence quantitative polymerase chain reaction (real-time PCR). The changes of cell proliferation, cell cycle, cell early apoptosis, cell migration and invasion capabilities were detected respectively by the technologies of methyl thiazolyl tetrazolium (MTT), flow cytometry, Annexin V cell early apoptosis assay, scratch assay and Transwell assay, to check AS-miR-21's effect on the biological characteristics of human TSCC cell lines.</p><p><b>RESULTS</b>For the TSCC cells, the antisense oligonucleotide of targeting miR-21 could effectively inhibit cell proliferation, promoted cell apoptosis, and inhibited the capability of cell's migration and invasion.</p><p><b>CONCLUSION</b>The expressions of miR-21 decrease after AS-miR-21 transfected into TSCC cells, and miR-21 can affect biological behavior of TSCC cells.</p>

In vivo study on antisense-micro ribonucleic acid-21 oligonucleotide inhibiting tongue squamous cell carcinoma growth / 华西口腔医学杂志

<p><b>OBJECTIVE</b>To detect antisense-micro ribonucleic acid-21 oligonucleotide (AS-miR-21)'s inhibiting effect to tongue squamous cell carcinoma.</p><p><b>METHODS</b>Living image and TUNEL experiments were performed, based upon the xenograft animal models set up by introduction of Tca8113-luc cells which were stably transfected with pGL6 luciferase report gene plasmid into nude mice, while the tumors were injected with AS-miR-21.</p><p><b>RESULTS</b>Tca8113-luc cell line which steadily expressed luciferase activity was constructed by transfecting pGL6 report gene plasmid. The subcutaneous tumor formation rate was much higher in nude mice introduced with the cells, and the tumors grew well. After injection of AS-miR-21 into mice tumors, it was obviously viewed that tumors grew slower, the volume of the tumors was smaller, the photon number in live body imaging was getting less, the necrosis in the tumor specimens was rare, cell nuclei was getting smaller, dyeing color was lighter, heteromorphism and new vessels were decreased, micro ribonucleic acid-21 expression in tumor cells was considerably lower, and apoptotic index was increased.</p><p><b>CONCLUSION</b>All the results indicate that the injection of AS-miR-21 can inhibit growth of tongue squamous cell carcinoma in nude mice model, and effectively promote cell apoptosis of tongue squamous cell carcinoma.</p>

Construction and Genetic Analysis of Murine Hepatitis Virus Strain A59 Nsp16 Temperature Sensitive Mutant and the Revertant Virus / 中国病毒学·英文版

Coronaviruses (CoVs) are generally associated with respiratory and enteric infections and have long been recognized as important pathogens of livestock and companion animals. Mouse hepatitis virus (MHV) is a widely studied model system for Coronavirus replication and pathogenesis. In this study, we created a MHV-A59 temperature sensitive (ts) mutant Wu"-ts18(cd) using the recombinant vaccinia reverse genetics system. Virus replication assay in 17C1-1 cells showed the plaque phenotype and replication characterization of constructed Wu"-ts18(cd) were indistinguishable from the reported ts mutant Wu"-ts 18. Then we cultured the ts mutant Wu"-ts 18(cd) at non-permissive temperature 39.5℃, which "forced" the ts recombinant virus to use second-site mutation to revert from a ts to a non-ts phenotype. Sequence analysis showed most of the revertants had the same single amino acid mutation at Nsp16 position 43. The single amino acid mutation at Nsp16 position 76 or position 130 could also revert the ts mutant Wu"-ts 18 (cd) to non-ts phenotype, an additional independent mutation in Nsp13 position 115 played an important role on plaque size. The results provided us with genetic information on the functional determinants of Nsp16. This allowed us to build up a more reasonable model of CoVs replication-transcription complex.

Determination of Zeranols in Plant Tissue by Rapid Resolution Liquid Chromatography-Mass Spectrometry/Mass Spectrometry / 分析化学

A method was developed for the determination of zeranols (α,β-zearalanol,α,β-zearalenol,zear-alanone,zearalenone) with RRLC-MS/MS in the plant tissue. The samples were extracted with acetonitrile and reextracted with aqueous alkaline solution,cleaned up with MAX column and then determined by RRLC-MS/MS using multiple reaction monitoring ( MRM) scan mode. The results showed that the working curves were linear in the range of 0 -20 μg/kg. The limits of detection ( LOD) of zeranols were 0.5 μg/kg and the limits of quantification (LOQ) was 1. 0μg/kg. Extraction recoveries for zeranols ranged from 75. 8% to 105.4% and RSD was between 2.4% and 12.1%. The method is suitable for the determination of zeranols in the plant tissue.

The specific killing of human melanoma cells by replication selective adenovirus / 中华肿瘤杂志

<p><b>OBJECTIVE</b>To construct replication selective adenovirus AdhepE1 targeting human melanoma and observe its specific killing of human melanoma cells in vitro.</p><p><b>METHODS</b>Adenovirus E1 region, the murine tyrosinase promoter and enhancer DNA sequences were acquired respectively by PCR cloning. The shuttle plasmid of replication-selective adenovirus targeting human melanoma was constructed by DNA recombination. Replication-selective adenovirus AdhepE1 was generated by homologous recombination. The human melanoma cell line SK-Mel-1 and hepatocellular carcinoma cell line HepG2 were attacked separately by lower dose of AdhepE1. Change of cell morphology was observed and the surviving cells were calculated. The expression of E1A was assayed by RT-PCR to verify the specific-replication of AdhepE1.</p><p><b>RESULTS</b>Replication selective adenovirus AdhepE1 targeting human melanoma was acquired by PCR. Human melanoma cell line SK-Mel-1 was sensitive to oncolytic killing of AdhepE1 whereas HepG2 was little responsive. The results of RT-PCR suggested that AdhepE1 replicated specifically in human melanoma cells.</p><p><b>CONCLUSION</b>AdhepE1 can selectively kill human melanoma cells.</p>

The protective effect of CGRP on ET-1 induced injury of human hepatocyte / 中国普通外科杂志

Objective To investigate the protective effect of Calcitonin Gene-related Peptide(CGRP)on ET-1 mediated injury of human hepatocyte.Methods Human liver tissues obtained from patients undergoing partial hepatectomies were randomly divided into five groups:control group,liver perfused with D-Hank's solution group;liver perfused with ET-1 group and three liver perfased with ET plus CGRP(10-6M,10-7M,10-8M)treated groups.Collagenase digestion method was used to isolate human hepatocytes,then hepatocytes were cultured,and the level of MDA and TNF-?,the viability and proliferation of hepatocyte,and the hepatocyte function(ALT,Alb,Urea and LDH)were determined.Results As compared with control group,in ET-1 group,the viability and proliferation of hepatocytes,the level of Alb and Urea declined significantly(P