OBJECTIVE To observe the anti-influenza effect of Jiawei Xiangru oral liquid through MDCK cells and mice, and predict its therapeutic mechanism for influenza based on network pharmacology. METHODS Observed the effect of different concentrations of Jiawei Xiangru oral liquid on MDCK cell damage caused by influenza A virus type 3 and its protective effect on influenza virus infected mice. The effective ingredients and action targets of each drug were collected through the Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform(TCMSP). Collect influenza disease related targets using GeneCards 5.14, OMMI and other databases, and construct a network diagram. Build PPI network diagrams, GO enrichment, and KEGG pathway analysis using STRING and Metascape databases. Complete molecular docking and 3D visualization display using AutoDockTools and PyMOL software. RESULTS In vitro experiments shown that the Jiawei Xiangru oral liquid had a significant inhibitory effect on MDCK cell damage and hemagglutination caused by influenza virus. In vivo experiments shown that it could prolong the survival time of mice infected with influenza A virus, improve the death protection rate of mice infected with the virus, and reduce the lung index and lung tissue virus titer of mice. The 82 active ingredients and 168 common targets were screened, with carotenoid, folic acid, 3'- methoxy daidzein, coellitin, and kaempferol as key components, and TNF, AKT1, EGFR, STAT3, SRC, MMP9, and PTGS2 as core targets. KEGG pathway enrichment analysis showed that the main mechanism of action maight exert anti-influenza virus effects through the TNF signaling pathway, chemokine signaling pathway, PI3K/Akt signaling pathway, and VEGF signaling pathway. CONCLUSION Pharmacodynamics have shown that the Jiawei Xiangru oral liquid can effectively treat influenza diseases, and its mechanism maybe related to the synergistic regulation of "multi components, multi targets, and multi pathways".

ObjectiveTo optimize and establish a multiplex polymerase chain reaction （PCR） system to simultaneously identify Ziziphi Spinosae Semen （ZSS），Hovenia acerba semen （HAS），and Ziziphi Mauritianae Semen （ZMS）， etermine their content to solve the problem of adulteration of ZSS pieces and its preparations. MethodAfter the analysis and comparison of the internal transcribed spacer （ITS） sequence differences of ZSS and its adulterants，specific single nucleotide polymorphism （SNP） sites were found，and specific primers for identification were designed. The samples of ZSS，HAS， and ZMS from different sources were specifically amplified under the conditions of optimized annealing temperature，the number of cycles， and concentration of primers，as well as different polymerases and PCR systems after evaluation. Identification was carried out according to the size of specific amplification bands，and the lower limit of detection （LOD） and adulteration LOD were studied. ResultWhen the annealing temperature was 63 ℃ and the number of cycles was 23，549，169，389 bp specific bands were amplified from ZSS，HAS， and ZMS. The lower LOD of this method was 0.24 ng and 1.2 ng for ZSS and HAS， respectively. The adulteration LOD for ZSS，HAS， and ZMS was 0.5%，2%， and 2% respectively. ConclusionThe established multiplex allele-specific PCR identification method can accurately identify ZSS，HAS， and ZMS at the same time，which can provide a basis for solving the problem of adulteration of ZSS and references for controlling the quality，security， and clinical application of ZSS.

Objective:To establish the reference for serum metabolomics profiles among healthy Han adults in China, and explore the variation on metabolomics profiles by geographic regions, sex, and age.Methods:Cross-sectional data and serum samples were obtained from the China National Health Survey. A total of 1 039 male and 1 032 female healthy adults(≥30 years) were included in this study. Serum metabolomics analyses were conducted with ultra-performance liquid chromatography-mass spectrometry(UPLC-MS). Orthogonal partial least squares discriminant analysis(OPLS-DA) was performed to compare the differences of metabolomics among different region, sex, and age.Results:Significant differences on metabolomics profiles were identified among region, sex, and age. A total of 114 region-related metabolites were spotted, including 53 metabolites that involved in human metabolic pathways, mainly peptides(20 metabolites) and glycerophospholipid metabolism-related(14 metabolites). Fifty-nine metabolites were pinned down to be sex-related, among which cotinine was significant in all 7 provinces. Age-related metabolites were only found in Shaanxi and Hainan, with 22 metabolites were recognized.Conclusion:Serum metabolomics varies by geographic regions, sex, and age. When metabolomics is applied for diagnosis or biomarker screening in various studies, it shall take into consideration of setting tailored references.

Primary hemifacial spasm is a motor disorder of facial muscles related to facial nerve. During the attack, the facial muscles present irregular and involuntary clonus, which can be induced or aggravated by emotional excitement, mental tension and random facial movement, seriously affecting daily work and life. The pathogenesis, diagnosis, differential diagnosis and treatment of the primary hemifacial spasm have been studied extensively in recent years. This article reviews the progress in these aspects.

Objective: To investigate the interference factors causing false-positive result of novel coronavirus IgM antibody (SARS-CoV-2 IgM) detected by gold immunochromatography assay (GICA) and enzyme-linked immunosorbent assay (ELISA). Methods: A total of 71 serum from different pathogen infections and related chronic diseases patients were collected from the Affiliated Hospital of North Sichuan Medical College from January 25, 2020 to February 15, 2020. GICA and ELISA were used to detect 2019-nCoV IgM in 71 serum, including 5 influenza A virus (Flu A) IgM positive serum, 5 influenza B virus (Flu B) IgM positive serum, 5 Mycoplasma pneumonia (MP) IgM positive serum, 5 Legionella pneumophila (LP) IgM positive serum, 29 rheumatoid factor (RF) IgM positive serum, 5 hypertension patients serum, 5 diabetes mellitus patients serum, 6 human immunodeficiency virus (HIV) infection patients serum and 6 Corona Virus Disease 2019 (COVID-19) patients serum. The interference factors causing false positive results of the two methods were analyzed, and urea dissociation test was employed to dissociate the 2019-nCoV IgM positive serum using the best dissociation concentration. Statistical analyses were performed by SPSS, version 19.0. Result: s 2019-nCoV IgM was positive in 18 cases of middle-high level RF-IgM positive serum and 6 cases of 2019-nCoV-infected serum detected by two methods, and the other 47 serum were negative. When the dissociation concentration of urea was 6 mol/L, 2019-nCoV IgM was negative in 17 cases of middle-high level RF-IgM positive serum and positive in 6 cases of 2019-nCoV-infected serum detected by GICA. When the urea dissociation concentration was 4 mol/L, dissociation time was 10 min and the avidity index<0.46 was set as negative, 2019-nCoV IgM was negative in 15 cases of middle-high level RF-IgM positive serum and positive in 6 cases of 2019-nCoV-infected serum detected by ELISA. Conclusion The middle-high level of RF-IgM could cause false positive results of SARS-CoV-2 IgM detected by GICA and ELISA, and the urea dissociation test would be helpful for reducing the probability of false-positive results of SARS-CoV-2 IgM test.

Growth plate,the developmental center of endochondral osteogenesis,can be divided morphologically and functionally into a resting zone,a proliferative zone,a prehypertrophic zone and a hypertrophic zone.Injuries to growth plate often lead to bone growth defects including limb length discrepancy and angulation deformity in children.Currently,their orthopedic corrective surgeries are invasive and limitedly effective and no effective biotherapy has been available.Previous studies on animal models of growth plate damage have investigated the related cellular and molecular events in the repair of damaged growth plates in the 4 distinct inflammatory,fibrogenic,osteogenic and remodeling phases.Related molecules involved in the regulation of the above processes,such as inflammatory cytokines tumor necrosis factor alpha,mitogenic platelet-derived growth factor and bone morphogenetic protein,are found to participate in the regulation of growth plate injury.Exploration of the mechanisms may provide new targets for biotherapy.In addition,development of cartilage tissue engineering,especially application of mesenchymal stem cells,also provides potential interventions for growth plate injury.

Objective To explore therapeutic effect of dipeptidyl peptidase IV inhibitor on type 2 diabetes mellitus with nonalcoholic fatty liver disease. Methods 120 patients suffering type 2 diabetes mellitus with nonal-coholic fatty liver disease from October 2014 to March 2016 in our hospital were randomly divided into two groups:treatment group and control group. Both groups were given type 2 diabetes conventional treatment ,and the treat-ment group was given dipeptidyl peptidase IV inhibitor in addition. Height,weight,waist circumference,hip cir-cumference were measured. Lipid metabolism and,function index,FPG,2 h PG,HbA1c,Ins,C peptide were detected. HOMA-IR was calculated. Results The total effective rate of clinical treatment of fatty liver in the treat-ment group(88.3%)was higher than the control group(78.3%). The difference was statistically significant. FPG , 2hPG,HbA1c,HOMA-IR,LDL-C,TC,TG,AST,ALT andγ-GT in the treatment group were lower than the con-trol group. The difference was also statistically significant. Conclusion The efficacy of sitagliptin in the treatment of type 2 diabetes with nonalcoholic fatty liver was significant. It can also significantly reduce blood glucose and in-sulin resistance. Furthermore ,it has a better effect on the patients with blood glucose control and lipid metabolism regulation.

OBJECTIVE:To develop a method for the determination of valsartan concentration in human plasma and urine. METHODS:Plasma sample were acidified and extracted with diethyl ether for analysis,and urine sample was diluted directly for analysis. The samples were all determined by LC-MS/MS,and the separation was performed on a Aglient ZORBAX SB-C18 column with mobile phase consisted of acetonitrile and 0.1% formic acid (gradient elution) at flow rate of 0.2 ml/min. Ion transition was determined ESI ion source under multiple ion reaction monitoring with quantitative pair m/z 436.4→253.2 and qualitative ion pair m/z 436.4→291.3 for valsartan,and quantitative pair m/z 423.4→207.1 and m/z 423.4→180.2 for internal standard losartan. RE-SULTS:The linear range of valsartan were 4-5 000 ng/ml in plasma and 20-50 000 ng/ml in urine;the limit of quantification were 4 ng/ml and 20 ng/ml;plasma extraction recovery of valsartan were 61.21%-70.30%. The variation coefficient of internal standard normalized matrix effect were 3.20% and 11.21%. The within-day and between-day RSDs were no more than 8.34%. CONCLU-SIONS:The method is proved to be rapid and sensitive,and suitable for the determination of valsartan in human plasma and urine and pharmacokinetics study.

Objective To explore the effect of particulate matter (PM) 2.5 on the expression of pigment epithelium-derived factor (PEDF) protein in bronchial epithelial cells (BEAS-2B). Methods Human BEAS-2B were subcultivated, followed by low, medium and high concentrations of PM2.5 (25μg/ml, 50μg/ml, 100μg/ml) stimulation for 24 hours. The expression of PEDF protein in supernatant was ana-lyzed by enzyme-linked immunosorbent assay (ELISA), and the expression in BEAS-2B cells was detected by Western blotting. Results Compared with the control group, the expression of PEDF protein in supernatant and BEAS-2B cells induced by PM2.5 (25 μg/ml) in-creased, but no significance was found (t=-0.730, t=-1.840, P>0.05), and the expression induced by PM2.5 with the concentrations of 50μg/ml and 100μg/ml significantly increased (t>5.798, P<0.05). Conclusion PM2.5 with the concentrations of 50μg/ml and 100μg/ml could increase the expression of PEDF protein in a concentration-dependent manner both in supernatant and BEAS-2B cells.

OBJECTIVETo analyze potential mutation of chloride channel 7(CLCN7) gene in a patient with autosomal dominant osteopetrosis II (ADO II).

METHODSGenomic DNA was extracted from peripheral blood samples from the patient and 100 healthy subjects. The DNA was used as template of polymerase chain reaction (PCR) to amplify the exons of CLCN7. Then the PCR products were sequenced to detect the mutation.

RESULTSA novel heterozygous deletional mutation (c.2460delA) was detected in exon 25 of the CLCN7 gene in the patient, which has resulted in substitution of Gly residue for Arg at position 784 of the CLCN7 protein and caused frame shift of the following 28 amino acids (Arg784GlyfsX29). The same mutation was not found in the healthy subjects.

CONCLUSIONThe ADO II in the patient probably results from a Arg784GlyfsX29 mutation in the CLCN7 gene.

Base Sequence ; Chloride Channels ; genetics ; DNA Mutational Analysis ; Genetic Predisposition to Disease ; genetics ; Humans ; Male ; Molecular Sequence Data ; Mutation ; Osteopetrosis ; diagnostic imaging ; genetics ; Radiography ; Young Adult