1.Salidroside Inhibits the Proliferation of Gastric Cancer Cells by Regulating the miR-1343-3p/SOX18 Signaling Axis
Zhendong ZHANG ; Xiaolan CAO ; Xinrui HOU ; Mingyuan CAO ; Yuxin DU ; Jie ZHANG ; Yanan SUN ; Xiaoping WANG
Journal of Sichuan University (Medical Sciences) 2025;56(4):1018-1026
Objective To investigate the molecular mechanism by which salidroside inhibits the proliferation of gastric cancer(GC)cells through upregulation of miR-1343-3p.Methods RNA databases were used to screen for mRNAs associated with tumor proliferation and with miR-1343-3p,and exhibiting significant changes in their expression levels after salidroside treatment of human GC cells.Gene matching and immunoprecipitation of RNA-binding proteins were conducted to analyze the association between miR-1343-3p and SOX18.Immunocytochemistry was performed to determine the localization of SOX18 protein.The effect of salidroside on the proliferation of human GC cells(MGC-803 and AGS)was determined by CCK-8 assay.Human GC cells were divided into a blank control group and low-and high-dose salidroside groups.The expression of miR-1343-3p and SOX18 mRNA was measured by real-time quantitative fluorescence PCR(qPCR).The protein expression of SOX18 was measured by Western blot.GC cells were co-transfected with miR-1343-3p mimic and miR-1343-3p inhibitor,respectively,via LipofectamineTM 2000 liposomes.The expression of miR-1343-3p and SOX18 mRNA was measured by qPCR,and the protein expression of SOX18 was measured by Western blot.Results Through bioinformatic analysis,SOX18 was identified as a downstream target of miR-1343-3p.Gene alignment confirmed the presence of specific binding sites between the two genes,and immunoprecipitation of RNA-binding proteins validated the targeting relationship between them(P<0.05).Immunocytochemistry demonstrated the nuclear localization of SOX18 protein.CCK-8 assay findings demonstrated that salidroside significantly inhibited the proliferation of GC cells in a time-and dose-dependent manner.Compared with the blank control group,salidroside-treated GC cells showed decreased expression of both SOX18 mRNA and protein(P<0.05)and an increased miR-1343-3p expression(P<0.05).Compared with the control group,GC cells in the miR-1343-3p mimic group exhibited increased expression of miR-1343-3p and decreased expression of SOX18 mRNA and protein.In contrast,GC cells in the miR-1343-3p inhibitor group showed decreased expression of miR-1343-3p and increased expression of SOX18 mRNA and protein(all P<0.05).Conclusion Salidroside may inhibit the proliferation of GC cells by regulating the miR-1343-3p/SOX18 signaling axis and these regulators may present new potential therapeutic targets or biomarkers for gastric cancer.
2.Pharmacodynamic and Mechanism of Anti-influenza Effect of Jiawei Xiangru Oral Liquid
Wei LI ; Qing DU ; Dan CHEN ; Xiaoqing WANG ; Xiaolan LI ; Ke LI
Chinese Journal of Modern Applied Pharmacy 2024;41(7):927-938
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".
3.Simultaneous Identification of Ziziphi Spinosae Semen and Its Adulterants by Multiplex Allele-specific PCR
Kefan LI ; Xiaoxiong SUO ; Xiaolan LI ; Chenhui DU ; Yan YAN ; Xiangping PEI
Chinese Journal of Experimental Traditional Medical Formulae 2023;29(2):141-148
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.
4.Variability in reference for serum metabolomics profiles among healthy Han people in different regions of China
Ye WANG ; Qianqian LIU ; Zhi ZHENG ; Feng LIU ; Jianwei DU ; Li PAN ; Xiaolan REN ; Hailing WANG ; Ze CUI ; Xia PENG ; Jingbo ZHAO ; Huijing HE ; Wei SUN ; Xiaoyan LIU ; Guangliang SHAN
Chinese Journal of Endocrinology and Metabolism 2022;38(6):475-482
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.
5.Advances in diagnosis and treatment of primary hemifacial spasm
Xiaolan LIU ; Shuangshuang XU ; Xindan DU ; Yajing CHEN ; Bing HUANG
Chinese Journal of General Practitioners 2022;21(12):1197-1201
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.
6.Research advances in repair of growth plate injury
Yangli XIE ; Qiaoyan TAN ; Fengtao LUO ; Can LI ; Junlan HUANG ; Xiaolan DU ; Lin CHEN
Chinese Journal of Orthopaedic Trauma 2020;22(1):88-92
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.
7. Performance of urea-mediated dissociation in reducing false-positive of 2019-nCoV IgM test
Qiang WANG ; Qin DU ; Bin GUO ; Yangliu GUO ; Li FANG ; Xiaolan GUO
Chinese Journal of Laboratory Medicine 2020;43(0):E014-E014
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.
8.Therapeutic effect of dipeptidyl peptidase-4 inhibitor on patients with type 2 diabetes mellitus with nonalco-holic fatty liver disease
Yeli ZHANG ; Leiqun LU ; Xiaoying MA ; Ying SHEN ; Ping GAO ; Xi GU ; Xiaolan DU ; Ling CHEN
The Journal of Practical Medicine 2017;33(13):2160-2164
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.
9.Determination of Valsartan in Human Plasma and Urine by LC-MS/MS and Its Pharmacokinetic Study
Shiyin FENG ; Xiaolan YONG ; Juan HUANG ; Xiaolin DU ; Nan LI ; Lantian WANG
China Pharmacy 2016;27(5):619-621
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.
10.Effects of PM2.5 on Expression of Pigment Epithelium-derived Factor in Bronchial Epithelial Cells
Chenghua ZHU ; Lijun SHI ; Lingfeng ZHANG ; Xiaolan JI ; Qiang DU
Chinese Journal of Rehabilitation Theory and Practice 2016;22(10):1151-1153
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.


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