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Palmitate induces fat accumulation by activating C/EBPβ-mediated G0S2 expression in HepG2 cells

World Journal of Gastroenterology 2017年第43期23卷 7705-7715页

作者： Nai-Qian Zhao Xiao-Yan Li Li Wang Zi-Ling Feng Xi-Fen Li Yan-Fang Wen jin-xiang han Department of Gerontology the Second Hospital of Shanxi Medical University Department of Infectious Diseases the First People’s Hospital of Jinzhong

AIM To determine the role of G0/G1 switch gene 2(G0 S2) and its transcriptional regulation in palmitate-induced hepatic lipid *** Hep G2 cells were treated with palmitate,or palmitate in combination with CCAAT/enhancer binding protein(C/EBP)β si RNA or G0 S2 si RNA. The m RNA expression of C/EBPβ,peroxisome proliferator-activated receptor(PPAR)γ and PPARγ target genes(G0 S2,GPR81,GPR109 A and Adipoq) was examined by q PCR. The protein expression of C/EBPβ,PPARγ,and G0 S2 was determined by Western blotting. Lipid accumulation was detected with Oil Red O staining and quantified by absorbance value of the extracted Oil Red O dye. Lipolysis was evaluated by measuring the amount of glycerol released into the medium. RESULTS Palmitate caused a dose-dependent increase in lipid accumulation and a dose-dependent decrease in lipolysis in Hep G2 cells. In addition,palmitate increased the m RNA expression of C/EBPβ,PPARγ,and PPARγ target genes(G0 S2,GPR81,GPR109 A,and Adipoq) and the protein expression of C/EBPβ,PPARγ,and G0 S2 in a dose-dependent manner. Knockdown of C/EBPβ decreased palmitate-induced PPARγ and its target genes(G0 S2,GPR81,GPR109 A,and Adipoq) m RNA expression and palmitate-induced PPARγ and G0 S2 protein expression in Hep G2 cells. Knockdown of C/EBPβ also attenuated lipid accumulation and augmented lipolysis in palmitate-treated Hep G2 cells. G0 S2 knockdown attenuated lipid accumulation and augmented lipolysis,while G0 S2 knockdown had no effects on the m RNA expression of C/EBPβ,PPARγ,and PPARγ target genes(GPR81,GPR109 A and Adipoq) in palmitate-treated Hep G2 cells. CONCLUSION Palmitate can induce lipid accumulation in Hep G2 cells by activating C/EBPβ-mediated G0 S2 expression.

关键词： Obesity Nonalcoholic fatty liver disease Saturated fatty acid G0/G1 switch gene 2 CCAAT/enhancer binding protein β Adipogenesis Lipolysis Proliferator-activated receptor γ

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Association Between Dentin Matrix Protein 1 （rs10019009） Polymorphism and Ankylosing Spondylitis in a Chinese han Population from Shandong Province

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Chinese Medical Journal 2016年第6期129卷 657-664页

作者： Jian-Min Liu Ya-Zhou Cui Geng-Lin Zhang Xiao-Yan Zhou jing-xiang Pang Xue-Zheng Wang jin-xiang han Allergy and Clinical Immunology Research Centre First Affiliated Hospital of Liaoning Medical University Jinzhou Liaoning 121001 China National Laborat'ory for Bio-Drugs of Mini'stry of Health Provincial Laboratory fo'r Modern Medicine and Technology of Shanclong Research Center for Medicinal Biotechnology Shandong Academy of Medical Sciences Jinan Shandong 250062 China

Background： Ankylosing spondylitis （AS） is the most common rheumatic condition that is slowly progressive and predominantly affects adolescents. Pathological bone formation associated with AS is an important cause of disability. The aim of the study was to investigate the possible involvement of the genes related to endochondral ossification and ectopia ossification in genetic susceptibility to AS in a Chinese han population. Methods： Sixty-eight single nucleotide polymorphisms （SNPs） from 13 genes were genotyped in discovery cohorts including 300 AS patients and 180 healthy controls. The rs10019009 in dentin matrix protein 1 （DMP1） gene shown as association with AS after multiple testing corrections in discovery cohorts was replicated in a validation independent cohort of 620 AS patients and 683 healthy controls. The rs 10019009 was assessed with bioin fomlatics including phylogenetic context, F-SNP and FastSNP functional predictions, secondary structure prediction, and molecular modeling. We performed a functional analysis of rs10019009 via reverse transcription-polymerase chain reaction, alkaline phosphatase （ALP） activity in human osteosarcoma U2OS cells. Results： Interestingly, the SNP rs10019009 was associated with AS in both the discovery cohort （P = 0.0012） and validation cohort （P - 0.0349）, as well as overall （P = 0.0004） in genetic case-control association analysis. After a multivariate logistic regression analysis, the effect of this genetic variant was observed to be independent of linkage disequilibrium. Via bioinformatics analysis, it was found that the amino acid change of the rs 10019009 led to changes of SNP function, secondary structure, tertiary confomlation, and splice mode. Finally, functional analysis ofrsl0019009 in U2OS cells demonstrated that the risk T allele of the rsl0019009 increased enzymatic activity of ALP, compared to that of the nonrisk allele （P = 0.0080）. Conclusions： These results suggested that the DMP1 gene seems to be involve

关键词： Alkaline Phosphatase Ankylosing Spondylitis Dentin Matrix Protein I Ectopic Mineralization Polymorphisms

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Rapid quantification of hepatitis B virus DNA by real-time PCR using efficient TaqMan probe and extraction of virus DNA

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World Journal of Gastroenterology 2006年第45期12卷 7365-7370页

作者： Yan-Qin Lu jin-xiang han Peng Qi Wei Xu Yan-Hui Zu Bo Zhu School of Medicine Shandong University Jinan 250012 China Shandong Medicinal Biotechnology Center Shandong Academy of Medical Science Key laboratory of Ministry of Health for Biotech-Drug Jinan 250062 Shandong Province China Shandong Medicinal Biotechnology Center Shandong Academy of Medical Sciences Key Laboratory of Ministry of Health for Biotech-Drugs Jinan 250062 Shandong Province China jinan Infectious Disease Hospital Jinan 250021 Shandong Province China

AIM： To rapidly quantify hepatitis B virus （HBV） DNA by real-time PCR using efficient TaqMan probe and extraction methods of virus DNA. METHODS： Three standards were prepared by cloning PCR products which targeted S, C and X region of HBV genome into pGEM-T vector respectively. A pair of primers and matched TaqMan probe were selected by comparing the copy number and the Ct values of HBV serum samples derived from the three different standard curves using certain serum DNA. Then the efficiency of six HBV DNA extraction methods including guanidinium isothiocyanate, proteinase K, NaI, NaOH lysis, alkaline lysis and simple boiling was analyzed in sample A, B and C by real-time PCR. Meanwhile, 8 clinical HBV serum samples were quantified. RESULTS： The copy number of the same HBV serum sample originated from the standard curve of S, C and X regions was 5.7 × 10^4/mL, 6.3 × 10^2/mL and 1.6 × 10^3/ mL respectively. The relative Ct value was 26.6, 31.8 and 29.5 respectively. Therefore, primers and matched probe from S region were chosen for further optimization of six extraction methods. The copy number of HBV serum samples A, B and C was 3.49 × 10^9/mL, 2.08 × 10^6/mL and 4.40 × 10^7/mL respectively, the relative Ct value was 19.9, 30 and 26.2 in the method of NaOH lysis, which was the efficientest among six methods. Simple boiling showed a slightly lower efficiency than NaOH lysis. Guanidinium isothiocyanate, proteinase K and NaI displayed that the copy number of HBV serum sample A, B and C was around 10^S/mL, meanwhile the Ct value was about 30. Alkaline failed to quantify the copy number of three HBV serum samples. Standard deviation （SD） and coefficient variation （CV） were very low in all 8 clinical HBV serum samples, showing that quantification of HBV DNA in triplicate was reliable and accurate. CONCLUSION： Real-time PCR based on optimized primers and TaqMan probe from S region in combination with NaOH lysis is a simple, rapid and accurate method for quantificati

关键词： Hepatitis B virus Serum DNA Real-time PCR Extraction method

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Effect of NS398 on metastasis-associated gene expression in a human colon cancer cell line

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World Journal of Gastroenterology 2005年第28期11卷 4337-4343页

作者： Xue-Qin Gao jin-xiang han Hai-Yan Huang Bao Song, Bo Zhu Chang-Zheng Song Key Laboratory of Ministry of Public Health for Biotech-Drug Shandong Medicinal and Biotechnology Center Shandong Academy of Medical Sciences Jinan 250062Shandong Province China Shandong University School of Medicine Jinan 250062 Shandong Province China

AIM： To investigate the effect of NS398 on the metastasis-associated gene expression in LoVo colorectal cancer cells.METHODS： LoVo cells were treated with NS398 at the concentration of 100 IJmol/L for 24 and 48 h respectively. Total RNA was extracted with TRIzol reagents and reverse transcribed with Superscript Ⅱ and hybridized with cDNA mlcroarray （containing oncogenes, tumor suppressor genes, signal transduction molecules, adhesive molecules, growth factors, and ESTs） fabricated in our laboratory. After normalization, the ratio of gene expression of NS398 treated to untreated LoVo cells was either 2-fold up or 0.5-fold down was defined as the differentially expressed genes. Semi-quantitative RT-PCR was used to validate the microarray results.RESULTS： Among the 447 metastasis-associated genes, 9 genes were upregulated and 8 genes were downregulated in LoVo cells treated with NS398 for 24 h compared to untreated cells. While 31 genes were upregulated and 14 genes were downregulated in LoVo cells treated with NS398 for 48 h. IGFBP-5, PAI-2, JUN,REL, BRCA1, and BRCA2 might be the new targets of NS398 in treatment of colorectal cancer.CONCLUSION： NS398 might exert its anti-metastasis effect on colorectal cancer by affecting several metastasis-associated gene expression.

关键词： NS398 Colorectal cancer gene expression Metastasis cDNA microarray

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A sensitive method to detect the hepatitis B virus mutations by using solid phase PCR on oligonucleotide array

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Journal of Microbiology and Immunology 2006年第4期4卷 265-271页

作者： YAN QIN LU jin xiang han ZHONG LIN SHEN CHUAN XI WANG Shandong Medicinal Biotechnology Research Centre Shandong Academy of Medical Sciences Key Laboratory for Biotech-Drugs Ministry of Health Ji-nan P. R. China Department of Detection Hospital of Yankuang Group Jining P. R. China

A sensitive method based on solid phase PCR on oligonucleotide array was established to detect two point mutations 1896G-A and 1901G-A in hepatitis B virus (HBV) DNA, in which 6 probes including these two point mutations were immobilized on modified glass slides through 5' terminal linker, while the 3' terminal was made to be free. The mutated loci were designed to locate on the last nucleotides of 3' terminal respectively, and the positive control probes lacked the last nucleotide of 3' terminal in comparison with the probes used for detection. Probes fixed on oligonucleotide array were also the solid phase amplification primers. One pair of liquid primers was used to amplify the short template product from whole HBV DNA. Using target DNA as template, the solid primers were extended under the action of Taq DNA polymerase and incorporated with Cy-3dCTP as marker. During the thermal cycling reaction, probes served as solid phase amplification primers and amplification products bound to the oligonucleotide array, which could be visualized by incorporation with fluorescent dyes. After amplification, the oligonucleotide array was washed, performed with laser scanning, and then used for quantitative analysis to obtain the information for mutations. The hybridization results were compared with DNA sequencing. It was demonstrated that in case of sample A, the ratios of fluorescence intensities in wide type and in the mutated types of 1896G-A and 1901G-A mutations in HBV were 3.81:1 and 1:3.79 respectively, while, in case of sample B, those were 1:2.89 and 1:3.03 respectively, indicating the presence of point mutations in these two loci. These results correlated to those obtained from DNA sequencing analysis in which the fluorescence intensity ratios in wide type and in the mutated types of 1996G-A and 1901D-A mutations in HBV were 1.26:1 and 1.67:1 respectively. From the above observations, it is evident that the method using solid phase PCR based on oligonucleotide array

关键词： Oligonucleotide array Solid phase PCR Hepatitis B virus Point mutation

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Identifi cation of the differential expressive tumor associated genes in rectal cancers by cDNA microarray

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World Journal of Gastroenterology 2007年第3期13卷 341-348页

作者： Xue-Qin Gao jin-xiang han Zhong-Fa Xu Wei-Dong Zhang Hua-Ning Zhang Hai-Yan Huang Key Laboratory for Biotcch-Drug Ministry of Health Key Laboratory of Modem Drugs and Technology of Shandong Province Shandong Medical and Biotechnology Center Shandong Academy of Medical Sciences Jinan 250062 Shandong Province China Department of Surgery Shandong Tumor Hospital Shandong Academy of Medical Sciences Jinan 250117 Shandong Province China Department of Experimental Pathology Institute of Basic Medicine Shandong Academy of Medical Sciences Jinan 250062 Shandong Province China

AIM： To identify tumor associated genes of rectal cancer and to probe the application possibility of gene expression profiles for the classification of tumors.METHODS： Rectal cancer tissues and their paired normal mucosa were obtained from patients undergoing surgical resection of rectal cancer. Total RNA was extracted using Trizol reagents. First strand cDNA synthesis was indirectly labeled with aminoallyl-dUTP and coupled with Cy3 or Cy5 dye NHS mono-functional ester. After normalization to total spots, the genes which background subtracted intensity did not exceed 2 SD above the mean blank were excluded. The data were then sorted to obtain genes differentially expressed by ≥ 2 fold up or down in at least 5 of the 21 patients.RESULTS： In the 21 rectal cancer patients, 23 genes were up-regulated in at least 5 samples and 15 genes were down-regulated in at least 5 patients. Hierachical cluster analysis classified the patients into two groups according to the clinicopathological stage, with one group being all above stage Ⅱ and one group all below stage Ⅱ.CONCLUSION： The up-regulated genes and downregulated genes may be molecular markers of rectal cancer. The expression profiles can be used for classification of rectal cancer.

关键词： Rectal cancer Tumor associated genes cDNA microarray Differential expression genes

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Inhibition of Yuyihe Powder on Tumor Growth in Mice Models of Sarcoma 180

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Chinese Herbal Medicines 2017年第2期9卷 183-187页

作者： Jun-ling Li Shi-li Wang jin-xiang han Hai Xu Xiao-bin Xu Jun-ying Xue Key Laboratory of Ministry of Public Health for Biotech-Drug Shandong Medicinal Biotechnology CenterShandong Academy of Medical Sciences State Key Laboratory of Heavy Oil Processing and Centre for Bioengineering and Biotechnology China University of Petroleum (East China)

Objective To explore the antitumor effect of Yuyihe Powder（Yu Yi He San,YYHS） and its antitumor *** After treatment,tumor weight,immune apparatus weight,the life span of transplanted animals,spleen lymphocyte proliferation assays,and IL-2 concentration in mouse serum were recorded or *** YYHS showed strong antitumor *** with control group,mid-dose YYHS（1.0g/kg） could inhibit the tumor growth,prolong the life span of S180-bearing mice to some extent,significantly increase the thymic and splenic indices of S180 mice,and strongly promote the secretion of IL-2 in blood;The inhibitory rate on tumor growth and life prolongation rate were 37.1%and 38.37%,*** YYHS could not only significantly inhibit the growth of S180 cells,but also markedly prolong the survival time of S180 bearing *** mechanism of antitumor effect could obviously enhance immunologic function of the S180 bearing mice to inhibit the growth of S180 cells.

关键词： antitumor effect mice model Sarcoma 180 traditional Chinese medicine recipe Yuyihe Powder

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Simultaneous detection of HBV and HCV by multiplex PCR normalization

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World Journal of Gastroenterology 2004年第16期10卷 2439-2443页

作者： Ning Wang Xue-Qin Gao jin-xiang han Shandong Medicinal Biotechnological Center,Shandong Academy of Medical Sciences Key Laboratory for Biotechdrugs,Ministry of Public Health,jinan 250062,Shandong Province,China Shandong Medicinal Biotechnological Center Shandong Academy of Medical Sciences Key Laboratory for Biotechdrugs Ministry of Public Health Jinan 250062 Shandong Province China

AIM: To design and establish a method of multiplex PCR normalization for simultaneously detecting HBV and ***: Two pairs of primers with a 20 bp joint sequence were used to amplify the target genes of HBV and HCV by two rounds of amplification. After the two rounds of amplification all the products had the joint sequence. Then the joint sequence was used as primers to finish the last amplification. Finally multiplex PCR was normalized to a single PCR system to eliminate multiplex factor interference. Four kinds of nucleic acid extraction methods were compared and screened. A multiplex PCR normalization method was established and optimized by orthogonal design of 6 key factors. Then twenty serum samples were detected to evaluate the validity and authenticity of this ***: The sensitivity, specificity, diagnostic index and efficiency were 83.3%, 70%, 153.3% and 72.2%,respectively for both HBsAg and anti-HCV positive patients,and were 78.6%, 80%, 258.6% and 79.2%, respectively for HBsAg positive patients, and were 75%, 90%, 165% and 83.3%, respectively for anti-HCV positive ***: The multiplex PCR normalization method shows a broad prospect in simultaneous amplification of multiple genes of different sources. It is practical, correct and authentic, and can be used to prevent and control HBV and HCV.

关键词： HBV HCV 多元PCR 基因型肝炎

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Effects of aspirin on metastasis-associated gene expression detected by cDNA microarray

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Acta Pharmacologica Sinica 2004年第10期 87-93页

作者： Xue-qin GAO, jin-xiang han, Hai-yan HUANG, Shi YAN, Chang-zheng SONG, Hai-nan HUANGKey Laboratory of Ministry of Health for Biotech-Drug, Shandong Medicinal Biotechnology Center, Shandong Academy of Medical Sciences, Ji-nan 250062, ChinaNow in Medical College of Shandong University, Ji-nan 250012, China.

AIM: To investigate the effect of aspirin on the metastasis-associated gene expression in 3AO ovarian cancer cells. METHODS: 3AO cells were treated with aspirin at the concentration of 1.2 mmol/L for 16 and 48 h, respectively. The total RNA was extracted with Trizol reagents and reverse transcribed with Superscript Ⅱand hybridized with cDNA microarray (containing oncogenes, tumor suppressor genes, signal transduction pathway molecules , adhesive molecules, growth factors and ESTs) fabricated in our lab. After normalization, the ratio of gene expression of aspirin treated to untreated 3AO cells being either 2 fold up higher or 0.5 fold down (lower) were defined as differential expression. Semi-quantitative RT-PCR was used to validate the microarray results. RESULTS: Among the 447 metastasis-associated genes, 4 genes were up-regulated and 14 genes were down-regulated in 3AO cells treated with aspirin for 16 h compared with untreated cells. While 24 genes were up-regulated and 10 genes were down-regulated

关键词： aspirin ovarian neoplasms gene expression neoplasm metastasis cDNA microarray

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FK506 contributes to peripheral nerve regeneration by inhibiting neuroinflammatory responses and promoting neuron survival

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Neural Regeneration Research 2025年第7期20卷 2108-2115页

作者： Yuhui Kou Zongxue jin Yusong Yuan Bo Ma Wenyong Xie Na han Department of Trauma and Orthopedics Peking University People’s HospitalBeijingChina National Center for Trauma Medicine Peking University People’s HospitalBeijingChina Key Laboratory of Trauma and Neural Regeneration(Peking University) Ministry of EducationBeijingChina Trauma Medicine Center Peking University People’s HospitalBeijingChina Department of Orthopedics China-Japan Friendship HospitalBeijingChina

FK506(Tacrolimus)is a systemic immunosuppressant approved by the *** and Drug ***506 has been shown to promote peripheral nerve regeneration,however,its precise mechanism of action and its pathways remain *** this study,we established a rat model of sciatic nerve injury and found that FK506 improved the morphology of the injured sciatic nerve,increased the numbers of motor and sensory neurons,reduced inflammatory responses,markedly improved the conduction function of the injured nerve,and promoted motor function *** findings suggest that FK506 promotes peripheral nerve structure recovery and functional regeneration by reducing the intensity of inflammation after neuronal injury and increasing the number of surviving neurons.

关键词： FK506 inflammation motor neurons nerve regeneration neuron peripheral nerve injury sensory neurons

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