2.Gene Expression of Cytokine Pathway,Jak-Stat Signal Pathway and Neuroactive Ligand-receptor Pathway in Patients with Primary Sjgren′s Syndrome.
Fang WANG ; Yongjing CHENG ; Cibo HUANG ; Yingjuan CHEN ; Bei LAI
Journal of Medical Research 2006;0(02):-
Objective This study is to investigate cytokine pathway,Jak-Stat signal pathway,neuroactive ligand-receptor pathway gene expression pattern of peripheral blood mononuclear cell(PBMC) of primary sjgren syndrome patients.Methods The PBMC sample of 3 patients with sjgren syndrome and 3 healthy volunteers with consistent age were collected.The total RNAs was extracted from the PBMC samples,and reverse transcripted in vitro transcription(IVT),labeled with Cy5/Cy3 and hybridized on the gene chips.After scanning and data extraction with LuxScan 3.0,differentially expressed genes were analyzed with SAM method.The online tool of molecule analysis system(MAS) was used for biological knowledge mining.Results Statistical difference was calculated between the patient and control group in the following three pathways: cytokine pathway,Jak-Stat signal pathway,neuroactive ligand-receptor pathway.Among these,genes of IL-2RA,IL-10 were up-regulated and genes of PF4,GZMA were down-regulated.Conclusion Understanding of differently expressed genes should help us disclose the potential molecular mechanism underlying the development process of pathogenesis of primary Sjgren′s syndrome.And the research may provide new target therapy for SS.
3.Study on anti-pyretic and anti-inflammator effect of Shufeng Jiebiao decoction
Baojin CHEN ; Hong WANG ; Bangjiang FANG ; Hao CHEN ; Quan GUO ; Jiaojiao LU ; Bei WANG
International Journal of Traditional Chinese Medicine 2012;34(7):613-616
Objective To observe the effect of anti-pyretic and anti-inflammatory of Shufeng Jiebiao decoction.Methods Intraperitoneal injection of Lipopolysaccharides (LPS) was made to cause fever in rats and then to observe the anti-pyretic effect of Shufeng Jiebiao decoction.Intraperitoneal injection of glacial acetic acid was made to led inflammatory exudate in rats and then to observe the anti-inflammatory effect of Shufeng Jiebiao decoction.Smearing cylene in auricles was done to cause inflammatory swelling in rats and then to observe the effect of the alleviation of the inflammatory swelling of Shufeng Jiebiao decoction.Results ①The temperature of rats in the group of the aspirin and the Shufeng Jiebiao decoction were become lower at each time.The basic temperature of the model control group was (37.14±0.39) ℃,the temperature in the first hour was (40.31±0.34) ℃,the second hour was (40.44±0.44) ℃,the fourth hour was (40.24±0.34) ℃,the sixth hour was (40.05 ±0.44)℃,and the eighth hour was (39.85 ±0.37)℃.The basic temperature of the aspirin group was (37.13±0.33)℃,the temperature in the first hour was (38.74±0.42)℃,the second hour was (38.86±0.33) ℃,the fourth hour was (39.05±0.36)℃,the sixth hour was (38.74±0.37)℃,and the eighth hour was 38.64±0.24) ℃.The basic temperature of the Shufeng Jiebiao decoction group was (37.03±0.46) ℃,the temperature in the first hour was (39.02±0.49) ℃,the second hour was (38.82±0.49) ℃,the fourth hour was (38.63±0.46)℃,the sixth hour was (38.62±0.52)℃,and the eighth hour was (38.42±0.44)℃.The differences were statistical significance compared with the model control group (P<0.01),the onset of anti-pyretic of the Shufeng Jiebiao decoction group was slower than the aspirin group,but it had a longer lasting effect.Moreover,the rats' temperature decrease of the Shufeng Jiebiao decoction group in the fourth hour had a statistical significance compared with the aspirin group.(P<0.05).② After the intevention of aspirin and the Shufeng Jiebiao decoction,the optical density of evans blue:the model control group was (0.221 ±0.045),the aspirin group was (0.162±0.053),the Shufeng Jiebiao decoction group was (0.176±0.049),the permeability of the abdominal capillary of the rats reduced significantly (P<0.01).The intervention of the aspirin and the Shufeng Jiebiao decoction had almost no difference.③ After the intervention of the dexamethasone and the Shufeng Jiebiao decoction,the weight of the auricals:the model control group was (1.94±0.55)mg,dexamethasone group was (1.18±0.40)mg,Shufeng Jiebiao decoction group was (1.04±0.41)mg,showing the degree of the swelling of auricals decreased obviously (P<0.01).The intervention of the dexamethasone and the Shufeng Jiebiao decoction had almost no difference.Conclusion Shufeng Jiebiao Decoction had anti-pyretic and anti-inflammatory effects.
4.Gene expression profiling reveals signal transduction and abnormal expression of cytokine gene in patients with primary Sjogren syndrome
Fang WANG ; Yongjing CHENG ; Cibo HUANG ; Xiuqing HUANG ; Bei LAI ; Yingjuan CHEN
Chinese Journal of Microbiology and Immunology 2009;29(2):186-190
Objective To investigate signal transduction and cytokine gene expression pattern in peripheral blood mononuclear cell (PBMC) between primary Sjogren syndrome patients and healthy controls. Methods The PBMC of 3 patients with Sjogren syndrome and 3 healthy volunteers with consistent age were collected. The total RNA extracted from the PBMC was carried the reverse transcription and in vitro transcription (IVT), then labeled with Cy5/Cy3 and hybridized on the gene chips. After scanning and data extraction with LuxScan 3.0, differentially expressed genes were analyzed with SAM method. The online tools of molecule analysis system (MAS) was used for biological knowledge mining. The difference of signal transduction and cytokine gene expression of each patient and control were compared. Results Statistical difference was calculated between the patient and control group in the following four pathway: the chemokine-cytokine receptor interaction pathway[(FASLG(Fas ligand), CCL5, CCR3, IL-4R, CXCL10 (CXC chemokine ligand 10), TNFRSF19L (tumor necrosis factor supeffamily 19 ligand), CX3CR1 (CX3C chemokine receptor 1)], adhension molecule pathway [interleukin adhesion molecules-1 (ICAM-1)], Jak-STAT signal pathway [STAT4 (signal transducer and activation of transcription 4), CISH (chromogenic in situ hybridization), IL-4R], Toll-like receptor signal pathway(CCL5, CXCL10). Among these, 4 genes were up-regulated and 6 genes were down-regulated. Conclusion Understanding of differently expressed genes should help us disclose the potential molecular mechanism underlying the development process of pathogenesis of primary Sjogren syndrome.
5.Molecular mechanism of indirubin-3'-monoxime and Matrine in the reversal of paclitaxel resistance in NCI-H520/TAX25 cell line.
Su-xia LUO ; Wen-ying DENG ; Xin-feng WANG ; Hui-fang LÜ ; Li-li HAN ; Bei-bei CHEN ; Xiao-bing CHEN ; Ning LI
Chinese Medical Journal 2013;126(5):925-929
BACKGROUNDMultidrug resistance (MDR) is a main reason for paclitaxel (TAX) treatment failure. Indirubin-3'-monoxime (IRO) and Matrine are traditional Chinese medicines, which may reverse the resistance of tumor cells to some chemotherapy drugs, but the relationship between paclitaxel resistance and Matrine is still unclear. The aim of this study was to explore the potential molecular mechanism of IRO and Matrine in reversal of TAX resistance.
METHODSIn this study, MTT assay was used to measure the non-cytotoxic dosage of IRO and Matrine on NCI-H520/TAX25 cells and determine the reversal extent of TAX resistance under non-toxic doses. In addition, RT-PCR and Western blotting were used to evaluate the mRNA expression and the protein level of survivin, Oct-4, and Sox-2 in NCI-H520/TAX25 cells using semi-quantitative methods.
RESULTSThere was no obvious inhibition on sensitive cell strains and drug-resistant strains, when the final concentration was at lest 4 µmol/L for IRO and 100 µmol/L for Matrine. So 4 µmol/L of IRO and 100 µmol/L of Matrine were considered as the reversal dosage. When 4 µmol/L of IRO or 100 µmol/L of Matrine were used together with TAX, the sensitivity to TAX increased evidently in NCI-H520/TAX2 cells; the reversal rate of IRO and Matrine was about 1.92 (43.56/22.6 nmol/L) and 1.74 (43.56/25.0 nmol/L), respectively. The mRNA expression and the protein level of survivin, Oct-4, and Sox-2 in NCI-H520/TAX25 decreased significantly (P < 0.05) after addition of IRO or Matrine in TAX treatment, compared to that of TAX treatment alone.
CONCLUSIONThe decrease in both mRNA expression and protein level of survivin, Oct-4, and Sox-2 might be the molecular mechanism, by which IRO and Matrine mediate the reversal of TAX resistance.
Alkaloids ; pharmacology ; Blotting, Western ; Cell Line, Tumor ; Drug Resistance, Neoplasm ; Humans ; Indoles ; pharmacology ; Inhibitor of Apoptosis Proteins ; genetics ; metabolism ; Octamer Transcription Factor-3 ; genetics ; metabolism ; Oximes ; pharmacology ; Paclitaxel ; pharmacology ; Quinolizines ; pharmacology ; SOXB1 Transcription Factors ; genetics ; metabolism
6.Effects of 5-Aza-dC on 5-Fu chemosensitivity by modulating TIP30 gene expression in human colorectal cancer cells.
Xiao-bing CHEN ; Bei-bei CHEN ; Jian LI ; Xin-feng WANG ; Yi-jie MA ; Su-xia LUO ; Hui-fang LÜ
Chinese Journal of Oncology 2013;35(11):824-827
OBJECTIVETo investigate the effect of 5-Aza-2'-deoxycytidine (5-Aza-dC) on TIP30 gene expression and the relationship between TIP30 expression and the sensitivity to 5-fluouracil (5-Fu) in colorectal cancer cells.
METHODSThe methylation profile of TIP30 gene in HCT116 colorectal cancer cells was determined by methylation-specific PCR. The levels of TIP30 mRNA and protein were determined by RT-PCR and Western blot after the 5-Aza-dC treatment. MTT assay was used to detect the chemosensitivity of HCT116 cells to 5-Fu.
RESULTSTIP30 gene displayed complete DNA methylation in the HCT116 cells without 5-Aza-dC pretreatment. After the 5-Aza-dC treatment for 3 days, only demethylating PCR amplification product was detected and TIP30 gene showed DNA demethylation. With the prolongation of the time of removal of 5-Aza-dC treatment, methylated and demethylated PCR amplification products were observed and TIP30 gene displayed both DNA methylation and DNA demethylation in the colorectal cancer cells. At the day 10 after removal of 5-Aza-dC, methylating PCR amplification product appeared and TIP30 gene showed DNA methylation. No expressions of TIP30 mRNA and protein were detected in the HCT116 cells untreated with 5-Aza-dC. After the treatment of 5-Aza-dC for 3 d and then removed the 5-Aza-dC, the expressions of TIP30 mRNA and protein were increased obviously. With the prolonged time after 5-Aza-dC removal, the expressions of TIP30 mRNA and protein decreased and reached the lowest level on day 10. The IC50 values of 5-Fu were 41.62, 33.17 and 4.96 µg/ml in the HCT116 cells pretreated with 5-Aza-dC, d0 and d10 with the drug removal after drug treatment for 3 d, respectively.
CONCLUSIONSThe results of this study show that the expression of TIP30 gene may be associated with its DNA methylation status and may affect the sensitivity of colorectal cancer cells to 5-Fu.
Acetyltransferases ; genetics ; metabolism ; Antimetabolites, Antineoplastic ; pharmacology ; Azacitidine ; analogs & derivatives ; pharmacology ; Cell Proliferation ; drug effects ; CpG Islands ; genetics ; DNA Methylation ; drug effects ; Drug Resistance, Neoplasm ; drug effects ; Fluorouracil ; pharmacology ; Gene Expression Regulation, Neoplastic ; HCT116 Cells ; Humans ; Inhibitory Concentration 50 ; RNA, Messenger ; metabolism ; Transcription Factors ; genetics ; metabolism
7.Potential effect of tumor necrosis factor-alpha and tumor necrosis factor receptor II gene polymorphisms on the pathogenesis of silicosis.
Lin LI ; Chen YU ; Fang QI ; De-hong LI ; Bei-chen LIU
Chinese Journal of Industrial Hygiene and Occupational Diseases 2004;22(5):323-326
OBJECTIVETo approach the role of tumor necrosis factor-alpha (TNF-alpha) and tumor necrosis factor receptor II (TNFR II) gene polymorphisms in genetic susceptibility to silicosis and their interaction with silica-dust exposure.
METHODSTwo hundred and fifty-nine cases of silicosis and three hundred and forty-one silica-dust exposure workers (control) were selected, and the cases of silicosis were divided into three subgroups based on the various stages of I, II and III. Exposure history, pneumoconiosis history and past history of each subject were obtained by questionnaire. 3 ml peripheral venous blood was drawn from each subject. Using polymerase chain reaction-restriction fragment length polymorphisms (PCR-RFLP) techniques, TNF-alpha and TNFRII gene polymorphisms were analyzed.
RESULTSIn both group matching and 1:1 paired matching, there was no significant difference between cases of silicosis and workers in control in distribution frequencies of G/A + A/A (TNF-alpha-308) and T/G + G/G (TNFRII 196) genotypes. The risk of silicosis in those with G/A + A/A genotype was 6.74-fold higher than G/G genotype (OR = 6.74, 95% CI: 1.01 approximately 44.99) in subjects whose exposure time was less than 15 years.
CONCLUSIONTNF-alpha and TNFR II gene polymorphisms did not play an important role in susceptibility to silicosis of Han race. There was interaction between polymorphism of TNF-alpha gene promoter and exposure time in the occurrence of silicosis. The risk of silicosis in those with G/A + A/A genotype was significantly higher than G/G genotype in low accumulative exposure.
Adult ; Aged ; Aged, 80 and over ; Genetic Predisposition to Disease ; Genotype ; Humans ; Male ; Middle Aged ; Polymerase Chain Reaction ; Polymorphism, Restriction Fragment Length ; Receptors, Tumor Necrosis Factor, Type II ; genetics ; Silicosis ; etiology ; genetics ; Tumor Necrosis Factor-alpha ; genetics
8.TH gene mutation in Chinese patients with autosomal recessive dopa-responsive dystonia.
Wei LIU ; Bei-sha TANG ; Gui-fang CAO ; Tao CHEN ; Hai-yan LI
Chinese Journal of Medical Genetics 2004;21(5):452-454
OBJECTIVETo explore the mutation of tyrosine hydroxylase(TH) gene in Chinese patients with autosomal recessive(AR) dopa-responsive dystonia(DRD) and to lay a solid basis for gene diagnosis of AR-DRD in China.
METHODSMutation analysis of TH gene was performed in 5 probands with AR-DRD and 2 sporadic patients with DRD by use of polymerase chain reaction-single strand conformation polymorphism (PCR-SSCP) combining DNA direct sequencing.
RESULTSThe PCR-SSCP analysis and DNA direct sequencing following PCR revealed no mutation in all the 14 exons of TH gene.
CONCLUSIONThe mutation rate of TH gene in Chinese patients with AR-DRD is low, hence suggesting the genetic heterogeneity and a new locus for AR-DRD.
Asian Continental Ancestry Group ; genetics ; China ; Dystonia ; ethnology ; genetics ; Female ; Genes, Recessive ; genetics ; Humans ; Male ; Mutation ; Polymerase Chain Reaction ; Polymorphism, Single-Stranded Conformational ; Sequence Analysis, DNA ; Tyrosine 3-Monooxygenase ; genetics
9.Multi-detector-row CT findings of ACTH-independent macronodular adrenal hyperplasia
Qi SONG ; Weiwei ZHOU ; Tingwei SU ; Lianjun DU ; Bei DING ; Huan HANG ; Zilai PAN ; Kemin CHEN ; Fuhua YAN ; Weiqing WANG ; Guang NING ; Wenqiang FANG
Chinese Journal of Endocrinology and Metabolism 2012;28(4):335-339
ObjectiveTo explore the characteristics multi-detector-row computed tomography (MDCT)findings of ACTH-independent macronodular adrenal hyperplasia ( AIMAH ).Methods The un-enhanced and contrast-enhanced MDCT features in 24 patients ( 14 males and 10 females) with clinically confirmed AIMAH were retrospectively assessed for the morphology and enhancement patterns.ResultsThe adrenal glands were involved bilaterally in all of the 24 cases( 100% ).24 patients had massively enlarged multinodular adrenal glands.Nodules were( 1.79 ± 1.02) cm (0.50 ~ 3.85 cm),which usually distorted and completely obscured the normal adrenal glands.The enlarged adrenal glands were still retained the adreniform contour,showed characteristic ginger-like.22 of the 24 ( 91.7 % ) hyperplastic nodular glands demonstrated mild homogeneous enhancement.Calcification was revealed in 1 adrenals ( 1/24,4.2% ).Conclusion MDCT reveals the characteristic morphology and CT attenuation in AIMAH.Combined with its clinical presentation and biochemical findings,AIMAH is able to be diagnosed with high specificity and accuracy on MDCT.