2.The expression of DNA methyltransferase,demethylase and tumor-associated genes in human gastric cancer
Zhonghua CHENG ; Jingyuan FANG ; Li YANG
Chinese Journal of Digestion 2001;0(04):-
Objective To investigate the relationship of the expression of DNA methyltransferase,demethylase(mbd2) and tumor-associated genes in human gastric cancer. Methods Tissue samples of cancerous,para-cancerous and normal gastric mucosa were obtained surgically from 28 primary gastric cancer patients. The transcription level of DNA methyltransferase 1 (DNMT1),mbd2,methyl-CpG binding protein (MeCP2),p16 INK4A and c-myc were determined by using real-time RT-PCR and RT-PCR. The relationship between the expression of DNA methylation-associated genes and tumor-associated genes was analyzed. Results The mRNA level of DNMT1 was higher and the mRNA level of mbd2 gene was lower in cancerous tissue than that in normal tissue. The expression of c-myc instead of p16 INK4A and MeCP2 was increased in cancer tissues. The mRNA level of c-myc related negatively to mbd2 when gastric cancer developed. However,there was no any close relation between the transcription level of all above genes and tumor biological behavior in human gastric cancer. Conclusion This study indicates that MeCP2 but not DNMT1 may contribute to the regulation of tumor-associated genes expression in human gastric cancer.
3.Observations on the Therapeutic Effect of Filiform Fire Needling on Tennis Elbow
Lingxing OUYANG ; Xianmin GUO ; Xinkai FANG ; Yong REN ; Zhonghua GUAN
Shanghai Journal of Acupuncture and Moxibustion 2015;(2):161-163
Objective To investigate the clinical therapeutic effect of filiform fire needling on tennis elbow.Method Eighty patients with tennis elbow were randomly allocated to a filiform fire needling group of 30 cases, a block group of 20 cases and an electroacupuncture group of 30 cases. The treatment group received. Pain severity was scored using the VAS after one and two weeks and one month of treatment. The therapeutic effects were evaluated and compared after one month of treatment.Result The total efficacy rate was 93.3% in the filiform fire needling group, 60.0% in the block group and 50.0% in the electroacupuncture group; there was a statistically significant difference among the three groups (P<0.05). There was a statistically significant difference between the filiform fire needling and block groups (P<0.05) and between the filiform fire needling and electroacupuncture groups (P<0.05) but no significant difference between the block and electroacupuncture groups (P>0.05). The VAS score decreased significantly in the block group (P<0.05) but did not in the filiform fire needling and electroacupuncture groups (P>0.05) after one week of treatment compared with before. The VAS score decreased significantly in the filiform fire needling and block groups (P<0.05) but did not in the electroacupuncture group (P>0.05) after two weeks of treatment compared with before. There was no statistically significant posttreatment difference in the VAS score between the filiform fire needling and block groups (P>0.05) but there was such a difference between the filiform fire needling or block group and the electroacupuncture group (P<0.05). There was a statistically significant difference in the VAS score in the filiform fire needling and block groups (P<0.01) and also in the electroacupuncture group (P<0.05) between before and after one month of treatment. There was a statistically significant difference in the VAS score after one month of treatment between the filiform fire needling group and the block or electroacupuncture group (P<0.05) and between the block and electroacupuncture groups (P<0.05). The results of the study showed that filiform fire needling, local drug blockage and electroacupuncture all had clinically a better therapeutic effect on tennis elbow and a better improving effect on the symptoms in the patients. Filiform fire needling produced a marked therapeutic effect. Its effect was superior to those of block therapy and electroacupuncture.Conclusion Filiform fire needling is a definitely effective way to treat tennis elbow.
4.Aldosterone modulates the production of matrix metalloproteinase-2 and -9 via transforming growth factor β1 signaling pathway in podocytes
Zhan FANG ; Xifeng SUN ; Fangfang HE ; Zhonghua ZHU ; Chun ZHANG
Chinese Journal of Nephrology 2009;25(7):503-508
objective To assess the effect of aldosterone on the production of matrix metalloproteinase-2 (MMP-2), matrix metalloproteinase-9 (MMP-9)and collagen Ⅳ in culture supematants of podocytes and the possible molecular mechanisms involved in the influence of aldosterone on the synthesis and degradation of extracellular matrix produced by podocytes. Methods Podecytes were treated with aldosterone at the concentration of 10-11, 10-9, 10-7 mol/L respectively. Cultured podocytes were examined at 24, 48 and 72 hours respectively. Spironolactone, a receptor antagonist of aldosterone, was added to observe the blocking effect on aldosterone. An inhibitor of TGF-β1 receptor was used to determine whether the effect of aldosterone on podocytes were mediated through TGF-β1 system. The enzymatic activities of MMP-2 and MMP-9 were assayed by gehtin zymography. Collagen Ⅳ 0.5 chain and TGF-β1 proteins released into culture supematants were assessed by Western blot and ELISA analysis. The adhesion rate of podocytes was monitored by flow cytometry. Results Aldosterone increased the activities of MMP-2 and MMP-9 in a dose- and time-dependent manner (P<0.05). Aldosterone decreased the level of collagen Ⅳ or5 chain protein in culture supernatants (P<0.05). Meanwhile, the expression of TGF-β1 was also increased (P<0.05). Spironolactone completely abolished the above-mentioned changes(P< 0.05). Blockage of TGF-β1 signaling with SB431542 prevented the aldosterone-induced upregulation of MMP-2 and MMP-9 as well as the downregulation of the collagen Ⅳ α5 chain protein and the adhesion rate of podocytes (P<0.05). Conclusions Aldosterone increases the activities of MMP-2 and MMP-9 but decreases the expression of collagen Ⅳ α5 chain and the adhension rate of podocytes possibly via TGF-β1 signaling pathway. Such alterations may contribute to glomerular podocyte injury associated with the GBM abnormality caused by the imbalance between matrix synthesis and degradation.
5.The mechanism research of Rhubarb phenol and P38 inhibitor suppression on light aging of human skin HaCaT induced by UVB
Hui XIONG ; Liu YANG ; Jianmin LI ; Yeqiu WANG ; Zhonghua HU ; Fang GENG ; Ning ZHANG
International Journal of Traditional Chinese Medicine 2017;39(3):234-238
Objective To study the rhubarb phenol and P38 inhibitor suppression on light aging of human skin HaCaT induced by UVB. Methods The potential of cell proliferation of the different concentrations of rhubarb phenol and P38 inhibitor (SB203580) on human skin HaCaT was detected by MTT method. The cells was divided into the control group, the model group, the rhubarb phenol group, the SB203580 group by random number table method after the 24 h incubation. The 10-6 mol/L rhubarb phenol and 10-7 mol/L SB203580 were added to the rhubarb phenol group and SB203580 group for 24h, The competence of cultured cell proliferation which was irradiated with UVB of intensity of 0.61mW/cm2, mutiply time of 7 min and distance of 10 cm for 24 h except the control group; Western Blot method detected rhubarb phenol and P38 inhibitor of the influence of P38, P-P38, TNF-α, IL-6 protein. Results Compared with control group, the cell proliferation in UVB group significantly reduced (P<0.01); Compared with UVB group, the expression of the P-P38 (0.419 ± 0.029, 0.398 ± 0.015 vs. 0.497 ± 0.051), TNF-α (0.435 ± 0.025, 0.411 ± 0.021 vs. 0.509 ± 0.040) and IL-6 (0.457 ± 0.027, 0.432 ± 0.018 vs. 0.478 ± 0.036) in rhubarb phenol and P38 inhibitor group significantly reduced (P<0.01). Conclusions The rhubarb phenol and P38 inhibitor could significantly suppress HaCaT cells light aging, and its mechanism may be related with inhibiting P38 signaling pathways, and inhibiting the secretion of inflammatory cytokines.
6.Regulative effect of pinoresinol diglucoside on gene of ESF-1 cells collagen secretion
Xiaobo GAO ; Hongdan XU ; Yonghua QI ; Hui XIONG ; Zhonghua HU ; Haiyang LIU ; Fang GENG
International Journal of Traditional Chinese Medicine 2017;39(2):131-135
Objective To study the pinoresinol diglucoside (PDG) on gene regulation role of ESF-1 cells in collagen secretion, to reveal PDG repair mechanisms on scalded skin.Methods The cells cultured in vitro were divided into the control group, the estradiol group and the three different PDG doses groups. The concentration of the high, medium and low dose groups were 100, 10, 1μmol/L, and that of estradiol group were 10-3μmol/L. The activity of proliferation was detected by MTT. Then collagen type I (Col I), collagen typeⅢ (ColⅢ), tissue inhibitors of metalloproteinase 1 (TIMP-1), tissue inhibitors of metalloproteinase 2 (TIMP-2) and matrix metalloproteinase 1 (MMP-1) expression levels of mRNA after administration of cells were detected by RT-PCR.Results Compared with the control group, the proliferation of ESF-1 cells (0.559 ± 0.027, 0.552 ± 0.034vs. 0.489 ± 0.027,P<0.05) in the estradiol and medium-dose PDG was significantly higher. The expression level of mRNA of ColⅠ(0.958 ± 0.021, 0.929 ± 0.031, 0.916 ± 0.015vs. 0.844 ± 0.022), ColⅢ (0.783 ± 0.038, 0.918 ± 0.021, 0.855 ± 0.017vs. 0.678 ± 0.024), TIMP-1 (0.939 ± 0.025, 0.889 ± 0.036, 0.853 ± 0.015 vs. 0.780 ± 0.023), TIMP-2 (0.507 ± 0.024, 0.655 ± 0.037, 0.572 ± 0.025vs. 0.405 ± 0.062) in the estradiol, low-, medium-dose PDG groups were significantly higher than those in the control group (P<0.05 or P<0.01). Besides, the MMP-1 (0.343 ± 0.038, 0.407 ± 0.046, 0.435 ± 0.037vs.0.519 ± 0.041) mRNA expression level in the middle and low dose PDG groups significantly decrease (P<0.05 orP<0.01). Conclusions The PDG could enhance the activity of ESF-1 cell proliferation, increase the expression of related collagen and tissue inhibitor of metalloproteinases and inhibit that of matrix metalloproteinases to repair scalded skin.
7.Effect of levcromakalim on ET-1 induced proliferation and expression of protein kinase C in vascular smooth muscle cells
Wenxin HUANG ; Biwen MO ; Yaozhong YANG ; Chunsheng FANG ; Zhonghua XIA ; Dihua PAN ; Tongtong XU ; Xinling MO
Chinese Journal of Clinical Pharmacology and Therapeutics 2002;0(06):-
AIM:To observe the effect of levcromakalim(Lev) on endthelin-1(ET-1) induced proliferation and the expression of protein kinase C(PKC) in cultured rat vascular smooth muscle cells(VSMCs).METHODS: VSMCs were isolated from rat aorta and were cultured with different concentrations of Lev in vitro after stimulated to proliferation by ET-1(10~(-8)(mol?L~(-1))).Vitality of VSMCs was detected by MTT;DNA replication was evaluated by 3H-TdR method.Flow cytometry was used to analyze cell cycle and apoptosis.The expression of PKC? protein and mRNA were determined by Western blot and reverse transcription-polymerase chain reaction(RT-PCR).RESULTS:Vitality of VSMCs and 3H-TdR intermingle rate were decreased(P
8. Analysis of occupational chronic carbon disulfide poisoning: a study of 372 cases
Zhonghua FANG ; Rongming MIAO ; Haiyan SONG
Chinese Journal of Industrial Hygiene and Occupational Diseases 2018;36(3):202-203
Objective:
To investigate the clinical features of occupational chronic carbon disulfide(CS2) poisoning.
Methods:
A total of 372 patients with occupational chronic CS2 poisoning were selected from a chemical fiber factory, and their clinical features were summarized and analyzed.
Results:
Major clinical manifestations of the 372 patients with occupational chronic CS2 poisoning included sleep disorders, dizziness, headache, and numbness of limbs, and the detection rates of these manifestations were 84.7%, 84.4%, 79.8%, and 72.8%, respectively. Electroneuromyography showed peripheral nerve injuries.
Conclusion
Occupational chronic CS2 poisoning can affect the central and peripheral nervous system.
9.Effects of genistein on the mRNA expressions of collagen, matrix metalloproteinase and tissue inhibitor of matrix metalloproteinase in human embryonic skin fibroblasts
Hongdan XU ; Meng LI ; Xiaobo GAO ; Zhonghua HU ; Hui XIONG ; Zhigang WANG ; Fang GENG
International Journal of Traditional Chinese Medicine 2015;(7):617-620
Objective To investigate the effects of genistein on the mRNA expressions of collagen (Col), matrix metalloproteinase (MMP ) and tissue inhibitor of matrix metalloproteinase (TIMP) in human embryonic skin fibroblasts (CCC-ESF-1).MethodsThe cultured CCC-ESF-1cells were divided into a black control group, an estradiol group and genistein groups of different doses. The mRNA expressions of ColⅠ, ColⅢ , MMP-1, TIMP-1 and TIMP-2 were detected by RT-PCR.Results Compared with the black group, estradiol and medium dose of genistein (0.451 ± 0.037, 0.446 ± 0.047vs.0.385 ± 0.061, allP<0.05) could promote the proliferation of the CCC-ESF-1 cells, estradiol and medium dose of genistein could up-regulate the mRNA expressions of ColⅠ (0.960 ± 0.012, 0.929 ± 0.015vs.0.812 ± 0.014, allP<0.01), ColⅢ (0.892 ± 0.009, 0.824 ± 0.022vs.0.768 ± 0.025, allP<0.01), TIMP-1 (0.841 ± 0.023, 0.838 ± 0.053vs.0.751 ± 0.027, allP<0.01) and TIMP-2 (0.456 ± 0.017, 0.448 ± 0.036vs.0.381 ± 0.029, allP<0.01), and down-regulate MMP-1 mRNAexpression (0.398 ± 0.043, 0.402 ± 0.044vs.0.525 ± 0.006, allP<0.01).Conclusions Genistein could promote the proliferation of the CCC-ESF-1 cells, and that may be related with up-regulating the mRNA expressions of ColⅠ, ColⅢ , MMP-1, TIMP-1and down-regulating MMP-1 mRNA expression.
10.The expression features of hydrolase genes related to the sec retion of thyroid hormone of H22 hepatoma mice with different symptoms in early stage
Wenli LU ; Zhaoqin FANG ; Zhiqiang PAN ; Xiaomei LIU ; Chao LIANG ; Zhonghua WU ; Dongyuan GUAN ; Yuanyuan ZHANG
International Journal of Traditional Chinese Medicine 2014;(7):623-627
Objective To study the expression features of hydrolase genes related to the secretion of thyroid hormone of H22 hepatoma mice with different symptoms in early stage. Methods Firstly, The quantitative diagnosis and syndrome differentiation methods were used in H22 tumor-bearing mice in early stage, the expression profile of Tg and related hydrolase genes in poisonous pathogenic factors syndrome group (PPFS) and qi-deficiency syndromes (QDS) were got, and the major differential expression were selected. Secondly, the experiment was repeated and ELISA were used to detect T3 and T4 in serum, RT-PCR were applied to detect gene transcription level of genes including Tg, Ctsb, Ctsd, Ctsl, Napsa and Tpp1. Results ① Based on gene chip, the expression of Tg, Ctsb, Ctsd, Ctsl, Napsa and Tpp1were decreased in the first batch of experiment, the exactly ratio was Tg(0.77 in PPFS;0.84 in QDS), Ctsb(0.83 in PPFS, 0.91 in QDS), Ctsd(0.79 in PPFS;no notable change in QDS), Ctsl(no notable change in PPFS; 0.65 in QDS), Napsa(0.78 in PPFS; no notable change in QDS), and Tpp1 (0.75 in PPFS; no notable change in QDS), respectively. ② T3 and T4 downregulated in PPFS (the T3 value was 1.519±0.162ng/ml, T4 value was 2.194±0.305mg/dl) and in QDS (the T4 value is 4.366±0.727μg/dl) in early stage (P<0.01), especially in PPFS, which was in accordance with the change of Tg in both batches. ③the same trend happened in the validation of Tg(0.22 in PPFS;0.38 in QDS), Ctsb(0.31 in PPFS;0.55 in QDS), Ctsd(0.36 in PPFS;0.78 in QDS) and Napsa(0.24 in PPFS;0.59 in QDS) ,while ctsl(1.24 in PPFS;2.11 in QDS) and Tpp1 (2.85 in PPFS;0.85 in QDS)werethe opposite;even this, the total trend of the expression in QDS was still higher than that in PPFS. Conclusion All the results showed that the thyroid function of H22 hepatoma mice was inhibited in early stage especially in PPFS.