3.Tumor necrosis factor-αupregulates MMP9 expression through site-specific DNA demethylation
Li LING ; Meng REN ; Feng LI ; Chuan YANG ; Li YAN
Chinese Journal of Endocrinology and Metabolism 2016;32(8):685-690
Objective To determine the involvement of DNA demethylation in tumor necrosis factor-α(TNF-α)-induced matrix metalloproteinase 9 ( MMP9) expression in human epidermal keratinocytes. Methods Real-time RT-PCR, Western blot, and enzyme-linked immuno sorbent assay (ELISA) were performed to determine the mRNA and protein levels of MMP9 after human keratinocyte cell line (HaCaT) cells were treated with 10 ng/ ml TNF-α or 2. 5 μmol/ L DAC/ 300 nmol/ L TSA. Bisulfite sequencing PCR ( BSP) and Methylation-sensitive high-resolution melt analysis ( Ms-HRM) were used to detect significantly differentially demethylated CpG sites in the human MMP9 promoter region in cells exposed to TNF-α. Different sites methylation constructs of promoter-luciferase reporter gene were made to detect the influences of site-specific DNA demethylation on transcription activity of MMP9 promoter. Results Compared with PBS-treated control, TNF-α significantly increased the expression of MMP9 in HaCaT cells for indicated culture duration ( P < 0. 05 ). Real time PCR, Western blot, and ELISA analysis demonstrated that the mRNA and protein levels of MMP9 were increased initially, followed by a decline with prolonged incubation time. After TNF-α treatment, varied degrees of DNA demethylation occurred at 10 CpG sites in the promoter of MMP9, and the changes at the -36 bp site were statistically significant (P<0. 05). The demethylation at the -36 bp site greatly increased the transcription activity of MMP9. Conclusion TNF-α promotes MMP9 expression in HaCaT cells through inducing -36 bp site DNA demethylation on the promoter of MMP9.
4.Role of itopride in the improvement of bowel preparation before colon endoscopy examination in constipated patients
Ling REN ; Liyang GU ; Haiyan LI ; Yan SONG ; Xiaobo LI
Chinese Journal of Digestion 2013;(2):93-96
Objective To investigate the role of prokinetic agent itopride in colonic preparation before colonoscopy examination in patients with constipation.Methods A total of 115 outpatients with history of chronic constipation who requested colonoscopy were collected.According to colonic preparation proposal,patients were divided into three groups.Group A (39 cases) took standard dosage of PEG-E solution six hours before colonoscopy examination.Group B (38 cases) took 150 mg itopride 30 minutes before administration of lavage solution.Group C (38 cases) took itopride 150 mg at 7 am,12 am and 8 pm the day before the examination and on the examination day took the same medicine as that of group B.The blood pressure,heart rate and blood electrolytes were monitored before and after taking medicine in the patients of three groups.Quality of colon cleansing of each group was observed and side effects were also observed.One-way analysis of variance (least significant difference,LSD) test was performed for pairwise comparisons among the three groups.Chi-square test was applied for count data.Results Both group A and group B excluded one patient because of malignant carcinoma with colon stricture under colonoscopy,and 113 patients completed the whole colon examination.There was no significant difference in the baseline patients' data of three groups.The colon cleaning score of group C (7.28±1.11) was higher than those of group A and B (6.55±1.18 and 6.51±1.16,LSD test,both P<0.05).The frequency of bowel movements defecation of group C (8.31± 1.32) was more than those of group A and group B (7.11± 1.41 and 6.94± 1.51,LSD,test,both P<0.05).There was no significant difference in terms of intestinal bubble scores,blood pressure,heart rate,blood electrolytes the uncomfortable degree of colonic preparation and rate of side effects of the three groups.Conclusion The colonic preparation can be safely and effectively improved by taking high dose of itopride one day before and on the day of colonoscopy examination.
5.Effect of homoharringtonine combined with AG490 on JAK2-STAT5 associated signal pathway in HEL cells.
Journal of Experimental Hematology 2011;19(5):1117-1120
This study was aimed to explore the effect of homoharringtonine in combination with AG490 on JAK2-STAT5 associated signal pathway in HEL cells, and analyze its mechanism so as to provide theoretical basis for therapy of chronic myeloproliferative neoplasma by new program. The cell survival rates were tested by MTT, apoptosis was tested by flow cytometry after HEL cells were treated by 20 ng/ml HHT, 100 µmol/L AG490 and 20 ng/ml HHT in combination with 100 µmol/L AG490, while the signal proteins such as P-JAK2, P-STAT5 and BCL-xL activated by abnormal activated JAK2 were tested by Western blot. The results showed that both HHT and AG490 could inhabit the HEL cell proliferation after being treated for 24 hours, and Annexin V-PI double staining confirmed early apoptosis while HHT effect was more obvious, Western blot showed that the expressions of P-JAK2 and P-STAT5 were down-regulated, while the total protein levels of JAK2 and STAT5 were stable. It is concluded that HHT combined with AG490 can obviously inhibit the proliferation and induce early apoptosis of HEL cells, and there is synergistic effect between the two drugs. HHT possibly acts as a broad-spectrum PTK inhibitor and synergistically with AG490 inhibits the phosphorylation of signal proteins caused by JAK2V617F, thus down-regulating the transcription of STAT5.
Cell Line, Tumor
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Harringtonines
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pharmacology
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Humans
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Janus Kinase 2
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metabolism
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STAT5 Transcription Factor
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metabolism
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Signal Transduction
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drug effects
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Tyrphostins
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pharmacology
7.Construction of OTX1 Lentiviral Vector and Overexpression Research
Ping REN ; Shu-Yan WANG ; Yun-Qian GUAN ; Yan-Ling XU ; Yu ZHANG ;
China Biotechnology 2006;0(01):-
OTX1 gene is one of the pivotal transcriptional factors involved in the neurogenesis.In order to overexpress the OTX1 gene in distinct cell types and find out its contribution to the proliferation and differentiation of stem cells in vitro,OTX1 cDNA was subcloned into lentiviral vectors.The resulting constructions pDUETOTX1,pDUETGFPOTX1 and pDUETGFP were packaged in 293 cells producing viral particles to transduce 293T cells,SY5Y cells,mouse embryonic stem cells and E15 neural stem cells.It was proved that the transferred OTX1 gene was located in the nuclei of the transduced cells in stead of plasma.Lentivirus is an ideal vector delivering gene to different cells.The overexpression of OTX1 in transduced 293T cells were validated by Western blot and immunofluorescence.
8.Anti-inflammatory mechanism research of flavonoid compounds in Dalbergiae Odoriferae Lignum by module-based network analysis.
Shi-chao ZHENG ; Zhen-zhen REN ; Yan-ling ZHANG ; Yan-jiang QIAO
China Journal of Chinese Materia Medica 2015;40(8):1565-1569
Dalbergiae Odoriferae Lignum as a traditional Chinese medicine (TCM) has been widely used for promoting blood circulation and removing blood stasis. Flavonoid compounds are main chemical constituents of Dalbergiae Odoriferae Lignum, which exert anti-inflammatory property. However, the underlying anti-inflammatory mechanisms of flavonoid compounds are incompletely understood. It has been reported that isoliquiritigenin, liquiritigenin, naringenin and butein possess anti-inflammatory property. The purpose of this study is to illuminate the anti-inflammatory mechanism of flavonoid compounds based on the protein interaction network (PIN) analysis on molecular network level. 130 targets of the main medicinal ingredients of flavonoid compounds were gained though database retrieval. A protein interaction network of flavonoid compounds was constructed with 589 nodes and 216 interactions. By a graph theoretic clustering algorithm Molecular Complex Detection (MCODE), 26 modules were identified and analyzed by Gene ontology (GO) enrichment. Two modules were associated with anti-inflammatory actions. The most interesting finding of this study was that the anti-inflammatory effect of flavonoid compounds may be partly attributable to inhibite FOS, PTGS2 expression, inhibite of IL-1beta release, and block the MAPK pathway and toll-like receptor pathway.
Anti-Inflammatory Agents
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pharmacology
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Dalbergia
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chemistry
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Drugs, Chinese Herbal
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pharmacology
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Flavonoids
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pharmacology
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Humans
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Protein Binding
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drug effects
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Protein Interaction Maps
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drug effects
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Proteins
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metabolism
9.Study on relations between transient receptor potential vanilloid 1 and pungent property of traditional Chinese medicines.
Xing WANG ; Yan-Ling ZHANG ; Yun WANG ; Zhen-Zhen REN ; Hong-Juan BAO ; Yan-Jiang QIAO
China Journal of Chinese Materia Medica 2014;39(13):2422-2427
The five-flavor theory of traditional Chinese medicines (TCM) and the flavor efficacy generation mechanism has long been focuses and difficulties in studies on traditional Chinese medicinal properties. In this paper, by using the pharmacophore-based virtual screening technique, the authors discussed the relations between the pungent property and transient receptor potential vanilloid 1 (TRPV1) by studying the TCM components' role in regulating TRPV1 ion channel. The results showed that the matching relationship between TRPV1 agonist pharmacophore model and TCM chemical components could identify the active ingredients from pungent herbs. Therefore, the authors proposed that TRPV1 is one of the potential targets for efficient pungent herbs. The pungent property of TCMs is decided by its chemical components, and consistent with the inherited and additive characteristics.
Drugs, Chinese Herbal
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chemistry
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pharmacology
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Humans
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Smell
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TRPV Cation Channels
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antagonists & inhibitors
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metabolism
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Taste
10.Clinical competence model of famous and veteran physicians of Chinese medicine
Jiang-Yan XU ; Yan-Ling LUO ; Xiao-Peng REN ; Yan-Xia GUO
Journal of Beijing University of Traditional Chinese Medicine 2009;32(9):581-585
Objective To investigate the various factors of clinical competence model of famous and veteran physicians of Chinese medicine, and to mine effectively the clinical experience of modern famous and veteran physicians of Chinese medicine.Methods The clinical experiences of famous and veteran physicians of Chinese medicine were determined by applying the methods of philology, psychology and statistics. The clinical competence model of famous and veteran physicians of Chinese medicine was established and its reliability and validity were tested with the expert experience evaluation.Results The codes of various typical behaviors in 42 medical records showed a high consistency. There were six competence factors confirmed according to the typical behaviors. The clinical competence model of famous and veteran physicians of Chinese medicine was established. The test result indicated that the model had a higher validity.Conclusion A complete clinical competence model of famous and veteran physicians of Chinese medicine contains six competence factors, including the abilities of clinical communication, clinical information collection, clinical thinking, patient orientation, diagnosis and treatment, and research and innovation. The model also includes standard operational definition of every competence factor, and clinical typical behaviors and explanation reflecting the competence factors.