1.Mutual effect between neuropeptides and inflammatory cytokines in neurogenic SMSCs of human temporomandibular joint.
Zhi-Ming, LIU ; You-Jian, PENG ; Xing, LONG ; Jian, LI ; Jin, KE ; Wei, FANG
Journal of Huazhong University of Science and Technology (Medical Sciences) 2014;34(4):602-7
In temporomandibular disorders (TMD), pain takes place when neuropeptides stimulate synovial tissue to produce several cytokines such as interleukin (IL)-1β, IL-6 and tumor necrosis factor (TNF)-α, which activate neurons and glia of synovial membrane at the bilaminar regions of temporomandibular joint (TMJ). It has been reported that, after neurogenic differentiation, the synovial mesenchymal stem cells (SMSCs), deriving from TMJ, possess the same cytological features as the neuronal cells. This study examined the ability of substance P (SP) and calcitonin gene-related peptide (CGRP) to stimulate SMSCs and neurogenic SMSCs secreting inflammatory cytokines during TMD, evaluated the mutual effects of inflammatory cytokines and neuropeptides and tested the analgesic effect of hyaluronic acid (HA). The levels of IL-1β, IL-6 and TNF-α in SMSCs and neurogenic SMSCs in the presence of neuropeptides were measured by ELISA. SP and CGRP produced by SMSCs and neurogenic SMSCs were determined by RT-PCR and Western blotting. The results showed that the expression of SP and CGRP was significantly enhanced in the neurogenic SMSCs in response to IL-1β, IL-6 and TNF-α, and the effect was remarkably inhibited by HA. IL-1β, IL-6 and TNF-α, in return, could be enhanced in the neurogenic SMSCs upon stimulation by SP and CGRP. Neuropeptides and inflammatory cytokines might work mutually on the TMD pain. The HA-mediated analgesic effect may be implicated in the inhibition of SP and CGRP expression in neurogenic SMSCs.
2.Inhibitive effect of troglitazone on TGF-beta(1) and fibronectin expression in human peritoneal mesothelial cells.
Hong LIU ; You-ming PENG ; Fu-you LIU ; Ying-hong LIU ; Ling-yan LI ; Jun LI ; Xing CHEN
Journal of Central South University(Medical Sciences) 2007;32(3):473-479
OBJECTIVE:
To investigate the effect of the peroxisome proliferator activated receptor-gamma (PPAR-gamma) agonist troglitazone on TGF-beta(1) and fibronectin (Fn) expression in human peritoneal mesothelial cells (HPMCs).
METHODS:
HPMCs were cultured from human omentum by an enzyme digestion method, growing in medium containing 30 mmol/L D-glucose. TGF-beta(1) and Fn expression were measured in HPMCs in the presence and absence of 15 micromol/L troglitazone. The mRNA expressions of PPAR-gamma,TGF-beta(1) and Fn were determined by semi-quantification reverse transcriptive PCR (RT-PCR). The protein of TGF-beta(1) was determined by enzyme-linked immunosorbent assay (ELISA) and proteins of PPAR-gamma and Fn were determined by Western blot.
RESULTS:
The mRNA and protein expression of TGF-beta(1) and Fn were significantly increased in HPMCs stimulated with 30 mmol/L D-glucose compared with the control group with F12 media (P<0.01). Obvious decrease of TGF-beta(1) was found in troglitazone(15 micromol/L) treated group compared with group stimulated with 30 mmol/L D-glucose (P<0.05). Exposure of HPMCs to troglitazone reduced the Fn secretion (P<0.05).
CONCLUSION
Troglitazone reduced the expression of TGF-beta(1) in HPMCs stimulated by 30mmol/L D-glucose, and reduced Fn production. PPAR-gamma agonists may have a specific role in ameliorating the course of progressive peritoneal fibrosis under long-term peritoneal dialysis states.
Blotting, Western
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Cells, Cultured
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Chromans
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pharmacology
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Dose-Response Relationship, Drug
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Enzyme-Linked Immunosorbent Assay
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Epithelial Cells
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cytology
;
drug effects
;
metabolism
;
Fibronectins
;
biosynthesis
;
genetics
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Glucose
;
pharmacology
;
Humans
;
PPAR gamma
;
biosynthesis
;
genetics
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Peritoneum
;
cytology
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RNA, Messenger
;
biosynthesis
;
genetics
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Reverse Transcriptase Polymerase Chain Reaction
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Thiazolidinediones
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pharmacology
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Transforming Growth Factor beta1
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biosynthesis
;
genetics
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Troglitazone
3.Hepatic veno-occlusive disease: report of two cases.
Wei-xing CHEN ; Ming YANG ; Chao-hui YU ; You-ming LI
Chinese Journal of Hepatology 2005;13(5):394-395
4.Heteroexpression of Rhizopus arrhizus delta6-fatty acid desaturase gene in Pichia pastoris.
Qi ZHANG ; Ming-Chun LI ; Ying SUN ; You-Wei CHEN ; Biao ZHANG ; Lai-Jun XING
Chinese Journal of Biotechnology 2005;21(6):871-877
Delta6-fatty acid desaturase is a membrane-bound enzyme, which is rate-limiting for the biosynthesis of polyunsaturated fatty acids. A cDNA sequence putatively encoding a delta6-fatty acid desaturase was isolated from Rhizopus arrhizus NK300037 using RT-PCR and RACE methods in our previous work. Sequence and function analysis indicated that this sequence was a novel delta6-fatty acid desaturase gene which had an open reading frame of 1377bp coding 458 amino acids of 52kD. The methylotrophic yeast Pichia pastoris, has been developed into a highly successful system for the production of a variety of heterologous proteins during the past 20 years. In this work, the Rhizopus arrhizus delta6-fatty acid desaturase gene (RAD6) was subcloned into expression vector pPIC3.5K to generate a recombinant plasmid pPICRAD6, which was subsequently transformed into Pichia pastoris strain GS115 for heterologous expression by electroporation method. Total fatty acids were extracted from the induced cells and methylated. The resultant fatty acid methyl esters (FAME) were analyzed by gas chromatography (GC) and gas chromatography-mass spectrometry (GC-MS). Fatty acids analysis showed that the coding product introduced a new double bond at delta6 position of appropriate fatty acid substrates including C16:1, C17:1, C18:1, linoleic acid and alpha-linolenic acid without chain length specificity of fatty acids. Furthermore, modification of sequence flanking AUG codon of this delta6-fatty acid desaturase gene increased the expression of target gene in P. pastoris. All of these results suggest that P. pastoris is an optimal expression system of delta6-fatty acid desaturase gene.
Cloning, Molecular
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Electroporation
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Fungal Proteins
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biosynthesis
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genetics
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Genetic Vectors
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Linoleic Acid
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metabolism
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Linoleoyl-CoA Desaturase
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biosynthesis
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genetics
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Pichia
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genetics
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metabolism
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Rhizopus
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enzymology
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genetics
5.Clinicopathological significance of altered metallothionein 2A expression in gastric cancer according to Lauren's classification.
Yuan-ming PAN ; Rui XING ; Jian-tao CUI ; Wen-mei LI ; You-yong LÜ
Chinese Medical Journal 2013;126(14):2681-2686
BACKGROUNDDysregulated metallothionein 2A (MT2A) has been implicated in carcinogenesis. The purpose of this study was to investigate the expression of MT2A in gastric cancer (GC) and its correlation with prognosis.
METHODSReverse transcription-polymerase chain reaction and real-time polymerase chain reaction were used to detect the mRNA expression of MT2A in 12 GC cell lines, normal gastric epithelial GES-1 cells, and 36 GC and adjacent normal tissues. MT2A protein expression was determined in 258 GC tissues and 171 adjacent normal tissues by immunohistochemistry.
RESULTSMT2A mRNA expression was lower in GC cells and primary tumors than in GES-1 cells and adjacent normal tissues, respectively. High protein expression of MT2A was present in 130 of 171 normal tissues (76.0%) and in 56 of 258 GC tissues (21.7%; P < 0.001). MT2A protein expression was higher in well/moderately differentiated GC (22/54; 40.7%) than in poorly differentiated GC (34/204; 16.7%; P < 0.001). Moreover, the protein expression of MT2A was lower in diffuse-type GC (6/82; 7.3%) than in intestinal-type GC (50/176; 28.4%; P = 0.0001). Importantly, MT2A expression was an independent prognostic factor for GC, and decreased MT2A expression was associated with poor clinical outcome (P < 0.001). The expression status of MT2A could predict prognosis in intestinal and diffuse-type GCs.
CONCLUSIONExpression status of MT2A might be a useful prognostic biomarker for GC, especially when used in combination with Lauren's classification.
Adult ; Aged ; Cell Line, Tumor ; Female ; Humans ; Logistic Models ; Male ; Metallothionein ; analysis ; genetics ; MicroRNAs ; analysis ; Middle Aged ; Prognosis ; Proportional Hazards Models ; Stomach Neoplasms ; chemistry ; classification ; pathology
6.Longbishu Capsule combined with mesylate doxazosin: an efficacious therapy for benign prostatic hyperplasia.
De-gui CHANG ; Guang-sen LI ; Cheng-hua PENG ; Xu-jun YU ; Pei-hai ZHANG ; Ming-shuai BI ; Di-ang CHEN ; Yao-dong YOU ; Xing-zhi YANG
National Journal of Andrology 2015;21(2):165-169
OBJECTIVETo assess the clinical effect and safety of the Chinese medicine Longbishu Capsule combined with mesylate doxazosin in the treatment of benign prostatic hyperplasia (BPH) of the kidney deficiency and blood stagnation type.
METHODSThis was a randomized, double-blind, double-simulation control study. We equally assigned 60 men diagnosed with BPH of the kidney deficiency and blood stagnation type to an experimental and a control group, the former treated with mesylate doxazosin plus Longbishu Capsule and the latter with mesylate doxazosin plus placebo. We compared the International Prostate Symptom Score (IPSS), quality of life (QOL), Chinese symptom score (CSS), maximal urinary flow rate (Qmax), and prostate volume between the two groups of patients before and after 6 months of medication.
RESULTSAfter treatment, there were 5 cured cases, 13 markedly effective cases, 9 effective cases, 1 ineffective case, and 2 eliminated cases in the experimental group, as compared with 2 cured cases, 8 markedly effective cases, 10 effective cases, 7 ineffective cases, and 3 eliminated cases in the control group. The total effectiveness rate was obviously higher in the former (96.4%) than in the latter (74.1%). IPSS, Qmax, and CSS were improved in both of the groups after medication, even more significantly in the experimental than in the control group (IPSS: 15.22 ± 2.98 vs 18.15 ± 5.88, P <0.05; Qmax: [13.56 ± 2.26] ml/s vs [11.78 ± 2.97] ml/s, P <0.05; CSS: 6.18 ± 2.13 vs 9.52 ± 3.15, P <0.05). Because of the difference in the QOL score between the two groups at the baseline (P = 0.038 <0.05), no more comparison was made in this aspect after treatment.
CONCLUSIONThe combination of Longbishu Capsule with mesylate doxazosin is safe and effective for the treatment of BPH.
Adrenergic alpha-Antagonists ; therapeutic use ; Capsules ; Double-Blind Method ; Doxazosin ; therapeutic use ; Drug Therapy, Combination ; Drugs, Chinese Herbal ; therapeutic use ; Humans ; Male ; Prostatic Hyperplasia ; drug therapy ; physiopathology ; Quality of Life ; Treatment Outcome ; Urination
7.Successful tubes treatment of esophageal fistula.
Ning ZHOU ; Wei-xing CHEN ; You-ming LI ; Zhun XIANG ; Ping GAO ; Ying FANG
Journal of Zhejiang University. Science. B 2007;8(10):709-714
AIMTo discuss the merits of "tubes treatment" for esophageal fistula (EF).
METHODSA 66-year-old female who suffered from a bronchoesophageal and esophagothoratic fistula underwent a successful "three tubes treatment" (close chest drainage, negative pressure suction at the leak, and nasojejunal feeding tube), combination of antibiotics, antacid drugs and nutritional support. Another 55-year-old male patient developed an esophagopleural fistula (EPF) after esophageal carcinoma operation. He too was treated conservatively with the three tubes strategy as mentioned above towards a favorable outcome.
RESULTSThe two patients recovered with the tubes treatment, felt well and became able to eat and drink, presenting no complaint.
CONCLUSIONTubes treatment is an effective basic way for EF. It may be an alternative treatment option.
Aged ; Chest Tubes ; Combined Modality Therapy ; Drainage ; instrumentation ; methods ; Enteral Nutrition ; methods ; Esophageal Fistula ; Female ; Humans ; Intubation, Intratracheal ; methods ; Male ; Middle Aged ; Suction ; instrumentation ; methods ; Treatment Outcome
8.Mutual effect between neuropeptides and inflammatory cytokines in neurogenic SMSCs of human temporomandibular joint.
Zhi-ming LIU ; You-jian PENG ; Xing LONG ; Jian LI ; Jin KE ; Wei FANG
Journal of Huazhong University of Science and Technology (Medical Sciences) 2014;34(4):602-607
In temporomandibular disorders (TMD), pain takes place when neuropeptides stimulate synovial tissue to produce several cytokines such as interleukin (IL)-1β, IL-6 and tumor necrosis factor (TNF)-α, which activate neurons and glia of synovial membrane at the bilaminar regions of temporomandibular joint (TMJ). It has been reported that, after neurogenic differentiation, the synovial mesenchymal stem cells (SMSCs), deriving from TMJ, possess the same cytological features as the neuronal cells. This study examined the ability of substance P (SP) and calcitonin gene-related peptide (CGRP) to stimulate SMSCs and neurogenic SMSCs secreting inflammatory cytokines during TMD, evaluated the mutual effects of inflammatory cytokines and neuropeptides and tested the analgesic effect of hyaluronic acid (HA). The levels of IL-1β, IL-6 and TNF-α in SMSCs and neurogenic SMSCs in the presence of neuropeptides were measured by ELISA. SP and CGRP produced by SMSCs and neurogenic SMSCs were determined by RT-PCR and Western blotting. The results showed that the expression of SP and CGRP was significantly enhanced in the neurogenic SMSCs in response to IL-1β, IL-6 and TNF-α, and the effect was remarkably inhibited by HA. IL-1β, IL-6 and TNF-α, in return, could be enhanced in the neurogenic SMSCs upon stimulation by SP and CGRP. Neuropeptides and inflammatory cytokines might work mutually on the TMD pain. The HA-mediated analgesic effect may be implicated in the inhibition of SP and CGRP expression in neurogenic SMSCs.
Calcitonin Gene-Related Peptide
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biosynthesis
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Cells, Cultured
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Cytokines
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biosynthesis
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Female
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Humans
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Male
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Mesenchymal Stromal Cells
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metabolism
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pathology
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Substance P
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biosynthesis
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Synovial Membrane
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metabolism
;
pathology
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Temporomandibular Joint
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metabolism
;
pathology
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Temporomandibular Joint Disorders
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metabolism
;
pathology
9.Genomic analysis of an unkown pap genotype from uropathogenic Escherichia coli 4030 strain
Qing-Gang WU ; Jing-Ping ZHANG ; Jin-Ying CHEN ; Ming-Huan ZHENG ; Feng-Xing YOU ; Yong JIANG
Chinese Journal of Epidemiology 2008;29(11):1123-1127
Objective To investigate the differences between the sequences of papA and papG of UPECA030 strain and the related genes, to better understand the genetic variation of UPEC4030 papA and its combination forms with papG so as to identify if it was a new genotype. Methods Cloning and sequencing methods were used to analyze the sequences of papG and papA of UPEC4030 strain and to compare their related sequences. Results Through sequence analysis of papA, it was revealed that there was a 722 bp gene,encoding 192 amino acid polypeptide. The overall homology of the papa genes between UPEC4030 and the standard strains of ten F types were 36.11%-77.95 % and 22.20%-78.34% at nueleotide and deduced amino acid levels. Homology between the sequences of reverse primers and the corresponding sequence of UPEC4030 papA was 10.00%-66.67%. The results confirmed that UPECA030 strain contained a novel papA variant. Through sequence analysis of UPEC4030 papG, we revealed a 1100 bp gene, encoding 337 amino acid polypeptide. The homology of the papG genes between UPEC4030 and UPEC IA2, the standard strain, was 99.00 % at nucleotide level and 99.11% at deduced amino acid and UPEC4030 strain carried class I] genotype of papG. Conclusion UPEC4030 strain contained an unknown papA variant or the new genotype and carried class II genotype of papG. The pathogenic mechanism and epidemiology call for further study.
10.Research on cytokines gene express of rats alcoholic liver disease affected by tea polyphenol.
Xing-Guo ZHANG ; Chao-Hui YU ; Qi JIANG ; Yu ZHANG ; Shao-Hua CHEN ; You-Ming LU
China Journal of Chinese Materia Medica 2005;30(11):847-850
OBJECTIVETo evaluate the effect of tea polyphenol(TP) on the rat with alcoholic liver damage.
METHODRats were divided into 3 groups, in which 2 groups were stomach perfused with alcohol to result in ALD, and 1 group of them stomach perfused with TP simultaneously. Another group was normal control groups (stomach perfused with drinking water). In the end of 12 weeks, the liver specimen of each rat was observed by anglicizing its tissue damage, and all data collected was performed by statistical analysis in quantum and semi-quantum. Meanwhile cytokines gene express of each group is determined.
RESULTIn the end of 12 weeks, alcoholic hepatitis appeared in rat liver. Hepatic injury in alcohol group and TP group were found, but could not be found in normal group. Compared with pure alcohol group, alcoholic liver damage mainly showing with steatosis in TP group were slight, in addition showing liver cellular swelling with small area, with less spot and focal necrosis, none bridging necrosis. Steatosis were slight relatively, mega-bubble steatosis were less found. Collagen deposition of TP group were less than those of pure alcohol group. Gene expression of. cytokine have diversity statistically such as IL-3, IL-4, IL-1R2, IL-6R, IL-7R2, IL-3Ra, IL-R1, IL-13, IL-1R1, IL-7R2, EPO-R, LIFR, IL-1R2, IL-5R2, CSF1, CD27, IL-6R.
CONCLUSIONTP is able to attenuate alcoholic liver damage. It's mechanism is possibly due to modulating cytokines gene expression of cytokine.
Animals ; Flavonoids ; isolation & purification ; pharmacology ; Gene Expression ; Interleukins ; biosynthesis ; genetics ; Liver ; metabolism ; pathology ; Liver Diseases, Alcoholic ; genetics ; metabolism ; pathology ; Oligonucleotide Array Sequence Analysis ; Phenols ; isolation & purification ; pharmacology ; Plants, Medicinal ; chemistry ; Polyphenols ; RNA, Messenger ; biosynthesis ; genetics ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Receptors, Interleukin ; biosynthesis ; genetics ; Tea ; chemistry