2.Repair of damaged intestinal mucosa in a mouse model of sepsis
Rui-Ming CHANG ; Li-Qiang WEN ; Jian-Xing CHANG ; Yu-Ru FU ; Zhi-Peng JIANG ; Shuang CHEN
World Journal of Emergency Medicine 2013;4(3):223-228
BACKGROUND:The intestine is not only the main target attacked by sepsis but also the vital organ which mediated sepsis. The recovery of the damaged intestinal barrier structure and function is related to the occurrence and outcome of multiple organ dysfunction syndrome (MODS). How to protect and reduce the damage of the intestinal mucosa and how to promote the reconstruction of the intestinal mucosa have been the important topics in sepsis for many years. This study aimed to investigate the influential factors of intestinal mucosal reconstruction after intestinal epithelial injuryin vivo in a mouse model of sepsis.METHODS:Mice were subjected to cecal ligation and puncture (CLP) for induction of sepsis to assess intestinal mucosal damage, epithelial cell apoptosis, and transformed number of goblet cells, and to detect the concentration of TNF-α, IL-1 and TGF-β1 and TFF3 (trefoil factor 3) expression in the small intestinal mucosa. All above were performed by HE staining, western blot, ELISA and immunohistochemistry respectively. The experimental animals were divided into a sepsis group and a sham-operation group. The animals with sepsis were separately killed at 6 (7 animals), 24 (7 animals) and 48 hours (7 animals) after CLP.RESULTS:Injured intestinal mucosa was observed in the 3 groups under a light microscope, in which damage scores in the 24-hour and 48-hour groups were higher than in the 6-hour group and no difference was found between the two groups. Moreover, less of goblet cells or other epithelial cells adjacent to the injured surface migrated into the wound to cover the denuded area. The number of goblet cells was substantially decreased in the three CLP groups compared with the sham-operation group. Protein levels of IL-1 and TNF-α were significantly increased by 3-4 fold at all time points when compared with the sham-operation group, and cleaved caspase-3 by 4 fold. Although TFF3 expression was modestly increased for 6 hours after the onset of CLP, it appeared to decline at 24 hours and 48 hours as shown by Western blot. A similar tendency was observed upon TGF-β1, i.e. the protein level was not elevated at 24 hours and 48 hours, but increased modestly at 6 hours.CONCLUSIONS:Sepsis from CLP shows less restitution on the surface of injured intestinal mucosa. There is evidence that both constant inflammatory reaction and epithelial cell apoptosis may affect mucosal reestablishment of the intestine at the onset of sepsis. Mucosa after severe sepsis showed the state of high inflammation, and declined goblet cell function and mucosal reconstruction, which affected the repair of damaged intestinal barrier. Constant inflammatory reaction, and declined goblet cell function and mucosal reconstruction ability may affect the reestablishment of intestinal mucosa at the onset of sepsis.
3.Construction of NIM sub-network regulated by Liuwei Dihuang decoction and its characteristics
Tong-Xing WANG ; Lu HAN ; Xiao-Rui CHENG ; Wen-Xia ZHOU ; Yong-Xiang ZHANG
Chinese Journal of Pharmacology and Toxicology 2018;32(4):306-307
OBJECTIVE To construct the neuroendocrine immunomodulation (NIM) sub-network regulated by Liuwei Dihuang decoction (LW) and analyze its characteristics. METHODS We took the GSE57273 in GEO database and screened the differentially expressed genes (DEGs)(P<0.01) by the GEO2R tool as gene expression signature of LW. The global PPI network was constructed in the context of whole PPI network through direct interaction algorithm and forest algorithm respectively.Then the enrichment and the topological characteristics of NIM signaling molecules were evaluated by permutation test. Finally, we abstracted the NIM sub-network by NIMNT, which combined the NIM molecular network and forest algorithm, and analyzed the topological characteristics of it by the Network Analyzer(release 2.7)plugin in Cytoscape v3.5.1.RESULTS We got 2468 DEGs in the gene expression signature of LW.After analyzing the global PPI network of LW got by two kinds of algorithms,we found that the NIM signaling molecules significantly enriched and located in important positions in the global PPI network. The NIM sub-network regulated by LW contained 1099 nodes and 1056 edges. We screened out 22 hub nodes (Degree>10). Among them, there were 19 NIM signaling molecules in which only ESR1 changed significantly and 3 non-DEGs(NFATC2,RARA,TP53).However,the down-stream of the hub nodes were significantly changes. CONCLUSION The results suggested that LW might mainly regulate the non-hub nodes to recovery of the network imbalance of the body in the state of disease.
4.Apparatus for the measurement of the oxygen uptake of rats subjected to hypobaric hypoxia.
Rui-Feng DUAN ; Wen-Kao NAN ; Yi-Ping XING ; Huai-Xin WANG ; Wen-Yu CUI ; Hai WANG
Chinese Journal of Applied Physiology 2011;27(4):507-509
OBJECTIVETo construct an apparatus for the oxygen uptake measurement of rats exposed to hypobaric hypoxia at different simulated altitude.
METHODSThe capacity of this apparatus was about 0.01 m3. It included animal experimental cabin, reference cabin, altimeter, altitude vertical velocity indicator, pressure difference inductor and oxygen compensator, low scale manometer, soda lime and calcium chloride, small fan, thermometer, circulating water system and vacuum pump. The oxygen uptake of the rats at 6 000 m, 4 000 m and 1 000 m simulated altitude was measured using this apparatus.
RESULTSThe oxygen uptake of the rats at 50 m, 4 000 m and 6 000 m simulated altitude was (24.4 +/- 2.1), (10.8 +/- 2.0) and (8.8 +/- 1.6) ml O2/(kg x min) respectively (average +/- s, n = 10). The oxygen uptake decreased as altitude increased.
CONCLUSIONThis apparatus can be used to measure the oxygen uptake of the rats at different simulated altitude.
Altitude ; Altitude Sickness ; physiopathology ; Animals ; Computer Simulation ; Equipment and Supplies ; Hypoxia ; physiopathology ; Male ; Oxygen ; metabolism ; Oxygen Consumption ; physiology ; Rats ; Rats, Sprague-Dawley
5.Determination concentration of duloxetine hydrochloride in human plasma by HPLC-MS/MS and study on its pharmacokinetics
Rui-Ke ZHAO ; Jia-Wen LIU ; Gang CHENG ; Jing TANG ; Juan SONG ; Wen-Xing PENG
The Chinese Journal of Clinical Pharmacology 2010;26(2):137-140
Objective To establish a liquid chromatography-tandem mass spectrometry(LC-MS/MS)method for the determination concentration of duloxetine in human plasma and study on its pharmacokinetics in healthy human.Methods The separation of duloxetine was performed on Phenomenex C_(18) column with fluoxetine as the internal standard.The mobile phase was composed of 5 mmol L~(-1) ammonium acetate with 0.02% formic acid acetonitrile(55:45).Electrospray ionization source was applied and operated in positive ion mode.A single dose of 60 mg duloxetine hydrochloride was given to 5 male and 5 female healthy volunteers and the plasma was separated.The concentration of duloxetine was determined by HPLC-MS/MS and the pharmacokinetic parameters were calculated.Results The linear range of duloxetine was 0.89-106.80 ng·mL~(-1)(γ=0.9977).The methodological recovery and the extraction recovery ranged between 93.19%-107.27% and 72.81%-89.96%,respectively.Both the inter-day RSD and intra-day RSD were less than 11%.The main pharmacokinetic parameters after a single dose of 60mg duloxetine are as follows: C_(max) was(44.40 ±17.78)ng·mL~(-1),t_(max) was(6.10±1.29)h,t_(1/2) was(12.81 ±2.31)h;AUC_(0-60) and AUC_(0-∞) were (696.04±337.82),(733.82±343.40)ng·h·mL~(-1),respectively.Conclusion The method is simple,accurate and reliable,and suitable for the determination of duloxe-tine in therapeutic drug monitor and its pharmacokinetics study.
6.Expression of IGF-1R in esophageal squamous cell carcinoma and the effect of its silencing by siRNA on the proliferation of esophageal cancer EC9706 cells in vitro.
Wang MA ; Wen LI ; Qing-Xia FAN ; Liu-Xing WANG ; Rui-Lin WANG ; Shi-Xin LU
Chinese Journal of Oncology 2011;33(8):609-612
OBJECTIVETo explore the correlation of IGF-1R expression with clinical features of esophageal squamous cell carcinoma (ESCC) and to investigate the effect of silencing IGF-1R by siRNA on the proliferation of esophageal cancer cell line EC9706 cells.
METHODSImmunohistochemistry was used to detect the expresion of IGF-1R in 80 specimens of ESCC and 18 specimens of normal esophageal mucosa. IGF-1R siRNA was transfected into esophageal squamous cell carcinoma EC9706 cells, and the effect of RNAi was assessed by Western blot. The proliferation of EC9706 cells was determined by drawing growth curve, MTT assay and plate colony-forming assay.
RESULTSThe total and strong positive rates of IGF-1R expression were 86.3% and 51.3% in ESCC, and 61.1% and 11.1% in normal esophageal epithelium, respectively. The total and strong positive rates of IGF-1R expression in patients with lymph node metastasis were 94.4% and 74.1%, significantly higher than 69.2% and 3.9%, respectively, in those without lymph node metastasis (P<0.01). A significantly higher IGF-1R expression was associated with lower histological grade (P<0.05). The total and strong rates of IGF-1R expression in 39 patients of stages III and IV were 97.4% and 71.8% , significantly higher than the 75.6% and 31.7%, respectively, in 41 cases of stages I and II (P<0.01). IGF-1R RNAi significantly inhibited IGF-1R expression and the growth of EC9706 cells. The clone formation rate of RNAi-IGF-1R transfected cells was 19.1%, significantly lower than that of 52.3% in non-transfected cells and 49.0% in empty vector-transfected EC9706 cells (P<0.05).
CONCLUSIONSThe overexpression of IGF-1R is colerated with lymph node metastasis, differentiation and clinical stage. Down-regulation of IGF-1R can inhibit the proliferation of esophageal cancer EC9706 cells in vitro.
Aged ; Carcinoma, Squamous Cell ; genetics ; metabolism ; pathology ; Cell Line, Tumor ; Cell Proliferation ; Down-Regulation ; Esophageal Neoplasms ; genetics ; metabolism ; pathology ; Female ; Gene Expression Regulation, Neoplastic ; Humans ; Lymphatic Metastasis ; Male ; Middle Aged ; Neoplasm Grading ; Neoplasm Staging ; RNA Interference ; RNA, Small Interfering ; genetics ; Receptor, IGF Type 1 ; genetics ; metabolism ; Transfection
7.Recent progress on anti-HIV research of traditional Chinese medicine and components.
Zhao-Mei LIU ; Yi-Shu YANG ; Xiao-Li WANG ; Rui-Xing WEN
China Journal of Chinese Materia Medica 2006;31(21):1753-1758
This paper summarized the recent 6 years' progress of anti-HIV compounds and traditional Chinese medicines by searching international network and reviewing the domestic and foreign literature. Traditional Chinese medicinal appeared to be a rich source of potentially useful materials for the treatment of human immunodeficiency virus infection. Some of them are much more potent in anti-HIV activity. And some components extracted from the herbs are even more tonic than the crude herb medicines. It has been proved that some active components such as alkaloids, proteins, flavonoids, quercetin, terpene, lignanoid are able to work on anti-HIV. People should pay more attention to the study of traditional Chinese medicine and the leading compounds on anti-HIV/AIDS in the clinic and in the laboratory. So searching for high efficacy and low toxicity anti-HIV drug from traditional Chinese medicine is an important and prospective research direction in the future.
Acquired Immunodeficiency Syndrome
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drug therapy
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Adjuvants, Immunologic
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isolation & purification
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pharmacology
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Animals
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Anti-HIV Agents
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isolation & purification
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pharmacology
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Drugs, Chinese Herbal
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isolation & purification
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pharmacology
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therapeutic use
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HIV
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drug effects
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Humans
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Medicine, Chinese Traditional
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Phytotherapy
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Plants, Medicinal
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chemistry
8.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
9.The effects of infrasound on the blood coagulation function of guinea pigs.
Wen-min MA ; Peng QI ; Jian-zhong ZHANG ; Yong YI ; Xing-ming CHEN ; Jun ZHANG ; Rui-gang HAN
Chinese Journal of Industrial Hygiene and Occupational Diseases 2011;29(3):209-211
OBJECTIVETo study the change of the blood coagulation function of guinea pigs exposed to 16 Hz/120 dB, 16 Hz/125 dB infrasound and to explore the mechanism of circulation system damage.
METHODSSeventy-two guinea pigs were divided into 3 groups: the control group, the group exposed to 16 Hz/120 dB infrasound for 1.5 h a day and the group exposed to 16 Hz/125 dB infrasound for 1.5 h a day. Each exposure group was divided into 4 sub-groups (8 guinea pigs a sub-group) which were exposed to infrasound for 1, 7, 14 and 21 d, respectively. The coagulation function and serum nitric oxide (NO) were measured for control group and all sub-groups after exposure to infrasound.
RESULTSThe prothrombin time (PT), international normalized ratio (INR) and serum NO of group exposed to 16 Hz/125 dB infrasound were (31.16 ± 3.05) s, 2.53 ± 1.21 and (88.304 ± 52.601) µmol/L, respectively, which were significantly higher than those [(21.36 ± 0.10) s, 1.65 ± 0.07 and (30.943 ± 26.864) µmol/L] of control group (P < 0.05). PT and INR of sub-groups exposed to 16 Hz/125 dB infrasound for 14 and 21 d were significantly higher than those of control group. NO of sub-groups exposed to 16 Hz/125 dB infrasound for 1 week and 2 weeks were significantly higher than that of control group (P < 0.05), but NO of sub-group exposed to 16 Hz/125 dB infrasound for 3 weeks decreased slightly.
CONCLUSIONThe blood coagulation function of guinea pigs exposed to 16 Hz/125 dB infrasound decreased, PT and INR may be used as the indexes to assess of blood coagulation function change induced by the infrasound exposure.
Animals ; Blood Coagulation ; Blood Physiological Phenomena ; Female ; Guinea Pigs ; Male ; Nitric Oxide ; blood ; Noise ; adverse effects ; Prothrombin Time
10.Sequence analysis of LEAFY homologous gene from Dendrobium moniliforme and application for identification of medicinal Dendrobium.
Wen-Rui XING ; Bei-Wei HOU ; Jing-Jiao GUAN ; Jing LUO ; Xiao-Yu DING
Acta Pharmaceutica Sinica 2013;48(4):597-603
The LEAFY (LFY) homologous gene of Dendrobium moniliforme (L.) Sw. was cloned by new primers which were designed based on the conservative region of known sequences of orchid LEAFY gene. Partial LFY homologous gene was cloned by common PCR, then we got the complete LFY homologous gene Den LFY by Tail-PCR. The complete sequence of DenLFY gene was 3 575 bp which contained three exons and two introns. Using BLAST method, comparison analysis among the exon of LFY homologous gene indicted that the DenLFY gene had high identity with orchids LFY homologous, including the related fragment of PhalLFY (84%) in Phalaenopsis hybrid cultivar, LFY homologous gene in Oncidium (90%) and in other orchid (over 80%). Using MP analysis, Dendrobium is found to be the sister to Oncidium and Phalaenopsis. Homologous analysis demonstrated that the C-terminal amino acids were highly conserved. When the exons and introns were separately considered, exons and the sequence of amino acid were good markers for the function research of DenLFY gene. The second intron can be used in authentication research of Dendrobium based on the length polymorphism between Dendrobium moniliforme and Dendrobium officinale.
Amino Acid Sequence
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Base Sequence
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DNA, Plant
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genetics
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Dendrobium
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genetics
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Exons
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Introns
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Orchidaceae
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genetics
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Phylogeny
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Plant Leaves
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genetics
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Plant Proteins
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genetics
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Plants, Medicinal
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genetics
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Sequence Alignment
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Sequence Homology, Amino Acid