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TLR4 signaling induced TLR2 expression in the process of mimic cerebral ischemia/reperfusion in vitro

中国科学：生命科学英文版 2010年第2期53卷 223-228页

作者： SUN Peng1,ZHANG Qing2,HAN JiYuan1,TIAN Yuan3.& ZHANG JingHui3.1department of emergency,Union hospital,{3. medical College,huazhong university of science and technology,wuhan 43.022,china 2department of Anesthesia,Union hospital,{3. medical College,huazhong university of science and technology,wuhan 43.022,china 3.0} of General Surgery,Union hospital,{3. medical College,huazhong university of science and technology,wuhan 43.022,china 1. department of emergency Union Hospital Tongji Medical College Huazhong University of Science and Technology Wuhan 430022 China2. Department of Anesthesia Union Hospital Tongji Medical College Huazhong University of Science and Technology Wuhan 430022 China3. Department of General Surgery Union Hospital Tongji Medical College Huazhong University of Science and Technology Wuhan 430022 China

Both TLR4 and TLR2 participated in the mediation of the inflammatory injury in the process of partial cerebral ischemia/***,it still remains unclear whether a crosstalk exists between TLR2 and TLR4 in ischemic cerebral *** the present study,we investigated the effect of TLR4 signaling on TLR2 expression during mimic cerebral I/R in ***-2 cells were cultured and treated with ischemia/reperfusion,then transfected with the plasmid pEGFP-H1/TLR4-siRNA,the plasmid pEGFP-H1/control sequence-siRNA and the blank plasmid,***,the expression of TLR2 and TLR4 mRNA and protein,NF-κB p65 mRNA and supernatant TNF-α level were significantly higher in ischemia/reperfusion treated cells than those lack of ischemia/reperfusion treatment,and as compared with those in ischemia/reperfusion treated cells without transfection,no significant differences about the above mentioned gene and protein expression were found in the blank plasmid tranfected cells and the plasmid pEGFP-H1/control sequence-siRNA transfected cells respectively,while the expression levels in the plasmid pEGFP-H1/TLR4-siRNA transfected cells were significantly ***,in order to determine the effects of pyrrolidinediethyldithiocarbamate (PDTC),an NF-κB inhibitor,on the TLR4-induced TLR2 expression in BV-2 cells treated with ischemia/reperfusion,it was found that TLR4 and TLR2 mRNA expressions in PDTC pretreated cells were significantly lower in comparison with normal saline pretreated cells and non-pretreated *** data suggested that TLR2 activation,signaled by TLR4 and regulated by NF-κB,might be directly involved play an important role in ischemia/reperfusion induced brain damage.

关键词： Toll-like receptor 4 Toll-like receptor 2 cerebral ischemia/reperfusion microglia

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Inhibition of ethanol-induced toxicity by tanshinone ⅡA in PC12 cells

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Acta Pharmacologica Sinica 2006年第6期27卷 659-664页

作者： Xian-fang MENG~2 Xiao-jing ZOU~3.Bin PENG~2 Jing SHI~2 Xin-min GUAN~2 Chun ZHANG~(4,5)2 department of Neurobiology,tongji medical College,{3.,wuhan 43.03.,china 3.department of Anesthesiology,4 department of Nephrology,Union hospital,tongji medical College,{3.,wuhan 43.022.china department of Neurobiology Tongji Medical College Huazhong University of Science and Technology Wuhan China department of Anesthesiology Union Hospital Tongji Medical College Huazhong University of Science and Technology Wuhan China department of Nephrology Union Hospital Tongji Medical College Huazhong University of Science and Technology Wuhan China

Aim:To observe the effects of tanshinone IIA(Tan IIA)on the neurotoxicityinduced by ethanol in PC 12 cells and to explore its protective ***:PC 12cell survival was measured by MTT *** formation of reactive oxygenspecies(ROS)and lactate dehydrogenase(LDH)release were detected by 2′,7′-dichlorofluorescin(DCF)fluorescence and calorimetric method,*** of cell apoptosis was monitored by flow *** expression ofp53.was detected by immuno-fluorescence and flow ***:Ethanolsignificantly impaired the survival of PC12 cells as demonstrated by MTT *** also induced significant ROS formation and increased LDH ***-incubation with Tan IIA in the culture medium significantly reversed these *** caused cell apoptosis and the upregulation of p53.*** anti-apoptosis effects of Tan IIA on ethanol-induced toxicity were accompanied by thedownregulation of pro-apoptotic p53.protein ***:Tan IIAcan protect neurons from apoptosis and might serve as a potential therapeuticdrug for neurological disorders induced by ethanol.

关键词： apoptosis ethanol PC12 cells p53 tanshinone ⅡA

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Hyaluronic acid fragments evoke Kupffer cells via TLR4 signaling pathway

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science china(Life sciences) 2009年第2期52卷 147-154页

作者： ZHANG JinXiang1, WANG Hui2, XIAO Qing3. LIANG HuiFang4, LI ZhuoYa4, JIANG ChunFang1, WU HeShui5 & ZHENG QiChang5 1 department of emergency Surgery, Union hospital Affiliated to {3., wuhan 43.022, china 2 department of medical Genetics, tongji medical College Affiliated to {3., wuhan 43.03., china {13. of Ophthalmology, Union hospital Affiliated to {3., wuhan 43.022, china 4 department of medical Immunology, tongji medical College Affiliated to {3., wuhan 43.03., china 5 department of General Surgery, Union hospital Affiliated to {3., wuhan 43.022, china 1. department of emergency Surgery Union Hospital Affiliated to Huazhong University of Science and Technology Wuhan 430022 China 2. department of medical Genetics Tongji Medical College Affiliated to Huazhong University of Science and Technology Wuhan 430030 China 3. department of Ophthalmology Union Hospital Affiliated to Huazhong University of Science and Technology Wuhan 430022 China 4. department of medical Immunology Tongji Medical College Affiliated to Huazhong University of Science and Technology Wuhan 430030 China 5. department of General Surgery Union Hospital Affiliated to Huazhong University of Science and Technology Wuhan 430022 China

Kupffer cells, expressing toll-like receptor 4 (TLR4), play a central role in hepatic ischemia/reperfusion (I/R) injury. Hyaluronic acid (HA) fragments, degradative products of high-molecular-weight HA (HMW-HA), acquire the ability to activate immune cells under inflammatory conditions. Here we inves- tigated whether HA fragments could activate Kupffer cells and analyzed the underlying mechanism. Kupffer cells were isolated from wild-type mice (WT, C3./HeN) and TLR4 mutant mice (C3./HeJ) and HA fragments were produced by the methods of enzyme digestion and chromatography. Then Kupffer cells were stimulated by HA fragments or other control stimuli. The activation of Kupffer cells was estimated as the release of pro-inflammatory cytokines. The activation of p3. MAPK pathway of Kupffer cells was checked and blocking experiments were done as well. The results indicated that HA fragments acquired the ability to activate Kupffer cells in vitro, which was TLR4 dependent and not due to contamination of lipopolysaccharide. Experiments of p3. MAPK kinase inhibition by SB-203.80 verified p3. MAPK was required in HA fragments induced Kupffer cells activation. This suggests that HA fragments, degradative products of one of the major glycosaminoglycans of the extracellular matrix, play critical roles in Kupffer cell activation mediated by TLR4 signaling pathway, which is, at least partially, de- pendent on p3. MAPK activation.

关键词： Kupffer cells hyaluronic acid fragments toll-like receptor 4 p3. MAPK lipoplysaccharide reperfusion injury liver

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Expression and signifi cance of TLR4 and HIF-1α in pancreatic ductal adenocarcinoma

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World Journal of Gastroenterology 2010年第23期16卷 2881-2888页

作者： Jian-Jun Zhang,He-Shui Wu,Lin Wang,Yuan Tian,Jing-Hui Zhang,Hai-Long Wu department of Pancreatic Surgery,Union hospital,tongji {3. College,huazhong university of science and technology,wuhan 43.022,Hubei Province,china department of Pediatrics,Union hospital,tongji {3. College,huazhong university of science and technology,wuhan 43.022,Hubei Province,china Laboratory of General Surgery,Union hospital,tongji {3. College,huazhong university of science and technology,wuhan 43.022,Hubei Province,china department of Pancreatic Surgery Union Hospital Tongji Medical College Huazhong University of Science and Technology Wuhan 430022 Hubei Province China. heshuiwu@***

AIM:To investigate the expression of toll-like receptor(TLR) 4,nuclear factor-κB(NF-κB) p65 and hypoxiainducible transcription factor 1α(HIF-1α) in pancreatic ductal adenocarcinoma and their clinical ***:The mRNA of TLR4 and HIF-1α were investigated by real-time polymerase chain reaction in 3. cases of pancreatic ductal adenocarcinoma and its adjacent tissues,and expression of TLR4,NF-κB p65 and HIF-1α protein were detected by immunohistochemistry in 65 cases of pancreatic ductal adenocarcinoma tissues and 3. cases of corresponding adjacent *** relationship between TLR4 or HIF-1α and pathologic features,as well as the association between TLR4 and HIF-1α,were also ***-Meier method was used to assess the impact of expression of TLR4 and HIF-1α on survival of patients with pancreatic ***:The relative quantif ication of TLR4 and HIF-1α mRNA in tumor tissues was 0.81±0.10 and 0.87±0.11,respectively,signif icantly higher than that in adjacent tissues(0.81±0.10 vs 0.70±0.16,P=0.002;0.87±0.11 vs 0.68±0.13.P=0.000).The protein expression of TLR4,NF-κB p65 and HIF-1α in tumor tissues was 69.20%,66.15% and 70.80%,respectively,being signif icantly higher than that in adjacent normal tissues(69.20% vs 3..50%,P=0.003.66.15% vs 3..58%,P=0.001;70.80% vs 3..80%,P=0.001).There was no signif icant correlation between TLR4 or HIF-1α expression and the age,gender,tumor location,the degree of tumor differentiation in the patients(P>0.05).However,there was signif icant correlation between the expression of TLR4 or HIF-1α and tumor size,lymph node metastasis,venous invasion and clinical staging(P<0.05).The expression of TLR4 and HIF-1α had a signif icant impact on survival of patients with pancreatic ***:TLR4,NF-κB p65 and HIF-1α are overexpressed in pancreatic adenocarcinoma,TLR4 may be partly involved in up-regulating HIF-1α,and both synergestically promote development of pancreatic adenocarcinoma.

关键词： Pancreatic ductal adenocarcinoma Toll-like receptor 4 Nuclear factor-κB p65 Hypoxia-inducible factor 1

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Effects of L-Tetrahydropalmatine on the Expressions of bcl-2 and bax in Rat after Acute Global Cerebral Ischemia and Reperfusion

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Journal of huazhong university of science and technology(medical sciences) 2004年第5期24卷 445-448页

作者：刘彬杨光田 department of emergency Tongji Hospital Tongji Medical College Huazhong University of Science and Technology Wuhan 430030 China

To investigate the effects of L-Tetrahydropalmatine (L-THP) on the expressions of bcl-2, bax and neuronal apoptosis after cerebral ischemia and reperfusion, 60 Wistars rats were randomly divided into 3.groups: sham-operation group (group S, n = 20), ischemic-reperfusion group treated with saline (group I, n=20) and ischemia-reperfusion group treated with L-THP (group T, n=20) .The rat model of global cerebral ischemia and reperfusion was induced by Pulsinelli's four-vessel occlusion method. The expression of bcl-2 and bax mRNA was detected by in situ hybridization and reverse transcriptional polymerase chain reaction (RT-PCR). The number of apoptotic neurons was examined by terminal deoxynucleotidyl-transferase (TdT)-mediated dUTP nick end-labeling (TUNEL) method. Compared with group S, the expression of bcl-2 and bax mRNA in group I was increased significantly (P<0.01), and the number of apoptotic neurons increased either (P< 0.01). After L-THP treatment, the expression of bcl-2 mRNA was up-regulated (P<0.01) and that of bax mRNA was down-regulated (P<0.01); the number of apoptotic neurons was decreased (P<0.01). Our results indicated that bcl-2 may suppress apoptosis and bax promote apoptosis after cerebral ischemia and reperfusion. L-THP could ameliorate cerebral ischemia and reperfusion damage by reducing the apoptosis through regulating bcl-2 and bax.

关键词： cerebral ischemia and reperfusion neuronal apoptosis expression of bcl-2 and bax L-Tetrahydropalmatine

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Influences of dibutyryl cyclic adenosine monophosphate and forskolin on human sperm motility in vitro

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Asian Journal of Andrology 2003年第2期5卷 113-115页

作者： Ji-Hong LIU, Yang LI, Zheng-Guo CAO, Zhang-Qun YEdepartment of Urology, tongji hospital, tongji {3. College, huazhong university of science and technology,wuhan 43.03., china department of Urology Tongji Hospital Tongji Medical College Huazhong University of Science and Technology Wuhan 430030 China

Aim: To study the influences of dibutyryl cyclic adenosine monophosphate (dbcAMP) and forskolin on human sperm motility in vitro. Methods: Semen samples, aseptically obtained by masturbation and prepared by swim-up technique from 20 fertile men, were incubated with different concentrations of dbcAMP and forskolin at 3. 癈. Measurements were carried out after 10 min, 20 min, 3. min and 60 min incubation. Motility parameters were estimated by using an automatic analyzing system. Results: Treatment with dbcAMP or forskolin resulted in a significant increase in sperm motility and progressive motility. The larger the concentrations of dbcAMP or forskolin, the greater the effect appeared. The straight linear velocity and curvilinear velocity were not affected by both agents. Conclusion: dbcAMP and forskolin increase the motility and progressive motility of human sperm in vitro.

关键词： dibutyryl cyclic AMP forskolin sperm motility

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Synthesis of cystine C_(60) derivative and its protective effects on hydrogen peroxide-induced apoptosis in PC12 cells

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Chinese Chemical Letters 2007年第2期18卷 145-148页

作者： Zhen Hu, Hai Ping Xing, Zhou Zhu, Wei Wang, Wen Chao Guan department of Chemistry, huazhong university of science and technology, wuhan 43.074, {3. department of Neurology, tongji hospital, tongji medical College, wuhan 43.03., {3. [a]department of Chemistry Huazhong University of Science and Technology Wuhan 430074 China [b]department of Neurology Tongji Hospital Tongji Medical College Wuhan 430030 China

Oxidative stress has been considered as a major cause of cellular injuries in a variety of clinical abnormalities, especially prominent in neural diseases. One of the usable ways to prevent the reactive oxygen species (ROS)-mediated cellular injury is dietary or pharmaceutical augmentation of some free radical scavenger. Water-soluble amino-fullerene is a novel compound that behaves as a free radical scavenger with excellent biology consistent. In the present study, we have synthesized and characterized a novel cystine C60 derivative for the first time, and investigated the effects on hydrogen peroxide-induced oxidative stress and apoptotic death in cultured rat pheochromocytoma (PC12) cells. PC12 cells treated with hydrogen peroxide underwent apoptotic death as determined by MTT, PI/Hoechst 3.3.2 staining and flow cytometry analysis. These results suggested that cystine C60 derivative has the potential to prevent oxidative stress-induced cell death and has no evident toxicity.

关键词： Cystine C_(60) derivative Reactive oxygen species H_2O_2 Apoptosis PC 12 cells

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PEGylated PLGA Nanoparticles as Tumor Ecrosis Factor-α Receptor Blocking Peptide Carriers:Preparation,Characterization and Release in vitro

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Journal of wuhan university of technology(Materials {3.) 2007年第1期22卷 112-116页

作者：刘卫杨祥良 College of Life science and technology Huazhong University of Science and Technology

To assess the merits of PEGylated poly （lactic-co-glycolic acid）（PEG-PLGA） nanoparticles as drug carriers for tumor necrosis factor-α receptor blocking peptide （TNFR-BP）, PEG-PLGA copolymer, which could be used to prepare the stealth nanoparticles, was synthesized with methoxypolyethyleneglycol, DL-lactide and glycolide. The structure of PEG-PLGA was confirmed with ^1H-NMR and FT-IR spectroscopy, and the molecular weight （MW） was determined by gel permeation chromatography. Fluorescent FITC-TNFR- BP was chosen as model protein and encapsulated within PEG-PLGA nanoparticles using the double emulsion method. Atomic force microscopy and photon correlation spectroscopy were employed to characterize the stealth nanoparticles fabricated for morphology, size with polydispersity index and zeta potential. Encapsulation efficiency （EE） and the release of FITC-TNFR-BP in nanopartieles in vitro were measured by the fluorescence measurement. The stealth nanoparticles were found to have the mean diameter less than 270 nm and zeta potential less than -20 mV. In all nanoparticle formulations, more than 45% of EE were obtained. FITC-TNFR-BP release from the PEG-PLGA nanoparticles exhibited a biphasic pattern, initial burst release and consequently sustained release. The experimental results show that PEG-PLGA nanoparticles possess the potential to develop as drug carriers for controlled release applications of TNFR-BP.

关键词： tumor necrosis factor-α receptor blocking peptide PEG-PLGA stealth nanoparticles ring-opening polymerization controlled and sustained drug release

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Comparison of Arrhythmias among Different Left Ventricular Geometric Patterns in Essential Hypertension

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Journal of huazhong university of science and technology(medical sciences) 2001年第2期21卷 93-96页

作者：郑智周荣梁黔生 department of emergency Medicine Tongji Hospital Tongji Medical College of Huazhong University of Science and Technology Wuhan 430030

The differences of arrhythmias among distinct left ventricular geometric patterns in the patients with essential hypertension were studied. 179 patients with essential hypertension received 24 h dynamic ECG recording, ambulatory blood pressure monitoring, echocardiography examination, etc. According to the examinations, left ventricular geometric patterns and arrhythmias were identified. The comparison of morbidity of arrhythmias between the left ventricular remodeling group and the normal geometric pattern group was performed. The multiple stepwise regression analysis was carried out to identify the independent determinants of arrhythmias. After these predictors were controlled or adjusted, the severity of arrhythmias among different left ventricular geometric patterns was compared. It was found that the morbidity of atrial arrhythmia, ventricular arrhythmia and complex ventricular arrhythmias in the left ventricular remodeling group was significantly higher than in the normal geometric pattern group respectively. There were many independent factors influencing on arrhythmias in essential hypertension. Of all these factors, some indices of left ventricular anatomic structure, grade of hypertension, left atrial inner dimension, E/A, diastolic blood pressure load value at night and day average heart rate and so on were very important. After the above mentioned factors were adjusted, the differences of the orders of arrhythmias between partial geometric patterns were reserved, which resulted from the differences of the geometric patterns. Many factors contributed to arrhythmias of essential hypertension, such as grade of hypertension, LVMI, LA, PWT and so on. The severity of arrhythmias was different in different left ventricular geometric patterns.

关键词： essential hypertension left ventricular remodeling arrhythmia echocardiography

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Imaging changes of the pancreas and the occasion of refeeding in patients with acute pancreatitis

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Hepatobiliary & Pancreatic Diseases International 2002年第2期1卷 290-293页

作者： Ren-Yi Qin Fa-Zu Qiu From the department of Surgery, tongji hospital, tongji {3. College, huazhong university of science and technology, wuhan 43.03., china department of Surgery Tongji Hospital Tongji Medical College Huazhong University of Science and Technology Wuhan 430030 China. ryqin@***

Objective: To explore the objective basis of the time and characterstics of refeeding of patients with acute pancreatitis (AP). Methods: AP patients were randomly divided into routine group (n=105) and special group (n=99). The refeeding time and characteristics of the routine group were based on their levels of blood and urine amylase, function of the gastrointestinal tract, and symptoms and signs, and those of the special group on their imaging changes of ultrasonography and CT of the pancreas. Clinical data from the groups were analyzed prospectively and statistically. Results: At the beginning of refeeding, patients of the two groups showed a high recurrence rate of ab- dominal pain (routine group, 9.1%; special group, 10.5%), but no AP relapse. Three days to 2 weeks after refeeding, the patients of the routine group had a higher recurrence rate of abdominal pain (11.1%) and a relapse rate of AP (14.1%) as compared with those of the special group (P3.days to 4 weeks after refeeding, the patients of the special group had no recurrence of abdominal pain and relapse of AP. Imaging changes of the pancreas and peripancreatic tissue were not consistent with the symptoms and signs of AP patients. The higher the Balthazar CT grading and APACHE-Ⅱ score, the higher the recurrence rate of abdominal pain and the relapse rate of AP after refeeding (P<0. 05). Conclusions: Symptoms and signs were usually dis- cordant to the imaging changes of the pancreas in AP patients. Imaging changes of the pancreas might serve as a basis for the best occasion of refeeding light semi-fluid or light food in AP patients.

关键词： acute pancreatitis pancreatic imaging refeeding

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