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Inosine inhibits apoptosis and cytochrome C mRNA expression in rat neurons after cerebral ischemia/reperfusion

Neural Regeneration Research 2006年第7期1卷 589-592页

作者： Jinrong Wang1, Mingjun Bi1, Qin Li2.1Department of neurology, Rongcheng Second People’s {2., Rongcheng 2.4309, Shandong province, china 2.epartment of neurology, affiliated union hospital, tongji medical college, huazhong university of science and Technology, wuhan 430030,{12., china Department of neurology Rongcheng Second People's Hospital Rongcheng 264309 Shandong Province China Department of neurology Union Hospital Huazhong University of Science ad Technology Wuhan 430030 Hubei Province China

BACKGROUND: It has been demonstrated that adenosine can induce glial cell to release cytochrome C, enhance expression of apoptotic gene bax, inhibit anti-apoptotic gene bcl-2. and activate caspase-3 to apoptosis; Whereas inosine can inhibit neuronal apoptosis which is similar to bcl-2. OBJECTIVE: To observe the effects of inosine on neuronal apoptosis and expression of cytochrome C mRNA in rats after focal cerebral ischemia/reperfusion, and analyze the pathway of its neuroprotective effect. DESIGN: A randomised controlled animal trial. SETTINGS: Department of neurology, Rongcheng Second People's {2.; Department of neurology, affiliated union hospital, tongji medical college, huazhong university of science and Technology. MATERIALS: Sixty-eight rats, weighing 2.0-2.0 g and clean grade, were used. TdT-mediated dUTP-biotin nick end labeling (TUNEL) and cytochrome C mRNA in situ hybridization kits and DAB staining kit were purchased from wuhan Boster Biological Co., Ltd.; Inosine injection [2.0 mg (2.mL) each] from Qingdao First Pharmaceutical Factory. METHODS: The experiment was accomplished in the animal experimental center in tongji medical college of huazhong university of science and Technology from December 2.03 to June 2.05. ① Sixty-four rats were made into focal ischemia by middle cerebral artery occlusion (MCAO) with a nylon monofilament suture. The successfully induced rats were assigned to inosine group (n =32. and model group (n =32. at random. Rats in the inosine group were intraperitoneally administrated with inosine in dose of 100 mg/kg preoperatively, twice a day, 7 days in all. The rats in the control group were injected with the same dose of saline solution by the similar way preoperatively. Each group was randomized into ischemia /reperfusion 2. 6, 12. 2. hours, 2. 3, 7 and 14 days subgroups consisted of 4 rats. The other 4 rats were taken as the sham-operated group, the rats were given the same treatment except for not introduced the filament in

关键词： mRNA Inosine inhibits apoptosis and cytochrome C mRNA expression in rat neurons after cerebral ischemia/reperfusion

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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 {2.,tongji medical college,huazhong university of science and Technology,wuhan 43002.,hubei province,{12. Department of Pediatrics,union {2.,tongji medical college,huazhong university of science and Technology,wuhan 43002.,hubei province,{12. Laboratory of General Surgery,union {2.,tongji medical college,huazhong university of science and Technology,wuhan 43002.,hubei province,{12. 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 30 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 38 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 69.2.%,66.15% and 70.80%,respectively,being signif icantly higher than that in adjacent normal tissues(69.2.% vs 39.50%,P=0.003;66.15% vs 31.58%,P=0.001;70.80% vs 36.80%,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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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 {2. college,huazhong university of science and Technology,wuhan 430030,china 3 Department of Anesthesiology,4 Department of Nephrology,{12. hospital,tongji {2. college,huazhong university of science and Technology,wuhan 43002..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 12.ell 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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The Study on BCR/ABL Fusion Gene in Adult Acute Lymphoblastic Leukemia

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wuhan university Journal of Natural {2.s 2001年第3期6卷 742-746页

作者： Li Hui yu 1,2.,Zuo Guo Lin 1, ,Zuo Ping 2.1.college of Life science, {2. university,{2. 430072.china 2.union hospital,tongji medical college,huazhong university of science and Technology,{2. 43002.,china wuhan university College of Life Science Wuhan China (GRID:grid.49470.3e) (ISNI:***) huazhong university of {2. and Technology Union Hospital Tongji Medical College Wuhan China (GRID:grid.33199.31) (ISNI:***) wuhan university College of Life Science Wuhan China (GRID:grid.49470.3e) (ISNI:***) huazhong university of {2. and Technology Union Hospital Tongji Medical College Wuhan China (GRID:grid.33199.31) (ISNI:***)

In order to assess the significance of BCR/ABC fusion gene in adult acute lymphoblastic Leukemia (ALL), 2. patients who were diagnosed as ALL were enrolled to detect BCR/ABC gene using nested-RT-PCR. The results showed that 9 cases (31.2.%) were BCR/ABL positive, and expressed P2.0 subtype. Among them 7 cases were B-ALL, and one was T-ALL. The diagnosis was proved by monoclonal antibodies recognition by indirect immunofluorescence. Adult patients with BCR/ABL positive ALL were significantly older (p0.05). The induction failure rate was high in BCR/ABL positive patients and those who achieved complete remission usually relapsed earlier. In conclusion, adult ALL patients with BCR/ABL-positive have poorer prognosis.

关键词： Key words BCR/ABL gene acute lymphoblastic leukemia nested RT PCR

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NMDA and AMPA receptors mediate intracellular calcium increase in rat cortical astrocytes

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Acta Pharmacologica Sinica 2004年第6期25卷 12-18页

作者： Bo HU, Sheng-gang SUN2. E-tang TONGneurology Department, {2. hospital, tongji college, huazhong university of science and Technology, wuhan 43002., china neurology Department Union Hospital Tongji College Huazhong University of Science and Technology Wuhan 430022 China

AIM: To study the effect of glutamate on the intracellular calcium signal of pure cultured rat astrocytes and the roleof N-methyl-D-aspartate (NMDA) and alpha-amino-3-hydroxy-5-methyl-4-isoxazole-propionic acid (AMPA) re-ceptors in the procedure. METHODS: The fluorescence of calcium was measured by Fura-2.AM (F345/F380).RESULTS: L-Glutamate induced [Ca2.]i increase in most of the cells in concentration- and time-dependent *** 50 mmol/L induced the fluorescence increase by almost three to four times, while the effect of AMPA 50mmol/L was just half of that of D-(-)-2.amino-5-phosphonopentanoic acid (D-AP-5; a selective antagonist of theNMDA receptor). 6-Cyano-7-nitroquinoxaline-2.3-dione (CNQX, a selective antagonist of the AMPA receptor)abolished the effects of NMDA and AMPA, respectively. D-AP-5 and CNQX simultaneously or respectively attenu-ated the effect of L-glutamate at different degrees, but could not abolish it entirely. CONCLUSION: Glutamatemodulated intracellular Ca2. of pure cultured rat astrocytes through different pathways. The activation of NMDAand AMPA receptors took part in the complex mechanisms.

关键词： glutamate astrocytes NMDA receptors AMPA receptors calcium

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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 {2. affiliated to huazhong university of science and Technology, wuhan 43002., 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 {2. affiliated to huazhong university of science and Technology, wuhan 43002., 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 {2. affiliated to huazhong university of science and Technology, wuhan 43002., 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, C3H/HeN) and TLR4 mutant mice (C3H/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 p38 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 p38 MAPK kinase inhibition by SB-2.3580 verified p38 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 p38 MAPK activation.

关键词： Kupffer cells hyaluronic acid fragments toll-like receptor 4 p38 MAPK lipoplysaccharide reperfusion injury liver

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

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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 {2.,tongji medical college,huazhong university of science and Technology,wuhan 43002.,china 2.epartment of Anesthesia,union {2.,tongji medical college,huazhong university of science and Technology,wuhan 43002.,china 3Department of General Surgery,union {2.,tongji medical college,huazhong university of science and Technology,wuhan 43002.,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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Therapeutic effects of berberine in impaired glucose tolerance rats and its influence on insulin secretion

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Acta Pharmacologica Sinica 2004年第4期25卷 98-104页

作者： San-hua LENG, Fu-er LU2. Li-jun XU Institute of Integrated Traditional Chinese & Western Medicine, tongji hospital, tongjimedical college, huazhong university of science & Technology, wuhan 430030, china Institute of Integrated Traditional Chinese & Western Medicine Tongji Hospital Tongji Medical College Huazhong University of Science & Technology China

AIM: To explore the anti-diabetic effects of berberine and its influence on insulin secretion. METHODS: Impaired glucose tolerance rats induced by iv injection of streptozotocin 30 mg/kgwere treated with berberine 187.5 and 562.5 mg/kg while fed with high fat laboratory chow. After rats were treated for 4 weeks, oral glucose tolerance was determined, and for 8 weeks, the fasting blood glucose, insulin, lipid series were determined. In insulin secretion experiments, berberine 93.75, 187.5, and 562.5 mg/kg was administered orally to BALB/c mice at a bolus. The murine serum was collected 2.h after the berberine administration for insulin determination. Insulin released from HIT-T15 cells and pancreatic islets incubated with berberine 1-100 μmol/L for 12.h was determined. RESULTS: The levels of fasting blood glucose (7.4±1.5 or 7.3±1.3 vs 9.3±1.3 mmol/L), triglycerides (0.61±0.2. or 0.63± 0.17 vs 1.8±0.7 mmol/L), total cholesterol (1.8±0.3 or 1.9±0.3 vs 2.2.0.2.mmol/L), free fatty acid (456±93 or 460±72.vs 550±113 μmol/L) and apolipoprotein B (0.37±0.02.or 0.42.0.05 vs 0.46±0.04 g/L) were reduced greatly in berberine-treated groups at doses of 187.5 and 562.5 mg·kg-1·d-1, respectively as compared with those in control group (P2..5±2.7 or 2.±4 or 2.±4 vs 2..3±2.8 pIU/L) was undoubtedly promoted and blood glucose (4.52.0.31 or 4.45±0.2. or 4.30±0.19 vs 4.87±0.2. mmol/L) was reduced after berberine administration at doses of 93.75, 187.5, and 562.5 mg/kg, respectively in the BALB/c mice. CONCLUSION: Berberine possesses anti-diabetic effec

关键词： berberine diabetes mellitus glucose tolerance test insulin HIT-T15 cells pancreatic islets lipids

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Risk Factors of Learning Disabilities in Chinese Children in wuhan

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Biomedical and Environmental sciences 2003年第4期16卷 392-397页

作者： BIN YAO AND HAN-RONG WUDepartment of Children and Adolescent Health, Maternity and Child Health Care, tongji medical college, {2. university of science and Technology, wuhan 430030, hubei, china Department of Children and Adolescent Health Maternity and Child Health Care Tongji Medical College Huazhong University of Science and Technology Wuhan 430030 Hubei Chin

Objective To investigate prevalence rate of learning disabilities (LD) in Chinese children, and to explore related risk factors, and to provide theoretical basis for preventing such disabilities. Methods One thousand and one hundred fifty one children were randomly selected in primary schools. According to criteria set by ICD-10, 118 children diagnosed as LD were classified into the study group. Four hundred and ninety one children were classified into the normal control group. Five hundred and forty two children were classified into the excellent control group. The study instruments included PRS (The pupil rating scale revised screening for learning disabilities), Conners' children behavior check-list taken by parents and YG-WR character check-list. Results The prevalence rate of LD in Chinese children was 10.3%. Significant differences were observed between LD and normally learning children, and between the LD group and the excellent group, in terms of scores of Conners' behavior check-list (P china. LD is resulted from a number of factors. Good studying and living environments should be created for LD children.

关键词： Learning disabilities (LD) Behavior problems Character Children Risk factors

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Rapid Inhibition of the Glutamate-induced Increase of Intracellular Free Calcium by Magnesium in Rat Hippocampal Neurons

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Journal of huazhong university of {2. and Technology(medical {2.s) 2004年第5期24卷 424-426页

作者：张蕲胡波孙圣刚邓学军梅元武童萼塘 Department of Phathophysiology School of Basic Medical Sciences Tongji Medical College Huazhong University of Science and Technology Wuhan 430030 China Department of neurology Union Hospital Tongji Medical College Huazhong University of Science and Technology Wuhan 430030 China

By using Fura-2.AM, the effects of magnesium (Mg 2.) on the glutamate-induced increase of intracellular free calcium ([Ca 2.]i) in the cultured hippocampal neurons and the features were investigated by integrated photoelectric detecting system. The experiments were designed to three groups (The drug was spit to the cells for 2. s): Group A receiving 1×10 —5 mol/L glutamate; Group B receiving 1×10 —5 mol/L glutamate and1×10 —5 mol/L Mg 2. simultaneously; Group C receiving 1×10 —5 mol/L glutamate again after [Ca 2.]i in group B back to the baseline. The results showed that in group A, [Ca 2.]i was obviously increased. In group B, the changes in [Ca 2.]i and the peak value were significantly decreased. Moreover, the elevation of Phase 1 was slowed down and Phase 2.was shortened to some extent, and the plateau phase between them was relatively prolonged. In group C, calcium oscillation similar to that in group A occurred, but both the Phase 1 and Phase 2.were shortened and the △[Ca 2.]i was slightly decreased. It was suggested that Mg 2. could quickly inhibit the rise of [Ca 2.]i induced by glutamate in the cultured hippocampal neurons in rats.

关键词： magnesium glutamate intracellular free calcium

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