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Acta Pharmacologica Sinica 2005年第9期26卷 1039-1044页

作者： Xiao-fei AN Hai-ping CHEN Shu-lan MA Yi FENG Jun-wei HAO Bo-ying CHEN~1 Department of Neurobiology and Integrative Medicine,shanghai medical college,Fudan university shanghai 200032,china Department of Neurobiology and Integrative Medicine Shanghai Medical College Fudan University Shanghai China

Aim:To investigate effect of the nociceptin/orphanin FQ(OFQ)on hypothala-mus gonadotropin-releasing hormone(GnRH)release in ovariectomized(OVX)***:GnRH radioimmunoassay(RIA)was used to study the effect ofOFQ on GnRH release in hypothalamus slices in ***-pull perfusion andintracerebroventicular(icy)injection were used to examine the effect of OFQ onGnRH release in the hypothalamus medial preoptic area(POA)in ***-mies were performed on female Sprague-Dawley rats,and their plasma luteinizinghormone(LH)levels were measured after icy injection of OFQ with or without[Nphe~1]NC(1-13)NH2,a competitive antagonist of opioid receptor-like 1 receptor(ORL1 receptor).Reverse transcription-polymerase chain reaction(RT-PCR)wasused to investigate the expression of the ORLI receptor in rat ***:GnRH release from hypothalamus slices was inhibited 90 min after the administra-tion of 2 mmol/L and 20 mmol/L OFQ(P3)NH2,that is,NC ***,plasma LH levels in OVX rats increased markedly 2 h after icv injec-tion of 100 nmol and 200 nmol NC ***-PCR analysis further revealed that theORL1 receptor was not expressed in the pituitary of OVX ***:Cen-tral administration of nociceptin/orphanin FQ might inhibit the release of hypo-thalamic GnRH and decrease the plasma LH levels through ORLI receptors inOVX rats.

关键词： nociceptin ORL1 receptor gonadorelin hypothalamus

Beneficial effects of melatonin in experimental models of Alzheimer disease

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Acta Pharmacologica Sinica 2006年第2期27卷 129-139页

作者： Yong CHENG~2 Zheng FENG~3 Qing-zhu ZHANG~4 Jun-tian ZHANG~(2.5) ~2Department of Pharmacology,Institute of Materia Medica,Chinese Academy of medical Sciences and Peking Union medical college,Beijing 100050,china:~3Department of Anesthesiology,university of California,San Diego,and Veterans Affairs medical Center,3350 La Jolla Village Drive,San Diego,CA 92161,USA:~4Department of Pharmacology,college of pharmacy,Shandong university,Jinan 250012,china Department of Pharmacology Institute of Materia Medica Chinese Academy of Medical Sciences and Peking Union Medical College Beijing China Department of Anesthesiology University of California San Diego Veterans Affairs medical Center San Diego USA Department of Pharmacology College of Pharmacy Shandong University Jinan China

Alzheimer’s disease(AD),a progressive degenerative disorder,is characterizedby the presence of amyloid deposits,neurofibrillary tangles and neuron *** evidence indicates that antioxidants could be useful either for the pre-vention or treatment of *** has been shown that melatonin is a potent antioxi-dant and free radical ***,melatonin stimulates severalantioxidative enzymes and improves mitochondrial energy *** find-ings led us to study amyloid precursor protein transgenic mice,ovariectomizedrats,and pheochromocytoma and astroglioma cell lines,to observe whether mela-tonin had any effect on Alzheimer’s symptoms or pathological *** foundthat melatonin had many beneficial effects in experimental models of AD,includ-ing improvement of cognitive function,anti-oxidative injury,anti-apoptosis,inhi-bition of β-amyloid(Aβ)deposition and Aβ fiber *** groups haveshown that melatonin has an inhibitory effect on tau protein *** actions may potentially slow down or stop the progression of dementia.

关键词： melatonin Alzheimer disease beta-amyloid protein calcium overload APP transgenic mice ovariectomized rats tau protein hyperphosphorylation

Transcriptional Regulation of Urokinase Receptor in High-(95D) and Low-metastatic (95C) Human Lung Cancer Cells

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Acta Biochimica et Biophysica Sinica 2004年第6期36卷 405-411页

作者： Chun-Yang LI, Li TAN, Guo-Jing ZHANG, Ping LI, Chang TONG, Jing FAN, and Yun-Song ZHU* Key Laboratory, the Education Ministry, Department of Biochemistry and Molecular Biology, shanghai medical college,Fudan university, shanghai 200032, china Key Laboratory the Education Ministry Department of Biochemistry and Molecular Biology Shanghai Medical College Fudan UniversityShanghai 200032 China

To study the transcriptional regulation of urokinase receptor (uPAR) in high- (95D) and low-metastatic (95C) human lung cancer cells, we performed PCR to amplify 2238 bp uPAR promoter from 95C and 95D cells. According to the results of sequencing, five different bases are found in uPAR promoter between 95C and 95D cells. The results of luciferase activity assay showed that these differences have no significant effect on the uPAR promoter activity. Based on a normal uPAR promoter, progressive truncated mutants were constructed. The transient transfection/reporter assay showed that the promoter region from –136 to +9 may interact with relevant nuclear factors, which result in different levels of uPAR expression between 95C and 95D cells.

关键词： uPAR promoter transcriptional regulation

Germline mutation analysis of hPMS2 gene in Chinese families with hereditary nonpolyposis colorectal cancer

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World Journal of Gastroenterology 2010年第30期16卷 3847-3852页

作者： Xia Sheng, Xiao-Yan Zhou, Xiang Du, Tai-Ming Zhang, WeiQi Sheng, Da-Ren Shi, Department of Pathology, shanghai Cancer Center, Fudan university, shanghai 200032, china Department of Oncology, shanghai medical college, Fudan university, shanghai 200032, china Heng-Hua Zhou, Department of Pathology, shanghai Ninth People’s Hospital Affi liated to shanghai Jiao Tong university School of Medicine, shanghai 200011, china San-Jun Cai, Department of Abdominal Surgery, Cancer Center, Fudan university, shanghai 200032, china Department of Pathology Shanghai Cancer Center Fudan University Shanghai 200032 China.

AIM: To study the germline mutation of hPMS2 gene in 26 unrelated Chinese hereditary nonpolyposis colorectal cancer (HNPCC) probands and to fulfill the screening strategy for HNPCC in Chinese. METHODS: Genomic DNA was extracted from the peripheral blood. To avoid the interference of pseudogene in detection of the remaining 11 exons (exon 1-5, 9, 11-15), long-range polymerase chain reaction (PCR) was conducted to amplify the complete coding region of hPMS2 gene firstly. Then 1/8 of the PCR productswere used as template to amplify the individual exon respectively and DNA sequencing was done. Direct DNA sequencing of the conventional PCR products of exon 6, 7, 8 and 10 of hPMS2 gene was performed. The same analysis was made in 130 healthy persons without family histories of HNPCC to further investigate the pathological effects of the detected missense mutation. RESULTS: One HNPCC proband fulf illed Bethesda guidelines and was found to carry the germline mutation of hPMS2 gene, which has not been reported in Chinese HNPCC families. It was a missense mutation at c.1532C>T of exon 11. It was detected in three controls as well with an occurrence rate of 2.3% (3/130). Since it could not be found in the PMS2-single nucleotide polymorphism (SNP) database, this missense mutation is a new SNP unreported up to date. Meanwhile, 260 reported SNPs of hPMS2 gene were detected in the 26 HNPCC probands. The 2nd and 5th exons were probably the hot SNP regions of hPMS2 gene in Chinese HNPCC families involving 53.1% of all reported SNP. CONCLUSION: The germline mutation of hPMS2 gene may be rare in Chinese HNPCC families. The 2nd and 5th exons are hot SNP regions of hPMS2 gene.

关键词： Hereditary nonpolyposis colorectal cancer hPMS2 Missense mutation Single nucleotide polymorphism Colorectal cancer

Electrophysiological evidences for the contribution of NMDA receptors to the inhibition of clonidine on the RVLM presympathetic neurons

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海军军医大学学报 2004年第12期25卷 1328-1328页

作者： Wang WZ,Yuan WJ,Tang CS,Su DF(Department of Physiology,college of Basic medical Sciences,second military medical university ,shanghai 200433,china) Department of Physiology College of Basic Medical SciencesSecond Military Medical UniversityShanghai 200433China

The main objective of this study is to test the hypothesis that N-methyl-D-aspartate(NMDA) receptors within the rostral ventrolateral medulla(RVLM) are involved in the inhibition of clonidine on the RVLM presympathetic neurons. Totally,22 presympathetic neurons were recorded in anesthetized and paralyzed rats. The majority of these neurons(n=16 of 22) were significantly inhibited by iontophoretic(30 nA) clonidine,the other 6 neurons were insensitive to clonidine. In seven clonidine-sensitive neurons,iontophoretic clonidine(30 nA) antagonized the neuronal excitation of iontophoretic NMDA receptor agonist NMDA(20 nA). In remaining nine clonidine-sensitive neurons,iontophoretic NMDA receptor antagonist MK801 (60 nA) significantly attenuated the neuronal inhibition of iontophoretic(30 nA) clonidine. In conclusion,these results suggest that NMDA receptors contribute to the inhibition of clonidine on the RVLM presympathetic neurons.

关键词： NMDA Electrophysiological evidences for the contribution of NMDA receptors to the inhibition of clonidine on the RVLM presympathetic neurons

Role of hypothalamus nociceptin/orphanin FQ in pre-ovulatory luteinizing hormone surge of estrogen and progesterone-primed,ovariectomized rats

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Acta Pharmacologica Sinica 2007年第8期28卷 1189-1197页

作者： Xiao-fei AN~(1,3) Jiang-yi YU~1 Yi FENG~2 Bo-ying CHEN~2 Su-lin ZHANG~1 ~1Department of Endocrinology,Jiangsu Province Hospital of Traditional Chinese Medicine,Nanjing 210029,china ~2Department of Neurobiology and Integrative Medicine,shanghai medical college,Fudan university,shanghai 200032,china Department of Endocrinology Jiangsu Province Hospital of Traditional Chinese Medicine Nanjing China Department of Neurobiology and Integrative Medicine Shanghai Medical College Fudan University Shanghai China

Aim:To investigate the role of hypothalamus nociceptin/orphanin FQ（OFQ）andits endogenous receptor,the opioid receptor-likel receptor（ORL1 receptor）in theestrus cycle of female ***:Radioimmunoassay was used to detect theeffect of the intracerebroventricular（icy）administration of OFQ and/or the ORL1receptor antagonist[Nphe1]Nociceptin（1-13）NH2,that is,NC13 on luteinizinghormone（LH）levels of estrogen- and progesterone（EBP）-primed,ovariectomized（OVX）rats（EBP-primed OVX rats）.RT-PCR,Western blotting,and immunohis-tochemistry techniques were adopted to observe the changes of OFQ and theORL1 receptor in the pre-optic area（POA）and the medial basal hypothalamus（MBH）of the estrus cycle of female ***:Pre-ovulatory LH surges in EBP-primed,OVX rats were significantly reduced by icv administration of 20 and 200nmol OFQ（P<0.05）,and the effect of 20 nmol OFQ could be abolished by pretreat-merit with 20 nmol *** OFQ mRNA level in the POA on pro-estrus waslowered markedly compared to diestrus and estrus（P<0.05）,while the mRNA andprotein levels of the ORL1 receptor showed no significant changes in the POAand MBH across the estrus ***,the number of OFQ-immunoreac-tive neurons in the medial POA,ventromedial hypothalamus,and the arcuatenucleus on pro-estrus was significantly decreased compared to diestrus and es-tins（P<0.05）.Conclusion:The inhibitory effect of OFQ on the LH surge of EBP-primed,OVX rats and its downregulation in POA and MBH on pro-estrus sug-gests that it might play a negative modulatory role in the estrus cycle.

关键词： orphanin FQ opioid receptor-like 1 receptor luteinizing hormone estrus cycle hypothalamus

A novel homospermidine conjugate inhibits growth and induces apoptosis in human hepatoma cells

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Acta Pharmacologica Sinica 2007年第11期28卷 1827-1834页

作者： Song-qiang XIE~(2,3),Ying-liang WU~2,Peng-fei CHENG~4,Min-wei WANG~2,Guang-chao LIU~5,Yuan-fang MA~5,Jin ZHAO~4,Chao-jie WANG~(4,6)~2 Department of Pharmacology,Shenyang Pharmaceutical university,Shenyang 110016,china ~3 pharmacy college,Henan university,Kaifeng 475001,china ~4 Institute of Natural Products and Medicinal Chemistry,Henan university,Kaifeng 475001,china:~5 medicalcollege,Henan university,Kaifeng 475001,china Department of Pharmacology Shenyang Pharmaceutical University Shenyang China pharmacy college Henan University Kaifeng China Institute of Natural Products and Medicinal Chemistry Henan University Kaifeng China medical college Henan University Kaifeng China

Aim:To elucidate the mechanism responsible for the antiproliferative effects of anovel homospermidine conjugate,anthracenylmethyl homospermidine(ANTMHspd),in the human hepatoma BEL-7402 cell ***:The viabilityof the cells was assessed by MTT assay and the trypan blue dye exclusion *** changes were observed by fluorescence microscopy with Hoechst33258 *** cycle distribution,apoptosis,and mitochondrial membranepotential were measured by flow *** expression was detected byWestern blot ***:ANTMHspd strongly decreased BEL-7402 cellproliferation in a dose-and time-dependent *** 33258 staining andthe flow cytometry assay showed that ANTMHspd induced cell apoptosis andcell cycle ***,ANTMHspd could induce mitochondrialmembrane potential loss and cytochrome c release and enhance cleaved caspase-3,cleaved caspase-9,and Bax protein expression without caspase-8 *** could also decrease the expression of Bcl-2 and cytochrome c *** addition,the specific inhibitors of caspase-9 and caspase-3almost abolished the ANTMHspd-induced caspase-9 and caspase-3 activation,***:ANTMHspd could induce BEL-7402 cell apoptosis viathe mitochondrial/caspase-dependent pathway and the Bcl-2 family was involvedin the control of apoptosis.

关键词： polyamine apoptosis cell cycle mitochondrial membrane potential caspase Bcl-2

Detection of germline mutations of hMLH1 and hMSH2 based on cDNA sequencing in china

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World Journal of Gastroenterology 2005年第42期11卷 6620-6623页

作者： Chao-Fu Wang, Xiao-Yan Zhou, Tai-Ming Zhang, Meng-Hong Sun, Da-Ren Shi, Laboratory of Molecular Pathology, Cancer Hospital of Fudan university Department of Oncology, shanghai medical college of Fudan university, shanghai 200032, china Laboratory of Molecular Pathology Cancer Hospital of Fudan University Shanghai 20032 China.

AIM： TO detect the germline mutations Of hMLH1 and hMSH2 based on mRNA sequencing to identify hereditary non polyposis oolorectal cancer （HNPCC） families. METHODS： Total RNA was extracted from peripberal blood of 14 members from 12 different families fulfilling Amsterdam criteria II. mRNA of hMLH1 and hMSH2 was reversed with special primers and heat-resistant reverse tmnscriptase, cDNA was amplified with expand long template PCR and cDNA sequendng analysis was followed. RESULT： Seven germline mutations were found in 6 families （6/12, 50%）, in 4 hMLH1 and 3 hMSH2 mutations （4/12, 33.3%）; （3/12, 25%）. The mutation types involved 4 missense, 1 silent and 1 frame shift mutations as well as 1 mutation in the non-coding area. Four out of the seven mutations have not been reported previously. The 4 hMLH1 mutations were distributed in exons 8, 12, 16, and 19. The 3 hMSH2 mutations were distributed in exons 1 and 2. Six out of the 7 mutations were pathological, which were dislTibuted in 5 HNPCC families. CONCLUSION： Germline mutations of hMLH1 and hMSH2 can be found based on cDNA sequencing so as to identify HNPCC family, which is highly sensitive and has the advantages of cost and time saving.

关键词： hMLH1 hMSH2 Colorectal cancer Hereditarynon-polyposis Reverse transcription Germline mutation

Trehalose is an important mediator of Cap1p oxidative stress response in Candida albicans

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Trehalose is an important mediator of Cap1p oxidative stress...

Formation of 4'-carboxyl acid metabolite of imrecoxib by rat liver microsomes

第三届中日双边药理学和临床药理学学术会议

作者： CAO Ying-Ying~1 JIANG Yuan-Ying~2 (1.Department of Biochemical pharmacy,2.Department of Pharmacology,School of pharmacy,the second military medical university,shanghai 200433,china)

AIM The nonreducing disaccharide treha- lose,which accumulates dramatically during stationary phase or under oxidative stress,enhances the resistance of the cells to *** we investigated the significance of trehalose and its putative correlation with Capl p,which is a basic region-leucine zipper (bZip) transcription factor participated in oxidative stress tolerance in Candida albi- *** *** strain CAI4 (CAP1/ CAP1) and *** strain CJD21 (caplΔ::hisG/ caplΔ::hisG) were *** content was measured by *** mRNA expression of the genes involved in trehalose biosynthesis was assayed by real time RT- *** Trehalose did not accumulate in the capl/capl mutant during stationary phase compared to the wild type strain *** the mutant was exposed to high concentration of HO,trehalose accumulation was still not *** both of the conditions above,the capl/capl mutant showed high sensitivity to HO,and the cell viability was rather ***,when exog- enous trehalose was added to the culture of the capl/capl mutant,the tolerance of this strain to oxidative stress was *** transcript levels of TPS2 and TPS3 were increased in the CAI4 strain compared to that in capl/ capl mutant when exposed to *** Trehalose accumulation is important to the oxidative stress tolerance mediated by Capl p in ***.

关键词： Candida albicans trehalose Capl p oxidative stress

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Acta Pharmacologica Sinica 2006年第4期27卷 506-512页

作者： Hai-yan XU~2 Peng ZHANG~2 Ai-shen GONG~3 Yu-ming SUN~2 Feng-ming CHU~4 Zong-ru GUO~4 Da-fang ZHONG~(2,5,6)2 Laboratory of Drug Metabolism and Pharmacokinetics,Shenyang Pharmaceutical university,Shenyang 110016,china 3 Research and Development Center,shanghai Hengrui Pharmaceutical Co,shanghai 200245,china:4 Institute of Materia Medica,Chinese Academy of medical Sciences and Peking Union medical college,Beijing 100050,china 5 shanghai Institute of Materia Medica,Chinese Academy of Sciences,shanghai 201203,china Laboratory of Drug Metabolism and Pharmacokinetics Shenyang Pharmaceutical University Shenyang China Research and Development Center Shanghai Hengrui Pharmaceutical Co Shanghai China Institute of Materia Medica Chinese Academy of Medical Sciences and Peking Union Medical College Beijing China shanghai Institute of Materia Medica Chinese Academy of Sciences Shanghai China

Aim:Imrecoxib is a novel and moderately selective COX-2 *** aim ofthe present in vitro investigation was to study the formation of the major metabo-lite 4′-carboxylic acid imrecoxib（M2）and identify the enzyme（s）involved in ***:The formation of M2 was studied in rat liver cytosol in theabsence or presence of liver *** formed metabolite was identifiedand quantified by LC/MS~*** addition,to characterize the cytochrome P450（CYP）isozymes involved in M2 formation,the effects of typical CYP inhibitors（such asketoconazle,quinine,α-naphthoflavone,methylpyrazole,and cimetidine）on theformation rate of M2 were ***:The formation of M2 from 4’-hydroxymethyl imrecoxib（M4）was completely dependent on rat liver microsomesand *** kinetic studies demonstrated that the formation rate of M2conformed to monophasic Michaelis-Menten *** experimentsshowed that the formation of M2 was induced significantly by dexamethasoneand lowered by ketoconazole strongly and ***,other CYP inhibitors,such as α-naphthoflavone,cimetidine,quinine,and methylpyrazole had no inhibitory effects on this metabolic ***:These biotransformation studies of M4 and imrecoxib in rat liver atthe subcellular level showed that the formation of M2 occurs in rat liver microsomesand is *** reaction was mainly catalyzed by CYP 3A inuntreated rats and in dexamethasone-induced *** CYP,such as CYP 1A,2C,2D,and 2E,seem unlikely to participate in this metabolic pathway.

关键词： imrecoxib cytochrome P450 metabolism liver microsomes rats