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Effect of Porphyromonas gingivalis PrtC on cytokine expression in ECV304 endothelial cells and its level in subgingival plaques from patients with chronic periodontitis

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Acta Pharmacologica Sinica 2007年第7期28卷 1015-1023页

作者： Yan-min WU~2,Li-li CHEN~(2,6),Jie YAN~(3,6),Chun-yan ZHUANG~4,Zhi-yuan GU~5~2 department of Stomatology,the Second affiliated hospital,College of medicine,Zhejiang university,Hangzhou 310009,china ~3 Depart-ment of Medical Microbiology and Parasitology,College of medicine,Zhejiang university,Hangzhou 310006,china ~4 department ofstomatology,the affiliated hospital of the school of medicine,ningbo university,ningbo 315020,china ~5 department of Oral andMaxillofacial Surgery,the affiliated hospital of Stomatology,College of medicine,Zhejiang university,Hangzhou 310006,china department of Stomatology the Second Affiliated Hospital College of Medicine Zhejiang University Hangzhou China department of Medical Microbiology and Parasitology College of Medicine Zhejiang University Hangzhou China department of Stomatology the Affiliated Hospital of the School of Medicine Ningbo University Ningbo China department of Oral and Maxillofacial Surgery the Affiliated Hospital of Stomatology College of Medicine Zhejiang University Hangzi China

Aim:To investigate the effect of the collagenase gene (prtC) product ofPorphyromonas gingivalis on inducing host cells to secrete inflammatorycytokines,and to discuss the correlation between the PrtC level in subgingivalplaque samples and clinical ***:A prokaryotic expression sys-tem pET32a-prtC-Escheria coli BL21 DE3 was *** and im-munoreactivity of the recombinant PrtC protein (rPrtC) was identified by *** was applied to detect interleukin (IL)-1α,IL-8,and TNF-α levelsin supernatants from rPrtC-induced human umbilical vein endothelial cells (HUVEC)originated ECV304 *** parameters recorded at baseline and after treat-ment included bleeding on probing (BOP),probing depth (PD),and attachmentloss (AL).ELISA was established to measure the PrtC level in 196 subgingivalplaque samples from 49 patients with chronic ***:After co-incubation with 1 μg/mL rPrtC for 24 h and with 5 or 10 μg/mL rPrtC for 12 h,thelevels of IL-1α,IL-8,and TNF-α secreted by the ECV304 cells increased signifi-candy (P6 mm PDsites was higher than that in the BOP-negative or the ≤2 mm AL or ≤6 mm PD sites(P6 mm PD sites,the PrtC levels changed insignifi-cantly (P>0.05).Conclusion:rPrtC is able to directly induce host cells to synthe-size and secrete IL-1α,IL-8,and TNF-α.The PrtC level in subgingival samples iscorrelated with BOP,AL,and PD.

关键词： Porphyromonas gingivalis collagenase cloning cytokine ELISA

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Anticancer effect of aloe-emodin on cervical cancer cells involves G2/M arrest and induction of differentiation

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Acta Pharmacologica Sinica 2007年第12期28卷 1991-1995页

作者： Jun-ming GUO~(2,3),Bing-xiu XIAO~2,Qiong LIU~2,Shun ZHANG~(2,4),Dong-hai LIU~(2,4),Zhao-hui GONG~2~2 ningbo university school of medicine,ningbo 315211,china ~4 affiliated hospital(ningbo № 2 hospital),ningbo university school ofmedicine,ningbo 315010,china ningbo university school of medicine Ningbo China affiliated hospital (ningbo No 2 hospital) Ningbo University School of Medicine Ningbo China

Aim:The aim of this study was to investigate the effects of aloe-emodin,a naturalcompound from the root and rhizome of Rheum palmatum,on the growth of hu-man cervical cancer cells,***:HeLa ceils were treated with variousconcentrations of aloe-emodin for 1-5 d,and cell growth was measured by 3-（4,5-dimethylthiazol-2-yl）-2,5-diphenyl tetrazolium bromide *** long-termgrowth effect was investigated by crystal violet *** distributions of thecell cycle and apoptosis were analyzed by flow *** alkaline phos-phatase （ALP） activity was analyzed by a chemical ***,Westernblotting was used to indicate the abundant changes of protein kinase C （PKC）,c-myc,cyclins,cyclin-dependent kinases （CDK）,and proliferating cell nuclear anti-gen （PCNA）.Results:Aloe-emodin inhibited the growth of HeLa cells in a dose-dependent manner at concentrations ranging between 2.5 and 40 μmol/*** cytometric analysis showed that HeLa cells were arrested at the G2/M *** effect was associated with the decrease in cyclin A and CDK2,and theincrease in cyclin B1 and *** importantly,the ALP activity was found tobe increased by aloe-emodin treatment,and accompanied by the inhibition ofPCNA *** addition,aloe-emodin suppressed the expression of PKCαand ***:These findings provide a possible mechanistic explana-tion for the growth inhibitory effect of aloe-emodin on HeLa,which includes cellcycle arrest and inducing differentiation.

关键词： HeLa cells cell cycle aloe-emodin alkaline phosphatase c-myc

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Effects of CpG oligodeoxynucleotide on transcription factors GATA-3 and T-bet mRNA expression in asthmatic mice

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Acta Pharmacologica Sinica 2005年第9期26卷 1117-1122页

作者： Zu-qun WU~(1,2) Yi-ping XU~3 Hua XIANG~4 Hua-hao SHEN~11 department of Respiratory medicine,the Second affiliated hospital,4 department of Pathology,the First affiliated hospital,school of medicine,Zhejiang university,Hangzhou 310009,china 3 department of Respiratory medicine,Renji hospital,Shanghai Second Medical university Shanghai 200001,china department of Respiratory medicine the Second Affiliated Hospital Hangzhou China department of Respiratory medicine Renji Hospital Shanghai Second Medical University Shanghai China department of Pathology the First Affiliated Hospital School of Medicine Zhejiang University Hangzhou China

Aim:To investigate effects ofCpG oligod'eoxynucleotide(CpG ODN)on the mRNAexpression of transcription factors GATA binding protein 3(GATA-3)and T-boxexpressed in T cells(T-bet)in asthmatic ***:An asthmatic mousemodel was established and treated with CpG *** inflammatory cells andeosinophils in bronchoalveolar lavage fluid(BALF)were counted and inflamma-tory cell infiltration in lung tissue was ***-y and interleukin-4concentrations in BALF and splenocyte culture supernatants were detected us-ing an enzyme-linked immunosorbent *** factor GATA-3 and T-bet mRNA expression in splenocytes and lung tissue were detected by reversetranscription-polymerase chain ***:Total inflammatory cells andeosinophils in BALF were reduced in the CpG ODN-treated group compared withthe asthma group,and inflammatory cell infiltration in lung tissue was also signifi-cantly *** ODN treatment increased the interferon-γ concentrationbut decreased the interleukin-4 concentration in both BALF and splenocyte cul-ture ***-3 mRNA expression was reduced in both lung tissueand splenocytes in the CpG ODN-treated group,while the mRNA ratio ofT-bet toGATA-3 in splenocytes was ***:CpG ODN treatment inhibitsairway inflammatory cell infiltration and regulates interferon-y/interleukin-4 syn-thesis in asthmatic mice,possibly through a mechanism of downregulation ofGATA-3 mRNA expression in both lung tissue and splenocytes.

关键词： asthma inflammation CpG oligodeoxy-nucleotide GATA-3 T-bet mice

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Distribution of cysteinyl leukotriene receptor 2 in human traumatic brain injury and brain tumors

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Acta Pharmacologica Sinica 2005年第6期26卷 685-690页

作者： Hua HU~(2,3), Gao CHEN~2, Jian-min ZHANG~2, Wei-ping ZHANG~3, Lei ZHANG~3, Qiu-fu GE~3, Hong-tian YAO~4, WeiDING~4, Zhong CHEN~3, Er-qing WEI~(3,5) ~2 department of Neurosurgery, the Second affiliated hospital, school of medicine, Zhejiang university, Hangzhou 310009 ~3 department of Pharmacology school of medicine, Zhejiang university, Hangzhou 310031 ~4 department of Pathology, the First affiliated hospital, school of medicine, Zhejiang university, Hangzhou 310003, china department of Neurosurgery the Second Affiliated Hospital School of Medicine Zhejiang University Hangzhou China department of Pharmacology School of Medicine Zhejiang University Hangzhou China department of Pathology the First Affiliated Hospital School of Medicine Zhejiang University Hangzhou China

Aim: To determine the distribution of cysteinyl leukotriene receptor 2 （CysLT2）, one of the cysteinyl leukotriene receptors, in human brains with traumatic injury and tumors. Methods: Brain specimens were obtained from patients who underwent brain surgery. CysLT2in brain tissues was examined using immunohis-tochemical analysis. Results: CysLT2was expressed in the smooth muscle cells （not in the endothelial cells） of arteries and veins. CysLT2was also expressed in the granulocytes in both vessels and in the brain parenchyma. In addition, CysLT2was detected in neuron- and glial-appearing cells in either the late stages of traumatic injury or in the area surrounding the tumors. Microvessels regenerated 8 d after trauma and CysLT2expression was recorded in their endothelial cells. Conclusion: CysLT2is distributed in vascular smooth muscle cells and granulocytes, and brain trauma and tumor can induce its expression in vascular endothelial cells and in a number of other cells.

关键词： cysteinyl leukotriene receptor 2 brain injuries brain neoplasms vascular smooth muscle cell

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Suppression of Type 1 Insulin-like Growth Factor Receptor Expression by Small Interfering RNA Inhibits A549 Human Lung Cancer Cell Invasion in vitro and Metastasis in Xenograft Nude Mice

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Acta Biochimica et Biophysica Sinica 2007年第2期39卷 137-147页

作者： Jianfang QIAN~1 Aiqiang DONG~1 Minjian KONG~1 Zhiyuan MA~2 Junqiang FAN~1 and Guanyu JIANG~3 ~1department of Cardiothoracic Surgery,Second affiliated hospital,school of medicine,Zhejiang university,Hangzhou 310009,china ~2department of Cardiovascular Surgery,Shanghai Jiaotong university affiliated First People's hospital,Shanghai 200080,china ~3department of Emergency,Second affiliated hospital school of medicine,Zhejiang university,Hangzhou 310009,china department of Cardiothoracic Surgery Second Affiliated Hospital School of Medicine Zhejiang UniversityHangzhou 310009 China department of Cardiovascular Surgery Shanghai Jiaotong University Affiliated First People's HospitalShanghai 200080 China department of Emergency Second Affiliated Hospital School of Medicine Zhejiang UniversityHangzhou 310009 China

Cancer invasion and metastasis, involving a variety of pathological processes andcytophysiological changes,contribute to the high mortality of lung *** type 1 insulin-like growthfactor receptor (IGF-1R),associated with cancer progression and invasion,is a potential anti-invasion andanti-metastasis target in lung *** inhibit the invasive properties of lung cancer cells,we successfullydown-regulated IGF-1R gene expression in A549 human lung cancer cells by small interfering RNA (siRNA)technology,and evaluated its effects on invasion-related gene expression,tumor cell in vitro invasion,andmetastasis in xenograft nude mice.A549 cells transfected with a plasmid expressing hairpin siRNA forIGF-1R showed a significantly decreased IGF-1R expression at the mRNA level as well as the *** biological assays,transfected A549 cells showed a significant reduction of cell-matrix adhesion,migration and *** with these results,we found that down-regulation of IGR-1Rconcomitantly accompanied by a large reduction in invasion-related gene expressions,including MMP-2,MMP-9,u-PA,and IGF-1R specific downstream *** tail vein injections of plasmid expressinghairpin siRNA for IGF- 1R significantly inhibited the formation of lung metastases in nude *** resultsshowed the therapeutic potential of siRNA as a method for gene therapy in inhibiting lung cancer invasionand metastasis.

关键词： lung cancer invasion metastasis type 1 insulin-like growth factor receptor small interfering RNA

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Shp2,a novel oncogenic tyrosine phosphatase and potential therapeutic target for human leukemia

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Cell Research 2007年第4期17卷 295-297页

作者： Rongzhen Xu~1 ~1department of Hematology,Second affiliated hospital,school of medicine,Zhejiang university,Hangzhou 310009,china department of Hematology Second Affiliated Hospital School of Medicine Zhejiang University Hangzhou China

Shp2,encoded by the PTPN11 gene in human,is aubiquitously expressed protein tyrosine phosphatase thatcontains two N-terminal Src homology 2(SH2)domains(N-SH2,C-SH2,respectively),a catalytic protein-tyrosinephosphatase(PTP)domain,and a C-terminal tail withtyrosyl phosphorylation sites and a prolyl-rich motif[1].The progress of our understanding of biological functionsof Shp2 has clearly shown that Shp2 plays an importantrole not only in biology of normal hematopoietic cells andother mammalian cells,but also in the development ofleukemia and other *** recently,PTPNll genehas been firmly established as the first proto-oncogene thatencodes a protein tyrosine phosphatase[1-3].In the hema-topoietic system,most if not all function of Shp2 is to actas a positive component that is essential for proliferationand/or survival of hematopoietic cells through regulationof signaling pathways involving Erk,Akt and STAT5[1-4].Over the past few years,a number of disease-associatedShp2 mutants have been identified in human leukemia andother malignancies[1,3,4].Recently,studies from ourlaboratories and others strongly suggest that dysregula-tion of wild-type Shp2 enzyme may be involved in the

关键词： SHP Shp2,a novel oncogenic tyrosine phosphatase and potential therapeutic target for human leukemia CML

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Characterization of Genes Associated with Different Phenotypes of Human Bladder Cancer Cells

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Acta Biochimica et Biophysica Sinica 2006年第9期38卷 602-610页

作者： Yu-Cong YANG Xu LI Wei CHEN Center for Experimental medicine,the First affiliated hospital,school of medicine,Xi’an Jiaotong university,Xi’an 710061,china Center for Experimental medicine the First Affiliated Hospital School of Medicine Xi'an Jiaotong UniversityXi'an 710061 China

To identify genes associated with morphological phenotypes of human bladder transitional cellcarcinoma,we used suppression subtractive hybridization (SSH) to create a subtractive cDNA library of twoestablished cell lines,BLZ-211 and BLS-211,derived from a patient with transitional cell carcinoma of thebladder,then to screen for differentially expressed ***-time reverse transcription-polymerase chainreaction was used to further confirm the selected differentially expressed *** and reverse subtractivecDNA libraries yielded 168 and 305 putative clones,and among them more than 90% contained the *** differential screening,36 different transcripts were obtained from 64 cDNA clones of a forward andreverse subtraction *** them,17 were identified as known genes by homology,for example,Vimentin,Keratin 7,DDH and *** remaining 19 were unknown expressed genes,and were collectedas new expressed sequence tags by the GenBank dbEST database with the accession numbers DR008207,DR010178,DR159652-DR159660,DY230447-DY230448,and *** function willbe studied ***,SSH appears to be a useful technique for identifying differentially expressed genesbetween cell lines or *** results,as revealed by SSH,also suggest that differences in gene expressionof cytoskeletal proteins might contribute to the different morphologies in BLZ-211 and BLS-211 cells.

关键词： bladder cancer phenotypes suppression subtractive hybridization

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Microtubule stability and MAP1B upregulation control neuritogenesis in CAD cells

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Acta Pharmacologica Sinica 2006年第9期27卷 1119-1126页

作者： Wen LI~(2,5)Jin-tang XIA~3 Yue FENG~(4,5)2 Laboratory of Surgery,The First affiliated hospital,Sun Yat-sen university,Guangzhou 510080,china 3 department of General Surgery,The First People's hospital of Guangzhou,Guangzhou 510180,china 4 department of Pharmacology,Emory university school of medicine,USA Laboratory of Surgery The First Affiliated Hospital Sun Yat-sen University Guangzhou China department of General Surgery The First People's Hospital of Guangzhou Guangzhou China department of Pharmacology Emory University School of Medicine USA

Aim:To study the role of microtubule dynamics and microtubule associated pro-tein 1B(MAP1B)in regulation of the neurite extension in CAD catecholaminergicneuronal cell ***:The neuritogenesis of the CAD cells was abolishedby inhibiting microtubule polymerization with nocodazole and by blocking micro-tubule depolymerization with ***1B and tubulin protein expression wasdetected by Western *** staining of tubulins was observedby fluorescent and confocal ***:Microtubule dynamics wasessential for CAD neurite *** analysis revealed that neurite exten-sion was much more sensitive to nocodazole than to taxol,suggesting a func-tional requirement for highly active microtubule assembly.A remarkableupregulation of MAP IB protein was detected during neurite extension accompa-nied with increased microtubule ***:Upregulation of MAP1Bleads to the stabilization of newly formed microtubules in the developing neurites,which in turn promotes neurite extension.

关键词： microtubule stability microtubule associated protein 1B neuritogenesis

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Immunosuppression in Human Peripheral Blood T Lymphocytes by Fluvastatin

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Acta Biochimica et Biophysica Sinica 2004年第10期36卷 649-655页

作者： Li-Hua WU, Yun-Le WAN1, Hai-Yang XIE, Wen-Jin ZHANG, and Shu-Sen ZHENG* Institute of Surgery, The First affiliated hospital, school of medicine, Zhejiang university, Hangzhou 310003, china 1Centre of organ transplantation, The Second affiliated hospital, school of medicine, Zhongshan university, Guangzhou 510000, china Institute of Surgery The First Affiliated Hospital School of Medicine Zhejiang UniversityHangzhou 310003 China Centre of organ transplantation The Second Affiliated Hospital School of Medicine Zhongshan UniversityGuangzhou 510000 China

To investigate the immunosuppressive effect of fluvastatin on the PHA-activated T lymphocytes. T lymphocytes were isolated from the blood of healthy volunteers, cell proliferation and the activation mar- kers expression were examined by flow cytometric analysis. Cytokine secretion was assayed by ELISA. LDH-release assay was used to detect activity of killer cells. NFAT activation was evaluated by TransAMTM ELISA kit. Results were as following. (1) Whereas no modification in CD25 expression was seen, fluvastatin at 5 μM caused a lower level of CD69 expression, accompanied by an essential suppression on proliferation, IL-2 production and cytotoxicity development in PHA-stimulated T cells. However, the level of secreted IL-10 had no change, and the level of IL-4 even experienced a significant increase. (2) Combined with cyclosporine A (CsA), fluvastatin would further repress CD69 expression, cells proliferation and activity of killer cells, meanwhile significantly induced the secretion of IL-4 and IL-10. (3) Fluvastatin treatment also resulted in a strong inhibition of NFAT activation. In conclusion, partly involving the blockage of activation of NFAT, fluvastatin exhibited an immunosuppressive effect in vitro.

关键词： fluvastatin T cell activation T cell proliferation cytokine nuclear factor of activated Tlymphocytes

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Retrorectal tumors in adults:Magnetic resonance imaging findings

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World Journal of Gastroenterology 2010年第46期16卷 5822-5829页

作者： Bo-Lin Yang,Yun-Fei Gu,Wan-Jin Shao,Hong-Jin Chen,Gui-Dong Sun,department of Coloproctology,The affiliated hospital of Nanjing university of Chinese medicine,Nanjing 210029,china Hei-Ying Jin,department of Coloproctology,The 3rd affiliated hospital of Nanjing university of Chinese medicine,Nanjing 210029,china Xin Zhu,department of Radiology,The affiliated hospital of Nanjing university of Chinese medicine,Nanjing 210029,china department of Radiology The Affiliated Hospital of Nanjing University of Chinese Medicine No.155 Hanzhong Road Nanjing 210029 China.

AIM:To retrospectively evaluate the magnetic resonance imaging (MRI) features of adult retrorectal tumors and compare with histopathologic ***:MRI features of 21 patients with preoperative suspicion of retrorectal tumors were analyzed based on the histopathological and clinical ***:Fourteen benign cystic lesions appeared hypointense on T1-weighted images,and hyperintense on T2-weighted images with regular peripheral *** or dermoid cysts were unilocular,and tailgut cysts were *** of intracystic intermediate signal intensity was observed in one case of tailgut cyst with a component of *** solid tumors were malignant lesions and showed heterogeneous intensity on *** adenocarcinomas showed high signal intensity on T2-weighted and mesh-like enhancing areas on fat-suppressed T2-weighted *** was a fistula between the mass and anus with an internal opening in mucinous adenocarcinomas arising from anal *** stromal tumors displayed low signal intensity on T1-weighted images,and intermediate to high signal intensity on T2-weighted *** necrosis could be seen as a high signal on T2-weighted ***:MRI is a helpful technique to define the extent of the retrorectal tumor and its relationship to the surrounding structures,and also to demonstrate possible complications so as to choose the best surgical approach.

关键词： Retrorectal tumor Presacral lesions Magnetic resonance imaging Congenital cyst Malignant tumor Diagnosis

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