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Human multi-drug resistant hepatocellular carcinoma induced in nude mice by B-ultrasonographically-directed orthotopic implantation:a new experimental model

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Hepatobiliary & Pancreatic Diseases International 2007年第4期6卷 393-398页

作者： Lei Ding,Xiao-Ping Chen,Zhi-Wei Zhang,Kai Jing and Wan-Guang Zhang Hepatic Surgery Center,Affiliated tongji hospital,tongji medical college,huazhong university of science and technology,wuhan 430030,china department of Oncology Center,Beijing Shijitan hospital,Beijing 100038,china Hepatic Surgery Center Affiliated Tongji Hospital Tongji Medical College Huazhong University of Science and Technology Wuhan 430030 China. dinglei1005@***

BACKGROUND:Present studies on the reversal of multi-drug resistance(MDR) are almost entirely limited to in vitro systems,and are seldom carried out in vivo. In order to study MDR under the in vivo situation,a MDR cell strain was used to establish a model in nude mice via orthotopic implantation directed by B-ultrasonography. This model is expected to provide a good platform for evaluating strategies to reverse the phenomenon of primary hepatocellular carcinoma(HCC) MDR. METHODS:An orthotopic MDR1 hepatoma was obtained by injecting the cell lines HepG2 and HepG2/ADM subserosally into the liver of nude mice(10 control and 20 MDR mice). The injections were made under B-ultrasonographic direction. Ultrasonography and laparotomy were used to assess tumor growth,and the long chain PCR technique was applied to sequence the MDR1 gene from multi-drug resistant human HCC cells,HepG2/ADM,and corresponding implanted tumor tissues. Furthermore,MDR1 mRNA and P-gp protein expression were evaluated by reverse transcriptase-polymerase chain reaction(RT-PCR),Western blotting,and immunohistochemical methods. RESULTS:The success rates of tumor implantation and induction of MDR were 100%(30/30) and 95%(19/20),respectively. The 3.8kbp MDR1 gene band was detected from the HepG2/ADM cell line and the correspondingimplanted tumor tissues,and the MDR1 gene sequence coincided with that reported in GenBank. The expressions of MDR1 mRNA and P-gp protein in MDR mice were significantly higher than those in the control group. There was a significant difference in P-gp protein expression between MDR and control mice. CONCLUSION:A MDR model has been successfully established in nude mice via orthotopic implantation of multi-drug resistant human HCC cells directed by ultrasonography.

关键词： nude mice orthotopic implantation carcinoma,hepatocellular drug resistance,multiple

The Effects of Cholecystojejunostomy and Biliary Drainage on Biliary Motor

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Journal of huazhong university of science and technology(medical sciences) 2002年第2期22卷 144-147页

作者： ZHENG Qichang(郑启昌) CHEN Yanglong(陈阳龙) department of General Surgery Xiehe Hospital Tongji Medical College Huazhong University of Science and Technology Wuhan 430022 Department of General Surgery Xiehe Hospital Tongji Medical College Huazhong University of Science and Technology Wuhan 430022

Summary: Simulating physiological neuronal and hormonal conditions during digestive and interdigestive periods, the study identified the changes of the motility of biliary system including bile duct and sphincter of Oddi (SO) before and after cholecystojejunostomy. Thirty-five rabbits were divided into five groups randomly. The experimental groups received the venous injection of CCK 10 ng/kg, erythromycin 10 mg/kg, atropine 3 μg/kg and L-NAME 10 mg/kg respectively. Each rabbit underwent manometry through introducing a three-lumen catheter via the papilla retrogradely, using the low-compliance papillary infusion system. Then the gallbladder and the upper segment of the jejunum was anastomosed and the manometric procedures repeated after one week. SO basal pressure was increased, contraction amplitude decreased, contraction time shortened after cholecystojejunostomy. L-NAME, CCK and erythromycin could all excite SO. L-NAME could increase basal pressure and contraction amplitude, CCK increase basal pressure contraction amplitude and frequency, and erythromycin increase contraction amplitude, respectively. But comparing with that before cholecystojejunostomy, the increasing extent was decreased. The tensional and spontaneous contractions of the SO were under the control of the neural and hormonal mechanism. The anastomosis of gallbladder and jejunum and the drainage of bile made the tensional contraction stronger, but the spontaneous contraction weakened after the operation due to the decreases of the sensitivity of SO to hormonal factors. The clinical symptoms may not be relieved when the patients with SO dysfunction accepted cholecystojejunostomy.

关键词： cholecystojejunostomy biliary motor manometry sphincter of Oddi dysfunction

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Acta Pharmacologica Sinica 2006年第9期27卷 1159-1164页

作者： Jin QIN Zheng-xiang LIU~1 department of Cardiology,tongji hospital,tongji medical college,huazhong university of science and technology,wuhan 430030,china department of Cardiology Tongji Hospital Tongji Medical College Huazhong University of Science and Technology Wuhan 430030 China. eds.ufl.edu

Aim:To examine the inhibitory effect of FAK-related nonkinase(FRNK)in cardiachypertrophy in vitro and investigate the possible ***:A func-tional fragment of FRNK cDNA was amplified by reverse transcription-polymerasechain reaction and cloned into the vector *** in neonatal ratcardiac myocytes was established with angiotensin-Ⅱ *** pcDNA3.1-FRNK or pcDNA3.1 was respectively transfected into cardiomyocytes byLipofectamine *** surface area and mRNA expression of atrial natriureticpeptide(ANP)of myocytes were employed to detect cardiac ***-κBp65 protein in nuclear extracts,phosphorylation levels of ERK1/2(p-ERK 1/2)andAKT(p-AKT),as well as total ERK1/2,and AKT in variant treated cardiomyocyteswere determined by Western ***:Under the stimulation of angiotensin Ⅱ,the surface area of myocytes and levels ofANP mRNA were significantly *** transient transfection with pcDNA3.1-FRNK in advance may reduce the sur-face area and expression of ANP mRNA of hypertrophic *** proteinlevels of NF-κB p65 in nuclear extracts and p-ERKI/2,p-AKT in FRNK treatedcardiomyocytes were significantly decreased compared with that in angiotensin-Ⅱ induced cardiomyocytes,while different treatments had little effect on totalERK1/2 and ***:FRNK may inhibit angiotensin-Ⅱ-inducedcardiomyocyte hypertrophy via decreasing phosphorylation levels at ERK1/2 andAKT,consequently downregulating nuclear translocation of NF-κB p65.

关键词： FAK-related nonkinase angiotensin Ⅱ cardiomyocyte hypertrophy extracellular signal-regulated kinase 1/2 protein kinase B NF-kappaB p65

Adenoma of the middle ear

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Journal of Otology 2007年第2期2卷 109-113页

作者： XIAO Hong-jun1, WEI Yong-hao1, NIE Xiu2 1 department of Otolaryngology, 2 department of Pathology, union hospital, tongji medical college, huazhong university of science and technology. wuhan 430022, Hubei. department of Otolaryngology Union Hospital Tongji Medical College Huazhong University of Science and Technology. Wuhan 430022 Hubei. department of Pathology Union Hospital Tongji Medical College Huazhong University of Science and Technology. Wuhan 430022 Hubei.

Adenoma of the middle ear is a primary tumor of the middle ear that can have exocrine (mucinous) and/or neuroendocrine differentiation. It is an uncommon tumor in the practice of otolaryngology. Based on the first des... 详细信息

关键词： Neck Adenoma of the middle ear

Inhibitory effect of triptolide on lymph node metastasis in patients with non-Hodgkin lymphoma by regulating SDF-1/CXCR4 axis in vitro

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Acta Pharmacologica Sinica 2006年第11期27卷 1438-1446页

作者： Chun ZHANG Guo-hui CUI Fang LIU Qiu-ling WU Yan CHEN~2 department of Hematology,union hospital,tongji medical college,huazhong university of science and technology,wuhan 430022.china departmentofHematology UnionHospitalTongjiMedicalCollegeHuazhongUniversityofScienceandTechnologyWuhan430022China

Aim:To investigate the antiproliferative effect of triptolide on B-NHL cell line Rajicells,to study its effect on lymph node metastasis in patients with non-Hodgkin’slymphoma(NHL)in vitro,and to explore the underlying mechanism regulatingSDF-1/CXCR4 ***:The effects of triptolide on the growth of Raji cellswere studied by 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium(MTT)*** effects of triptolide on SDF-1 mRNA expression in lymph node stromalcells from patients with NHL were determined by reverse transcriptase-polymerasechain reaction(RT-PCR).The effects of triptolide on CXCR4 expression on lym-phoma cells freshly isolated from the lymph nodes of these patients were studiedby flow cytometric *** assays were performed to observe theeffects of triptolide on migration of primary lymphoma cells towards recombinanthuman SDF-1α(rhSDF-1α)or cultured lymph node stromal cells in ***:Triptolide inhibited the proliferation of B-NHL cell line Raji cells in a dose-andtime-dependent manner with a 24-h IC50value of 43.06 nmol/L and a 36-h IC50valueof 25.08 nmol/*** expression of SDF-1α mRNA in lymph node stromal cellsobtained from patients with NHL was decreased after treatment by triptolide atconcentrations of 25 and 50 nrnol/L for 24 *** cytometry analysis showed thatthe CXCR4 expression on primary lymphoma cells were downregulated graduallyin a dose-dependent manner following triptolide *** assaysrevealed that the migration of freshly isolated lymphoma cells towards either rhSDF-1or cultured lymph node stromal cells was markedly inhibited by the addition oftriptolide in vitro,and the inhibition was ***:Triptolidecan inhibit the proliferation of B-NHL cell line Raji ***,triptolide isable to inhibit the migration of lymphoma cells via lymph nodes in *** antitumor mechanisms of triptolide are related to the antiproliferativeeffect and the blockage of SDF-1/CXCR4

关键词： triptolide Raji metastasis non-Hodgkin’s lymphoma stromal cell-derived factor-1 CXCR4

Two-dimensional electrophoresis for comparative proteomic analysis of human bile

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Hepatobiliary & Pancreatic Diseases International 2007年第4期6卷 402-406页

作者： Bo Chen,Jing-Qing Dong,Yong-Jun Chen,Jian-Ming Wang,Jun Tian,Chun-Ben Wang and Sheng-Quan Zou department of General Surgery,tongji hospital and department of Proteomics Research Center,tongji medical college,huazhong university of science and technology,wuhan 430030,china department of General Surgery Tongji Hospital Tongji Medical College Huazhong University of Science and Technology Wuhan 430030 China.

BACKGROUND:Proteomic analysis of bile fluid holds promise as a method to identify biomarkers of bile tract diseases,especially for tumors. Two-dimensional electrophoresis(2-DE) is a popular and proven separation technique for proteome analysis,but using this strategy for bile fluid analysis is still not fully developed. This study was undertaken to(a) establish a reliable method for general clean-up to make bile fluid samples suitable for 2-DE;(b) obtain 2-D biliary maps with high reproducibility and resolution;and(c) identify protein patterns present in 2-D biliary maps for potential tumor biomarker discovery,with the intention of distinguishing malignant from benign causes of bile duct obstruction. METHODS:Bile fluid samples were obtained from two patients suffering from malignant and benign bile tract obstruction(one patient with cholangiocarcinoma as the experimental case,the other with cholelithiasis as control). A variety of sample preparation options,including delipidation,desalination and nucleic acid removal,were adopted to remove contaminants that affect 2-DE results. After that,each 350 μg purified sample was loaded onto nonlinear IPG strips(18 cm,pH 3-10 and pH 4-7) for first-dimension isoelectric focusing,and 12.5% SDS-PAGE electrophoresis for second dimension separation. Then 2-D maps were visualized after silver staining and analyzed with the Image Master 2-D software. RESULTS:A large number of protein spots were separated in 2-D maps from the experimental and control groups,with means of 250 and 216 spots on pH 3-10 IPG strips,and 182 and 176 spots on pH 4-7 strips,respectively. Approximately 16 and 23 spots were differentially expressed in matched pairs from the experimental and control cases using pH 3-10 and pH 4-7 strips. CONCLUSIONS:This study established a reliable sample preparation process suitable for 2-DE of bile fluid. By this method,2-D biliary maps with high reproducibility and resolution were obtained. The differentially displayed prote

关键词： proteome electrophoresis,gel,two-dimensional bile

The Role of Angiopoietin-1 and Thrombospondin-1 in the Kidney of Rats Subject to 5/6 Nephrectomy

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Journal of huazhong university of science and technology(medical sciences) 2009年第5期29卷 557-562页

作者：杨晓刘兰香 department of nephrology Union HospitalTongji Medical CollegeHuazhong University of Science and Technology

The expression of angiopoietin- 1 （Ang- 1） and thrombospondin- 1 （TSP- 1） in 5/6 subtotal nephrectomy （STN） rats model, and its correlation to the renal microvasculature injury were investigated. Rat 5/6 STN model was established in adult male SD rats, and the sham-operated group and 5/6 STN group were set up. The renal function and histopathological changes were examined at the 1st, 2nd, 4th, 8th and 12th week after operation. The expression orAng-1, TSP-1 and CD31 in renal tissues was detected by using immunohistochemistry. From 2nd to 8th week after operation, Ang-1 was significantly expressed in glomeruli of rats with STN. Ang-1 staining in glomeruli of STN group was increased significantly as compared with that in sham-operated group at 4th and 8th week after operation, and subsequently decreased after the 12th week. The expression of TSP-1 was increased significantly in STN group. As compared with sham-operated group, the CD31 expression was significantly down-regulated from the 2nd week. The expression of Ang-1 mRNA was detected by using RT-PCR at the same time points. The expression of Ang-1 mRNA in renal tissue of rats with STN was significantly up-regulated at the 2nd, 4th and 8th week after operation as compared with that in STN group at other time points or in sham-operated group at the same time points, while decreased evidently at the 12th week as compared with that in sham-operated group. It is concluded that there are changes in the mRNA expression of Ang-1, and the significant up-regulation of the expression of TSP-1 in renal tissue of rats with STN, which may be involved in the remnant renal microvasculature injury.

关键词： 5/6 nephrectomy angiopoietin- 1 thrombospondin- 1 CD31 microvasculature injury

Characteristics of fMRI BOLD signal and its neurophysiological mechanism

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Progress in Natural science:Materials International 2007年第6期17卷 630-638页

作者： Zhao Xiaohu1,2, Wu Yigen3 and Guo Shengli3(1. college of Life sciences, Zhejiang university, Hangzhou 310058, china 2. Imaging department of tongji hospital of tongji university, Shanghai 200065, china 3. college of Mathematics and Physics, Nanjing university of Information science and technology, Nanjing 210044, china) college of Life sciences Zhejiang University Hangzhou 310058 China Imaging department Tongji Hospital Tongji University Shanghai 200065 China college of Mathematics and Physics Nanjing University of Information Science and Technology Nanjing 210044 China

The functional magnetic resonance imaging (fMRI) based on blood oxygen level dependent (BOLD) contrast has emerged as one of the most potent noninvasive tools for mapping brain function and has been widely used to explore physiological, pathological changes and mental activity in the brain. Exploring the nature and property of BOLD signal has recently attracted more attentions. Despite that great progress has been made in investigation of the characteristics and neurophysiological basis, the exact nature of BOLD signal remains unclear. In this paper we discuss the characteristics of BOLD signals, the nonlinear BOLD response to external stimuli and the relation between BOLD signals and neural electrophysiological recordings. Furthermore, we develop our new opinions regarding nonlinear BOLD response and make some perspectives on future study.

关键词： functional magnetic resonance imaging blood oxygen level dependent nonlinear response neural electrophysiological activity.

Passive sensitization increases histamine-stimulated calcium signaling and NF-κB transcription activity in bronchial epithelial cells

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Acta Pharmacologica Sinica 2006年第6期27卷 708-714页

作者： Si JIN~(2,5)Dan TIAN~3 Jian-guo CHEN~4 Li-ping ZHU~2 Sheng-yuan LIU~2 Di-xun WANG~2 2 department of Pathophysiology,tongji medical college,huazhong university of science and technology Key Laboratory of Pulmonary Disease of Ministry of Health of china,wuhan 430030,china 3 department of Pathology,tongji medical college.huazhong university of science and technology.wuhan 430030,china 4 department of Pharmacology,tongji medical college,huazhong university of science and technology,wuhan 430030,china department of Pathophysiology Tongji Medical College Huazhong University of Science and Technology Key Laboratory of Pulmonary Disease of Ministry of Health of china Wuhan 430030 China department of Pathology Tongji Medical College Huazhong University of Science and Technology Wuhan 430030 China department of Pharmacology Tongji Medical College Huazhong University of Science and Technology Wuhan 430030 China

Aim:To find out if the two aspects of asthma（chronic airway inflammation andbronchial hyperresponsiveness）are related to hypersensitivity of calcium signal-ing in bronchial epithelial ***:Porcine bronchial epithelial cells（PBEC）were divided into sensitized（S）and nonsesitized（N）*** group S,the cellswere preincubated with serum from ovalbumin sensitized guinea *** group N,the cells were preincubated with serum from nonsensitized guinea *** calcium imaging and ELISA-based NF-κ3 activity were used to evaluate thehistamine-stimulated intracellular free calcium level and NF-κB activity,***:First,0.1μmol/L histamine could induce [Ca2+]ioscillationsin PBEC of group S,but not in group ***,1/Jmol/L histamine could induce[Ca2+]ioscillations of PBEC in both group S and group *** [Ca2+]ioscillationfrequency of PBEC was significantly higher in group S than in group N,thoughthe[Ca2+]ioscillation amplitude showed no difference between the two ***,when 10 μmol/L histamine was used to stimulate PBEC a transient initialincrease followed by a sustained elevation（FSE）of [Ca2+]iwas observed in PBECin both *** amplitude of the FSE of [Ca2+]iin PBEC was significantlyhigher in group S than in group *** subsequent NF-κB activity was in accor-dance to the calcium oscillation frequency evoked by histamine,but not to ***:It was suggested that the increased sensitivity of calciumsignaling in bronchial epithelial cells might contribute to the exorbitant inflamma-tion or increased susceptibility in asthmatic airway epithelial cells.

关键词： asthma histamine calcium signaling bronchi epithelial cell nuclear factor of kappaB