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ESTABLISHMENT OF A MAMMARY CANCER CELL LINE Ca 761-86 AND ITS BIOLOGICAL CHARACTERISTICS IN INBRED 615 MICE

Chinese Journal of Cancer Research 1989年第4期1卷 43-48页

作者：程一棹黄敏王明俭郑怀祖袁保和许广源程佳钱振超 Dalian Medical College Dalian Medical College cell line designated as Ca 761-86 has been established from the solid mouse mammary cancer (Ca 761) by suspension culture. It has been passaged for more than 212 generations. Moderate round cells were predominant and most of them were mononuclear. Some characteristics of malignant cells and A-type viral-like particles were observed by electron microscopy. The results of cytochemical studies (DNA RNA SDH 5' AMPase ACP etc.) were comparable to the ultramicroscopic results. It multiplied approximately 27.4 fold on day 5 with mitotic index reaching 1.8% on day 3. This cell line was a hyperdiploid with karyotype of 45 or 45 -2X tril2 tri17 +M1-5. Cell agglutination was observed when treated with ConA (≥7 fig ml). Spontaneous agglutination might also take place without adding any ConA. After 5×106 cells of Ca 761-86 suspension were transplanted into the normal inbred 615 mice by different ways (subcutan eous intrafoot-pad or intraperitoneal) the transplan lability rate reached 100%. Spontaneous remission was never observed and its metastatic ability reserved. PPLO were not detected. Ca 761-86 may be of value for practical purposes.

A cell line designated as Ca 761-86 has been established from the solid mouse mammary cancer (Ca 761) by suspension culture. It has been passaged for more than 212 generations. Moderate round cells were predominant and most of them were mononuclear. Some characteristics of malignant cells and A-type viral-like particles were observed by electron microscopy. The results of cytochemical studies (DNA, RNA, SDH, 5' AMPase, ACP etc.) were comparable to the ultramicroscopic results. It multiplied approximately 27.4 fold on day 5 with mitotic index reaching 1.8% on day 3. This cell line was a hyperdiploid with karyotype of 45 or 45, -2X, tril2, tri17, +M1-5. Cell agglutination was observed when treated with ConA (≥7 fig, ml). Spontaneous agglutination might also take place without adding any ConA. After 5×106 cells of Ca 761-86 suspension were transplanted into the normal inbred 615 mice by different ways (subcutan eous, intrafoot-pad or intraperitoneal), the transplan lability rate reached 100%. Spontaneous remission was never observed and its metastatic ability reserved. PPLO were not detected. Ca 761-86 may be of value for practical purposes.

关键词： line ESTABLISHMENT OF A MAMMARY CANCER CELL LINE Ca 761-86 AND ITS BIOLOGICAL CHARACTERISTICS IN INBRED 615 MICE RER ConA Ca

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Long-Term Expression of Human Factor Ⅸ cDNA in Rabbits

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Science China Chemistry 1993年第11期36卷 1333-1341页

作者：周洁民邱信芳卢大儒陆建英薛京伦 Institute of Genetics Fudan University Shanghai 200433 PRC Institute of Genetics Fudan University Shanghai 200433 PRC In this study rabbits were used as a model for gene therapy for hemophilia B Human factor ⅨcDNA was transferred to cultured normal rabbit skin fibroblasts (RSF) by a recombinant plasmid (pCMVIX) or retrovirus(XL-IX or N2CMVIX) constructed in our laboratoy. Infected fibroblasts capable of synthesizing and secreting high levels of biologically active human factor Ⅸprotein were selected and embedded in a collagen matrix. The latter was surgically implanted into rabbits as autografts or allografts. Human factor Ⅸprotein was detected in the plasma of all the grafted rabbits and its expression has been maintained for more than 10 months at the time of writing. In addition we have improved and simplified the method of implantation from surgically grafting the tissue-like matrix to the injection of the infected cell-collagen mixture subcutaneously. Using the latter method human factor Ⅸin rabbits injected with RSF-N2CMVIX reached a peak of 480 ng/ml plasma and its expression has continued for more than 3 months at the time of writing. We suggest that the simplified method of transplantation by subcutaneous injection would offer an effective and acceptable approach to somatic cell gene therapy and may be practical for human trials.

In this study, rabbits were used as a model for gene therapy for hemophilia B, Human factor Ⅸ cDNA was transferred to cultured normal rabbit skin fibroblasts (RSF) by a recombinant plasmid (pCMVIX) or retrovirus(XL-IX or N2CMVIX) constructed in our laboratoy. Infected fibroblasts capable of synthesizing and secreting high levels of biologically active human factor Ⅸ protein were selected and embedded in a collagen matrix. The latter was surgically implanted into rabbits as autografts or allografts. Human factor Ⅸ protein was detected in the plasma of all the grafted rabbits, and its expression has been maintained for more than 10 months at the time of writing. In addition, we have improved and simplified the method of implantation from surgically grafting the tissue-like matrix to the injection of the infected cell-collagen mixture subcutaneously. Using the latter method, human factor Ⅸ in rabbits injected with RSF-N2CMVIX reached a peak of 480 ng/ml plasma, and its expression has continued for more than 3 months at the time of writing. We suggest that the simplified method of transplantation by subcutaneous injection would offer an effective and acceptable approach to somatic cell gene therapy and may be practical for human trials.

关键词： gene transfer gene therapy human clotting factor Ⅸ cell-collagen injection expression in rabbit.

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基于孕激素受体基因的类人猿亚目的分子系统学研究

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四川动物 2011年第1期30卷 11-15页

作者：陈曹逸 Derek Wildman 朱新宇刘伟王康乐苏敏谭湘陵南通大学生命科学院江苏南通226007 Perinatology Research Branch National Institute of Child Health and Human Development/National Institutes of HealthDepartment of Health and HumanServicesBethesdamdusa20892 Center for Molecular Medicine and Genetics Wayne State University Departments of Obstetrics and Gynecology Wayne State University 复旦大学上海医学院医学分子病毒学重点实验室上海200032 复旦大学生物医学研究院上海200032 南通大学附属医院妇产科江苏南通226001

通过对类人猿亚目中部分种类的孕激素受体基因进行分析,重建类人猿亚目的系统发育关系。扩增并测定了来源于14个属的类人猿亚目物种的孕激素受体编码区序列,并基于这一序列数据,分别采用邻接法、最大简约法和最大似然法重建了系统发育... 详细信息

通过对类人猿亚目中部分种类的孕激素受体基因进行分析,重建类人猿亚目的系统发育关系。扩增并测定了来源于14个属的类人猿亚目物种的孕激素受体编码区序列,并基于这一序列数据,分别采用邻接法、最大简约法和最大似然法重建了系统发育关系。除了阔鼻下目,3种方法构建的系统发生树的拓扑结构类似且各节点支持率高。重建的人猿超科和猴超科内部亲缘关系支持多数人所认可的分类系统。本研究为黑猩猩和人的姐妹群关系提供了证据,提示黑猩猩比大猩猩或其他猿猴更接近人类。阔鼻下目中蜘蛛猴科、卷尾猴科和僧面猴科三者之间的系统发育关系在本研究中未得到很好辨析。

关键词：孕激素受体基因系统发育关系类人猿亚目

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Oral Gd-DTPA as a negative gastrointestinal contrast agent in magnetic resonance cholangiopancreatography

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Journal of Medical Colleges of PLA(China) 2003年第5期18卷 325-330页

作者：陈燕萍张雪林昌仁民成官迅 Imaging Diagnostic Center Nanfang HospitalFirst Military Medical UniversityGuangzhou 510515China Imaging Diagnostic Center Nanfang HospitalFirst Military Medical UniversityGuangzhou 510515Chinabjective : To evaluate the value of oral Gd-DTPA as a negative contrast agent during magnetic resonance cholangiopancreatography (MRCP) to eliminate the high signals of the gastrointestinal tract. Methods: To select the optimal concentration of oral Gd-DTPA for MRCP a phantom study was performed followed by clinical trial in 15 cases undergoing MRCP before and after oral Gd-DTPA (in a total volume of 250 ml 1:5 diluted Gd-DTPA 1.488 g/L). MRCP images were acquired using two-dimensional single slice fast spin-echo ( SSTSE) sequence and half-Fourier acquisition single slice fast spin-echo (HASTE) sequence. Results: The phantom study showed that the 1:5 diluted oral Gd-DTPA was best in decreasing the signal intensity both in T2-weighted imaging (59. 5% ) and in HASTE sequence (82.45% ). The high signal intensity of the stomach and intestinal fluid was completely suppressed in all the cases. The depictions of the common bile duct and pancreatic duct were markedly improved by using the oral contrast agent (P<0.05). Conclusion: Oral Gd-DTPA is effective and safe for eliminating the high signal of the gastrointestinal tract to improve the depiction of the biliary system by MRCP.

Objective: To evaluate the value of oral Gd-DTPA as a negative contrast agent during magnetic resonance cholangiopancreatography (MRCP) to eliminate the high signals of the gastrointestinal tract. Methods: To select the optimal concentration of oral Gd-DTPA for MRCP, a phantom study was performed followed by clinical trial in 15 cases undergoing MRCP before and after oral Gd-DTPA (in a total volume of 250 ml 1∶5 diluted Gd-DTPA, 1.488 g/L). MRCP images were acquired using two-dimensional single slice fast spin-echo (SSTSE) sequence and half-Fourier acquisition single slice fast spin-echo (HASTE) sequence. Results: The phantom study showed that the 1∶5 diluted oral Gd-DTPA was best in decreasing the signal intensity both in T2-weighted imaging (59.5%) and in HASTE sequence (82.45%). The high signal intensity of the stomach and intestinal fluid was completely suppressed in all the cases. The depictions of the common bile duct and pancreatic duct were markedly improved by using the oral contrast agent (P<0.05). Conclusion: Oral Gd-DTPA is effective and safe for eliminating the high signal of the gastrointestinal tract to improve the depiction of the biliary system by MRCP.

关键词： magnetic resonance cholangiopancreatography oral contrast agent pancreaticobiliary system gadolini-um-DTPA

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Transcatheter Aspiration of Coronary Thrombus in Patients with Acute Myocardial Infarction

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South China Journal of Cardiology 2003年第2期4卷 72-74,109页

作者：马礼坤余华褚俊冯克福韩晓萍严激顾统元 Department of Cardiology Anhui Provincial Hospital Hefei 230001 China Department of Cardiology Anhui Provincial Hospital Hefei 230001 Chinabjectives We introduced a simple method of transcatheter aspiration of coronary thrombus during emergency PCI in acute myocardial infarction (AMI) patients. Methods AMI patients presenting proximal coronary lesions with intracoronary thrombus were selected. We used the common coronary wire (0. 014") across the lesion after 7F guiding catheter to the ostium of coronary arteries. Aspiration catheter (Export PercuSurge) was directly advanced over the wire to the lesion and manually aspirated the thrombus with a 20 - ml syringe at the same time. After thrombus disappeared and occluded arteries reopened stents were directly implanted. Results 12 cases of AMI received the treatment. After aspiration the thrombus disappeared in 8 cases and decreased in another 4 cases. The total occluded artery was directly re- opened and the antegrade flow improved markedly after aspiration in 7 out of 8 cases. 9 patients achieved TIMI 3 flow and TMP 3 grade perfusion flow after stenting and the other 3 cases were TMP 2. No major cardiac events took place during hospitalization. The LVEF was 64.1 ±9. 3% before discharge. Conclusions Transcatheter aspiration of thrombus is a simple and effective method to manage the intracoronary thrombus in AMI patients.

Objectives We introduced a simple method of transcatheter aspiration of coronary thrombus during emergency PCI in acute myocardial infarction (AMI) patients. Methods AMI patients presenting proximal coronary lesions with intracoronary thrombus were selected. We used the common coronary wire (0. 014') across the lesion after 7F guiding catheter to the ostium of coronary arteries. Aspiration catheter (Export, PercuSurge) was directly advanced over the wire to the lesion and manually aspirated the thrombus with a 20 - ml syringe at the same time. After thrombus disappeared and occluded arteries reopened, stents were directly implanted. Results 12 cases of AMI received the treatment. After aspiration, the thrombus disappeared in 8 cases, and decreased in another 4 cases. The total occluded artery was directly re- opened and the antegrade flow improved markedly after aspiration in 7 out of 8 cases. 9 patients achieved TIMI 3 flow and TMP 3 grade perfusion flow after stenting, and the other 3 cases were TMP 2. No major cardiac events took place during hospitalization. The LVEF was 64.1 ± 9. 3% before discharge. Conclusions Transcatheter aspiration of thrombus is a simple and effective method to manage the intracoronary thrombus in AMI patients.

关键词： Transcatheter thrombus aspiration Myocardial infarction Stent Thrombus

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1H-MRS测量肝脏代谢产物的浓度

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国际医学放射学杂志 2013年第1期 68-68页

作者： R.Ouwerkerk R.I.Pettigrew A.M.Gharib 赵健仪 Biomedical and Metabolic Imaging Branch National Institute of Diabetes and Digestive and Kidney DiseasesNational Institutes of Health10 Center DrCRC Building 10Room 3-5340MSC 1263BethesdaMD 20892-1263

目的确定1H-MRS活体测量人肝脏胆碱和糖原浓度的可行性。材料与方法研究获得受试者签字同意和单位审查委员会许可,符合HIPAA要求,在285d内共46名受试者[平均年龄(46±17)岁;女性24名]参与研究。采用导航门控,以TE35ms,选择8ml的容积进... 详细信息

目的确定1H-MRS活体测量人肝脏胆碱和糖原浓度的可行性。材料与方法研究获得受试者签字同意和单位审查委员会许可,符合HIPAA要求,在285d内共46名受试者[平均年龄(46±17)岁;女性24名]参与研究。采用导航门控,以TE35ms,选择8ml的容积进行点解析(PRESS)MRS。为减小谱线宽度,测定快速屏气的B0场梯度并进一步手动匀场。分别记录水抑制和无水抑制的导航门控波谱,确定代谢产物的浓度,而以水的信号作为内部参照。在3名受试者中,改变TE时间以确定肝糖原和胆碱的T2值。采用线性回归分析胆碱、肝脏脂肪含量、体质量指数、肝糖原含量和年龄之间的相关性。结果在48次实验中,46次可确定胆碱浓度,结果为(8.6±3.1)mmol/kg(3.8～17.6mmol/kg;n=44)。25名受试者的27个波谱具有窄的峰宽和较低的脂肪含量,能够对肝糖原进行定量。肝糖原(葡萄糖基单位)浓度为(38.1±14.4)mmol/kg。3名受试者肝脏内糖原峰的T2值为(36±8)ms。胆碱峰较低,但与肝糖原相关(r2=0.15;P<0.05),而与脂质含量无相关性。结论导航门控和梯度回波匀场的PRESS1H-MRS可以定量评价肝脏内的代谢产物,这对于了解和发现肝脏的糖与脂质代谢异常有重要意义。

关键词： H-MRS 代谢产物胆碱肝脏代谢物三甲基烷基铵肝糖原肝醣

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Effect of adenovirus-mediated gene transfection of vascular endothelial growth factor on survival of random flaps in rats

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Chinese Journal of Traumatology 2003年第4期6卷 199-204页

作者：崔磊李发成张群钱云良关文祥 Department of Plastic Surgery Ninth People’s Hospital Shanghai Second Medical UniversityShanghai 200011China Department of Plastic Surgery Ninth People’s Hospital Shanghai Second Medical UniversityShanghai 200011Chinabjective: To evaluate the effect of local application of vascular endothelial growth factor ( VEGF ) via adenovirus-mediated gene transfer on survival of full thickness flaps selected randomly in rats.Methods: Thirty Sprague-Dawley rats weighing 480-520 g were used in this study. A dorsal flap (8 cm ×2 cm) in full thickness with the pedicle located at the level of the iliac crest was designed. Then the rats received 1 012 pfu replication-deficient recombinant adenovirus carrying VEGF ( AdCMV-VEGF group n = 10 ) 1012 pfu recombinant β-galactosidase adenovirus ( AdCMV-Gal group n = 10) and 1 ml saline (saline group n = 10) respectively in the distal two thirds of the proposed flap by means of subdermal injection at 8 different locations. Three days after treatment the flaps were elevated as originally designed and sutured back in situ. The survival rate of the flaps was evaluated on day 7 after operation.Results: The survival rate of the flaps in the AdCMV-VEGF group increased significantly as compared with those of the AdCMV-Gal group (P < 0.01) and the saline group ( P < 0.01). Immunohistochemical staining showed that VEGF was expressed in the survival flaps injected with AdCMV-VEGF. Histological analysis showed that more granulation tissues and angiogenesis were observed in the AdCMV-VEGF group than those in the AdCMV-Gal and the saline groups.Conclusions: Local application of adenovims-mediated VEGF165 cDNA may efficiently improve the survival of ischemic skin flaps.

Objective: To evaluate the effect of local application of vascular endothelial growth factor ( VEGF ) via adenovirus-mediated gene transfer on survival of full thickness flaps selected randomly in rats.Methods: Thirty Sprague-Dawley rats weighing 480-520 g were used in this study. A dorsal flap (8 cm × 2 cm) in full thickness with the pedicle located at the level of the iliac crest was designed. Then the rats received 1 012 pfu replication-deficient recombinant adenovirus carrying VEGF ( AdCMV-VEGF group, n = 10 ), 1012 pfu recombinant β-galactosidase adenovirus ( AdCMV-Gal group, n = 10) and 1 ml saline (saline group, n = 10), respectively, in the distal two thirds of the proposed flap by means of subdermal injection at 8 different locations. Three days after treatment, the flaps were elevated as originally designed and sutured back in situ. The survival rate of the flaps was evaluated on day 7 after operation.Results: The survival rate of the flaps in the AdCMV-VEGF group increased significantly as compared with those of the AdCMV-Gal group (P < 0.01) and the saline group ( P < 0.01). Immunohistochemical staining showed that VEGF was expressed in the survival flaps injected with AdCMV-VEGF. Histological analysis showed that more granulation tissues and angiogenesis were observed in the AdCMV-VEGF group than those in the AdCMV-Gal and the saline groups.Conclusions: Local application of adenovims-mediated VEGF165 cDNA may efficiently improve the survival of ischemic skin flaps.

关键词： Adenoviruses, human Gene expression Surgical flaps Survival rate

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