逆转录病毒载体介导双自杀基因治疗肝细胞肝癌的研究

我们将 2种最重要的自杀基因ＴＫ与ＣＤ基因整合在一起 ,以脂质体和逆转录病毒为载体 ,介导转入肝癌细胞ＨＣＣ 990 3[1] ,最大限度地消除肿瘤细胞对治疗药物的耐药性 ;同时 ,降低前药的毒副作用。1 材料与方法 :质粒ｐＷＺＬｎｅｏＣＤｇｌｙＴＫ、ｐＷＺＬｎｅｏＴＫ由澳大利亚布里斯班皇家医院基因治疗中心惠赠。ＨＣＣ 990 3肝癌细胞系由本课题组建立。ＣＤ与ＴＫ基因转染ＰＡ317包装细胞和肝癌细胞系ＨＣＣ 990 3,脂质体介导的ＣＤ与ＴＫ基因转染ＰＡ317包装细胞。测定逆转录病毒滴度 ,逆转录病毒转染ＨＣＣ 990 3细胞 ,ＲＴ ＰＣＲ测…     

nm23-H1抑制原发性肝癌细胞转移的初步机理研究

为了更深入阐明ｎｍ２３－Ｈ１抑制原发性肝癌转移的作用机理，观察ｎｍ２３－Ｈ１表达状况对肝癌细胞浸润相关因素的影响。本实验采用基因转染手段将外源ｎｍ２３－Ｈ１全长ｃＤＮＡ导入肝癌细胞并以此观察细胞体外浸润能力，细胞内游离Ｃａ＾２＋以及Ｎ－ｒａｓ基因ｍＲＮＡ表达的变化。     

原发性肝癌的手术治疗进展

原发性肝癌 (ＨＣＣ)是世界上最常见的恶性肿瘤之一 ,预后差 ,全球每年超过 10 0万人死于原发性肝癌。在每年新增加病例中 ,约 45 %在中国[1] 。ＨＣＣ由于易转移 ,且经常合并肝硬化 ,死亡率、复发率均高 ,目前采取的是以手术切除为主的综合治疗。 1891年Ｌｕｃｋｅ成功切除 1例肝恶性肿瘤 ,195 2年Ｌｏｒ ｔａｔ Ｊａｃｏｂ用解剖肝门技术行大肝癌规则切肝 ,196 3年Ｓｔａｒｔｚ进行首例肝癌肝移植 ,肝癌外科经历了巨大的跨越。我国原发性肝癌治疗经历了大肝癌规则切除、小肝癌局部切除 ,局部切除加综合治疗、生物治疗兴起及复发转移控制研…     

肝癌转移和复发的分子机制

肝癌的转移和复发是肝癌治疗一大难题,直接影响肝癌的预后,有报道指出肝癌根治切除术后5年复发率为61.5%,小肝癌的复发率仍高达43.5%[1],因此,如何控制肝癌术后转移和复发已成为当今研究的热点,也是目前肝癌治疗的关键性课题。近年来对转移和复发的分子机制进行了大量研究,取得了丰硕成果,本文对其研究进展总结如下。1　侵袭相关基因侵袭相关基因的研究可在分子水平阐明肝癌转移复发的机制,目前发现大约有10多种基因与之有关,主要通过以下两大类机制引起肝癌的转移和复发:1.1　抑癌基因的失活以nm23-H1基因最为重要,Yamaguchi等[2]研究发现…     

nm23-H1杂合性等位基因丢失与肝癌转移的关系

目的从ＤＮＡ水平揭示ｎｍ２３－Ｈ１杂合性基因丢失与人原发性肝癌肝内转移、门静脉形成癌栓等的临床病理特征的关系。方法应用Ｓｏｕｔｈｅｒｎ印迹杂交技术,对２５例肝癌及癌旁肝组织ＤＮＡ进行分析。结果ｎｍ２３－Ｈ１两条等位基因分别为７．６Ｋｂ、２．３Ｋｂ,杂合性等位基因丢失率为３１．２５％（５／１６）。ｎｍ２３－Ｈ１杂合性基因丢失多见于分化较差的ＥｄｍｏｎｄｓｏｎⅢ、Ⅳ级和伴肝内转移或门静脉形成癌栓的肝癌。结论ｎｍ２３－Ｈ１可能诱发肝细胞肝癌的转移潜能,有助于预测肝癌复发和转移。     

转移抑制基因Kai1在前列腺癌中的表达

目的了解转移抑制基因Ｋａｉ１蛋白在前列腺癌（ＰＣａ）中的表达。方法采用免疫组化ＬＳＡＢ法分别测定５例正常前列腺、１０例前列腺增生症（ＢＰＨ）和３４例ＰＣａ（均为腺癌,临床Ｃ期或Ｄ期）新鲜前列腺组织中Ｋａｉ１蛋白的表达。结果Ｋａｉ１蛋白分布在腺上皮细胞膜的细胞与细胞连接部,正常前列腺和ＢＰＨ的Ｋａｉ１蛋白染色连续均匀一致,而在ＰＣａ则染色分布不连续,并且染色强度较ＢＰＨ明显降低,差异有显著性（Ｐ＜０．０１）。癌细胞Ｋａｉ１蛋白染色强度与病理分级呈负相关（Ｐ＜０．０５）,与临床分期无相关（Ｐ＞０．０５）。结论Ｋａｉ１蛋白表达下降可能预示ＰＣａ转移,成为临床判断ＰＣａ预后的分子指标。     

肝癌肝外转移复发的临床研究进展

随着肝癌诊疗技术的进步 ,肝癌患者的生存期不断延长 ,但肝外转移复发的发生率也呈增高趋势。目前 ,国内外关于肝癌肝外转移复发的发生、发展和预后 ,以及诊断和治疗等方面的研究报道较少 ,本文就这些方面进行综述 ,以期为临床工作提供帮助。一、肝癌肝外转移复发的发生率及分布对肝癌患者的尸体检查表明 ,肝外转移发生率达6 4 % [1] 。根治性切除术后肝癌肝外复发的发生率约为9 7%～ 2 5 8%不等 ,可能与随访时间的差异及样本选择有关。术后平均肝外复发时间为 7～ 13个月 ,其中 38%伴有肝内复发[2 4] 。肺是肝外转移最常见的脏器 ,约占所…     

肝癌中p16、PRb 及PCNA蛋白的表达及意义

肿瘤组织的过度生长有赖于细胞周期素 (Ｃｙｃｌｉｎ)的蛋白及Ｃｙｃｌｉｎ的蛋白激酶ＣＤＫ (Ｃｙｃｌｉｎｄｅｐｅｎｄｅｎｔｋｉｎａｓｅ)的调控。其中ＣｙｃｌｉｎＤ/ＣＤＫ4 所形成的复合物能够磷酸化抑癌基因Ｒｂ(ｒｅｔｉｎｏｂｌａｓｔｏｍａ)的产物ＰＲｂ(成视网膜细胞瘤蛋白ｒｅｔｉｎｏｂｌａｓ ｔｏｍａｐｒｏｔｅｉｎ) ;启动基因转录及细胞增殖[1] 。ｐ16基因是ＣＤＫ4的抑制因子 ,可阻止ＰＲｂ磷酸化、阻止细胞分裂[2 ] 。ＰＲｂ及ｐ16基因的变异与肿瘤的发生发展有关 ,但二者间的相互关系尚不清楚。增殖细胞核抗原[3 ] (ｐｒ…     

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1　概述近 30年来 ,原发性肝癌 (ＨＣＣ ,简称肝癌 )的诊治取得了很大进展 ,以手术为主的综合治疗使部分病人获得治愈。但肝癌有侵犯血管尤其是门静脉的生物学特性 ,根据尸检、影像学及肝癌病理学检查 ,2 0 %～ 70 %的ＨＣＣ合并有门静脉癌栓 (ｐｏｒｔａｌｖｅｉｎｔｕｍｏｒｔｈｒｏｍｂｕｓ,ＰＶＴＴ) [1] 。门静脉癌栓的出现使大部分病人失去手术或其它有效治疗措施的机会 ,也是导致肝癌转移复发、影响预后的主要因素。门静脉主支癌栓导致门静脉压力明显升高 ,随之而来的食道胃底静脉曲张破裂出血、腹水及肝功能衰竭等并发症严重影响肝…     

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肝癌细胞要完成转移这一复杂的过程是肝癌细胞本身的生物学特性、宿主微环境和全身状态的改变共同作用的结果。1　肝癌转移基因和转移抑制基因　　肿瘤细胞自身与宿主之间错综复杂的关系 ,受多种基因调控 ,肝癌转移基因的激活和转移抑制基因的失活均可诱发肝癌细胞转移表型的改变而导致转移的发生。免疫组化发现ｐ5 3蛋白的阳性率在有转移的肝癌为 85 7%(12 / 14) ,无转移肝癌为 43 75 % (7/ 16 ) ;有血管侵犯的肝癌为 10 0 % ,无血管侵犯者为 6 6 7% ,并与肝癌病人的生存期密切相关[1～ 3] 。ｍｄｍ2基因的表达与 ｐ5 3基因突变相关 ,二…     

Hazards and complications of endoscopic transurethral ureterolithotripsy and lithoextraction and means of prevention

Complications related to ureterolithotomy and ultrasonic ureterolithotripsy performed under the control of visual endoscope were analyzed in 86 ureterolithiasis patients, methods of their prevention discussed. All the aforementioned complications were distributed into three groups: inapplicability of surgery due to anatomic and functional defects of lower and upper urinary tracts, intraoperative, and postoperative complications. The commonest ones were ureteral abruption and perforation, acute pyelonephritis, temporary vesicoureteral reflux. Their control measures were considered as relative methods of treatment: immediate surgical intervention in case of ureteral abruption, renal catheterization in patients with insignificant ureteral perforation or acute pyelonephritis. Adequate ureteroscopy, careful consideration of pro- and contraindications, catheterization of renal pelvis and urinary bladder performed within 2-3 days after the surgery and adequate antibacterial therapy are the most decisive steps in the control of aforementioned complications.     

牙体牙弓颌骨阻力中心及其临床意义

牙体、牙弓及颌骨的阻力中心在正畸矫治力系统中具有重要的意义,也是正畸学领域争论较多的一个问题。Dermaut等研究表明,当力作用于物体阻力中心时,物体将发生平动,否则将发生平动和转动的复合运动。目前,国内外多数学者认为牙体、牙弓及颌骨存在阻力中心,但其位置存在争议。本文就牙体、牙弓及颌骨的阻力中心及其临床意义作一综述。     

Characterisation and differentiation of chondroblastomas and chondromyxoidfibromas - presence and expression of collagen types I and II

AIM: Chondroblastomas and chondromyxoidfiibromas are rare benign skeletal neoplasms with reported overlapping histology. Aim of this study was to analyse the biochemical composition of the matrix of these tumour entities in order to further characterise the cellular phenotypes of these neoplasms using typical cell biological marker genes. METHODS: The matrix compositions of chondroblastomas and chondromyxoidfibromas were analyzed by HE-histology, histochemistry, and immunolocalization techniques. Cellular gene expression patterns were detected by mRNA in situ hybridization. RESULTS: Chondroblastomas are rich in collagen type I and show foci of an osteoid-like matrix, whereas collagen type II as a typical marker of chondrocytic differentiation was not detected in any of the specimens. Chondromyxoidfiibromas had foci of chondroid appearance with chondroblastic cellular differentiation characterised by collagen type II expression. CONCLUSION: These results characterise chondroblastomas and chondromyxoidfiibromas as skeletal neoplasms that have a different biology and which can be distinguished by matrix protein expression products: collagen type II, the typical marker of chondroblast differentiation, could only be detected in chondromyxoidfibromas, but not in chondroblastomas. Thus, both neoplasms are clearly different on the cell biological level.     

Histochemical and contractile properties of striated muscles of urethra and levator ani of dogs and sheep

AIMS: To understand their possible importance in long- and short-term control of continence, some properties of the striated muscles of the urethra and pelvic floor (levator ani) of dogs and sheep were investigated, especially fiber types and contractile characteristics. MATERIALS AND METHODS: Striated muscles of urethra and levator ani of 29 male and 6 female dogs and 11 male and 6 female sheep were removed and cut into strips. Some strips were frozen and stained for ATPase at pH 9.4 and 4.3 for fiber typing; others were set up in an organ bath to study contractile responses to nerve stimulation. RESULTS: All muscles contained both type I (slow) and type II fibers, ranging from 97% type II in female greyhound urethra to 60% in female sheep levator ani. For each muscle, there were fewer type II muscles in sheep than in dog. The diameters of the urethral fibers were about 60% of the levator ani in dogs and 34% in sheep. Contraction of the urethral muscle was faster than for levator ani and declined to about 80% of the peak, 500 msec after the beginning of stimulation at 20 Hz. The levator ani contraction rose to a steady level as long as stimulation continued. CONCLUSIONS: Both the levator ani and urethral striated muscles contain slow and fast fiber types. The levator ani muscles are capable of sustained contraction with rapid onset which will produce long-term closure of the urethra. The circular urethral muscle contraction was faster but less well maintained.     

Incorporation and release of85Sr and14C-proline-hydroxyproline in mineral and collagen of bone and incisor teeth of growing rats

The extent to which exchange and reutilization processes of mineral tracers affect skeletal mineral accretion and resorption measurements was evaluated by comparing the rates of appearance and disappearance of⁸⁵Sr and¹⁴C-proline-hydroxyproline in bones and teeth in growing rats for 12 days following simultaneous parenteral injection of these tracers. Expressions for the relative rates of collagen synthesis and breakdown, which unlike mineral metabolism are considered not to be complicated by exchange phenomena, were based on¹⁴C-proline conversion to¹⁴C-hydroxyproline; the specific activity of the latter was determined. Both the mineral and the collagen specific activities reflected the rates and patterns of growth of the samples assayed; rapid growth and a short interval of time between formation and resorption of tissue in themetaphyseal bone which contains the cartilagineous growth plate, slow growth and an interval of time between formation and resorption of tissue indiaphyseal bone and incisor teeth which is longer than the 12 days of the experiment. However, in metaphyseal bone the specific activity collagen/mineral ratio dropped by one half during the 4–12 day interval in contrast to diaphyseal bone and incisor teeth in which no change in this ratio was observed during this period of time. The data indicate that collagen in the metaphyseal growth zone is removed by resorption before it has become fully mineralized, and that exchange is a relatively unimportant factor in the long term kinetics of bone mineral.

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Zusammenfassung Das Ausmaß, bis zu welchem Austausch- und Wiederverwendungsprozesse der mineralen Tracer die Messungen des mineralen Skelett-Auf- und Abbaues beeinflussen können, wurde ausgewertet; zu diesem Zweck wurde die Geschwindigkeit des Auftretens und Verschwindens von⁸⁵Sr und von¹⁴C-Prolin-Hydroxyprolin in Knochen und Zähnen von wachsenden Ratten während der 12 auf die simultane parenterale Injektion dieser Tracer folgenden Tage verglichen.Der Ausdruck für die relative Geschwindigkeit des Kollagen-Auf- und Abbaues, bei welchem im Gegensatz zum Mineralmetabolismus kein Mitwirken des Austauschphänomens vermutet wird, basiert auf der Umwandlung von¹⁴C-Prolin zu¹⁴C-Hydroxyprolin; die spezifische Aktivität des letzteren wurde bestimmt.Aus der spezifischen Aktivität des Minerals sowie jener des Kollagens konnten die Geschwindigkeit und die Art des Wachstums der untersuchten Proben ersehen werden, d.h.schnelles Wachstum und ein kurzes Zeitintervall zwischen Bildung und Resorption des Gewebes imKnochen der Metaphyse, die auch die knorpelige Wachstumsplatte enthält, und andererseitslangsames Wachstum und längeres Zeitintervall (länger als die 12 Tage des Experimentes) zwischen Bildung und Resorption des Gewebes imKnochen der Diaphyse und in den Schneidezähnen. Immerhin fiel die spezifische Aktivität des Kollagen/Mineral-Anteils im Knochen der Metaphyse während dem 4–12tägigen Zeitintervall auf die Hälfte, im Gegensatz zum Knochen der Diaphyse und der Schneidezähne, bei welchen während dieser Zeitspanne kein Unterschied in diesem Verhältnis beobachtet wurde.Diese Ergebnisse zeigen, daß Kollagen in der Wachstumszone der Metaphyse durch Resorption verschwindet, bevor es ganz mineralisiert ist, und daß der Austausch ein relativ unwichtiger Faktor in der Kinetik auf lange Sicht des Knochenminerals ist.