原位PCR和免疫组化检测垂体腺癌P16基因

目的研究人垂体腺瘤P16基因的改变,同时探索原位PCR技术的适宜条件和结果确认,以及用于基因缺失检测的可行性。方法原位PCR和免疫组化技术。结界绝大部分间质细胞为P16蛋白免疫组化阴性,而一部分垂体肿瘤细胞呈P16阳性;针对P16基因的原位PCR信号则可见于垂体肿瘤细胞和间质细胞等各种细胞,即P16组化阴性的细胞同样表明有P16基因存在。结论原位PCR可以是P16基因研究的一种有效手段,提示垂体腺癌的P16基因改变形式可能主要是表达过度而不是基因缺失。     

CARD In Situ Hybridization: Sights and Signals

During the last decade, several strategies have been developed to improve the detection sensitivity ofin situ hybridization (ISH) by amplification of either target nucleic acid sequences prior to ISH (e.g.,in situ PCR), or the detection signals after the hybridization procedures (signal amplification). Here we outline the principles of tyramide signal amplification using the catalyzed reporter deposition (CARD) technique, summarize applications as well as possible limitations of CARD ISH, and discuss some future directions ofin situ nucleic acid detection using this amplification strategy.     

Receptor tyrosine kinase tie 1 mRNA is upregulated on cerebral microvessels after embolic middle cerebral artery occlusion in rat

Tie 1 is an endothelial specific transmembrane receptor tyrosine kinase and may be required during angiogenesis. Using in situ hybridization, we measured tie 1 mRNA in ischemic brain (n=15). Rats were subjected to middle cerebral artery (MCA) occlusion by a single fibrin rich clot. Expression of tie 1 was not detected in non ischemic brain. Cerebral microvessels expressed tie 1 in the ischemic lesion as early as 2 h after MCA occlusion. The number of microvessels containing tie 1 mRNA decreased in the ischemic lesion at 8 h after MCA occlusion. However, expression of tie 1 increased on microvessels at 24 h and 14 days after ischemia and tie 1 was primarily localized to the microvessels bordering pan necrotic tissue. Ninety-seven percent of cerebral vessels which expressed tie 1 mRNA had diameters of 3.7+/-0.17 microm. Our findings suggest a role for tie 1 in cerebral microvascular remodeling after embolic stroke.     

三种镍化合物诱发转化细胞恶性度的鉴别与分析

目的 :利用已建立的三种镍化合物体外转化细胞进行裸鼠体内成瘤实验研究 ,比较它们恶性度的差异。方法 :三种镍化合物转化细胞接种BALB/C裸鼠 ,进行肿瘤细胞和转化细胞的透射电镜及扫描电镜观察。结果 :硫化镍 ,氯化镍 ,硫酸镍分别诱发的转化细胞都能在BALB/C裸鼠皮下形成肿瘤 ,病理组织学检查确定为纤维肉瘤。经扫描电镜与透射电镜观察瘤细胞形态与转化细胞有一定差别。结论 :本文进一步说明了三种镍化合物所诱发的细胞转化为恶性转化 ,且都具有体内致瘤性。     

Fibroblast growth factor induces primitive streak formation in rabbit pre-implantation embryos in vitro

Culturing of rabbit pre-implantation embryos was performed in Ham's F10 medium supplemented with polyvinylpyrrolidone. Under these culture conditions, day 6 post coitum blastocysts increased their diameter within 24 h to 80% of that of day 7 blastocysts grown in vivo. Despite this substained growth, the embryonic disc remained undifferentiated with clear signs of degeneration after 24 h of culture. Basic fibroblast growth factor (bFGF) was able to overcome this developmental block. After 12 h of culture, day 6 blastocysts showed pear-shaped embryonic discs, and after 24 h, the primitive streak with Hensen's node was visible. The bFGF had no comparable effects on day 5 and day 7 blastocysts. The embryonic discs of day 5 blastocysts degenerated, even in the presence of bFGF, whereas day 7 blastocysts were able to form their primitive streak, also in the absence of bFGF. TGF ₁ did not promote embryonic development in vitro. The data indicate that the onset of mesoderm formation in the rabbit is controlled by a growth factor of the FGF-family.     

The feasibility of demonstration of GM- and InV-systems in decaying organs

Summary -globulin factors (Gm and InV-systems) were determined in 12 cadavers in blood and various organ exprimates (kidney, liver, spleen and muscle). Also checked was the time interval up to which these factors could still be demonstrated in decaying organs.For this purpose blood and portions of the organs were left to decay in plastic containers at an average temperature of 19.1°C. Examinations were done with the agglutination-inhibition-test. The exprimates were employed in dilutions 1:10 and 1:20. The serum factors Gm (1), (2), (4), (10) and InV (1) could be demonstrated for varying periods of time.In organs they could be demonstrated for between one and eight weeks. Only factors Gm(1) and Gm(2) could be demonstrated beyond this time. As a rule these serum factors were demonstrable in blood for a longer period of time than in organs. Factor Gm(4) proved to be the most stable one, as it could be demonstrated up to 11 weeks in decaying serum.In 2200 individual tests with a dilution 1:20 no false positive results were obtained, with the dilution 1:10, however, 5 false positive tests were found. No explanation can be given for the different periods of time for which these factors can be demonstrated under conditions of decay.

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Zusammenfassung Bei zwölf Leichen wurden im Blut und in verschiedenen Organpreßsäften (Niere, Leber, Milz und Muskel) die -Globulineigenschaften (Gm- und InV-Systeme) bestimmt und ihre Nachweiszeit bei Fäulnis überprüft. Blut und Organteile wurden hierbei in verschließbaren Plastikgefäßen bei einer mittleren Temperatur von 19,1°C faulen gelassen. Die Untersuchungen erfolgten mit dem Agglutinationshemmtest.Die Preß- bzw. Fäulnissäfte wurden in 1:10 und 1:20 Verdünnungen verwendet. Die Serumfaktoren Gm(1), (2), (4), (10) und Inv(1) konnten über unterschiedlich lange Zeiträume nachgewiesen werden.In den Organen betrug die Nachweiszeit zwischen einer und acht Wochen, wobei nur die Faktoren Gm(1) und Gm(2) über diese Wochen nachweisbar blieben. Die Serumeigenschaften waren im Blut allgemein länger nachweisbar als in den Organen. Am stabilsten erwies sich der Faktor Gm(4), der bis zu elf Wochen im faulenden Serum festgestellt werden konnte.Bei 2200 Einzelbestimmungen wurden mit der 1:20 Verdünnung in keinem Fall falsch positive Ergebnisse erzielt, mit der 1:10 Verdünnung hingegen fünf falsch positive Ergebnisse.Eine Erklärung für die unterschiedlich langen Nachweiszeiten unter Fäulnisbedingungen kann nicht gegeben werden.

     

外阴癌中HPV感染与p53、MDM2表达的关系

目的 :探讨HPV6/ 11、16/ 18在外阴癌组织中的感染情况及与p5 3、MDM2蛋白表达的关系。方法 :用原位杂交法 (ISH)检测HPV6/ 11、16/ 18在 30例外阴癌、2 1例外阴上皮内瘤变 (VIN)及 10例外阴正常皮肤组织中的表达。同时用免疫组化SP法检测p5 3、MDM2蛋白的表达。结果 :HPV6/ 11、16/ 18在外阴癌、VIN中的阳性表达率分别为 60 % (18/30 )、33.33% (10 / 30 ) ,4 2 .86% (9/ 2 1)、2 8.5 7% (6/ 2 1) ,正常对照组没有表达。p5 3、MDM2蛋白在外阴癌、VIN、正常对照组中的阳性表达率分别为 63.33% (19/ 30 )、4 0 .0 0 % (12 / 30 ) ,4 7.62 %(10 / 2 1)、5 2 .38% (11/ 2 1) ,0 % (0 / 10 )、0 % (0 / 10 )。HPV6/ 11的表达在外阴癌组、VIN组与正常组差异有显著性 (P <0 .0 5 ) ,外阴癌组HPV16/ 18表达与正常组差异有显著性 (P <0 .0 5 )。外阴病变各组p5 3、MDM2与正常组差异有显著性 (P <0 .0 5 )。结论 :HPV6/ 11、HPV16/ 18、p5 3、MDM2蛋白在外阴组织的不同病变中表达均差异有显著性 ,在外阴癌的发生发展中HPV感染、p5 3突变和MDM2表达可能起一定的作用     

体外受精胚胎移植后妊娠结局分析

目的　探讨通过体外受精 -胚胎移植妊娠妇女的妊娠结局。方法　对自 2 0 0 0年 9月～ 2 0 0 2年1 1月在我院实施IVF -ET的 75例患者 ,80个妊娠周期的临床资料进行回顾性分析。结果　 80个周期中 ,宫内妊娠率 96 2 5 % (77/ 80 ) ,其中自然流产率 2 3 75 % (1 9/ 80 ) ;妊娠 <1 6周流产率 1 6 2 5 % (1 3/ 80 ) ;妊娠 >1 6周流产率 7 5 0 % (6 / 80 ) ,其中 5周期均发生双胎妊娠 ;分娩率 72 5 0 % (5 8/ 80 ) ,早产率 1 5 0 0 % (1 2 / 80 ) ,宫外孕发生率 3 75 % (3/ 80 )。新生儿死亡率 3 6 1 % (3/ 83) ,均死于三胎妊娠早产。结论　减少IVF -ET后多胎妊娠率 ,加强多胎妊娠的整个孕期监护 ,以利于减少多胎妊娠 >1 6周流产率及早产率 ,是降低IVF -ET后妊娠胎儿丢失的有效方法