荧光原位分子杂交技术检测乳腺癌组织中HER-2基因的表达

目的荧光原位杂交(FISH)方法检测乳腺癌组织中人表皮生长因子2(HER-2)的扩增情况,并与免疫组化(IHC)结果相比较,优化乳腺癌HER-2检测方法。方法采用相关质控措施,利用FISH法及IHC法检测乳腺癌HER-2的表达,分析相关性。结果 FISH检测Her-2基因扩增40例,阳性率30.5%(40/131),在IHC(-)标本中,Her-2基因未扩增19例,阴性符合率为95.0%;在IHC(+)标本中,Her-2基因扩增7例,阳性符合率为20.0%;在IHC(++)标本中,Her-2基因扩增15例,阳性符合率26.8%;在IHC(+++)标本中,Her-2基因扩增17例,阳性符合率89.5%。结论 FISH检测准确率较高,IHC阴性和(+++)时与FISH结果一致性较好,可作为治疗依据,IHC(+)和(++)时仅能作初步筛查,明确Her-2情况仍需结合FISH检测。     

弥漫性大B细胞淋巴瘤中bcl-2蛋白表达及其与基因异常的相关性

目的探讨弥漫性大B细胞淋巴瘤(DLBCL)中bcl-2蛋白表达情况及其与bcl-2基因易位和扩增的相关性。方法应用免疫组织化学SP法检测59例DLBCL中bcl-2蛋白表达情况,并应用IgH/bcl-2双色双融合探针荧光原位杂交(FISH)检测bcl-2基因的易位和扩增。结果 59例DLBCL中bcl-2蛋白的表达率为67.8%(40/59),bcl-2基因易位和扩增的发生率分别为1.7%(1/59)和22.0%(13/59),bcl-2基因异常(易位或扩增)的总检出率为23.7%(14/59),bcl-2蛋白表达与基因异常之间无相关性(P=1.000)。结论 bcl-2基因可能在DLBCL的发病机制中起重要作用,bcl-2蛋白表达的调控机制有待进一步阐明。     

乳腺癌中HER-2异常表达的临床病理学意义

目的探讨免疫组织化学(IHC)和荧光原位杂交(FISH)检测乳腺癌组织中人表皮生长因子受体2(HER-2)异常表达的关系。方法分别采用IHC和FISH方法检测155例乳腺癌标本中的HER-2表达情况。结果 155例乳腺癌标本中,IHC与FISH结果符合率为80.6%,存在一致性(Kappa=0405,P=0000)。结论 IHC和FISH能有效地检测乳腺癌中HER-2的表达,两者可以取长补短,提高HER-2检测的准确率。     

氯硝基安定抗惊厥作用耐受及停药前后对大鼠下丘脑室旁核CRHmRNA表达的影响

氯硝基安定抗惊厥作用耐受性及依赖性机制目前尚不清楚。近来发现促肾上腺皮质激素释放激素（ＣＲＨ）具有促惊厥作用，并参与应激过程。我们用原位杂交方法观察了氯硝基安定抗大鼠点燃惊厥耐受及停药前后下丘脑室旁核ＣＲＨｍＲＮＡ表达的变化，发现耐受时ＣＲＨｍＲＮＡ表达增强，停药７ｄ时不再变化。提示ＣＲＨ参与了耐受性与依赖性过程。     

34例乳腺癌HER-2基因扩增与蛋白表达相关性研究

目的了解乳腺癌组织中人表皮生长因子受体2（HER-2）基因扩增与Her-2／neu蛋白表达的一致性与相关性。方法采用荧光原位杂交（FISH）法及免疫组化（IHC）法分别检测349例浸润性乳腺癌患者的HER-2基因扩增与蛋白表达情况,分析二者检测结果的一致性与相关性。结果FISH与IHC符合率为55．3％,结果存在一致性（Kappa=0．189,P=0．000）,呈正相关（r=0．434,P=0．000）。结论FISH与IHC两种检测结果存在一致性,但IHC（＋～＋＋＋）患者均应以FISH检测作为评价HER-2基因是否扩增的标准方法。     

Snail mRNA与环氧化酶-2mRNA在乳腺癌中的表达及临床意义

目的 探讨Snail mRNA及环氧化酶-2(COX-2) mRNA在乳腺浸润性导管癌中的表达及其临床意义.方法 应用原位杂交技术测定30例乳腺单纯性增生、30例乳腺导管内癌和70例乳腺浸润性导管癌组织中Snail mRNA及COX-2 mRNA的表达.结果 ①Snail mRNA及COX-2 mRNA在乳腺单纯性增生...     

Extraendothelial and constitutive COX‐2 expression is involved in the contractile effect of angiotensin II in the rat aorta

1 The role of the extraendothelial and constitutive isoforms of cyclo‐oxygenase‐2 (COX‐2) in the contractile effect of angiotensin II (Ang II) was investigated using thoracic and abdominal aortic rings without endothelium from young Wistar rats. 2 Ang II elicited similar contractions in both aortic segments, and the effect was inhibited by pretreatment with NS398 (a selective COX‐2 inhibitor) but not SC‐560 [selective cyclo‐oxygenase‐1 (COX‐1) inhibitor]. 3 COX‐2 mRNA was expressed under basal conditions in both aortic segments. Additionally, Ang II increased COX‐2 mRNA expression in the abdominal but not the thoracic segment, while cycloheximide (a protein synthesis inhibitor) did not affect the contractile response to Ang II in either of the two segments; this suggests that the effect is not associated with de novo COX‐2 synthesis. 4 In conclusion, the basal amount of COX‐2 found in aortic smooth muscle cells is sufficient to explain the production of the prostanoids related to the contractile effect of Ang II. The production of these prostanoids, which are derived from constitutive COX‐2, occurs independently of the endothelium vascular system.     

Effect of pregnancy on the roles of nitric oxide and prostaglandins in 5-hydroxytryptamine-induced contractions in rat isolated thoracic and abdominal aorta

1. Vascular resistance and sensitivity to circulating pressor and vasoconstrictor substances are blunted during pregnancy. This has been attributed mainly to an increased production of endothelium-derived mediators. The aim of the present study was to determine whether pregnancy changes the relative participation of nitric oxide (NO) and prostaglandins (PG) in the modulation of the contractile response to 5-hydroxytryptamine (5-HT) in two anatomically distint segments of the rat aorta. 2. Full concentration-response curves to 5-HT were obtained in isolated rings from the thoracic and abdominal portion of the aorta from pregnant and non-pregnant rats in the presence and absence of the NO synthase (NOS) inhibitor N(G)-nitro-l-arginine methyl ester (L-NAME; 10 micromol/L) or the PG synthesis inhibitor indomethacin (10 micromol/L). Cyclo-oxygenase (COX)-1, COX-2 and endothelial (e) NOS protein expression were determined in the same tissues by immunoblot. 3. The effects of pregnancy were accentuated in the abdominal compared with the thoracic aorta. In addition, the relative participation of the NO and PG pathways seems to be changed during pregnancy. Although NO seems to be the mediator mainly responsible for the effect of pregnancy in the thoracic aorta, our results suggest a complex interaction between NO and PG in the abdominal aorta. Indomethacin significantly reduced the contractile response of both segments of the aorta, whereas expression of COX-1, COX-2 and eNOS were increased only in the abdominal segment of pregnant animals. 4. These results show that the effect of pregnancy is not homogeneous along the aorta. There seems to be a mutual interaction between PG and NO in the abdominal, but not in the thoracic, aorta from pregnant rats: the role of NO becomes evident in the absence of vasodilatory PG, whereas the participation of the latter increases in the absence of NO working as a compensatory mechanism.     

吗啡依赖及戒断大鼠相关脑区多巴胺D2受体基因表达的变化

目的:研究吗啡依赖及戒断大鼠相关脑区D2R基因表达的变化.方法:将24只雄性Sprague-Dawley大鼠随机等分为吗啡依赖组(吗啡组:在大鼠腹腔内注射盐酸吗啡,2次/d,起始剂量为5mg/kg,逐日递增5 mg,至第10天为50 mg/kg)及生理盐水对照组(对照组:用相同方式注射同体积的生理盐水),于末次注射后3 h及72 h处死(每组每时间点各6只),取中脑腹侧被盖区(VTA)、伏隔核(Nac)、中脑导水管灰质(PAG)、尾壳核(CPU)、海马CA1区(HIPCA1)等脑组织.利用组织原位杂交技术检测各脑区的D2R mRNA的吸光度(A)值,并作同期平行比较.结果:末次注射3 h,吗啡组大鼠五个被检脑区D2R mRNA A值均低于同期对照;末次注射72 h,尾壳核高于同期对照,其它四个被检脑区仍低于同期对照,差异具有显著性(P<0.05或0.01).结论:慢性吗啡处理可抑制大鼠相关脑区D2R基因表达,戒断后有不同程度的恢复.     

Effects of chronic inflammation and morphine tolerance on the expression of phospho-ERK 1/2 and phospho-P38 in the injured tissue

Opioids are used in humans in the treatment of chronic osteoarticular pain, but the development of tolerance to the analgesic effects after continuous administration is still not well understood. The aim of the present study was to evaluate the expression of phospho-ERK 1/2 and phospho-p38 in mice with monoarthritis chronically exposed to morphine as a possible explanation for the development of tolerance. Inflammation was induced by intraplantar injection of complete Freund’s adjuvant (CFA) and the tolerance by implantation of 75 mg morphine pellets. The results of the present study show that ERKs phosphorylation is unaltered by inflammation or morphine tolerance, each one individually, in the plantar tissue. In contrast, phospho-p38 is similarly decreased by inflammation or morphine tolerance. In naïve but not in tolerant animals, acute injection of morphine induces significant increase in phospho-p38 without any changes in phospho-ERK 1/2 expression. During inflammation, the acute injection of morphine induces a significant increase in the expression of ERK 1/2, but not in phospho-p38, in naïve animals. Phospho-ERK 1/2 expression was significantly decreased in the presence of inflammation plus tolerance. In contrast, no significant differences in phospho-p38 expression were observed between naïve and tolerant animals acutely injected with saline or morphine in presence of CFA inflammation. These results suggest that ERK but not p38 could be implicated in the development of morphine tolerance during peripheral inflammation. These experiments could contribute to establish the mechanisms implicated in the development of morphine tolerance in presence of inflammatory pain.     

胃癌组织HER-2基因的检测用于靶向治疗

目的 荧光原位杂交（FISH）法与免疫组织化学（IHC）染色法检测胃癌组织HER-2基因状态,用于靶向治疗。方法 应用FISH法检测77例IHC染色法CerbB-2蛋白表达为（2＋）及（3＋）的胃腺癌标本中HER-2基因扩增情况。结果 FISH结果,22例HER（3＋）标本中,17例存在HER-2基因扩增,符合率77.3%：5例阴性病例中2例为多倍体,占40%。55例HER-2（2＋）的病例中,9例（16.3%）存在HER-2基因扩增,46例阴性病例中19例为多倍体,占41.3%。结论 对于IHC筛查HER-2（3＋）及HER-2（2＋）拟行靶向治疗的病例明确HER2基因状态,需进一步行FISH检测,同时要考虑l7号染色体多体的影响。     

左旋千金藤立定诱导6-羟基多巴胺损毁大鼠的纹状体Fos、前脑啡肽mRNA、前强啡肽mRNA表达与旋转行为的关系(英文)

目的:研究左旋千金藤立定(SPD)调节6-羟基多巴胺(6-OHDA)损毁大鼠纹状体的Fos、前脑腓肽(PENK)和前强啡肽(PDYN)mRNA表达水平与旋转行为的关系.方法:用免疫细胞化学法观察Fos表达,用地高辛非同位素法标记的寡核苷酸探针检测纹状体PENK和PDYN mRNA,进行图像处理作半定量分析.结果:(1)SPD给予1,3,7 d后,损毁大鼠旋转行为仍维持在高水平;(2)SPD诱导双侧纹状体Fos显著表达,尤以损侧为甚.SPD重复应用使双侧纹状体的Fos诱导表达下降,尤以健侧为显著;(3)与健侧相比,损毁侧纹状体的PENK mRNA表达水平增加非常显著.SPD重复应用7 d,使这种增加的PENK mRNA水平明显下降.同时,SPD也使健侧PENK mRNA水平降低.然而,6-OHDA损毁和SPD处理对双侧纹状体的PDYN mRNA水平无明显影响.结论:SPD激发6-OHDA损毁大鼠旋转行为维持在高水平,与损侧纹状体的Fos表达和PENK mRNA水平下降是同步的.但是,6-OHDA损毁和SPD均未显示出对PDYN mRNA表达的影响.     

胃癌组织环氧合酶-2异常表达与螺旋CT征象相关性研究

目的探讨胃癌组织环氧合酶-2(COX-2)异常表达与螺旋CT的关系。方法用免疫组化(SP)方法检测胃癌手术切除67例标本中上述基因表达。结果胃癌COX-2过表达率为70.1%(47/67),且过表达与肿瘤大小、大体类型、组织学类型、淋巴结转移(LN)及临床病理分期密切相关(P<0.05)。结论胃癌COX-2异常表达与螺旋CT征象密切相关。     

乙型和丙型肝炎病毒双重感染肝组织病毒标志物的检测

为观察乙型肝炎和丙型肝炎双重感染的尸体肝组织中两种病毒核酸及抗原的分布，对１５例ＨＢＶ与ＨＣＶ双重感染的尸检肝组织用免疫组织化学法检测ＨＢｓＡｇ、ＨＢｃＡｇ和ＨＣＡｇ，Ｄｉｇｏｘｉｇｅｎｉｎ标记的探针原位杂交法分别检测ＨＢＶ ＤＮＡ和ＨＣＶ ＲＮＡ。结果：ＨＢｓＡｇ、ＨＢｃＡｇ、ＨＢＶ ＤＮＡ、ＨＣＡｇ和ＨＣＶ ＲＮＡ的阳性数分别为１２／１５（８０．０％）、１０／１５（６６．７％）、９／１５（６０     

人TIMP-2基因的克隆及其原核表达

目的 构建人TIMP-2重组表达载体,并在大肠杆菌BL21 (DE3)中表达.方法 提取人肝癌细胞(Hep G2)总RNA后,经RT-PCR扩增获得目的片段,插入克隆载体pMD18-T;酶切鉴定和测序后将TIMP-2导入原核表达载体pGEX-4T-1中,构建重组质粒pGEX-TIMP-2.将重组质粒转化至大肠杆菌菌株BL21 (DE3)中,IPTG诱导融合蛋白表达后用SDS-PAGE电泳检测并用western blot验证(蛋白质印迹法).结果 成功构建原核重组表达载体pGEX-TIMP-2,并在原核宿主BL21中大量表达融合蛋白GST-TIMP-2.结论 克隆人TIMP-2基因并在原核生物中大量表达.     

氟西汀对大鼠海马脑源性神经营养因子基因表达的作用

目的　为了进一步探讨抗抑郁药作用的分子机制。方法　采用原位杂交技术 (地高辛标记的BDNFcDNA探针 )检测长期给予抗抑郁药氟西汀及生理盐水大鼠海马神经元BDNFmRNA的表达。结果　与生理盐水组相比 ,氟西汀组表达增强 ,尤以CA3、CA1区为甚。结论　长期给药后脑内有关区域BDNF表达改变可能是抗抑郁效应发生的受体后分子作用机制之一     

Effect of chronic treatment with typical and atypical neuroleptics on the expression of dopamine D2 and D3 receptors in rat brain

The effect of chronic treatment (21 days) with typical and atypical neuroleptics on the expression of striatal and limbic D₂ and D₃ dopamine receptors was investigated in rat brain by in situ hybridization and receptor autoradiography. Haloperidol and sulpiride increased D₂ receptor expression in striatal and limbic areas. In contrast, clozapine had no effect on D₂ receptor expression. Haloperidol decreased D₃ receptor expression in limbic areas, with the exception of the islands of Calleja where an increase occurred. Sulpiride and clozapine increased D₃ receptor expression in limbic and striatal regions but decreased D₃ receptor expression in the islands of Calleja. This study demonstrates that chronic treatment with typical and atypical neuroleptics produces different regionally specific changes in limbic and striatal D₂ and D₃ receptor expression. The alterations in dopamine receptor expression were different for each drug, but a distinction between the effects of atypical and typical neuroleptics could be made. Comparison of mRNA levels in animals which were not withdrawn from drug treatment with those that were withdrawn, demonstrated that some changes in receptor expression occurred during drug treatment, whilst others only manifested when drug treatment had ceased. The different regulation of dopamine D₂ and D₃ receptor expression by typical and atypical neuroleptics may have relevance to the ability of these drugs to cause extrapyramidal side-effects. Received: 26 February 1996 /Final version: 28 July 1996     

慢性肝病肝组织中TNF-α、TNFR及Bcl-2家族的表达

目的：探讨乙型肝炎病毒相关性慢性肝病肝损伤时TNF－α、TNFR及Bcl-2家族对肝细胞凋亡的调节作用。方法：采用HE染色、免疫组织化学方法检测28例慢性乙型肝炎、22例乙型肝炎后肝硬化患者肝组织中Fas、FasL、Bax、Bcl-2、Bcl-α及Bcl-XL的表达，用分子杂交法检测肝组织中TNF－αmRNA、TNF受体（TNFR) 表达的阳性率及表达程度。结果：各种凋亡蛋白在慢性肝炎组和肝硬化组中的表达无显著差别，但促凋亡因子与抑凋亡因子相比，有显著或极显著差异（P＜0．05，P＜0．01）。TNF－α、TNFR阳性表达率在肝硬化组为90．9％、95．5％，明显高于慢性肝炎组（P＜0．05，P＜0．01）。结论：抑制促凋亡因子、阻止肝细胞大量凋亡对防治慢性肝炎重症化有重要的临床意义。     

多奈哌齐对血管性痴呆模型小鼠海马p38 MAPK mRNA表达的影响

目的探讨多奈哌齐对血管性痴呆模型小鼠海马p38 MAPK mRNA表达的影响。方法双侧颈总动脉线结反复缺血—再灌注法制备血管性痴呆小鼠模型,设多奈哌齐治疗组,采用多奈哌齐连续治疗4 wk,并设假手术组及模型组为对照组,利用跳台试验和水迷宫试验观测其行为学改变,用原位杂交技术观测其p38 MAPK mRNA的表达变化。结果多奈哌齐组小鼠学习、记忆成绩明显优于血管性痴呆模型组(P<0.05),其海马p38 MAPK mRNA表达明显增强(P<0.05,P<0.01)。结论多奈哌齐影响血管性痴呆小鼠海马p38MAPK mRNA表达,改善血管性痴呆小鼠学习、记忆功能。