208例内镜活检胃癌组织中HER-2蛋白的表达及意义

目的：分析内镜活检胃癌组织中HER-2蛋白的表达及意义。方法应用免疫组织化学( immunohistochemistry, IHC)法检测208例内镜活检胃癌组织中HER-2蛋白表达,采用荧光原位杂交( fluorescence in situ hybridization, FISH)技术检测IHC 2+胃镜活检组织中HER-2基因扩增,并对相应的临床病理特征(性别、年龄、肿瘤发生部位及Lauren分型)进行综合分析。结果208例内镜活检胃癌组织中HER-2 IHC 3+阳性率为7．7%(16/208),IHC 2+为13．5%(28/208),IHC 1+为45．7%(95/208),IHC 0为33．2%(69/208)。 IHC 2+的胃癌活检组织中 FISH 阳性率为14．2%(4/28)。 HER-2总体阳性率为9．6%；HER-2蛋白过表达与患者性别、年龄均无关( P>0．05),与发生部位及Lauren分型密切相关(P<0．05)。结论内镜活检胃癌组织中HER-2蛋白过表达与胃癌发生部位、Lauren分型密切相关。     

新旧两版指南对浸润性乳腺癌HER-2基因扩增的判读影响

目的探讨2014版与2019版《乳腺癌HER-2检测指南》对浸润性乳腺癌HER-2基因扩增的判读影响。方法收集698例浸润性乳腺癌石蜡标本,采用免疫组化(immunohistochemistry, IHC)法和荧光原位杂交(fluorescence in situ hybridization, FISH)法对HER-2蛋白表达和基因扩增进行检测,并用2014和2019版指南分别对检测结果进行判读。结果根据2014版指南FISH判读法检测HER-2基因扩增的阳性检出率为40.7%(284/698),阴性检出率为52.1%(364/698),不确定为7.2%(50/698)。根据2019版指南FISH判读法检测HER-2基因扩增的阳性检出率为40.0%(279/698),阴性检出率为60.0%(419/698)。两版指南中IHC与FISH法检测HER-2蛋白表达与基因扩增的总一致率分别为54.7%、55.7%;两版检测结果具有一致性(Kappa=0.277,P0.001;Kappa=0.251,P0.001),且均呈正相关(r=0.608,P0.001;r=0.641,P0.001)。结论 2019版指南取消FISH法检测HER-2基因扩增的不确定结果,使其阴性率有所增加,更加明确了HER-2基因状态,为临床靶向治疗提供参考。     

乳腺Paget病HER2基因扩增及其临床意义

目的 探讨乳腺Paget病组织中HER2基因的表达、扩增及其关系.方法 分别应用免疫组织化学染色及FISH技术检测19例乳腺Paget病癌组织中HER2基因的蛋白表达及扩增.结果 19例乳腺Paget病癌组织中HER2蛋白表达为6例,7例,5例+,1例-.19例乳腺癌中FISH检测HER2基因扩增率68.42%(13/19),其中6例无HER2基因扩增.HER2基因扩增病例中7例为簇状扩增,占扩增病例的53.84%.7例簇状扩增病例中6例HER2蛋白表达,1例.HER2蛋白表达为"-和+"的病例中HER2基因扩增均为"-"(比值小于2).结论 回顾性分析后进一步证明乳腺Paget病的HER2基因扩增率高于其在其他乳腺癌症中的平均表达率,近似于白种人的表达结果.     

胃癌组织中HER-2基因扩增和K-ras基因突变的关系

目的探讨胃癌组织中HER-2蛋白表达和基因扩增与K-ras基因突变的关系及其意义。方法采用免疫组化、FISH和焦磷酸测序技术对67例胃癌组织中HER-2蛋白表达、HER-2基因扩增与K-ras基因的突变率进行了检测。结果 HER-2蛋白阳性率为40.3%(27/67),其中HER-2蛋白3+者9.0%(6/67),HER-2蛋白2+者13.4%(9/67),HER-2蛋白1+者17.9%(12/67)。FISH检测HER-2基因扩增率为18.5%(5/27),HER-2基因拷贝数增加和基因扩增者共48.1%(13/27)。K-ras基因突变定量检测为7.5%(5/67),均为K-ras基因第12密码子突变,其中低于10%低丰度突变2例,高于10%高丰度突变3例(突变数值分别为:17、29、30)。除1例为GGT→GAT突变型外,其它均为GGT→GTT突变型。本组K-ras基因突变5例中除1例既有K-ras基因突变,又有HER-2基因扩增,另外4例HER-2基因均无扩增。结论检测胃癌中HER-2扩增时选用抗肿瘤药物治疗的靶点曲妥珠单抗,同时可选用K-ras基因突变的抗肿瘤药物治疗的靶点西妥昔单抗;联合检测胃癌组织中HER-2基因扩增和K-ras基因突变为靶向抗肿瘤药物治疗过程中受益提供参考指标。     

微滴式数字PCR与IHC联合FISH对乳腺癌新辅助化疗前后HER-2检测的分析

目的分析乳腺癌新辅助化疗前后标本行微滴式数字PCR(droplet digital PCR,ddPCR)技术与免疫组织化学(immunohistochemistry,IHC)联合荧光原位杂交技术(fluorescence in situ hybridization,FISH)检测HER-2表达的一致性,探讨ddPCR技术在HER-2检测中的可行性。方法采用ddPCR技术对62例乳腺癌新辅助化疗前后的标本进行HER-2基因检测,同时运用IHC检测HER-2蛋白表达,并对IHC HER-22+的标本行FISH检测,比较两种方法检测的一致性。结果化疗前穿刺标本中ddPCR检测25例为阳性,37例为阴性;IHC联合FISH检测20例为阳性,42例为阴性;两种方法一致性较好(κ=0.758,P<0.01)。化疗后手术根治标本中ddPCR检测22例为阳性,40例为阴性;IHC联合FISH检测20例为阳性,42例为阴性;两种方法一致性较好(κ=0.784,P<0.01)。结论乳腺癌新辅助化疗前后的标本行ddPCR检测与IHC联合FISH检测HER-2的结果一致性好,证实ddPCR检测乳腺癌新辅助化疗前后HER-2的表达是可靠的,可弥补传统检测方法的局限性。     

胃癌中HER-2表达异质性的病理学分析

目的检测胃癌组织中HER-2蛋白表达水平和基因扩增情况,定量分析HER-2表达和分布特征的异质性,探讨其在判读中的影响。方法应用免疫组化检测并观察胃癌组织中HER-2蛋白表达水平及分布特征,应用荧光原位杂交(fluorescence in situ hybridization,FISH)技术检测HER-2基因扩增情况,并运用欧式距离等进行统计学处理。结果 373例胃癌手术切除标本中HER-2蛋白阳性率(3+)为12.33%,与基因扩增情况显著相关(P0.001)。98例有HER-2蛋白表达的标本总异质性、组内异质性及组间异质性大小差异均有统计学意义(P值分别为0.025、0.001、0.040);组内异质性大小明显低于组间异质性。根据标准判读为HER-2蛋白为2+及3+的病例异质性最大,1+病例最小。HER-2蛋白表达分布较均一的病例近乎为零,而同时具有阴阳脸及花斑样特征的病例最多。根据标准判读HER-2蛋白为0的病例中,有7例具有明显异质性。结论胃癌组织中HER-2蛋白表达的异质性较大且类型多样,判读时首先要增加观察视野,选择至少2个组织块进行HER-2蛋白检测;其次无论HER-2蛋白表达水平高低,均需同时检测基因扩增情况,以确保判读结果的准确性,更好的指导临床用药。     

双色银染原位杂交与荧光原位杂交检测胃癌患者HER2基因状态

目的 比较双色银染原位杂交(DSISH)与荧光原位杂交(FISH)两种技术在胃癌HER2检测中的优缺点,评价DSISH用于胃癌患者HER2基因扩增状态检测的可行性.方法 收集2009年1月至3月在四川大学华西医院行根治性手术的原发性胃或胃食管交界处腺癌80例,行全自动免疫组织化学(IHC)染色检测HER2蛋白表达,所有标本行FISH和全自动DSISH检测HER2基因状态,比较不同检测方法之间的符合率.结果 DSISH和FISH初检均有5例失败,经重复检测后结果满意.IHC检测3+的13例中DSISH检测12例、FISH检测11例为HEB2基因扩增;IHC检测2+的6例中DSISH、FISH检测均有1例为基因扩增;IHC检测1+的18例中DSISH、FISH检测均有2例为基因扩增;IHC检测为0的43例中DSISH、FISH检测均无基因扩增.80例原位杂交病例中,仅1例检测结果不一致(DSISH有基因扩增而FISH无基因扩增),两种方法的总体符合率为98.8%(79/80,κ=0.958,P＜0.01).结论 DSISH技术用于胃癌HER2基因检测结果与FISH符合率高.DSISH与FISH检测各有优缺点,DSISH更具有可行性和实际应用价值.

Abstract：

Objective To investigate the advantages and disadvantages of dual-color silverenhanced in-situ hybridization(DSISH)and fluorescence in-situ hybridization(FISH)for determination of HER2 gene status in gastric carcinoma and to evaluate tlle feasibility of DSISH.Methods Eighty cases of primary gastric or gastroesophageal junction adenecarcinomag diagnosed and treated surgically from January to March.2009 at the West China Hospital were enrolled in the study.Automated immunohistochemistry (IHC)staining,FISH and automated DSISH were carried out to detect the HER2 status,respectively,and the concordance ofthe three techniques was then evaluated.Results DSISH and FISH failed initially,but repeated detection Was successful in 5 eases.Gene amplification was detected in 12/13 IHC 3+ cases in DSISH and in 11/13 IHC 3+ cases in FISH.In 6 IHC 2+cages, the amplification rate was both 1/6;in 18 IHC 1+cases.the amplifieation rate was both 2/18.No amplification Was observed in 43 IHC 0 cases.Only one of the 80 cases showed discrepancy.and tIlerefore the overall concordance between FISH and DSISH Wag 98.8%(κ=0.958,P＜0.01).Conclusions DSISH represents a novel approach for the determination of HER2 status in gastric carcinoma, and the overall concordance between DSISH and FISH is excellent.Despite their advantages and disadvantages.DSISH is more feasible and practical for routine application in surgical pathology.     

免疫组织化学和荧光原位杂交检测肺腺癌ROS1表达的对比

目的比较免疫组织化学(immunohistochemistry,IHC)和荧光原位杂交(fluorescent in situhybridization,FISH)技术在检测肺腺癌中ROS1蛋白表达和融合基因的差异性和一致性,评估IHC在临床上的应用价值。方法应用IHC和FISH技术对332例肺腺癌中ROS1蛋白表达和融合基因进行检测,比较两者检测的结果和相关性,并探讨ROS1融合基因与临床病理特征之间的关系。结果 332例肺腺癌中FISH阳性者13例,阴性者319例,ROS1融合基因阳性率为3.9%;ROS1蛋白表达结果:0者312例,1+者2例,2+者5例,3+者13例,ROS1蛋白过表达率为6.0%;经FISH检测312例ROS1蛋白表达为0的标本无ROS1融合基因,2例1+的标本中也无ROS1融合基因;5例2+的标本中有2例ROS1融合基因,13例3+的标本中11例有ROS1融合基因;IHC检测0和1+、2+、3+标本FISH检测阳性符合率分别为0(0/314)、40%(2/5)和84.6%(11/13);IHC检测ROS1蛋白为2+~3+的标本中72.2%(13/18)显示ROS1融合基因,0~1+患者中无1例有ROS1融合基因(Kappa系数=0.831,P0.01);IHC检测ROS1蛋白表达的敏感性和特异性分别100%和98.4%。结论 IHC检测ROS1蛋白表达和FISH检测ROS1融合基因,两种方法有较高的符合率,一致性较好,IHC有较高的敏感性和特异性,可作为临床检测肺腺癌ROS1融合基因的简单而有效的方法。     

显色原位杂交和免疫组织化学检测乳腺癌HER2/neu基因状况和蛋白表达的对照性研究

目的探讨显色原位杂交（CISH）在检测乳腺癌中HER2／neu基因扩增上的作用。方法挑选乳腺浸润性导管癌患者组织石蜡蜡块（回顾性255例,前瞻性271例）,进行免疫组织化学（IHC）、CISH检测。15例回顾性标本送往德国HERA检测中心进行FISH检测。结果（1）回顾性病例中IHC阳性3＋者CISH基因扩增率为91．6％（120／131）,IHC2＋者CISH基因扩增率为56．5％（39／69）,IHC与CISH检测结果符合率为81．2％（207／255）,两者明显相关（P〈0．01）。（2）前瞻性病例中IHC蛋白过表达率31．7％．CISH基因扩增率27．3％。IHC3＋者CISH基因扩增率为91．4％（53／58）,IHC2＋者CISH基因扩增率为46．4％（13／28）,IHC与CISH检测结果符合率为89．7％（243／271）,两者明显相关（P〈0．01）。（3）经德国检测中心荧光原位杂交（FISH）检测的15例中14例和CISH结果完全一致,1例检测失败,而CISH为无扩增。（4）CISH检测基因扩增与雌激素受体（ER）、孕激素受体（PR）表达明显负相关（P值均〈0．01）。结论CISH检测HER2基因扩增结果与IHC检测蛋白过表达及FISH结果高度一致,CISH是检测HER2基因扩增的一项新技术。     

结直肠癌组织中EGFR及HER-2的表达及临床意义

目的：探讨结直肠癌组织中表皮生长因子受体（ epiderma1 growth factor receptor,EGFR）和人类表皮生长因子受体-2（ hu-man epiderma1 receptor-2,HER-2）蛋白的表达,并分析二者与临床病理特征的关系。方法采用免疫组化PV-9000两步法检测78例结直肠癌组织中EGFR和HER-2的表达,分析二者表达与结直肠癌临床病理特征的关系;应用银染原位杂交（ si1ver in situ hybridization,SISH）法检测结直肠癌组织中HER-2基因扩增情况。结果 EGFR在结直肠癌组织中的阳性率为69.23%（54/78）,EGFR表达与结直肠癌浸润深度、淋巴结转移呈正相关,与患者性别、年龄、肿瘤大小、细胞分化程度、Dukes分期无关;HER-2在结直肠癌组织中的阳性率为25.64%（20/78）,HER-2表达与结直肠癌浸润深度、淋巴结转移呈正相关,与患者性别、年龄、肿瘤大小、细胞分化程度、Dukes分期无关。EGFR与HER-2蛋白表达呈正相关。20例HER-2蛋白阳性者中,10例HER-2基因扩增;其中15例HER-2蛋白（~）中有10例HER-2基因扩增,占高表达组的66.67%;5例HER-2蛋白（+）者无HER-2基因扩增。结论 EGFR和HER-2蛋白在结直肠癌中均呈高表达,EGFR和HER-2的表达与结直肠癌侵袭、转移密切相关,二者可能存在协同作用。HER-2蛋白（~）与HER-2基因扩增密切相关,故该类患者可考虑先行免疫组化初步筛选,再行SISH法检测确认,从而为结直肠癌分子的靶向治疗提供有价值的信息。     

Secular change in menarche in women in Madrid

The age at menarche was estimated by recollection in 1617 women between the ages of 18 and 60 in Madrid and a nearby suburb, Pinto. The population of Pinto is working-class and the Madrid group, taken from residential neighbourhoods , belongs to the upper middle class. In both groups we found a diminution in average age at menarche, from 14.04 to 13.02 years in Madrid and from 14.55 to 13.16 years from about 1935 to about 1965 in Pinto. These changes have been more intense in the group which is less well-off economically, where living conditions have varied much more drastically.     

Intestinal helminthiasis in school children in Haiti in 2002

A survey on intestinal helminths in school children was conducted in Haiti in 2002. This first nationwide study involving the entire country was stratified by department according to urban and rural zones using the cluster method. Focusing on elementary school children (n=5792; age range 3 to 20 years), it involved 26 urban and 49 rural schools randomly selected. Stools were preserved in formalin and examined by the Ritchie technique. Thirty-four per cent of stools (1981/5792) tested positive for intestinal helminths with the following parasites identified: Ascaris lumbricoides (27.3%), Trichuris trichiura (7.3%), Necator americanus (3.8%), Hymenolepsis nana (2%), Taenia sp. (0.3%) and Strongyloides stercoralis (0.2%). The helminth prevalence was higher in rural (38.4%) compared to urban areas (30%). There was no significant difference in prevalence by sex and age. The importance of geohelminths changed from one department to another with the highest prevalence found in the Southern department of Grande Anse (73.7%) and the lowest prevalence in the Center department (20.6%). Five out of the country's nine departments had a similar prevalence varying from 25.5% to 28.2%. Intestinal helminthic polyparasitism was observed in a percentage of infested school children comprise between 3.4% and 28.6% according in relation to the geographical area. A program to fight against geohelminths in school children should be initiated as a public health priority. Albendazole is the drug of choice. Frequency of drug distribution should be based on the prevalence of geohelminths in each department.     

Changes in the hypothalamic nuclei in in experimental allergy

Summary In rabbits subjected to prolonged sensitization and in which the Arthus phenomenon was induced there was a marked reaction of the hypothalamic nuclei. Staining by Gomori's method indicated a cellular swelling, loss of granules, and protoplasmic vacuolization in the supraoptic nucleus. There was a considerable increase in the size of the cross-sectional area of the cells. The same effects were much less well shown in the paraventricular nucleus. These results show that marked signs of increased neurosecretion developed in the animals at the height of the Arthus phenomenon.(Presented by Active Member AMN SSSR V. V. Parin) Translated from Byulleten Éksperimental'noi biologii i Meditsiny, Vol. 55, No. 4, pp. 110–113, April, 1963.     

Experience in adult population in dengue outbreak in Delhi

A dengue outbreak has recently hit the Indian capital. We studied the clinical profile of adult patients. Five hundred and sixty patients of dengue infection were admitted in a specially created ward according to the criteria laid down by WHO. Haematemesis (28.28%), epistaxis (26.78%) and malena (14.28%) were some of the common presentations. Similarly lymphadenopathy, especially cervical (30.89%), palatal rashes (26.96%) and hepatomegaly (23.75%) were the most commonly encountered findings on physical examination. Most of the cases were of dengue fever with haemorrhage and only 2.5% cases were classified under dengue haemorrhagic fever or dengue shock syndrome. The average hospital stay was 3.4 days but only 9.8 hours in the eleven patients who died, suggesting their late arrival in preterminal situation giving little time for resuscitation. Thrombocytopenia was not a feature and only 12.85% patients had platelet count less than 70,000/cmm. Most of the patients who were admitted with thrombocytopenia, showed normalization in their platelet counts in next few days. Serological examination demonstrated evidence of recent dengue infection in 41.17% patients. Few patients required blood or platelet concentrate transfusion. Eleven patients died, three due to DIC, one of intracranial haemorrhage and seven due to massive gastric haemorrhage. Rest of the patients recovered completely. Thus we can conclude that recent outbreak in Delhi was of dengue fever with haemorrhage and mortality was very low in patients who came early to the hospital.     

Development in in vitro toxicology

There are three principal pressures driving the development of in vitro toxicology: (1) the need for more efficient testing systems to cope with the large number of xenobiotics currently being developed; (2) public pressure to reduce animal experimentation; and (3) a need for a better understanding of the mechanisms of toxicity. Within this, in vitro toxicology is focused on local, systemic, and target-organ toxicity. It is becoming increasingly apparent that a step or decision-tree approach using input of a variety of experimental data (physicochemical properties, biokinetics, cytotoxicity) provides the most efficient system for predicting toxicity. Examples of the use of in vitro toxicity systems for prediction of systemic toxicity and target-organ (liver) toxicity are presented.