胰腺癌抑癌基因P16、DPC4寡核苷酸芯片点突变及缺失的应用

目的建立P16、DPC基因寡核苷酸芯片对胰腺癌基因突变的检测系统。评估16例胰腺癌组织P16、DPC基因突变和缺失。方法采用双重或单独不对称PCR扩增标本中的目的DNA。扩增产物加杂交液后与芯片进行杂交、清洗、扫描。结果16例胰腺组织标本中有7例检测出存在着P16基因的改变，4例标本为点突变，2例标本为缺失型改变，还有1例患者存在着野生型与缺失型同时存在的杂和状态。有7例标本发生了D即4基因的改变，有6例突变，1例为缺失，基因异常比例为43.75％。结论P16、DPCA基因芯片可同时检测胰腺癌多个突变位点基因。     

k-ras基因寡核苷酸芯片的研制及在胰腺癌组织中检测应用研究

目的建立ras基因寡核苷酸芯片对胰腺癌基因突变的检测系统。评估16例胰腺癌组织k-ras基因12、13、61位密码子变突检测。方法采用双重或单独不对称PCR扩增标本中的目的DNA。扩增产物加杂交液后与芯片进行杂交、清洗、扫描。结果16例胰腺癌组织中k-ras为75%,突变都发生在12密码子,k-ras以12密码子第2位核苷酸突变发生率高(8/12)。结论k-ras基因芯片系统具有高度的灵敏性和准确性、快速简便、自动化程度高等优点,可同时检测胰腺癌k-ras多个突变位点基因,有利于临床应用。     

石蜡包埋胰腺癌组织中DPC4第5/6、7、9、10外显子基因改变的检测

目的研究石蜡包埋胰腺癌组织中DPC4第5/6、7、9、10外显子基因的改变.方法用聚合酶链反应单链构象多态性分析(PCR-SSCP)银染技术检测46例石蜡包埋胰腺癌组织及癌旁正常胰腺组织中DPC4基因的缺失和突变.结果 5例胰腺癌未扩增出DPC4条带,DPC4基因第5/6、7、9、10外显子的纯合性缺失率为10.9%(5/46);1例胰腺癌可见异常泳动带,基因突变率为2.2%(1/46).结合以前实验,DPC4基因所有外显子的纯合性缺失率为39.1%(18/46),基因突变率为23.9%(11/46),总改变率为58.7%(27/46).27例有基因改变的胰腺癌除1例外均为中、低分化腺癌,中、低分化组与高分化组之间的差异有显著性(P<0.01).结论 DPC4作为一种抑癌基因,其改变在胰腺癌的形成中可能起重要的作用.DPC4基因改变与胰腺癌细胞分化程度密切相关.     

肺活检和支气管肺泡灌洗标本p53、K-ras和p16基因检测在肺癌诊断中的意义

目的 探讨肺活检和支气管肺泡灌洗液(BALF)标本p53、K-ras和p16基因检测对肺癌诊断的意义。方法 应用多聚合酶链反应(PCR)一单链构象多肽性(SSCP)—银染法和多重PCR分析，检测60例肺癌患者和18例良性肺疾病患者支气管肺活检组织和BALF标本p53基因和k-ras基因突变及p16基因变异(突变 缺失)状况。结果 肺癌组支气管肺活检标本中p53基因和k-ras基因突变、p16基因变异及三基因联合检测阳性率分别是60％、33．3％、41．7％和81．3％；相应的BALF标本阳性率为43．3％、33．3％、38．3％和73．3％。两种标本相应基因异常检出率比较差异无显著性(P值均＞0．05)。良性肺疾病组仅活检标本检出p53和K-ras基因突变各1例，而BALF标本良性疾病组三种基因异常的检出率均为0。良恶性疾病组比较差异有显著性(P＜0．05)。肺癌组BALF标本不论单基因检测还是联合检测异常率均高于癌细胞的检出率(P＜0.05)。BALF标本细胞学联合三种基因异常检测可将诊断率从单纯细胞学的16．6％提高到81．7％。结论 BALF标本基因检测可提高纤支镜检查对肺癌的诊断率。     

石蜡包埋胰腺癌组织中DPC4第5／6、7、9、10外显子基因改变的检测

目的研究石蜡包埋胰腺癌组织中DPC4第5／6、7、9、10外显子基因的改变。方法用聚合酶链反应单链构象多态性分析(PCR-SSCP)银染技术检测46例石蜡包埋胰腺癌组织及癌旁正常胰腺组织中DPC4基因的缺失和突变。结果 5例胰腺癌未扩增出DPC4条带,DPC4基因第75／6、7、9、10外显子的纯合性缺失率为10．9％(5／46)；1例胰腺癌可见异常泳动带,基因突变率为2．2％(1／46)。结合以前实验,DPC4基因所有外显子的纯合性缺失率为39．1％(18／46),基因突变率为23．9％(11／46),总改变率为58．7％(27／46)。27例有基因改变的胰腺癌除1例外均为中、低分化腺癌,中、低分化组与高分化组之间的差异有显著性(P<0．01)。结论 DPC4作为一种抑癌基因,其改变在胰腺癌的形成中可能起重要的作用。DPC4基因改变与胰腺癌细胞分化程度密切相关。     

k-ras基因突变的定量检测在胰腺疾病诊断中的应用

目的:探讨k-ras基因第12密码子突变的定量检测在胰腺疾病鉴别诊断中的价值及其意义.方法:应用肽核酸钳制实时荧光定量PCR方法检测143例胰腺癌、110例慢性胰腺炎以及28名正常人的血液中k-ras基因第12密码子的突变量,并分析其与相关临床指标的关系.结果:胰腺癌、慢性胰腺炎和正常人血液中k-ras基因的突变阳性率分别为51.75%、37.27%和7.14%.k-ras基因的突变比例分别为0.821%±0.287%、0.200%±0.064%和0.080%±0.056%.慢性胰腺炎和正常人即使存在k-ras突变,其突变比例通常也较胰腺癌低.胰腺癌患者血液的k-ras突变情况与性别、年龄、吸烟、饮酒、肿瘤TNM分期与临床分期无明显相关.结论:肽核酸钳制实时荧光定量PCR方法可用于胰腺疾病的鉴别诊断.     

变性梯度凝胶电泳分析检测结直肠癌APC和p53基因突变

目的: 探讨APC和p53基因突变在结直肠癌中的意义.方法:采用变性梯度凝胶电泳(DGGE), DNA测序法分析15例正常人和60例散发性结直肠癌标本的APC基因15外显子和p53基因第5, 7外显子的基因突变.结果: 在结直肠癌组检出14例APC和16例p53基因突变, 测序证实其中13/14例APC发生在突变集中区域;9/16例p53基因突变位于第5外显子, 7/16例p53基因突变位于第7外显子, 2例同时检出了APC基因和p53基因突变.结论:DGGE是一种快速、简便、高效、灵敏的突变检测技术. 同时也证明APC基因突变和p53基因突变均参与了结直肠癌的发生、发展过程.     

K-ras基因点突变对胰腺癌的诊断价值

背景传统的放射或超声检查早期诊断胰腺癌非常困难.目的研究胰腺良、恶性病变中的K-ras基因12密码子点突变情况,探讨K-ras基因点突变对早期胰腺癌的诊断价值.方法取23例胰腺癌和12例胰腺良性病变标本,经DNA提取后,行半巢式聚合酶链反应(PCR),扩增产物借助聚丙烯酰胺凝胶电泳检测有无K-ras基因点突变.结果胰腺癌和胰腺良性病变石蜡包埋组织的K-ras基因12密码子点突变率分别为65.2%(15/23)和33.3%(4/12)(P＜0.05).结论K-ras基因12密码子点突变的检测可能有助于胰腺癌,尤其是早期胰腺癌的诊断.     

痰液脱落细胞k-ras基因检测对肺癌诊断的临床价值

目的通过痰脱落细胞k-ras基因点突变的检测探讨其对肺癌诊断的临床应用价值。方法应用多聚合酶链反应结合限制性长度片段多态性分析法(PCR-RFLP),检测50例肺癌(其中腺癌22例,鳞癌24例,小细胞癌4例)和48例肺部良性疾病患者痰标本中k-ras基因第12密码子的突变情况,检测结果进行对比分析。结果k-ras基因在肺癌组的突变检出率为24.00%(12/50),非肺癌组突变检出率为2.08%(1/48),二者有显著性差异(P<0.01)。其中腺癌突变率40.90%(9/22),鳞癌突变率12.50%(3/24),小细胞癌未检出突变。在肺癌早期(Ⅰ期和Ⅱ期)中,k-ras基因突变明显高于肺良性病变组,且肺癌组中,吸烟者k-ras基因突变率34.48%(10/29)高于非吸烟者9.52%(2/21),两者有统计学差异(P<0.05)。k-ras基因突变与年龄、性别和临床分期无明显关系。结论k-ras基因突变与肺癌的发生存在一定的相关性,痰液脱落细胞中R-ras基因检测突变有助于肺癌的临床诊断,对早期诊断亦有一定的意义。     

脑胶质瘤细胞polβ基因突变及其与Fas、p53基因mRNA水平表达的研究

目的研究脑胶质瘤细胞中DNA聚合酶β(polβ)基因突变及其与Fas、p53基因mRNA 水平表达的关系.方法应用RNA提取、逆转录、PCR扩增、基因克隆及 DNA序列分析法,对2 2例脑胶质瘤、7例良性脑肿瘤及1例肺部转移性鳞状细胞癌标本中的polβ基因进行检测 ;半定量分析法对脑胶质瘤和良性脑肿瘤标本及其周围正常脑组织中的polβ、Fas、p53基因进行mRNA水平检测及对比.结果 30例脑肿瘤标本中,polβ基因突变7例,其中胶质瘤6例,突变率为2 7.3%(6/22),肺部转移性鳞状细胞癌1例.2例polβ基因二级结构图有明显变异.脑胶质瘤组织中均存在polβ和Fas基因mRNA水平高表达及p53基因低表达,有显著性差异(P＜0. 01),良性脑肿瘤与正常脑组织p53基因表达无显著性差异(P＞0.05);各种脑肿瘤中pol β与Fas基因表达具有正相关性,而polβ、Fas与p53基因表达呈负相关性.结论脑胶质瘤细胞存在polβ基因突变,其突变率为27.3%.脑胶质瘤组织中均存在polβ和Fas基因mRNA 水平高表达及p53基因低表达,提示脑胶质瘤的发生和发展可能与polβ基因突变有关,且恶性程度越高,突变越明显;另外,p53基因低表达及polβ、Fas基因高表达可能与脑胶质瘤的发生发展及恶性程度相关.     

Pathogenesis of carcinoma of the papilla of Vater

Most adenomas and carcinomas of the small intestine and extrahepatic bile ducts arise in the region of the papilla of Vater. In familial adenomatous polyposis (FAP) it is the main location for carcinomas after proctocolectomy. In many cases symptoms due to stenosis lead to diagnosis at an early tumor stage. In about 80%, curative intended resection is possible. Operability is the most relevant prognostic factor. Most ampullary carcinomas resp. carcinomas of the papilla of Vater develop from adenomatous or flat dysplastic precursor lesions. They can be sited in the ampulloduodenal part of the papilla of Vater, which is lined by intestinal mucosa. They also can develop in deeper parts of the ampulla, which are lined by pancreaticobiliary duct mucosa. Intestinal-type adenocarcinoma and pancreaticobiliary-type adenocarcinoma represent the main histological types of ampullary carcinoma. Furthermore, there exist unusual types and undifferentiated carcinomas. Many carcinomas of intestinal type express the immunohistochemical marker profile of intestinal mucosa (keratin 7?, keratin 20+, MUC2+). Carcinomas of pancreaticobiliary type usually show the immunohistochemical profile of pancreaticobiliary duct mucosa (keratin 7+, keratin 20?, MUC2?). Even poorly differentiated carcinomas, as well as unusual histological types, may conserve the marker profile of the mucosa they developed from. These findings underline the concept of histogenetically different carcinomas of the papilla of Vater which develop either from intestinal- or from pancreaticobiliary-type mucosa of the papilla of Vater. Molecular alterations in ampullary carcinomas are similar to those of colorectal as well as pancreatic carcinomas, although they appear at different frequencies. In future studies, molecular alterations in ampullary carcinomas should be correlated closely with the different histologic tumor types. Consequently, the histologic classification should reflect the histogenesis of ampullary tumors from the two different types of papillary mucosa.     

Demonstration of the mechanism of action of biguanides

Summary Palmitic acid oxidation in rat diaphragm homogenate is depressed by biguanide concentrations that are still incapable of inhibiting oxidative phosphorylation. Glucose oxidation is not directly effected by the same biguanide concentrations: however, the inhibitory effect of palmitic acid on glucose oxidation is partly removed by biguanides. Inhibition of fatty acid oxidation, which accounts for most of the metabolic effects caused by these drugs, can be regarded as the fundamental mechanism of action of biguanides. There is some evidence suggesting that these drugs might interact with carnitine, thus preventing long-chain fatty acids from being transported across the mitochondrial membrane to the site of oxidation. Traduzione a cura degli AA.     

胰岛素瘤的解剖分布特点分析

目的胰岛素瘤是最常见的胰腺神经内分泌肿瘤，因其临床表现多样，导致诊断困难。影像学诊断尤其是超声内镜(EUS)在胰岛素瘤的诊断中起着重要作用，拥有较高的敏感性和特异性。本研究拟通过明确胰岛素瘤的解剖分布特点，以期有助于提高影像学的诊断准确率和降低漏诊率，尤其是在教育和培训实践中对于EUS的学习者更具有指导价值。 方法回顾性分析解放军总医院第一医学中心病案资料数据库1993年1月至2019年11月经外科手术、病理确诊为胰岛素瘤的患者的临床资料，检索方法采取搜索术后病理诊断为"胰岛素瘤"的病例，通过查阅病例的方法，提取出胰岛素瘤的大小和解剖分布等数据，进一步分析其特点。 结果共检索到确诊为胰岛素瘤的患者116例，其中，男45例、女71例，年龄13～76岁，平均年龄(44.4±14.85)岁。胰岛素瘤单发110例(94.8%)、多发6例(5.2%)。位置分布：头颈部46例(39.7%)，单发45例、多发1例；体尾部68例(58.6%)，单发65例、多发3例；全胰腺多发2例(1.7%)。病变大小特点：最大径0.4～3.4 cm，平均大小(1.53±0.58)cm。≤1 cm 29例、>1 cm而≤1.5 cm41例、>1.5 cm而≤2.0 cm28例，≤3 cm 15例，>3 cm 3例。年龄与肿瘤的大小相关，≤44岁患者肿瘤平均大小为(1.36±0.51)cm、>44岁患者肿瘤平均大小为(1.70±0.60)cm，P<0.05。头颈部的肿瘤大于体尾部的肿瘤，头颈部肿瘤平均大小(1.66±0.63)cm，体尾部(1.42±0.52)cm，P<0.05。 结论胰岛素瘤在胰腺体尾部较头颈部更好发；绝大多数单发，但可以全胰腺多发；多数小于1.5 cm，肿瘤的大小与患者年龄和肿瘤的解剖分布相关。     

氯硝柳胺悬浮剂的毒性评价

目的评价氯硝柳胺悬浮剂的毒性，为现场大规模应用灭螺提供依据。方法按照中华人民共和国国家标准GB 15670-1995《农药登记毒理学试验方法》和鱼类毒性试验方法进行。结果经口、经皮肤的LDso雌、雄性大鼠均>5 000 mg/kg，经呼吸道的LCso雌、雄性大鼠均>5 000mg/m3，该药经口、经皮肤、经呼吸道毒性均属微毒类药物；兔眼用药后，观察期内无不良反应，对眼无刺激性；皮肤用药后对皮肤无刺激性。与氯硝柳胺原药、氯硝柳胺乙醇胺盐原药和氯硝柳胺乙醇胺盐可湿性粉剂相比，氯硝柳胺悬浮剂对鱼急性毒性最低。结论氯硝柳胺悬浮剂属微毒类药物，对鱼的毒性低于其乙醇胺盐可湿性粉剂，适合于现场应用。     

Lack of correlation of degree of villous atrophy with severity of clinical presentation of coeliac disease

BACKGROUND AND AIM: Both the clinical presentation and the degree of mucosal damage in coeliac disease vary greatly. In view of conflicting information as to whether the mode of presentation correlates with the degree of villous atrophy, we reviewed a large cohort of patients with coeliac disease. PATIENTS AND METHODS: We correlated mode of presentation (classical, diarrhoea predominant or atypical/silent) with histology of duodenal biopsies and examined their trends over time. RESULTS: The cohort consisted of 499 adults, mean age 44.1 years, 68% females. The majority had silent coeliac disease (56%) and total villous atrophy (65%). There was no correlation of mode of presentation with the degree of villous atrophy (p=0.25). Sixty-eight percent of females and 58% of males had a severe villous atrophy (p=0.052). There was a significant trend over time for a greater proportion of patients presenting as atypical/silent coeliac disease and having partial villous atrophy, though the majority still had total villous atrophy. CONCLUSIONS: Among our patients the degree of villous atrophy in duodenal biopsies did not correlate with the mode of presentation, indicating that factors other than the degree of villous atrophy must account for diarrhoea in coeliac disease.     

CagA-Hp抗体IgG与胃炎的组织学分析

研究幽门螺杆菌(Hp)感染与胃炎的关系。方法对204例慢性胃炎患者胃粘膜进行观察分析,并测定其中137例Hp阳性患者血清CagA-Hp抗体IgG水平,与组织学对照。结果慢性萎缩性胃炎伴肠上皮化生患者血清CagA抗体IgG明显高于对照组(P＜0.01);其他类型胃炎患者血清CagA抗体IgG水平无明显增高(P＞0.05)。结论CagA-Hp可能是导致慢性萎缩性胃炎伴肠上皮化生的因素之一,对这类患者应密切随访观察。     

慢阻肺急性加重期患者预后的影响因素分析

目的探讨慢性阻塞性肺病急性加重期(AECOPD)患者预后的相关危险因素。方法回顾性调查、收集58例AECOPD患者可能影响其预后的相关因素,并对其分别进行单因素分析。并进行Logistic多元逐步回归进行多因素分析,筛选影响AECOPD患者预后的独立危险因素。结果单因素分析后将结果 P0.1的因素纳入多因素Logistic回归,分析发现是否合并呼吸衰竭、气促程度、白细胞计数、APACHEⅡ、应用抗氧化剂、慢阻肺治疗依从性为影响AECOPD患者预后不佳的独立因素(P0.05)。结论根据AECOPD患者预后的独立危险因素,及早判断,选择合适的后续治疗方案,对提高其生存率及生存质量具有重要意义。     

血吸虫童虫生长发育及其机制的研究进展

血吸虫童虫是宿主免疫系统攻击的重要靶标,包括皮肤型、肺型和肝门型童虫。宿主分子对童虫生长发育具有重要作用。童虫生长发育机制包括免疫调节、信号转导、性别发育及凋亡等。肌动蛋白、组织蛋白酶、烯醇化酶和葡萄糖基转移酶等分子为血吸虫童虫生长发育的重要分子。本文对血吸虫童虫生长发育及其机制的研究进展做一综述。     

Results of treatment of fistula-in-ano

To evaluate the application of Parks' classification in the management of patients with fistula-in-ano, a study was undertaken to assess the outcome of surgery, especially with respect to the recurrence rate and alteration of continence. A retrospective analysis of 160 consecutive patients who were classified at the time of operation was conducted. The distribution of fistulas was as follows: intersphincteric, 41.9 percent, transsphincteric, 52.1 percent, suprasphincteric, 1.3 percent, extrasphincteric, 0. A horseshoe extension occurred in 8.8 percent of the fistulas and 3.8 percent did not exactly conform to the classification as they were either complex or combinations of more than one type of fistula. The sole immediate postoperative complication was bleeding, which occurred one week postoperatively and ceased spontaneously (0.7 percent). Alteration in continence occurred in 6 percent of patients with 2.6 percent experiencing temporary incontinence to flatus, 1.3 percent to liquid stool, and 0.7 percent to solid stool. Permanent loss of control for flatus occurred in one patient (0.7 percent) and for liquid stool in one patient (0.7 percent). No patients suffered loss of control for solid stool. Recurrence developed in 6.3 percent of patients, all between five and 25 months postoperatively. Classifcation was found to be a useful guide in the operative management of patients with fistula-in-ano. Read at the joint meeting of the American Society of Colon and Rectal Surgeons with the Section of Colo-Proctology, Royal Society of Medicine, and the Section of Colonic and Rectal Surgery, Royal Australasian College of Surgeons, New Orleans, Louisiana, May 6 to 11, 1984.     

Intake of energy drinks in association with alcoholic beverages in a cohort of students of the School of Medicine of the University of Messina

BACKGROUND: Energy drinks (ED) are a widely used group of beverages known for their stimulant effects on central nervous system (CNS). The main components of ED are caffeine, taurine, carbohydrates, glucuronolactone, inositol, niacin, pantenol, and beta-complex vitamins. The studies evaluating the effects of ED describe improvements in attention and/or reaction times and indices of alertness. It has been also shown that combination of caffeine and glucose, fundamental constituents of ED, can ameliorate deficits in cognitive performance and subjective fatigue during extended periods of cognitive demand. Moreover, the associated ingestion of alcohol and ED has recently been observed to be becoming more and more widespread. METHODS: With the aim to know the habits and uses of students, we administered a questionnaire containing questions regarding ED drinking alone or in association with alcoholic beverages. Five hundred students of the School of Medicine of the University of Messina were interviewed, and 450 filled the questionnaire. RESULTS: A total of 56.9% of students declared using ED. A great part of users (48.4%) associate frequently ED and alcohol. In particular, 35.8% of ED + alcohol users have used ED + alcohol more than 3 times in the last month. Distinguishing the users into 2 groups (users of ED + alcohol and users of both ED and ED + alcohol), we observed in the second group a major use of cocktail containing a mix of ED and alcoholic beverages. This difference between the 2 groups is less represented about the ingestion of ED + alcohol in the night. CONCLUSIONS: Our data indicate that association of ED + alcohol is very popular among students. This behavior can be dangerous. In fact, the combination of ED + alcoholic drinks can reduce adversive symptoms of alcohol intoxication including the depressant effects. As consequence, users of ED + alcoholic beverages might not feel the signs of alcohol intoxication, thus increasing the probability of accidents and/or favoring the possibility of development of alcohol dependence.