Acetaminophen (APAP) is the most important common cause of acute liver failure; however, the exact mechanism of toxicity of this analgesic drug is not elucidated. In the present study, we generated APAP-resistant liver cell line and determined their basic properties. AML-12 mouse hepatocytes were cultured with clonal cell density to generate APAP-resistant cells by means of gradually increasing the concentration of the APAP drug. Cell proliferation was determined by cell counting. The levels of reactive oxygen species (ROS) and mitochondrial membrane potential (MMP) were determined by H2DCF-DA and MitoTracker detection, respectively. The levels of mRNA and protein were determined by qPCR and Western blot analysis, respectively. We successfully established 1.25 and 2.50 mmol/L APAP-resistant AML-12 cell lines (ARCs). Cell proliferation analysis revealed that, compared to the control group, ARCs had high proliferation capacity, low ROS levels, and high MMP and GSH/ GSSH levels. Furthermore, Western blot and qPCR analysis showed that the expression level of the Nrf2 signaling pathway was up-regulated, whereas the level of the JNK signaling pathway was down-regulated. In addition, ARCs retained normal hepatocytic phenotypes, except for the high expressions of Foxa1 and Foxa2 genes, which were related to chromatin remodeling. We successfully established APAP-resistant hepatocyte lines with low oxidative stress levels but with retained normal hepatocytic phenotypes. Activation of Nrf2 and inactivation of JNK signaling pathway appeared to be involved in the protection of hepatocytes against APAP-induced cell apoptosis.

目的 检测INHBA(inhibin-βA)在乳腺癌及癌旁乳腺组织中的表达,探讨其与乳腺癌临床病理特征的关系及对乳腺癌患者预后的意义.方法 应用免疫组化法检测85例乳腺浸润性导管癌及35例癌旁乳腺组织中INHBA的表达.结果 INHBA在乳腺癌和癌旁组织中的表达差异有统计学意义(P＜0.05).INHBA高表达和肿瘤较低的分化程度、淋巴结转移及较高的Ki～67增殖指数具有相关性(P＜0.05). INHBA高表达乳腺癌患者的生存期明显短于低表达组(P＜0.05).多因素Cox比例风险回归模型分析显示,INHBA表达可以作为判断乳腺癌患者的独立危险因素(P＜0.05).结论 INHBA表达与肿瘤分化程度、淋巴结转移及Ki-67增殖指数密切相关,可能参与乳腺癌的发生发展,可以作为乳腺癌患者预后的判断指标.

Objective To investigate the common problems of immunohistochemistry in skin pathology and the influence of quality control on the quality of immunohistochemical preparation.Methods Twenty male BALB/c rats aged from 6 to 8 weeks old were divided into observation group and the control group by random number table,with 10 rats in each group.The observation group was used by the making slices method after quality control and adopted the complete immunohistochemical procedures and the control group with the traditional production method and the simple im munohistochemical procedures.The making slices quality and immunohistochemistry of two groups were compared.Results The making slices quality in observation group was better than control group,the difference was statistically significant (P ＜ 0.05).The quality of the immunohistochemical pictures in observation group was significantly better than the control group.Conclusion Immunohistochemistry can affect the results of immunohistochemistry in any aspect.Quality control can enhance the comprehensive effect of immunohistochemistry,which is helpful to further improve the clinical diagnosis of the disease and it is worth promoting.

目的 分析前列腺导管腺癌的免疫组化染色特点及其鉴别诊断.方法 收集10例前列腺导管腺癌,应用EnVision两步法观察PAX8、GATA3、P40、P63、CK5/6、CK7、CK20、PSA、P504s、Villin、CDX2、CEA、Ki67等标志物的表达.结果 10例发病年龄57～79岁,平均(63.2±2.6)岁,术前PSA水平15.9～104.8ng/mL,平均(43.9±2.5)ng/mL.镜下导管腺癌均可见与腺泡性腺癌混合,导管腺癌占比20％～60％,组织学上呈复杂分支的管乳头状结构排列,3例可见局灶的筛状结构,瘤细胞被覆假复层柱状上皮,核仁明显,4例可见局灶的粉刺样坏死,与之混合的腺泡性腺癌的Gleason分级≥7分.免疫组化染色,10例导管腺癌均弥漫强表达PSA和P504s,5例可见CK7局灶强表达,4例可见CK20局灶中等强度表达,2例可见CDX2弥漫弱表达,2例可见Villin局灶强表达,1例可见CEA局灶表达,Ki67增值指数平均约10％.其余免疫组化染色标志物(包括PAX8、GATA3、P40、P63、CK5/6等)瘤细胞均阴性.结论 前列腺导管腺癌侵袭性较强,在免疫组化染色特点上显示异质性,可局灶或部分表达尿路上皮癌和结直肠腺癌相关标志物,掌握前列腺导管腺癌的组织学和免疫表型特点可避免误诊.

Objective To investigate the feasibility and clinical effect of the application of nano-carbon particle as a trace agent in radical gastrectomy for gastric cancer.Methods The clinical data of 111 patients with gastric cancer undergoing D2 (D2+) radical gastrectomy with intraoperative injection of nano-carbon particles (study group) and 106 patients undergoing D2 (D2+) radical resection without injection of nano-carbon particles (control group) were retrospectively analyzed.The operation time,intraoperative blood loss,hospitalization time,dissected lymph node number(N1,N2,partial N3 lymph node) and metastatic lymph node number were compared between two groups.Results There was no significant difference in operation time,blood loss and hospitalization time between the two groups (P＜0.05).The number of dissected and metastatic lymph nodes in study group was significantly higher than that of control group,except N1 lymph nodes (P＜0.05).Conclusion The intraoperative application of nano-carbon particle is simple and feasible for guiding the dissection of individual lymph nodes,which can increase the detection rate of lymph node metastasis,and is beneficial to accurate clinical staging and postoperative chemotherapy.

Objective To explore the relationship between Fascin,c-Met and clinicopathologic characteristics of breast carcinoma.Methods The expression of Fascin and c-Met was examined in 95 cases of human breast carcinoma tissues and non-cancerous adjacent breast tissues using immunohistochemical method.Results The expression rate of Fascin was 44.21％ in breast carcinoma tissues,and 10.53％ in non-cancerous adjacent breast tissues (P ＜ 0.01).The expression of Fascin was related breast carcinoma to histology grade,TNM stage and lymph node metastasis (P ＜ 0.05).The expression rate of c-Met was 55.79％ in breast carcinoma tissues,and 8.42％ in non-cancerous adjacent breast tissues (P ＜0.01),and there was a close relation between the expression of c-Met and tumor size,TNM stage and lymph node metastasis (P ＜ 0.05).Conclusion The expression of Fascin and c-Met may help to judge the abilities t of malignancy and invasion of breast carcinoma.

胸腹水的细胞学检查是病理的日常工作之一.常规涂片存在厚薄不均、细胞重叠、结构不清、细胞量少而阳性率低,以及难以进行免疫细胞化学染色等缺点.本次研究探讨细胞块石蜡切片及免疫细胞化学染色在胸、腹水检测中的应用价值.现报道如下. 1 资料与方法

[Objective] To study the relationship between the endoscopic and pathological diagnosis of intestinal metaplasia(IM).[Methods] We observed the gastric mucosal patterns with electronic gastroscope in each patient.The main pattern of IM were defined.The endoscopic diagnostic rate was compared with the pathologic result as control.Two mucin histochemical stains,AB/PAS and HID/AB,were applied to differentiate the type of IM.[Results] 285 cases were diagnosed IM by pathology.The specificity of endoscopic diagnosis as controlled with pathology was 91.5%(119/130),and the general sensitivity was 41.8%(119/285).The sensitivity in serve group was significantly higher than in mild group(P 0.01).In chronic atrophic gastritis,the more the gastric mucosas were atrophyed,the more type Ⅲ IM occurred.The endoscopic diagnosis accuracy of type Ⅲ IM was much higher than type Ⅰand type Ⅱ IM.[Conclusion] Endoscopic patterns of IM are specific markers for endoscopic diagnosis.The endoscopic diagnosis rate is correlated to the degree and type of IM.

Objective To study the effect of Helicobacter pylori(Hp) eradication on the precancerous lesion of gastric mucosa such as atrophy and intestinal metaplasia,and on the expression of ki-67 protein.Methods Sixty-seven chronic gastritis patients with Hp infection and glandula atrophy and/or intestinal metaplasia in the gastric mucosa were randomly divided into two groups: experimental group of 37 cases and control group of 30 cases.The former was given anti-Hp medication,while the latter,non anti-Hp medication.An examination was repeated one year later to compare the effect of Hp eradication on precancerous lesions such as atrophy and intestinal metaplasia.Immunohistochemical stain was used to detect the expression of ki-67 before and after the treatment,and the relationship between Hp eradication and expression of ki-67 was also investigated.Results In the experimental group,the degree of chronic inflammation of gastric mucosa ameliorated (34/37,(91.9%),P(0.01)).The number of active inflammation cases was reduced(P(0.01)).Atrophy and intestinal metaplasia were ameliorated or reversed(21/32,(65.6%) vs 15/26,(57.7%),All P(0.05)),and the expression of ki-67 was reduced,positive cell labeling index before and after the treatment (46.5)±(27.7) vs (15.5)±(18.7)(P(0.01)).Conclusion Hp eradication may relieve chronic inflammation and precancerous lesions such as atrophy and intestinal metaplasia of gastric mucosa,stop active inflammation and reduce the expression of ki-67.It may help to prevent gastric carcinoma.

胃黏膜异型增生是一种重要的癌前病变,在Padova国际分类中称非浸润性新生物,即由限制在基底膜内的肿瘤性上皮代替了正常的胃黏膜上皮[1].我们应用免疫组化技术检测c-myc、bcl-2、P53基因蛋白在胃黏膜异型增生中的表达,探讨它们与胃癌发生的关系.

近年来国外对幽门螺杆菌(Hp)和胃食管反流病(GERD)的关系研究较多,但结论一直不统一.而国内有关资料较少,现收集我院7年间经胃镜检查确诊的反流性食管炎(RE)病例,对Hp感染与RE的关系进行初步探讨.

Objective To study effects and mechanism of peptic ulcer(PU) on reflux esophagitis(RE).Methods 756 cases of primary RE were divided into two group,69 cases of reflux esophagitis with peptic ulcer(RE+PU),another group included 687 cases with reflux esophagitis without peptic ulcer(RE-PU). Histological examination and helicobacter pylori (Hp) positive rate of mucous membrane of gastric pylorus(756 cases) and lower segment of esophagus(174 cases) in two groups were analysed.Results Rate of grade Ⅱ and Ⅲ of RE in the RE+PU group (25/69,36.23%)was significantly difference than RE-PU group (124/687,18.05%,P0.01). Hp positive rate of mucous membrane of gastric pylorus in RE+PU group(70.59%) was significantly higher than RE-PU group(43.62%,P0.01).Hp positive rate of lower segment of esophagus in 174 cases of RE were 0, 8.57% and 39.53% in grade Ⅰ,Ⅱ and Ⅲ,respectively. Also there were significantly difference in three grades(P0.01). Conclusion Compared with RE-PU group, RE+PU group is year older, Hp positive rate of mucous membrane of gastric pylorus is higher ,with more severe lesions of lower segment of esophagus.

目的研究胃粘膜异型增生的诊断、转归及与胃癌的关系.方法对60例具有随访资料的胃粘膜异型增生病例进行胃镜、病理形态特征分析,并作PCNA免疫组化染色,HID-AB-PAS组织化学染色、Hp检测.结果高度异型增生胃镜形态表现为灰白色、粗糙、光泽消失、病灶＞0.5cm的与癌变关系密切(P＜0.01或0.05).中、重度萎缩性胃炎伴高度异型增生、不完全大肠型化生癌变率高.Hp感染阳性组PCNA增殖指数高于Hp感染阴性组(P＜0.01或0.05).结论高度异型增生应做胃部分切除或近期胃镜下活检随访,低度异型增生必须密切随访.Hp感染、PCNA增殖指数与胃粘膜增殖、恶化有关.

目的:研究胃上皮化生、幽门螺杆菌(HP)与十二指肠溃疡之间的关系.方法:应用特殊染色方法对60例十二指肠溃疡球部活检标本进行形态学观察及HP检测,并以50例正常十二指肠粘膜作对照.结果:胃上皮化生率球溃组(81.7%)高于对照组(26%)(P＜0.005);球部HP检出率球溃组(50%)高于对照组(14%)(P＜0.005);球部HP94.6%(35/37)生长在胃上皮化生区,5.4%(2/37)生长在无化生区(P＜0.005);轻度、中度、重度胃上皮化生区HP检出率分别为21.4%、64.3%、70.0%,中～重度胃上皮化生区HP检出率高于轻度化生区(P＜0.05).结论:胃上皮化生与HP相继作用,形成十二指肠溃疡.

目的探讨提高胃粘膜活检中印戒细胞癌的诊断率.方法对72例胃粘膜活检中印戒细胞癌进行胃镜、病理特征分析,并作HP检查,部分病例作AB/PAS染色和免疫组化染色.结果72例胃镜检查诊断胃癌65例,7例仅表现为粘膜糜烂、浅表溃疡而误诊.病理活检首次确诊64例,3例癌疑经连续切片多张证实,5例活检阴性的可疑病例,经2～3次胃镜复查再次活检确诊,2例切片中癌细胞少,散在分布而漏诊.HP感染阳性率52.8%,7例AB/PAS染色细胞内阳性率100%,15例CEA免疫组化阳性率100%.结论胃镜检查对可疑病变取材要充分,多点活检,尽量取到深处组织,一次活检阴性,短期内复查.不易确诊时作AB/PAS染色和CEA免疫组化染色帮助诊断.

目的探讨p16、p53蛋白表达与胃癌细胞增殖、浸润、转移及预后的关系.方法应用免疫组化S-P法研究100例胃癌,20例中、重度异型增生,16例萎缩性胃炎和10例正常胃黏膜p16 、p53蛋白和增殖细胞核抗原(PCNA)的表达.结果正常胃黏膜p16阳性率为80%,p53未表达 .萎缩性胃炎、异型增生和胃癌组织中p16阳性率分别为68.8%、35%和43%;p53阳性率分别为12.5%、25%和56%.p16在胃癌中的阳性率与预后明显相关(P<0.05);p53阳性率与肿瘤病理分级、Lanren分型有显著相关(P<0.05),与预后关系密切(P<0.01).从正常胃黏膜到病变组织PCNA指数逐渐上升,以异型增生、胃癌细胞增殖显著 (P<0.01),癌细胞PCNA指数p16阴性组,p53阳性组高于对照组,与肿瘤大小、浸润深度密切相关(P<0.05).p16、p53阳性表达具有协同性(P<0 .05).结论胃黏膜中、重度异型增生有较高的增殖活性,基因水平上已表现出癌变特性, 是癌变发生过程中的一个重要阶段.p16、p53蛋白表达在胃癌的发生发展中起重要作用,可作为判断预后的可靠指标.

To approach the relationship of p16,p53,C erbB 2 expression with the clinicopathological characteristics of gastric cancer.p16,p53,C erbB 2 and proliferating cell nuclear antigen( PCNA ) expressions of 100 cases of gastric cancer tissues were investigated by immunohistochemical method. Of 100 cases with gastric cancer,positive expression rates of p16,p53 and C erbB 2 were 43%, 56% and 50%,respectively.p53,C erbB 2 positive expression and p16 negative expressions indicated bad prognosis (P0.01 and0.05). p53 positive expression was closely related to cell proliferation,pathological types and differential degree .C erbB 2 postive expression was correlated to lymph node metastasis and infiltration degree.The protein expression mentioned above occurred in 83% of gastric cancer,the multiple protein expressioned simultaneously in 47%.[Conclusions] The analysis of multi gene is more valuable than that of single gene.The abnormal expressions of p16,p53 and C erbB 2 and their mutual actions play an important role in gastric carcinogenesis.These protein expressions are effective indicators for prognosis of gastric cancer.