不同病变胃粘膜DNA含量与癌基因表达的检测

用图像分析技术检测143例胃的慢性炎症、肠上皮化生、异型增生、癌各组细胞核DNA含量及倍体分布情况，并同时进行nm23、PCNA、 C-erbB-2抗体染色。结果显示nm23的阳性率在４种病变中无明显差异， DNA含量、PCNA、C－erbB-2则呈递增表达（P＜0.01）；nm23阴性表达、 PCNA与C-erbB-2阳性表达的细胞核DNA含量与５倍体(5c)及非整倍体(AN)检出率显著高于nm23阳性、PCNA、C-erbB－2阴性表达者（P＜0.05）。提示胃良恶性病变细胞核中DNA含量及倍体分布形式不同，nm23失表达、 PCNA与C－erbB-2过度表达与5c和AN细胞形成有关，形态定量分析及癌基因检测可作为胃粘膜上皮病变诊断和分类的综合性指标。     

c—erbB—2表达及DNA含量与乳腺癌组织学类型的相关性研究

目的：了解乳腺癌组织学类型与ｃ－ｅｒｂＢ－２表达和ＤＮＡ异倍体的关系。方法：用免疫组化法（ＬＳＡＢ），经典的Ｆｅｕｌｇｅｎ染色法及图像分析技术对８４例乳腺癌，２２例乳腺良性病变进行ｃ－ｅｒｂＢ－２免疫组化及ＤＮＡ含量检测。结果：（１）乳腺良性病变全部为整倍体，ｃ－ｅｒｂＢ－２表达阴性。乳腺癌异倍体７０．２％，ｃ－ｅｒｂＢ－２阳性表达率为３３％。（２）浸润性导管癌ｃ－ｅｒｂＢ－２阳性表达率（４１．６％）、异倍体比例（７７％）均高于特殊类型浸润癌（９．６％和５４．８％）。而Ｐａｇｅｔ病阳性表达率和异倍体比例最高（分别为１００％和８０％）。（３）浸润性导管癌ｃ－ｅｒｂＢ－２表达率随合并导管原位癌成分或合并明显坏死而增高（Ｐ＜０．０５）。粉刺型占优势，其表达率高于非粉刺型（Ｐ＜０．０５）。（４）ｃ－ｅｒｂＢ－２阳性肿瘤异倍体高于ｃ－ｅｒｂＢ－２阴性肿瘤异倍体（Ｐ＜０．０５）。结论：ｃ－ｅｒｂＢ－２激活和异倍体的出现主要与导管上皮细胞异型增生有关。可作为乳腺癌一个特殊亚群的诊断依据     

青年人大肠癌细胞p53蛋白表达和DNA的定量研究

目的：研究青年人大肠癌抑癌基因ｐ５３蛋白的表达和细胞ＤＮＡ含量。方法：采用流式细胞光度术（ＦＣＭ）。结果：青年组大肠癌细胞ＤＩ值（１．３０±０．１７）显著大于老年癌组（１．１０±０．０９）（Ｐ＜０．０１），且细胞增殖指数（ＰＩ）亦明显大于后者（２４．９％±６．５％ｖｓ２０．２％±４．７％）（Ｐ＜０．０５）。青年组大肠癌中ＤＮＡ异倍体癌发生率为８７．１％（２７／３１），而老年癌组仅为２８．６％（４／１４），两者之间差别有高度显著性（Ｐ＜０．００１）。ｐ５３蛋白表达量青年患者（ＦＩ＝１．３４±０．２６）显著大于老年患者（ＦＩ＝１．１５±０．２５）（Ｐ＜０．０１）。在青年大肠癌中，粘液癌和侵犯周围软组织者，其ｐ５３蛋白的阳性表达率（１００％，９５％）分别高于腺癌和浸润较浅者（６７％，５８％）（Ｐ＜０．０５），而且低分化癌和有局部淋巴转移者ｐ５３蛋白的表达阳性率（９３％，１００％）也较高、中分化癌和无局部淋巴结转移者（６９％，６８％）为高。结论：在ＤＮＡ水平上，青年大肠癌的恶性程度高于老年大肠癌；ｐ５３蛋白的高表达可能是造成青年大肠癌恶性度高的原因之一。     

前列腺癌与增生性病变的流式细胞分析

应用流式细胞分析术（ＦＣＭ）对２０例前列腺癌（ＰＣ）和１５例良性前列腺组织作了ＤＮＡ含量检测，着重分析了ＰＣ及增生性病变的细胞增殖指数（ＰＩ）与ＤＮＡ指数（ＤＩ）。结果表明在良恶性前列腺组织中有明显差异（Ｐ＜０．０１）。２０例ＰＣ的组织学分级与ＦＣＭ检测结果有关，ＰＣ－Ⅲ与ＰＣ１－Ⅰ、ＰＣ－Ⅱ与ＰＣ－Ⅰ、ＰＣ－Ⅲ与ＢＰＨ的ＰＩ、ＤＩ相差显著（Ｐ＜０．０１）。本研究结果提示良恶性前列腺病变的ＰＩ与ＤＩ能较客观地反应不同细胞的生物活性，并对分化低、分级高的ＰＣ的恶性程度判断及其预后的推测，以及癌前病变的分析有意义。     

进展期胃癌DNA含量与生物学特性和预后的关系

应用流式细胞术测定了５７例进展期胃癌细胞ＤＮＡ倍体及Ｓ期细胞比，并就ＤＮＡ倍体及Ｓ期细胞比，与病理学因素及预后的关系进行了分析。结果表明，非二倍体癌淋巴结转移阳性率明显高于二倍体癌（Ｐ＜０．０５）。Ｓ期细胞低比者多为Ⅱ期、淋巴结转移阴性、呈膨胀性生长和未侵及浆膜的癌（分别Ｐ＜０．０５）。ＤＮＡ倍体对病人术后５年生存率影响不明显（Ｐ＞０．０５）。低Ｓ期细胞比癌患者术后５年生存率明显优于高Ｓ期细胞比者（Ｐ＜０．０１）。Ｃｏｘ模型多因素分析表明，Ｓ期细胞比为最佳的预后因素。     

Ki-67在胃癌及癌旁组织中的表达

目的:探讨Ki-67在胃癌和癌旁黏膜上皮中的表达、分布特征及其在胃癌组织发生学上的意义。方法:采用免疫组化方法对36例胃癌及癌旁组织进行检测。结果:胃腺癌与癌旁伴异型增生慢性萎缩性胃炎Ki-67阳性率无显著性差异;胃腺癌与癌旁伴高、中增殖型肠上皮化生慢性萎缩性胃炎Ki-67阳性率无显著性差异(P>0.05)。结论:Ki-67是一种较好的癌前标志物,胃黏膜上皮异型增生和高、中增殖型肠上皮化生萎缩性胃炎与胃癌的发生密切相关。     

喉癌细胞DNA含量、增殖与T细胞亚群关系的研究

运用流式细胞术（ＦＣＭ）对１０例喉癌患者的肿瘤细胞ＤＮＡ含量及增殖进行了分析，同时对喉癌患者手术前、后的外周血Ｔ淋巴细胞亚群ＣＤ３、ＣＤ４、ＣＤ８的百分比进行测定。结果表明：喉癌组、良性病变组和正常对照组之间细胞的ＤＩ值无显著差异（Ｐ＞０．０５），喉癌组和良性病变组细胞的ＰＩ值显著高于正常对照组（Ｐ＜０．０５）；手术前，喉癌患者ＣＤ８升高比良性病变组显著（Ｐ＜０．０１），而ＣＤ４／ＣＤ８比值则比良性病变组下降明显（Ｐ＜０．０５）；喉癌患者术后ＣＤ８要比术前显著下降（Ｐ＜０．０１），ＣＤ４／ＣＤ８比值术后比术前明显升高（Ｐ＜０．０５）；并且发现ＣＤ４／ＣＤ８比值与ＰＩ值呈直线负相关（ｒ＝－０．８９）。本实验的结果说明肿瘤细胞增殖程度与患者机体免疫功能之间所存在的关系，用这一关系来推断肿瘤的恶性程度、以及对预后的估计、疗效的监测等都有一定临床意义。     

鼻咽癌多阶段形态发生中血管密度改变

采用免疫组化法检测第Ⅷ因子相关抗原、层粘连蛋白和细胞角蛋白的表达，研究血管密度（ＢＶＤ）在９６例鼻咽癌（ＮＰＣ）多阶段形态发生中的改变，结果显示．在癌旁异型增生上皮下的ＢＶＤ显著高于癌旁增生和（或）化生上皮下的ＢＶＤ（Ｐ＜０．０１）；癌细胞呈膨胀型（Ｒ）生长时癌间质中ＢＶＤ以低密度血管（ＬＤ）为主，虽弥散型（Ｓ）和混合形（Ｃ）生长时以高密度血管（ＨＤ）为主（Ｐ＜０．０５）；转移病例原发灶中的ＢＶＤ显著高于无转移的病灶（Ｐ＜０．０１）；临床晚期ＮＰＣ中ＢＶＤ显著高于早期病例，前者以ＨＤ为主，后者ＬＤ血管为主（Ｐ＜０．０１）．癌生长的大小与ＢＶＤ之间差异无显著性，这些结果提示在ＮＰＣ多阶段形态发生中血管起重要的作用。     

胃癌组织HLA－I类和DR分子免疫组化研究

本文应用免疫组化法对６４例胃癌、癌旁组织和６例胃溃疡大致正常胃粘膜冰冻和石蜡切片进行了染色．结果表明，正常胃粘膜和癌旁胃粘膜上皮细胞ＨＬＡ－Ｉ类分子表达阳性，其着色较均一，ＨＬＡ－ＤＲ染色均阴性．胃癌细胞Ｉ类分子表达缺失（２７／６４例），与癌旁上皮比较差异显著（Ｐ＜０．０１）。粘液细胞癌和低分化癌Ｉ类分子缺失率显著高于高分化癌（Ｐ＜０．０２５）．此外，发生肿瘤转移的病例Ｉ类分子缺失率（１２／１５例）显著高于无转移组（１／５例，Ｐ＜０．０２５）．ＤＲ分子在癌组织表达阳性，其阳性率高达５３．１％（３４／６４例）．低分化癌ＤＲ分子阳性率亦显著高于高分化癌和中分化癌，未分化癌ＤＲ分子阳性率亦显著高于高分化癌（Ｐ＜０．０１～０．０５）．提示（１）ＨＬＡ－Ｉ类分子表达缺失可能与癌细胞逃避宿主免疫监视发生润浸生长和转移有关；（２）分化程度不同的癌组织ＨＬＡ－Ｉ类分子表达差异显著，提示癌细胞分化可能影响Ｉ、Ⅱ类分子表达和肿癌抗原呈递；（３）ＨＬＡ－Ｉ类和ＤＲ分子表达异常可能是上皮恶性转变的标志之一．     

子宫肌瘤ER,PR与其细胞核体定量关系的探讨

应用ＡＢＣ法和计算机图象分析系统（ＩＡＳ）定量分析３９例子宫肌瘤患者子宫组织雌、孕激素受体（ＥＲ．ＰＲ）含量，对其中８例子宦肌瘤及肌层细胞核体作定量分析，并探讨它们之间的关系。结果为子宫肌瘤ＥＲ、ＰＲ含量显著高于同一子宫肌层的含量（Ｐ＜０．０１）：子宫肌瘤细胞核体数密度、体密度亦显著高于相应子宫肌层的对应值（Ｐ＜０．０５，Ｐ＜０．０１）：子宫肌瘤ＥＲ与其细胞核体数密度呈正相关（ｒ＝０．８３，Ｐ＜０．０５）。提示子宫肌瘤的发生，发展与雌、孕激素及其受体含量有关。     

Pre-cancerous lesions of stomach

Total 39 cases of carcinoma stomach were noticed out of 142 malignant tumours of GIT (27.46 percent). Histologically maximum cases were of diffuse type (56.41 percent) followed by intestinal type (35.89 percent) and indolent mucoid carcinoma (7.69 percent) of the stomach. The surrounding epithelium showed lot of changes in the intestinal type of carcinoma stomach. About 78.57 percent showed intestinal metaplasia, 14.28 percent of these cases showed chronic gastric ulcer and severe dysplasia (carcinoma in situ) and another 14.28 percent revealed villous adenoma with carcinoma in situ. In contrast to this, in diffuse variety, only 13.63 percent cases revealed intestinal metaplasia, 27.27 percent showed basal cell hyperplasia, stratification of the epithelium of crypts and diffuse infiltration of mucosa by malignant cells and 4.54 percent showed atrophic gastritis also. In mucoid carcinoma all cases had basal cell hyperplasia and stratification of crypts. Hence these conditions should be taken as premalignant lesions of stomach and should be cured in proper time.     

Deoxyribonucleic acid ploidy and cell cycle events in benign colonic epithelium peripheral to carcinoma

DNA ploidy and cell cycle phases of benign human colonic epithelium peripheral to adenocarcinoma were analyzed by flow cytometry in 188 prospective cases. Human colonic epithelium was shown to be diploid with a mean DNA index (DI) of 1.01. The G0G1 compartment accounted for nearly 93% of the cells with the remainder in the S and G2+M phases. Parallel [3H]thymidine uptake on selected cases confirmed the relatively low proliferative activity of colonic mucosa. The DNA index and the cell cycle compartments exhibited no correlation to the ploidy, Dukes' stage, size, and anatomical location of the corresponding malignant tumors. Approximately 25% of the benign samples possessed DI values outside of the diploid range (defined as the mean +/- sd). Analysis of these apparently hypo- (less than 0.92) and hyper- (greater than 1.09) diploid, histologically normal samples in terms of cell cycle kinetics and their relationship to Dukes' stage, location, distance, and the ploidy of the tumor showed no correlation. The only characteristic differentiating these "aberrant" samples from diploid benign tissue was variation in DI possibly due to differences in fluorochrome binding or accessibility to DNA. Whereas these results indicate some degree of variability in the DNA content of benign colonic epithelia, neither DI nor cell cycle kinetics appear to be affected by the presence of a malignant tumor and are not representative of either the ploidy or pathologic stage of the corresponding colonic carcinoma.     

Toll-like receptors TLR4, TLR5 and TLR9 on gastric carcinoma cells: an implication for interaction with Helicobacter pylori

In the human stomach Toll-like receptors (TLRs) expressed by the gastric epithelium interact with Helicobacter pylori and mediate production of proinflammatory cytokines and chemokines during H. pylori infection. This results in chronic active gastritis, the background from which gastric carcinoma arises via the epithelial precursor lesions, intestinal metaplasia and dysplasia. Therefore, the question is arising whether gastric carcinoma cells are also able to interact with H. pylori. In this study, TLR4, TLR5 and TLR9 expression was investigated on tumor cells of gastric carcinoma and on its precursor lesions, intestinal metaplasia and dysplasia, by immunohistochemistry. Gastric epithelium with intestinal metaplasia (n=10) and dysplasia (n=3) expressed TLR4 and TLR5. TLR4 was strongly expressed by tumor cells of 17 out of 22 and TLR5 by tumor cells of all 22 patients with gastric carcinoma. TLR9, however, was not detectable in intestinal metaplasia or dysplasia and only focally in 6 out of 22 gastric carcinomas. In contrast to H. pylori gastritis, epithelial TLR expression in intestinal metaplasia, dysplasia and gastric carcinoma was diffusely distributed without subcellular polarization as demonstrated by confocal microscopy. This is the first study describing TLR expression on tumor cells of gastric carcinoma and its precursor lesions. Expression of TLRs enables gastric carcinoma cells to interact with H. pylori. As H. pylori can induce gastric carcinoma-promoting factors, such as IL-8, via epithelial TLR expression, TLR expression by gastric carcinoma cells may have a dangerous potential.     

Operative Link for Gastritis Assessment gastritis staging incorporates intestinal metaplasia subtyping

Both sulfomucin-type intestinal metaplasia (ie, types II and III intestinal metaplasia, colonic-type intestinal metaplasia) and gastritis in Operative Link for Gastritis Assessment stages III and IV are associated with an increased risk of intestinal-type gastric cancer. This study aimed to verify the hypothesis that gastritis in Operative Link for Gastritis Assessment stages III and IV (both consistently associated with an increased cancer risk) is associated per se with types II and III intestinal metaplasia. Two hundred consecutive cases of atrophic gastritis (Operative Link for Gastritis Assessment stages I, II, III, and IV) were considered (50 cases for each stage). All cases were stained with high iron diamine, and intestinal metaplasia was subtyped accordingly (type I [ie, small-intestinal type] and types II and III). Helicobacter pylori status was also considered, distinguishing H pylori–positive versus H pylori–negative versus H pylori–eradicated patients. A significant association was found between intestinal metaplasia subtype and the Operative Link for Gastritis Assessment stage of gastritis (the higher the stage, the more the colonic-type of intestinal metaplasia, and vice versa; Wilcoxon, P = .001). The strength of the association between Operative Link for Gastritis Assessment stages and the 3 intestinal metaplasia subtypes was confirmed by logistic regression analysis (P < .001; odds ratio, 4.84; 95% confidence interval, 2.97-7.88). Intestinal metaplasia subtyping also correlated with the patient's age (Kruskal-Wallis, P = .001) and H pylori status (Fisher exact, P < .001). Operative Link for Gastritis Assessment staging incorporates the prognostic message obtainable from histochemical gastric mucin subtyping.     

不同年龄组人群胃粘膜病变的观察(3258例胃非恶性肿瘤标本的分析研究)

本文对我国3258例非胃恶性肿瘤标本不同年龄组人群的胃粘膜良性病变进行了观察,发现CSG、CAG、IM、ATP(异型增生)、胃和十二指肠溃疡的检出率均随年龄增长有增高的趋势,高发年龄除CSG为30～39岁外,余者均为40～49岁,此与我国胃癌高发年龄(50～59岁)相比均提前10～20年,符合文献上记载从癌前状态发展为癌前病变直至癌的所需时间。各种病变检出率为IM1914例(58.75%)、CAG1850例(56.78%)、CSG1356例(41.62%)、GU1335例(40.98%)、DU 1221例(37.48%)。ATP1113例(34.16%)。在CAG中IM和ATP均高,在伴ATP中,肠化型CAG(37.35%)又高于非肠化型(5.72%),因此伴异型肠化的萎缩性胃炎可能与我国胃癌高发密切相关。     

Intestinal metaplasia in endoscopic biopsy specimens of gastric mucosa

A total of 1412 consecutive cases of endoscopic gastric biopsy, carried out over a four year period, were reviewed and specimens were examined histochemically to determine the prevalence of intestinal metaplasia and its variants. Three types were characterised: complete intestinal metaplasia and two classes of incomplete intestinal metaplasia (type IIa and type IIb) depending on the absence or presence, respectively, of sulphomucins within mucin secreting columnar cells. Type IIb intestinal metaplasia was significantly more common in patients with gastric carcinoma (p less than 0.001) and in those with dysplasia (p less than 0.001) than in patients with benign gastric pathology. No such association was found with either type I or type IIa intestinal metaplasia. In addition to those present in the columnar cells of type IIb intestinal metaplasia, sulphomucins were also commonly found in goblet cells of all three types of metaplasia. The presence of sulphomucins in goblet cells, however, was not significantly associated with gastric carcinoma or dysplasia. The significance of the different types of intestinal metaplasia in relation to the pathological findings is discussed.     

DNA index determination with Automated Cellular Imaging System (ACIS) in Barrett's esophagus: Comparison with CAS 200

Background

For solid tumors, image cytometry has been shown to be more sensitive for diagnosing DNA content abnormalities (aneuploidy) than flow cytometry. Image cytometry has often been performed using the semi-automated CAS 200 system. Recently, an Automated Cellular Imaging System (ACIS) was introduced to determine DNA content (DNA index), but it has not been validated.

Methods

Using the CAS 200 system and ACIS, we compared the DNA index (DI) obtained from the same archived formalin-fixed and paraffin embedded tissue samples from Barrett's esophagus related lesions, including samples with specialized intestinal metaplasia without dysplasia, low-grade dysplasia, high-grade dysplasia and adenocarcinoma.

Results

Although there was a very good correlation between the DI values determined by ACIS and CAS 200, the former was 25% more sensitive in detecting aneuploidy. ACIS yielded a mean DI value 18% higher than that obtained by CAS 200 (p < 0.001; paired t test). In addition, the average time required to perform a DNA ploidy analysis was shorter with the ACIS (30–40 min) than with the CAS 200 (40–70 min). Results obtained by ACIS gave excellent inter-and intra-observer variability (coefficient of correlation >0.9 for both, p < 0.0001).

Conclusion

Compared with the CAS 200, the ACIS is a more sensitive and less time consuming technique for determining DNA ploidy. Results obtained by ACIS are also highly reproducible.     

Helicobacter pylori regulates TLR4 and TLR9 during gastric carcinogenesis

Objective: To investigated the influence of H. pylori on TLR4 and TLR9 in gastric mucosa during gastric carcinogenesis. Methods: Gastric biopsy specimens were taken from 148 patients and divided into five groups, including normal group (n = 10), chronic superficial gastritis group (n = 35), atrophy/intestinal metaplasia group (n = 35), dysplasia group (n = 34) and gastric carcinoma group (n = 34). Immunohistochemistry was used to detect the expression of TLR4 and TLR9. Geimsa staining and rapid urea test were used for determine H. pylori infection. Results: TLR4 was detected in gastric epithelium and monocytes/macrophages in superficial gastritis, atrophy/intestinal metaplasia, dysplasia or carcinoma. TLR9 was mainly accentuated in monocytes/macrophages. TLR4 positive cells in epithelium and in monocytes/macrophages with H. pylori infection were much more than those without H. pylori infection. Similar results were also found in TLR9. When gastric epithelium was accompanied with H. pylori infection, TLR4 was significant higher in superficial gastritis and atrophy/intestinal metaplasia groups compared with dysplasia and carcinoma groups. When gastric epithelium was infected by H. pylori, TLR9 was significant higher in carcinoma group compared with superficial gastritis, atrophy/intestinal metaplasia and dysplasia. TLR4 and TLR9 show significant correlation with the severity of inflammation. Conclusions: H. pylori infection was associated with increased expression of TLR4 and TLR9 in gastric mucosa. In superficial gastritis and atrophy/intestinal metaplasia the inflammation was predominately mediated by TLR4, while in gastric cancer the inflammation was mainly mediated by TLR9.