p21和增殖细胞核抗原在肝硬化组织的表达及其与肝细胞不典型增生的关系

目的探讨p21和增殖细胞核抗原（proliferatingcellnuclearantigen,PCNA）的相互关系及它们在肝硬化、肝细胞不典型增生（LCD）、肝细胞癌(HCC)形成中的作用。方法运用S-P法对60例单纯肝硬化、30例癌旁肝硬化及27例HCC的p21、PCNA及HBsAg表达情况进行实验研究。结果癌旁肝硬化、单纯肝硬化、癌组织p21阳性率分别为90%、68.33%、62.96%。癌旁肝硬化与单纯肝硬化及癌组织之间,差别有显著性意义(P＜0.05)。伴LCD改变的肝硬化,p21、PCNA及HBsAg阳性率分别为92.45%、73.59%、64.92%均高于不伴LCD改变的肝硬化组织（P＜0.05）。HBsAg阳性及PCNA表达阳性的肝硬化组织,其p21阳性表达率分别为87.32%、86.21%,均明显高于HBsAg阴性及PCNA阴性的肝硬化组织（P＜0.01）。结论(1)p21的过度表达可能与HCC的起始过程有关,在HCC形成发展的早期阶段发挥重要作用。(2)LCD是一群具有肿瘤性增殖潜能的癌前细胞群,尤其是伴有HBV感染,且有p21及PCNA共同过量表达者,有发生HCC的高度危险。(3)HBV感染及PCNA过量表达与p21表达密切相关。     

Immunohistochemical study on p53, H-rasp21, c-erbB-2 protein and PCNA expression in HCC tissues of Han and minority ethnic patients

AIM To find out the difference of human primary liver carcinogenesis between Han and minority ethnic patients in Xinjiang.METHODS Expression of p53, c-erbB-2, Hrssp21 protein and proliferating cell nuclear antigen (PCNA) in tumor tissues of 50 patients (Han 38, minority 12 ) with primary hepatic carcinoma was detected by immunohistochemistry (LSAB). RESULTS The positive frequency of p53, cerbB-2, H-rasp21 and PCNA expression was 46.0% (23/50), 70.0% (35/50), 68.0% (34/50)and 82.0% (41/50) in tumor tissues; 4.0% (2/50), 22.0% (11/50), 64.0% (32/50) and 52.0%(26/ 50 ) in peritumors respectively and a significant difference, except for H-rasp21, of oncogene alteration was found (P＜0.05)between tumor and non- tumorous tissues.Combined the three oncogenes alteration, 26%(13/50)tumor tissues had positive immunoreactivity, but in peritumor and normal livers it was negative. The positive rate of p53,c-erbB-2 and H-rasp21 protein expression was 39.5% (15/38), 60.5% (23/38) and 39.5% (15/38) in tumors of Han patients; 66.7% (8/12),100% (12/12) and 75.0% (9/12) in minorities respectively, with statistical difference (P＜0.05). CONCLUSION Overexpression of p53, c-erbB-2 and H-rasp21 in human primary liver carcinoma is an important biomarker of genetic alteration.The different frequency of these oncogenetic changes may reflect some environmental or/and ethnic hereditary factors affecting the liver carcinogenesis. The special life style of Han,Uygur, Kazak and Mongolia nationalities in Xinjiang may also be related to the etiopathogenesis of this disease.     

Lactogenic hormones rapidly activate p21( ras )/mitogen-activated protein kinase in Nb2-11C rat lymphoma cells

Lactogenic hormone-dependent Nb2-11C cells proliferate in response to prolactin (PRL) or human growth hormone (hGH). We have investigated the activation of p21 ^ras and mitogen-activated protein kinase (MAP-kinase) by hGH in lactogen-dependent Nb2-11C and in autonomous hormone-independent Nb2-SP rat lymphoma cells. Exposure of Nb2-11C cells to hGH resulted in a dose-dependent activation of p21 ^ras and of MAP-kinase. Activation occurs at physiological hGH concentration and with a rapid onset (∼1 min) reaching maximal level at 10–20 min. In contrast, in Nb2-SP autonomous lactogen-independent cells, p21 ^ras and MAP-kinase are constitutively activated and a challenge with lactogenic hormone had a modest additional activating effect. TPA, an activator of protein kinase C, enhanced p21 ^ras and MAP-kinase activity in Nb2-11C cells but failed to induce proliferation. The mechanism of activation of p21 ^ras in Nb2-11C cells by lactogenic hormones involves both an increased binding of guanine nucleotides to p21 ^ras as well as an increase in GTP/GDP+GTP ratio. In summary, we have demonstrated here that activation of the p21 ^ras /MAP-kinase pathway follows PRL receptor activation but is not sufficient for the lactogenic hormone-dependent mitogenesis.     

Comparative Studies of Superoxide Production by Microbial Wall Product-Primed Neutrophils in Ulcerative Colitis

A diminished tolerance to the normal gut bacterial flora has been suggested to be pathogenic in ulcerative colitis (UC) and the aim of this study was to evaluate the priming effect of selected bacterial wall products on UC neutrophil granulocytes. Neutrophils from 10 UC patients and 10 healthy controls were primed with bacterial lipoprotein (BLP) or lipopolysaccharide (LPS) and subsequently activated. Extracellular superoxide production was measured by the cytochrome c reduction assay. Priming neutrophils with BLP or LPS dose dependently increased the superoxide production in both UC and controls (P < 0.01), and BLP was more potent than LPS (P < 0.05). No differences were found between UC and controls. UC neutrophils do not seem to have an intrinsic abnormality with reduced tolerance to bacterial substances. However, bacterial wall products such as BLP modify neutrophil tissue-destruction mechanisms and might be pivotal for perpetuation of chronic colonic inflammation.     

原发性小肠腺癌p16蛋白及PCNA的表达

目的检测p16蛋白和PCNA在原发性小肠腺癌组织、癌旁组织及正常小肠组织中的表达。方法采用SP免疫组化法对36例小肠腺癌及其相应癌旁组织、6例正常小肠组织进行p16蛋白及PCNA的定位观察。结果①原发性小肠腺癌组织中p16蛋白阳性率为88.89％，相应癌旁组织及正常小肠组织中p16蛋白未见表达；原发性小肠腺癌组织中p16蛋白的表达与其分化程度及有无淋巴结转移相关（P〈0.05）；②原发性小肠腺癌组织中PCNA阳性率为61.11％，相应癌旁组织及正常组织PCNA阳性率为28.57％，二者比较P〈0.05；原发性小肠腺癌组织中PCNA的表达与肿瘤部位、浸润深度、分化程度相关（P〈0.05）；③原发性小肠腺癌组织中p16蛋白的表达优于PCNA的表达（P〈0.05）。结论①小肠腺癌中p16蛋白的高表达可能是其癌变过程中的早期事件，有望作为小肠腺癌的免疫标志物；②PCNA可作为判断预后的有效指标；③原发性小肠腺癌组织中p16蛋白的表达优于PCNA的表达，两者联合检测可早期诊断小肠腺癌，判断预后。     

非小细胞肺癌组织中MinaS3、p53、PCNA的表达及意义

目的 探讨Mina53在非小细胞肺癌中的作用及Mina53、p53、增殖细胞核抗原（PCNA）在其中联合表达的意义。方法用免疫组化S-P法检测34例非小细胞肺癌组织,10例正常肺组织中Mina53、p53、PCNA的表达。结果在正常肺组织中Mina53、p53、PCNA表达量都很少,而在非小细胞肺癌组织中均高度表达。在非小细胞肺癌组织中,Mina53的阳性表达率是52．94％（18／34）,其表达与临床病理分型、淋巴结转移有明显的相关性（P〈0．05）。p53、PCNA的阳性表达率分别是64．71％（22／34）、76．47％（26／34）,其与各临床病理参数均无相关性（P〉0．05）。Mina53、p53、PCNA在非小细胞肺癌组织中的表达两两之间均有相关性（P均〈0．01）。结论Mina53有可能成为非小细胞肺癌的标记物,Mina53与p53、PCNA在非小细胞肺癌中联合检测,可起到互补作用,较全面地反映非小细胞肺癌肿瘤细胞的增殖情况。     

NACT前后宫颈癌组织中PCNA、p53、P—gp的表达变化及意义

目的探讨宫颈癌新辅助化疗（NACT）前后增殖细胞核抗原（PCNA）、抑癌基因p53和P-糖蛋白（P-gp）在宫颈癌组织中的表达变化及其与化疗疗效的关系。方法对60例局部晚期宫颈癌患者行NACT,采用免疫组化法检测化疗前后宫颈癌组织中的PCNA、p53、P-gp。结果化疗前宫颈鳞癌组织中PCNA、p53和P—gp阳性表达率分别为86．7％、60．0％、56．7％;化疗后分别为93．3％、83．3％、83．3％,化疗前后p53、P—gp阳性率相比,P均〈0．05。NACT前宫颈癌组织p53、P-gp表达阴性者NACT有效率与p53、P—gp表达阳性者相比,P均〈0．05。结论NACT前后宫颈癌组织中p53、P-gp的表达水平有显著差异,p53、P-gp的表达水平可作为宫颈癌化疗疗效敏感性的预测指标．     

Gastric Mucosa Epithelial Cell Kinetics Are Differentiated by Anatomic Site and Helicobacter Pylori Infection

Changes in epithelial cell turnover related to Helicobacter pylori infection may contribute to gastric cancer development. The response of different anatomic sites of the gastric mucosa to H. pylori is not known. We studied apoptosis and cell proliferation at the grater and lesser curvature of the antrum and corpus, the fundus, and the cardia from 9 H. pylori gastritis patients and 11 H. pylori-negative controls with normal histology. Proliferation was highest at the major curve of the antrum and lowest at the fundus, and apoptosis was highest at the cardia and lowest at the major curve of the antrum in both H. pylori gastritis and normal mucosa. Proliferation was significantly higher at all anatomic sites, while apoptosis was significantly lower only at the major and lesser curve of the corpus in H. pylori gastritis compared with normal controls. Our data suggest that gastric mucosa epithelial cell kinetics is differentiated by the anatomic site and H. pylori infection.     

Stem Cell Factor Expressed in Human Gastric Mucosa in Relation to Mast Cell Increase in Helicobacter pylori-Infected Gastritis

We previously reported mast cell increases in H. pylori gastritis. To determine the mechanism, we investigated the kinetics of mast cells and mast cell growth factor (stem cell factor, SCF) in H. pylori-positive and -negative gastric mucosa. Biopsy specimens from 12 H. pylori-negative and 28 positive subjects were examined. Sections were stained for mast cells, proliferating cell nuclear antigen (PCNA), and SCF. Densities of mast cells, PCNA-positive mast cells, and SCF-positive cells were significantly greater in H. pylori-positive than -negative subjects. SCF was expressed in mast cells and fibroblasts. The density of SCF-positive fibroblasts increased in H. pylori-positive gastritis and decreased after cure of infection. SCF mRNA was detected in H. pylori-positive gastric mucosa. Fibroblasts isolated from the normal gastric mucosa expressed SCF mRNA after incubation with H. pylori water extract. SCF may be one of the factors for mast cell increase. Fibroblasts may participate in mast cell increase and inflammation in H. pylori infection.     

黄苔胃病患者胃粘膜凋亡基因及相关蛋白的表达

目的:探讨胃病患者黄苔与胃粘膜组织细胞凋亡之间的关系.方法:选取黄苔胃病患者62例,采用免疫组化法与63例白苔胃病患者进行对照,采用免疫组化法检测胃粘膜增殖细胞核抗原(PCNA)、p53、bcl-2、fas凋亡基因相关蛋白.结果:黄苔患者p53、fas表达均明显高于白苔患者(均P<0.05),而PCNA、bc1-2表达则与白苔患者差异均无统计学意义(均P>0.05).结论:黄苔胃病患者伴有促凋亡基因相关蛋白的过度表达,黄苔是胃内有活动性炎症的较灵敏的指标.     

Expression of p53 and p21 Are Independent Prognostic Factors in Patients with Serosal Invasion by Gastric Carcinoma

To evaluate whether the expression of p53 andthat of p21 are independent prognostic factors inpatients with advanced gastric cancer, we investigatedclinicopathological factors and the expression of p53 and p21 in 158 patients with gastric cancerthat had invaded the serosa and who had undergonecurative gastrectomy. In multivariate survival analysisof 156 surviving patients, we evaluated the size of the tumor, lymph node metastasis, venousinvasion, lymph node dissection, expression of p53, andexpression of p21 as independent prognostic factors.Moreover, we divided patients into four groups according to the expression of p53 and p21 in theirtumors [group A, p53^-/p21^-, N = 40(one died within 30 days of surgery); group B, p53^-/p21^-, N = 23; group C,p53⁺/p21⁺, N = 58; and group D,p53⁺/p21⁺, N = 37 (one died within 30 days of surgery)]. The 5- and 10-yearsurvival rates of 39 patients in group A were 71.7% and64.3%, those of 23 patients in group B were 81.4% and81.4%, those of 58 patients in group C were 35.6% and 30.2%, and those of 36 patients ingroup D were 67.9% and 60.7%. The prognosis of patientsin group C was poorer than that of patients in the otherthree groups. This result indicates that the evaluation of the expression of both p53 andp21 expression might provide prognostic information thatis more accurate than that provided by evaluation of theexpression of p53 alone.     

胃复春对胃癌癌前病变的治疗及对p21ras、p53表达的调节作用研究

目的：研究胃复春片对胃黏膜中、重度异型增生的治疗作用及对P21^ras、P53蛋白表达的调节作用，探讨其逆转胃癌癌前病变的治疗价值及机制。方法：内镜下病理活检证实为中、重度异型增生患者共58例，随机分为胃复春治疗组32例，维酶素对照组26例。治疗前后比较临床疗效，镜下改变，病理改变及胃黏膜固定部位活检标本P21^ras、P53蛋白S-P法免疫组化染色变化情况。结果：胃复春组临床症状的总有效率81．25％，内镜下改变总有效率71．88％，病理学改变总有效率65．63％，均优于维酶素对照组(分为53．85％、46．15％、34．61％)。P21^ras、P53蛋白在中、重度异型增生组织中有过度表达，胃复春能使其表达明显减弱。结论：胃复春对胃黏膜中、重度异型增生有良好的治疗作用，能促进病变胃黏膜的逆转，并对P21^ras、P53蛋白的表达有一定的调节作用，降低癌变危险性，可用于胃癌癌前病变的治疗。     

Gastric epithelial proliferation and p53 and p21 expression in a general population sample: relations to age,sex, and mucosal changes associated with H. pylori infection

Helicobacter pylori infection is the main cause of chronic gastritis. The infection has been linked to altered proliferative activity and changes in various cell cycle regulating proteins. To determine, in a general population sample, the proliferative activity and expression of p53 and p21 in males and females of different age groups with and without H. pylori-associated chronic gastritis, gastric biopsies from 273 subjects (188 with and 85 without H. pylori infection) randomly selected from a general population were examined immunohistochemically for Ki-67, p53, and p21. One thousand epithelial cells, including the surface, neck, and glandular areas, were counted in both the corpus and the antrum. Results are expressed as the percentage of positive cells. Subjects with H. pylori infection showed significantly increased proliferative activity and expression of p53 compared to uninfected individuals. Regarding the expression of p21, no difference was detected. Multiple linear regression analysis showed significant associations between chronic inflammation or inflammatory activity, on the one hand, and the degree of proliferation in both the corpus and the antrum, on the other hand. In the antrum, the degree of H. pylori colonization was related to the expression of p53. H. pylori seems to cause increased proliferation and increased expression of p53 (but not p21) in the gastric mucosa, neither of which is age or sex dependent. The proliferative activity is related mainly to events associated with inflammation, while the expression of p53 in the antrum is associated with the degree of H. pylori infection. The action of p53 appears to be independent of p21 activity.     

血管内皮生长因子与结肠及直肠癌淋巴结转移的关系

目的 :研究结肠及直肠癌活体组织中血管内皮生长因子 (VEGF)、p53蛋白和增殖细胞核抗原 (PCNA)表达与淋巴结转移的关系。方法 :应用病理学诊断的方法检查淋巴结有无癌细胞转移。直肠癌组织标本做VEGF、p53蛋白和PCNA免疫组织化学染色。结果 :VEGF阳性表达的结肠及直肠癌患者淋巴结转移率为 65 .91 % (2 9/ 4 4 ) ,VEGF阴性的患者淋巴结转移率为 1 9.2 8% (1 6/ 83) ,差异有极显著意义 (χ2 =1 2 .0 777,P <0 .0 0 1 ) ;而p53及PCNA阳性表达的患者淋巴结转移和阴性表达患者的淋巴结转移之间差异无显著意义。结论 :结肠及直肠癌组织中VEGF的表达可为患者的淋巴结转移情况的判断及预后提供重要的参考依据     

直流电场对兔缺血心肌组织p27~(Kip1)及增殖细胞核抗原表达的影响

目的探讨低压稳恒直流电对兔缺血心肌组织p27~(Kip1)及增殖细胞核抗原(PCNA)表达的影响及作用。方法日本大耳白兔70只,随机分为假手术组(5只)、心肌梗死组(MI组,5只)、2.0 V/cm电治疗组(2EFs组,30只)和4.0 V/cm电治疗组(4EFs组,30只)。后2组建立急性心肌梗死模型后,在冠状动脉左回旋支两侧心外膜上,缝合一对铂金电极,予电刺激30 min/d,分别于电刺激第1、3、5天、1、2和4周6个时相点,检测梗死边缘区毛细血管密度,Masson染色测心肌梗死面积,免疫组织化学检测缺血心肌组织p27~(Kip1)及PCNA的表达。结果与假手术组比较,MI组毛细血管密度和PCNA明显降低,p27~(Kip1)明显升高(P0.01);与M1组比较,2EFs组和4EFs组第5天时毛细血管密度和PCNA明显升高,心肌组织p27~(Kip1)表达下降(P0.05,P0.01),心肌梗死面积减少(P0.05);2EFs组和4EFs组各时相点比较,差异无统计学意义(P0.05)。结论低压稳恒直流电场在一定作用时间内,可能通过下调缺血心肌组织p27~(Kip1)的表达,刺激血管生成,缩小心肌梗死面积。     

胃癌、胃黏膜不典型增生和胃炎组织中p185和p21蛋白的表达及其临床意义

目的研究胃癌、胃黏膜不典型增生和胃炎组织中c-erbB-2和ras癌基因产物p185和p21蛋白的表达,以探讨p185和p21蛋白在胃癌发生、发展过程中的作用。方法采用免疫组化S-P法检测39例胃癌组织、24例胃黏膜不典型增生组织和33例胃炎组织p185和p21蛋白的表达。结果p185蛋白在慢性胃炎、轻度胃黏膜不典型增生组织中不表达;在中、重度胃黏膜不典型增生组织中的表达率分别为11.8%和66.7%,差异有显著性(P<0.05),表达程度明显增强(P<0.05);在高、中、低分化胃癌组织中的表达率分别为77.8%、50.0%和29.2%,差异有显著性(P<0.05),高分化胃癌组织中p185蛋白的表达程度明显高于中、低分化胃癌组(P<0.05)。p21蛋白在胃黏膜不典型增生及胃癌组织中的表达率和表达程度较慢性胃炎组明显增高,差异有显著性(P<0.01);在轻、中、重度胃黏膜不典型增生组织中的表达率分别为25.0%、82.4%和100.0%,差异有显著性(P<0.05),表达程度明显增强(P<0.05);在高、中、低分化胃癌组织中的表达率分别为44.4%、66.7%和95.8%,差异有显著性(P<0.01),表达程度明显增强(P<0.05)。p185和p21蛋白在胃癌组织中的表达呈负相关(r=-0.450,P<0.01)。结论c-erbB-2和ras癌基因产物p185和p21蛋白在胃癌致病机制中起重要作用,p21蛋白作用于胃癌发生的早期阶段,p185蛋白可能作用于胃癌发生的较晚期。     

Salt and Stress Synergize <Emphasis Type="Italic">H. pylori</Emphasis>-Induced Gastric Lesions,Cell Proliferation,and p21 Expression in Mongolian Gerbils

Our aim was to determine if salt and stress enhance Helicobacter pylori (Hp) lesions in Meriones unguiculatus. Two hundred seventy-eight pathogen-free gerbils were allocated to seven groups: Hp-Sydney strain (45), 8% higher-salt diet (38), stress (60% space reduction/water immersion; 36), Hp + salt (33), Hp + stress (34), N-methyl-N-nitro-N-nitrosoguanidine (34), and sham (58). Gerbils were sacrificed at 1 week (67), 12 weeks (73), 52 weeks (65), and 68 weeks (73). Sydney, Padova, and Lauren classifications were blindly used. Proliferation, p53, p21, and apoptosis were assessed. Follicular active gastritis (grade 2/3) was observed in 10% of Hp gerbils, 38% of Hp + salt gerbils, and 29% of Hp + stress gerbils at 52 weeks and 67%, 83%, and 43% at 68 weeks (P < 0.05). Heterotopic proliferative glands were identified in synergy groups from 52 weeks, with increases in their number and size by 68 weeks. Higher proliferative rates were observed in Hp+salt gerbils (P < 0.0001), and p21 overexpression in Hp+salt and Hp+stress gerbils (both P's < 0.0001), by 68 weeks, without p53 increases. We conclude that salt and stress synergize Hp damage and increase pseudo-invasive gland foci.     

A High Degree of Aneuploidy,Loss of p53 Gene,and Low Soluble p53 Protein Serum Levels Are Detected in Ulcerative Colitis Patients

PURPOSE The causes for the increased risk of colorectal cancer associated with ulcerative colitis have not been fully defined. Colonic tissue of ulcerative colitis patients was examined for changes in chromosome-17-centromere copy number, loss of the p53 gene, and alterations in serum levels of the 53-kDa protein. This study was performed under the assumption that these molecular events correlate with ulcerative colitis status and duration.METHODS Ulcerative colitis patients (n = 42) and healthy controls (n = 37) participated in the study. All participants were histopathologically and medically diagnosed. The stage of ulcerative colitis patients was stratified according to increasing risk factors for the development of colorectal cancer: left-sided colitis, pancolitis, sclerosing cholangitis, and dysplasia-associated lesions or masses. Changes in centromere number of chromosome 17 alone or in association with changes in copy number of the p53 gene were analyzed in colon tissue biopsies by fluorescence in situ hybridization. Serum p53 level was determined in blood samples by immunoprecipitation followed by separation using high-pressure liquid chromatography.RESULTS Changes in chromosome 17 and p53 copy number and lower levels of serum p53 protein in ulcerative colitis patients directly correlated with colorectal cancer risk factors. All values significantly differed from controls. Significant direct correlations were obtained for ulcerative colitis disease duration, levels of p53 in the serum, and extent of aneuploidy.CONCLUSIONS We demonstrate that in the colonic mucosa of ulcerative colitis patients, high levels of genomic instability, changes in p53 gene copy number, and lower levels of p53 in the serum directly correlate with the extent of disease duration and increased risk factors for colorectal cancer. Any of the measurements described herein can provide an acceptable prognostic tool in the assessment of colorectal cancer risk in ulcerative colitis patients.