抗肺肿瘤转基因小鼠模型的建立

目的建立抗肿瘤转基因小鼠模型。方法用ＢａｍＨＩ酶切表达质粒ｐＵＬＢ３２３８获取小鼠细小病毒非结构基因,通过显微注射法将其接种入小鼠受精卵内制备转基因小鼠。转基因鼠尾部组织ＤＮＡ以ＰＣＲ检测靶基因整合;整合转基因的Ｇ０Ａ６小鼠与正常Ｃ５７ＢＬ／６Ｊ小鼠配种所产Ｇ１代转基因鼠,通过ＲＴ┐ＰＣＲ检测目的基因在其肺脏等组织的转录;Ｇ１代鼠根据ＰＣＲ检测结果分整合和未整合非结构基因两组,两组均腹腔注射致肺肿瘤化学致癌剂氨基甲酸乙酯水溶液。结果Ｇ０代有４只鼠整合靶基因;Ｇ１代（Ｇ０Ａ６子代）转基因鼠的肺组织有靶基因转录;Ｇ１代整合转基因实验组小鼠被诱导肺瘤结节平均数显著少于未整合转基因的对照组（Ｐ＜０．０１）。结论抗肺肿瘤转基因小鼠模型已建立。     

肿瘤小鼠模型建立应用进展

近百余年中,小鼠模型在医学研究中扮演着重要的角色,在肿瘤基础研究以及转化医学研究中具有重要的意义.随着肿瘤研究进展的不断加深,从最初的肿瘤移植模型到近年的基因编辑模型,模型小鼠的应用日益广泛.在人体环境中,不同的肿瘤在发生、发展及转移等方面均存在着差异.因此,建立合适的小鼠模型是开展肿瘤研究的必要条件.本研究总结目前肿...     

绿色荧光蛋白转基因小鼠诱导肝癌模型的建立

目的建立绿色荧光蛋白（green fluorescent protein,GFP）转基因小鼠肝癌模型,观察GFP蛋白在诱癌过程中荧光的变化。方法采用二乙基亚硝胺（diethylnitrosamine,DENA）／四氯化碳（CCk,乙醇／DENA诱导肝癌共20周,实验组50只和对照组10只均为GFP转基因小鼠。每周监测小鼠体重的变化;常规HE染色动态观察病理组织学;第4、12、16周处死小鼠取肝组织制作冷冻切片观察荧光表达,并以石蜡HE染色观察病理变化;第20周时处死小鼠取其肝肿物进行原代培养,观察肿瘤细胞中荧光表达和分布。结果实验组GFP转基因小鼠死亡15只,死亡率30％（15／50）。对照组10只小鼠未发生死亡。GFP转基因小鼠在诱癌的第4、12、16、20周依次出现肝炎,肝硬化、癌前病变和癌变等。荧光显微镜下观察到诱癌开始前、第4周、第12周和第16周肝脏组织的冷冻切片有GFP蛋白的表达,诱癌第20周肝肿物原代培养中肿瘤细胞的细胞质和细胞核中有GFP蛋白表达。结论本实验成功建立GFP转基因小鼠肝癌发生的动物模型,可动态观察肝癌细胞中GFP蛋白的表达。     

磁场对肿瘤影响的实验病理研究

目的探讨磁场作用对肿瘤生长的影响。方法用免疫组织化学和ＨＥ染色法对１７只磁场作用下的荷瘤小鼠进行病理学观察。结果１５ｍＴ的５０Ｈｚ交变磁场和８０ｍＴ的稳恒磁场均能提高机体的免疫功能,对肿瘤组织的供血情况有明显的抑制作用,且磁场使肿瘤组织内形成丰富间质纤维,肿瘤的坏死明显增多。而肿瘤的分化程度和ＰＣＮＡ的表达无明显差异。结论磁场能抑制肿瘤的生长。     

建立转基因小鼠突变研究模型的研究

建立转基因小鼠突变研究模型的研究黄建，成国辉，陈耀富，印木泉（上海第二军医大学卫生毒理教研室２００４３３扬州大学农学院生物所扬州２５０００９）本研究的目的是建立一个以ＸｙｌＥ（邻苯二酚氧化酶）基因为诱变靶基因，ｐＥＳｎｘ（１２．７ｋｂ）穿梭质粒为载体...     

xylE-ICR转基因小鼠模型体内致突变试验的规范化研究

本文通过ＭＮＮＧ作用于ｘｙｌＥＩＣＲ转基因小鼠,按１／１０ＬＤ５０给药,每天１次,连续５ｄ,最后一次给药后２１ｄ,测小鼠肝组织突变率为２１．４６×１０－５,与对照组（＜４．３８×１０－５）比相差极其显著（Ｐ＜０．０１）,表明该检测方法可作为规范化程序,对诱变物进行致突变研究的系统分析。     

前列腺癌干细胞相关肿瘤标志物的研究进展

前列腺癌(prostate cancer,PCa)是男性常见的恶性肿瘤。内分泌治疗是晚期前列腺癌的主要治疗方法,但该方法易使其发展成为激素难治性前列腺癌,且暂无切实有效的治疗方法。近年来的研究发现,前列腺癌干细胞在前列腺癌的发生、发展和转移中起着关键作用,因此前列腺癌干细胞的靶向治疗可能是根治前列腺癌的有效途径。靶向前列腺癌干细胞治疗需首先明确前列腺癌干细胞标志物,尤其是其特异标志物,才能更好地开展前列腺癌根治方案的研究。目前前列腺癌干细胞标志物的研究主要集中于CD44和CD133,但随着研究的不断深入其开始受到质疑,且发现了更多新的标志物,本文主要对前列腺癌干细胞领域研究较广和较新发现的肿瘤标志物进行综述。      

基因修饰小鼠肿瘤模型研究进展

动物模型在肿瘤病因的揭示、发病机理的探索以及治疗措施的评估中有着不可替代的重要作用。继常规转基因之后，可诱导表达转基因、基因打靶和条件性基因打靶等技术革新及其在肿瘤模型建立中的应用为我们提供了大量能较好模拟人体相应肿瘤的动物模型。而对它们的分析极大地深化了我们对肿瘤生物学行为的认识，并有助于人们找到攻克肿瘤的办法而造福人类。     

基于质粒载体的转基因小鼠突变研究模型的建立

基于质粒载体的转基因小鼠突变研究模型的建立黎怀星，李建秀，杨桦，胡以平，王肖鹏，傅继粱（上海第二军医大学生物教研室２００４３３华西医科大学分子生物学研究室成都６１００４１）本研究拟建立一个以ＬａｃＩ作为诱变靶基因，ｐＳＰＯＲＴＩ（４．１ｋｂ）作为载体...     

Origin of Androgen-Insensitive Poorly Differentiated Tumors in the Transgenic Adenocarcinoma of Mouse Prostate Model

Following castration, the transgenic adenocarcinoma of mouse prostate (TRAMP) model demonstrates rapid development of SV40-Tag-driven poorly differentiated tumors that express neuroendocrine cell markers. The cell population dynamics within the prostates of castrated TRAMP mice were characterized by analyzing the incorporation of 5-bromodeoxyuridine (BrdUrd) and the expression of SV40-Tag, synaptophysin, and androgen receptor (AR). Fourteen days postcastration, the remaining epithelial cells and adenocarcinoma cells were nonproliferative and lacked detectable SV40-Tag or synaptophysin expression. In contrast, morphologically distinct intraglandular foci were identified which expressed SV40-Tag, synaptophysin, and Ki67, but that lacked AR expression. These proliferative SV40-Tag and synaptophysin-expressing intraglandular foci were associated with the rare BrdUrd-retaining cells. These foci expanded rapidly in the postcastration prostate environment, in contrast to the AR- and SV40-Tag-expressing adenocarcinoma cells that lost SV40-Tag expression and underwent apoptosis after castration. Intraglandular foci of synaptophysin-expressing cells were also observed in the prostates of intact TRAMP mice at a comparable frequency; however, they did not progress to rapidly expanding tumors until much later in the life of the mice. This suggests that the foci of neuroendocrine-like cells that express SV40-Tag and synaptophysin, but lack AR, arise independent of androgen-deprivation and represent the source of the poorly differentiated tumors that are the lethal phenotype in the TRAMP model.     

Suppression of Prostate Cancer Growth by Resveratrol in The Transgenic Rat for Adenocarcinoma of Prostate (TRAP) Model.

Research into actions of resveratrol, abundantly present in red grape skin, has been greatly stimulated by its reported beneficial health influence. Since it was recently proposed as a potential prostate cancer chemopreventive agent, we here performed an in vivo experiment to explore its effect in the Transgenic Rat for Adenocarcinoma of Prostate (TRAP) model, featuring the rat probasin promoter/SV 40 T antigen. Resveratrol suppressed prostate cancer growth and induction of apoptosis through androgen receptor (AR) down-regulation, without any sign of toxicity. Resveratrol not only downregulated androgen receptor (AR) expression but also suppressed the androgen responsive glandular kallikrein 11 (Gk11), known to be an ortholog of the human prostate specific antigen (PSA), at the mRNA level. The data provide a mechanistic basis for resveratrol chemopreventive efficacy against prostate cancer.     

57例宫颈腺癌病理及免疫组织化学分析

 目的:57例宫颈腺癌的研究旨在分析宫颈腺癌的病理诊断要点及鉴别诊断。方法:采用回顾性方法总结57例宫颈腺癌的病理组织学特点,部分病例作了免疫组织化学和特殊染色。其中33例进行了长期随访并分析了预后与组织类型和临床分期的关系。结果:宫颈腺癌与非癌性增生的区别要根据多标准综合分析。PAS、Vimentin免疫标记在鉴别宫颈子宫内膜样腺癌和宫体子宫内膜腺癌方面有意义。结论:宫颈腺癌的5年生存率与宫颈腺癌类型和临床分期有关。     

Inhibitory Effect of Orally Administered 5-Aminolevulinic Acid on Prostate Carcinogenesis in the FVB-Transgenic Adenocarcinoma of a Mouse Prostate (FVB-TRAMP) Model

Background: 5-aminolevulinic acid (5-ALA) is a constituent of mitochondrial electron carriers, heme and cytochrome c, which are crucial for aerobic energy metabolism and cell apoptosis. We investigated the chemopreventive efficacy of 5-ALA against prostate cancer using the FVB-transgenic adenocarcinoma of mouse prostate (FVB-TRAMP) model. Methods: Samples were collected from 24 FVB-TRAMP mice at 12 and 20 weeks of age (named the first and second sets, respectively). Sixteen mice (from the first set) were randomly allocated into 3 treatment groups: 1) control (no treatment), 2) low dose of 5-ALA (30 mg/kg/day), and 3) high dose of 5-ALA (300 mg/kg/day). Similarly, 8 mice were divided into 2 treatment groups: 1) control and 2) high dose of 5-ALA (300 mg/kg/day). 5-ALA was orally administered to mice before cancer onset, from 6 weeks of age. Results: In the control group, prostate cancer was pathologically detected in 33 and 50 % of mice at 12 and 20 weeks, respectively, while 25% of 12-week old mice in the low-dose group were affected and none of the high-dose group mice developed prostate cancer. Immunohistochemical analysis showed higher expression of cytochrome c oxidase subunit 4 (COX4) in the prostate gland of the high-dose group compared to the control (P = 0.018). Similarly, enzyme-linked immunosorbent assay using lysed prostate tissue revealed higher amounts of cytochrome c in the prostate of the high-dose group compared to the control (P = 0.021). Furthermore, western blot analysis showed higher level of cleaved caspase-3 in mice in the high-dose group diagnosed with high-grade prostatic intraepithelial neoplasia. Conclusion: Our results suggest that oral 5-ALA may support the functional expression of mitochondrial cytochrome c and COX4, leading to caspase 3-dependent apoptosis in carcinogenesis in FVB-TRAMP mice. Future clinical studies are warranted to confirm the chemopreventive value of 5-ALA in prostate carcinogenesis.     

胃腺癌和癌旁腺体形态改变的病理学观察

我们对１０７例胃腺癌和癌旁腺体的形态改变，结合粘液组织化学和免疫酶组织化学的方法进行了观察，发现不同性质的腺体扩张和组织学类型有关。单纯性腺体扩张见于各种类型的腺癌；异型性腺体扩张多见于管状腺癌。尤其早期管状腺癌出现各级、多量异型腺体扩张，并与管状腺癌的腺体交叉，有向管状腺癌由轻度到重度异型增生上皮过渡的形态。异型性腺体扩张的组织结构、细胞形态、粘液组织化学和免疫酶组织化学都与管状腺癌相一致，只是程度的差异。     

结肠黏液腺癌临床病理特征分析

目的：探讨结肠黏液腺癌的临床病理特征及预后特点。方法回顾性分析672例初治的接受根治性手术的结肠癌患者,其中黏液腺癌为41例,非黏液腺癌为631例,比较两者临床病理特征和复发方式。结果黏液腺癌患者比非黏液腺癌患者更年轻（<50岁比例：48.8豫 vs 31.4豫,P <0.05）,肿瘤直径更大（6.0 cm vs 4.5 cm,P <0.05）,更多位于右半结肠（右半结肠比例：44.0豫 vs 17.3豫,P <0.05）和病理 T 分期更高（T3、T4比例：90.2豫 vs 72.7豫,P <0.05）,3 a 生存率更短（39.2豫 vs 68.5豫,P <0.05）。复发情况中,黏液腺癌腹膜转移率比非黏液腺癌更高（57.1豫 vs 1.5豫,P <0.05）。结论与非黏液腺癌比较,结肠黏液腺癌发病年龄更年轻,更多处于晚期和右半结肠,腹膜转移可能性更大,治疗预后更差,提示结肠黏液腺癌患者需要特异的治疗方案。     

Incidentally Detected Adenocarcinoma Prostate in Transurethral Resection of Prostate Specimens: a Hospital Based Study from India

Background: Awareness about prostate cancer has increased in the community, and prostate cancer screening examinations, including prostate specific antigen (PSA) assays, are now widely available. Prior to the PSA era, up to 27% of prostate cancers were detected incidentally at the time of transurethral resection of prostate (TURP). After PSA testing became widely available, the incidence of incidentally detected carcinoma prostate in TURP specimens without prior diagnosis reduced to 5-13%. However, the incidence of incidentally detected carcinoma prostate has been reported to vary across the globe since various factors can influence the identification of this malignancy in TURP specimens. In this paper, we focus on rates of incidentally detected prostate cancer in TURP specimens in our hospital and correlate it with various parameters. Materials and Methods: This retrospective study of histopathological findings of biopsy specimens was conducted for patients undergoing TURP during a period of 5 years from April 2010. The inclusion criteria were patients diagnosed with benign prostatic hyperplasia (BPH) (digital rectal examination (DRE) not showing any abnormally hard areas and normal age adjusted PSA values). Patients with elevated PSA, abnormal DRE, documented urinary tract infection and proved adenocarcinoma prostate (CaP) were excluded from the study. The total weight of prostatectomy specimen, occurrence of carcinoma prostate in the chips, percentage of total tissue resected showing malignancy and Gleason's scores were recorded. Results: A total of 597 patients belonging to the inclusion criteria were studied. The incidence of occult CaP in the study group was 5.2 % (31/597). Out of these, 8 belonged to T1a and 23 belonged to T1b stages. The age group 70 - 79 years had the maximum incidence of occult CaP. It was observed that the clinical grading of prostate did not have a bearing on the incidence of occult CaP whereas the weight of resected specimen correlated with the incidence of CaP. The incidence of occult CaP was greater with low volume prostates ( < 20 g). (P=0.15). Conclusions: The rate of incidentally detected adenocarcinoma prostate in patients undergoing TURP for clinically diagnosed BPH was found to be only 5.2 % in our study which is low when compared with similar studies done elsewhere. The age of the patient and weight of the resected specimen correlated with incidence of occult prostate cancer. The clinical grading of prostate by DRE however, demonstrated no correlation.     

Stromal Activation Associated with Development of Prostate Cancer in Prostate-Targeted Fibroblast Growth Factor 8b Transgenic Mice

Expression of fibroblast growth factor 8 (FGF-8) is commonly increased in prostate cancer. Experimental studies have provided evidence that it plays a role in prostate tumorigenesis and tumor progression. To study how increased FGF-8 affects the prostate, we generated and analyzed transgenic (TG) mice expressing FGF-8b under the probasin promoter that targets expression to prostate epithelium. Prostates of the TG mice showed an increased size and changes in stromal and epithelialmorphology progressing fromatypia and prostatic intraepithelial neoplasia (mouse PIN, mPIN) lesions to tumors with highly variable phenotype bearing features of adenocarcinoma, carcinosarcoma, and sarcoma. The development of mPIN lesions was preceded by formation of activated stroma containing increased proportion of fibroblastic cells, rich vasculature, and inflammation. The association between advancing stromal and epithelial alterations was statistically significant. Microarray analysis and validation with quantitative polymerase chain reaction revealed that expression of osteopontin and connective tissue growth factor was markedly upregulated in TG mouse prostates compared with wild type prostates. Androgen receptor staining was decreased in transformed epithelium and in hypercellular stroma but strongly increased in the sarcoma-like lesions. In conclusion, our data demonstrate that disruption of FGF signaling pathways by increased epithelial production of FGF-8b leads to strongly activated and atypical stroma, which precedes development of mPIN lesions and prostate cancer with mixed features of adenocarcinoma and sarcoma in the prostates of TG mice. The results suggest that increased FGF-8 in human prostate may also contribute to prostate tumorigenesis by stromal activation.