D-半乳糖致衰老模型小鼠皮肤衰老的观察

目的　观察D 半乳糖连续皮下注射对小鼠皮肤衰老指标的影响。　方法　 3月龄KM雌性小鼠 6 0只 ,随机分为空白对照组 (注射生理盐水 )、D 半乳糖低剂量组 (80mg/kg)和D 半乳糖高剂量组 (10 0 0mg/kg) ,连续每日背部皮下注射 4 2天后 ,观察小鼠皮肤中与衰老相关的生化指标及组织病理学变化 ,并用计算机图像分析系统定量分析。　结果　高剂量组小鼠真皮厚度〔(6 2 4 5±4 8 5 ) μm〕较对照组〔(839 3± 5 8 4 ) μm〕显著变薄 (P <0 0 5 ) ,胶原纤维减少 ,排列疏松 ,弹力纤维总面积亦显著减少 (P <0 0 5 ) ;高剂量组皮肤中超氧化物歧化酶 (SOD)活性〔(131 2± 11 5 )U/ml〕、羟脯氨酸含量〔(0 5 7± 0 13) μg/mg〕显著低于对照组的〔(178 1± 2 0 7)U/ml、(0 74± 0 17) μg /mg ,P <0 0 5〕 ,丙二醛 (MDA)含量增加〔(9 4± 1 5 )nmol/ g和 (6 8± 1 5 )nmol/ g ,P <0 0 5〕 ;而低剂量组上述改变则不显著。　结论　每日 10 0 0mg/kgD 半乳糖连续皮下注射 4 2天 ,可导致小鼠皮肤的明显衰老 ,为抗皮肤衰老研究提供一个简便、稳定的实验模型。     

利用不同浓度D(+)半乳糖建立卵巢早衰动物模型

目的:评价不同浓度半乳糖代谢建立小鼠卵巢早衰模型的方法及其可行性。方法:选择8-12周龄SPF级昆明小鼠60只,分为高浓度实验组,低浓度实验组,正常对照组,各20只,给予35%、28%半乳糖食物丸和标准食物丸。在饲养的第70天,取血检测小鼠体内半乳糖、E2、FSH、总蛋白、清蛋白水平,并行促排卵检查卵巢功能,光镜观察卵巢和肝组织形态学变化。结果:与对照组及低浓度实验组相比,高浓度实验组卵巢重量显著降低(P<0.001);光镜下,高浓度实验组小鼠肝组织呈变性、坏死,卵巢组织可见大量初级卵泡,但生长卵泡和黄体数量减少;血清总蛋白和清蛋白浓度下降,血清半乳糖、FSH水平明显高于对照组(P均<0.001),E2水平明显低于对照组(P<0.001),促排卵后检查小鼠输卵管中成熟卵母细胞数目,明显低于对照组(P<0.001)。结论:利用35%半乳糖代谢作用建立卵巢早衰动物模型是可行的,可为解决卵巢早衰患者生育问题提供研究之手段。     

Antimetastatic Effect of Hepatotropic Liposomal Adriamycin on Human Metastatic Liver Tumors

Adriamycin (ADM) was encapsulated in a galactose-conjugated hepatotropic liposome (hLip-ADM) and its ability to enhance the antitumor effect while reducing toxicity was compared with that of free ADM and a control Lip-ADM (cLip-ADM), in two in vitro animal models. Liver metastases were induced in rats by an intravenous injection of 8 × 10⁶ AH-130 rat hepatoma tumor cells. All forms of the ADM completely inhibited liver metastasis when given at a dose of 5 mg/kg on day 14 after tumor implantation, whereas liver metastatic foci were observed in six of ten rats in the control group. The reduction in ADM toxicity by liposomalization was remarkable, as significant body weight loss was observed only in the free ADM group, in which four of ten rats died. Additional experiments utilized a human colon cancer xenograft (TK-4) to induce the growth of the liver metastases in mice by orthotopic implantation. The hLip-ADM completely inhibited liver metastasis in rats (0/11), whereas liver metastases developed in 10 of 12 mice in the control group and in 5 of 12 mice given cLip-ADM. Interestingly, liposomal ADM did not have a significant inhibitory effect on transplanted tumor growth assessed 6 weeks after transplantation. These findings indicate that hLip-ADM may be an effective strategy for inhibiting liver metastases from human colon cancer. Received: June 15, 2000 / Accepted: November 20, 2000     

半乳糖受体介导的增殖细胞核抗原反义核酸对肝癌Bel-7402细胞增殖的靶向抑制作用

目的探讨半乳糖受体介导的增殖细胞核抗原（PCNA）反义核酸（ASODN）对肝癌Bel-7402细胞增殖的靶向抑制作用。方法选择来源于3个不同系统的肿瘤细胞：肝癌Bel-7402细胞、肺腺癌GLC-82细胞、慢性髓系白血病K562细胞，将半乳糖-聚乙烯亚胺（Gal-PEI）与PCNA反义核酸结合形成交联复合物Gal-PEI-ASODN，台盼兰拒染计数法观察不同浓度Gal-PEI-ASODN对这3种细胞增殖的影响；流式细胞仪检测3种细胞对羧基荧光素（FAM）标记的Gal-PEI-ASODN的摄取率及细胞内平均荧光强度；相差／荧光显微镜观察FAM标记的Gal-PEI-ASODN及ASODN进入3种不同细胞的情况。结果浓度从10μmol／L到40μmol／L的ASODN作用于Bel-7402细胞48h后，可抑制其增殖，IC50为29．3μmol／L。浓度从0．05μmol／L到0．25μmol／L的ASODN作用于Bel-7402细胞48h后，均未显著抑制其增殖；相同浓度的Gal-PEI-ASODN作用于3种不同细胞48h后，对GLC-82细胞、K562细胞的增殖未产生显著抑制作用，而Bel-7402细胞的增殖却受到明显抑制。对于Bel-7402细胞，Gal-PEI-ASODN的IC50为0．06μmol／L，与单纯ASODN组相比。增敏倍数为488倍。FAM标记的Gal-PEI-ASODN或ASODN与不同细胞孵育0．5h到2．0h内，Gal-PEI-ASODN-Bel-7402细胞组所有时间点的细胞荧光摄取率、胞内平均荧光强度均显著高于A-SODN-Bel-7402细胞组、Gal-PEI-ASODN-GLC-82细胞组和Gal-PEI-ASODN-K562细胞组。FAM标记的Gal-PEI-ASODN或ASODN与Bel-7402、GLC-82、K562细胞作用1h后．Gal-PEI-ASODN-Bel-7402组的细胞荧光摄取率较高（反义核酸大量分布于细胞的胞浆内），其他各组细胞荧光摄取率较低。结论半乳糖受体介导的PCNA反义核酸能被肝癌Bel-7402细胞高效地摄取。抑制Bel-7402细胞的增殖，增强反义核酸的作用效果，具有靶向性。     

二苯乙烯苷对D-半乳糖衰老模型小鼠衰老指征的影响

目的:探讨二苯乙烯苷对D-半乳糖所致亚急性衰老的影响。方法:将174只小鼠随机分为三组,即对照组、衰老模型组和二苯乙烯苷组,分别给予生理盐水、D-半乳糖和D-半乳糖+二苯乙烯苷,六周后部分小鼠测定游泳时间,耐缺氧、耐寒和耐高温能力。另一部分小鼠测定不同组织脂褐素(LPF)、Na+/K+-ATP酶、超氧化物歧化酶(SOD)和丙二醛(MDA)含量。测定二苯乙烯苷对人红细胞膜的MDA的生成量的影响。结果:二苯乙烯苷明显延长游泳时间、耐缺氧和耐高温能力,二苯乙烯苷可降低小鼠脑组织LPF含量,升高心肌Na+/K+-ATP酶和肝脏SOD活性。二苯乙烯苷可减少邻苯三酚所致的人红细胞膜MDA的生成。结论:二苯乙烯苷具有抗衰老作用,其机制是通过多种途径而实现的。     

Galectin-9在卵巢癌组织中的表达及其临床意义

目的 观察半乳糖凝集素-9(Galectin-9)在卵巢癌组织中的表达及其临床意义。方法 收集我院外科手术切除的卵巢癌组织、癌旁组织(距离癌灶＜2cm)及卵巢良性组织(距离癌灶>2cm)各60例,采用免疫组化法及RT-PCR法检测卵巢组织中Galectin-9蛋白及mRNA表达。结果 免疫组化证实卵巢癌组织中Galectin-9阳性表达率显著低于癌旁组织及良性卵巢组织,癌旁组织中Galectin-9阳性表达率低于卵巢良性组织,差异均具有统计学意义(P＜0.05)。RT-PCR证实卵巢癌组织中Galectin-9 mRNA表达显著低于癌旁组织及良性卵巢组织,癌旁组织中的Galectin-9mRNA表达低于卵巢良性组织,差异均具有统计学意义(P＜0.05)。免疫组化亦证实有淋巴结转移卵巢癌组织中的Galectin-9阳性表达率低于无淋巴结转移卵巢癌组织,差异具有统计学意义(P＜0.05)。卵巢癌组织中Galectin-9阳性表达率与病理类型,病理分级,TNM分期无关(P>0.05)。结论 Galectin-9低表达可能与卵巢癌的发生及淋巴结转移密切相关,可能成为卵巢癌预后的标志及基因治疗的靶点。     

Influence of Early Postoperative Enteral Nutrition versus Placebo on Cell-Mediated Immunity,As Measured with the Multitest® CMI

Background: Patients who become anergic after surgery have a very high death rate, mainly due to infectious complications. It has earlier been shown that preoperative skin testing has predictive value for the clinical outcome in gastrointestinal surgery. This study investigated the effect of early postoperative enteral nutrition versus placebo on cell-mediated immunity with the Multitest® CMI and correlated the results to the clinical outcome and to the preoperative nutritional status. Methods: Sixty patients were randomized to receive either enteral nutrition or placebo. All patients were stratified in accordance with preoperative nutritional status. CMI tests were applied 2 days before surgery and days 1 and 5 postoperatively. All tests were applied and recorded by the same investigator. Results: No significant differences in the score were seen between the groups. Nine patients were malnourished. No significant difference was seen preoperatively with regard to nutritional status, but the malnourished patients had a lower median value on the 3rd postoperative day. In patients with complications the preoperative median CMI score was 17.0 mm (95% confidence limits, 10.5-24 mm), and in patients without complications 9.5 mm (95% confidence limits, 5.5-16 mm). This result is significant and unexpected. Ten patients were anergic preoperatively, and three of these developed complications (30%). The overall complication rate was 27 of 60 (45%). A change in immunologic status from responding preoperatively to anergy postoperatively was significantly associated with severe complications (P = 0.043). Conclusions: Early enteral nutrition does not influence the CMI score. Preoperative anergy was not related to increased mortality/morbidity, whereas the change in CMI response towards anergy was significant.     

半乳糖性白内障晶体上皮细胞游离钙的实验研究

目的研究半乳糖性白内障晶体上皮细胞中游离钙浓度变化规律。方法给２８天鼠龄的Ｗｉｓｔａｒ大白鼠投喂５０％的半乳糖饮食复制白内障模型后,分别于给药后第１,３,７,１０,１４及１９天测量晶体上皮细胞中游离钙的浓度。结果半乳糖性白内障各时期晶体上皮细胞中游离钙的浓度均明显升高,第１０天游离钙浓度最高,是正常对照组的４～５倍。结论半乳糖性白内障晶体上皮细胞中游离钙的浓度升高,各期上升的程度不同,以皮质混浊期升高幅度最大。     

半乳糖基多聚赖氨酸携带HSV1-TK对HepG2细胞体外效应研究

目的为肝癌的自杀基因治疗寻找新的靶向分子方法通过还原氨化法进行乳糖（Lac）与聚赖氨酸（PLL）共价连接；经Sephadex G-10枉分离纯化；以苯酚-硫酸比色法测定n值；与真核表达质粒r-pAs16Dr按比例混匀，得到GlanPLL-r-pAs16Dr基因转移系统；分别转染不同的细胞株进行体外效应研究：人肝癌细胞株HepC2、人肺癌细胞株549，观察报告基因红色荧光蛋白能否在肝细胞中表达；RT—PCR检测HSV—tk基因的表达情况；观察给予不同浓度的更昔洛韦（GCV）后随时间延长细胞改变情况，用MTT法检测GCV对不同细胞的杀伤作用。结果化学合成得到34个半乳糖基的PLL；转染后48h在HepG2可见红色荧光蛋白表达，A549细胞未见红色荧光蛋白表达；RT-PCR证实HSV—tk仅在HepG2细胞中表达：体外杀伤实验显示：经GlanPLL-r-pAs16Dr基因转移系统处理的两种细胞对GCV表现出不同的敏感性HepG2细胞对GCV很敏感，低浓度的GCV（1mg／L）处理3d．即可使细胞生长抑制率达到70．5％，A549对GCV不敏感结论半乳糖基多聚赖氨酸能够与ASGPR有效的结合，合成配体来源丰富、制备简单，适合于作为自杀基因HSV—tk肝靶向运送的导向配体。     

D-半乳糖亚急性中毒大鼠拟衰老生理生化改变

目的　D 半乳糖皮下注射致Wistar大鼠亚急性衰老模型 ,观察大鼠衰老指标的变化。方法　 2 0只雄性大鼠随机分为 2组 ,每组 10只。对照组每日一次皮下注射生理盐水 10ml kg,衰老模型组每日一次皮下注射D 半乳糖 4 8mg kg。各组大鼠均连续给药 4 2d ,采血后分别测定各项衰老指标。结果　与对照组比较 ,衰老模型组体重增加比例降低 ,胸腺明显萎缩 ,脏器系数减少 (P <0 .0 1) ,血清中超氧化物歧化酶 (SOD)活性明显下降 (P <0 .0 1)。结论　D 半乳糖诱发的致大鼠亚急性衰老是探讨自由基衰老学说有用的实验动物造模技术。