Chemopreventive Effect of Dietary Maranta arundinacea L. Against DMBA-Induced Mammary Cancer in Sprague Dawley Rats Through the Regulation of Autophagy Expression

Background: Breast cancer prevention still needs to be improved. Calorie restriction is thought to prevent breast cancer through the induction of autophagy. Maranta arundinacea L. (MA) has the potential for calorie restriction because it contains high fiber. This research aimed to observe the effect of dietary MA against dimethylbenz(a)anthracene (DMBA)-induced mammary cancer in Sprague Dawley rats related to autophagy. Methods: Twenty-five Sprague Dawley rats were randomly divided into five groups: 1) control group without DMBA-induced with a standard diet, 2) 20 mg/kg BW of DMBA two times a week for five weeks with a standard diet, 3) DMBA and diet modification with 30% of MA, 4) DMBA and diet modification with 45% of MA, and 5) DMBA and diet modification with 60% of MA. Examination of the nodule was conducted once every week for 22 weeks. Breast tissue/tumor examination underwent histology examination with hematoxylin-eosin. Examinations of immunohistochemical staining against Beclin1, LC3B, and SQSTM1 were conducted to reveal autophagy. The difference of autophagy protein expression was analyzed using One way ANOVA with 95% confidence level and significance set as p<0.05. Results: Cancer was detected in four rats of DMBA standard diet, two rats of 30% MA, one rat of 45% MA. No cancer was detected in the rats of control and rats with 60% of MA group. The Beclin1 expressions showed that the 60% of MA group had the highest score (2.5±0.52) followed by the 45% of MA group (1.87±0.49), control group (1.77±0.11), 30% of MA group (1.28±0.75), and DMBA with standard diet had the lowest score (1.28±0.91). The difference of Beclin1 expressions was statistically significant (p-value=0.03). However, the difference of the LC3B expressions (p-value=0.11) and SQSTM1 expressions (p-value=0.225) were not statistically significant. Conclusion: Dietary modifications with MA potentially prevent breast cancer and induce initiation of autophagy.     

食管癌组织中肿瘤浸润性树突状细胞表型改变及功能分析

目的:探讨食管癌患者肿瘤组织中肿瘤浸润性树突状细胞表型改变及功能。方法:收集2017年01月至2018年09月我院收治的食管癌患者92例作为观察组,选择同时期我院收治的92例食管良性肿物患者作为对照组。术中分别取食管癌标本和良性肿物标本。检测相应组织中肿瘤浸润性树突状细胞表达水平及表型,同时检测T细胞亚群表达情况,分析肿瘤浸润性树突状细胞表达情况和食管癌患者临床特征的相关性。结果:与对照组比较,观察组肿瘤浸润性树突状细胞密度、MHC-Ⅱ阳性树突状细胞和CD54阳性树突状细胞百分比均显著降低(P＜0.05);观察组CD4+T细胞增高［(24.81±3.72）% vs （20.77±3.63）%,P=0.000］;CD8+T细胞降低［(20.90±4.12）% vs （23.08±4.42）%,P=0.001］。食管癌组织中肿瘤浸润性树突状细胞密度、MHC-Ⅱ阳性树突状细胞和CD54阳性树突状细胞百分比与肿瘤直径、TNM分期和淋巴结转移有关(P＜0.05)。食管癌组织中肿瘤浸润性树突状细胞密度、MHC-Ⅱ阳性树突状细胞和CD54阳性树突状细胞与CD4+T细胞显著负相关,与CD8+T细胞显著正相关(P＜0.05)。结论:肿瘤浸润性树突状细胞在食管癌组织中低表达,功能低下,与T细胞亚群失衡和预后不良有关。     

Anti-tumor Activity of Phyllanthus niruri (a Medicinal Plant) on Chemical-induced Skin Carcinogenesis in Mice

Chemoprevention is an important strategy to control the process of carcinogenesis. The potential of usingmedicinal herbs as cancer chemopreventive nutraceuticals and functional food is promising. Thus, there is aneed for exploring drugs/agents which act as chemopreventive agents. Phyllanthus niruri is a well known medicinalplant which has been used in Ayurvedic medicine as hepatoprotective, antiviral, antibacterial, analgesic,antispasmodic and antidiabetic. The present study was carried out to evaluate the anti-tumor activity of ahydro-alcoholic extract of the whole plant, in 7-9 week old male Swiss albino mice, on the two stage process ofskin carcinogenesis induced by a single topical application of 7, 12-dimethylbenz (a)anthracene (100μg/100μlacetone) and two weeks later promoted by repeated application of croton oil (1% in acetone/three times a week)till the end of experiment (16 weeks). The oral administration of P. niruri at a dose of 1000 mg/kg/b.wt. at peri-(i.e. 7 days before & 7 days after DMBA application) and post- (i.e. starting from the croton oil application)initiational phase of papillomagenesis caused significant reduction in tumor incidence, tumor yield, tumor burdenand cumulative number of papillomas as compared to carcinogen-treated controls. Furthermore, the averagelatent period was significantly increased in the PNE treated group. The results thus suggest that P. niruri extractexhibits significant anti-tumor activity, which supports the traditional medicinal utilization of this plant.     

Mammary carcinoma induced by oral administration of ethyl methanesulphonate. Determination of some of the parameters affecting tumor induction

The carcinogenic activity of ethyl methanesulphonate (EMS),an alkylating agent and a potent mutagen, was examined in WistarKing A and Sprague Dawley rats following oral administration.In the Wistar King A rats, mammary carcinomas were detectedin all of the young female rats by the 32nd week after initiationof the experiment. To determine the optimal experimental conditionsfor the rapid and invariable induction of mammary carcinomas,the relationship among age, sex, strain of the rats and concentrationand duration of EMS administration, and tumor production wasinvestigated. The tumor incidence was higher in the youngerfemale rats and the Wistar rats were more susceptible than Sprague-Dawleyrats. Mammary carcinomas were also induced in the younger malerats. In addition, concommitant production of renal tumors occurredby the 40th week in young rats administered 10^–2 M or2 x 10^–2 M of EMS solution, while renal and uterine tumorsdeveloped eoncommitantly in the older female rats given 3 x10^–3 EMS by the 56th week. Histologically, the prevailingfeatures of tumors were infiltrating ductal adenocarcinoma inthe mammary glands, mesenchymal tumor in the kidney, and leiomyosarcomain the uterus. Methyl methanesulphonate, an analogue of EMS,did not induce tumors in any organs.     

外周血粒淋比联合肿瘤相关中性粒细胞对术后胃癌患者预后的影响

  目的  探讨胃癌患者外周血中性粒细胞与淋巴细胞比值（neutrophil to lymphocyte ratio,NLR）和胃癌间质肿瘤相关中性粒细胞（tumor-associated neutrophils,TANs）对胃癌患者预后的预测价值。  方法  选取第二军医大学附属长海医院2006年6月至2011年5月收治的126例胃癌患者,根据术前外周血NLR及胃癌患者TANs的浸润情况,将其分为4个组,比较全组胃癌患者的术后生存率,并分析其与临床病理学特征及预后的相关性。  结果  NLR高合并TANs低浸润组肿瘤分化程度较低（P < 0.001）且肿瘤体积较大（P=0.026）。4个组中NLR高合并TANs低浸润组生存率最低,NLR低合并TANs高浸润组生存率最高,且差异具有统计学意义（P < 0.05）。单因素及多因素Cox回归分析均显示:NLR高合并TANs低浸润（P < 0.05）是影响胃癌预后的独立危险因素。  结论  术前外周血NLR联合胃癌组织中TANs浸润可作为判断胃癌患者预后的指标,且术前外周血NLR高联合胃癌组织中TANs低浸润胃癌患者的预后较差。      

卡培他滨片联合介入治疗大肠癌肝转移疗效分析

目的:观察口服卡培他滨片联合介入治疗大肠癌肝转移的疗效。方法:选择我院2010年1月-2010年10月76例大肠癌肝转移患者,采用隐匿数字随机法分为两组,对照组38例患者先行肝总动脉和腹腔干动脉造影,根据造影提示行肝动脉灌注化疗和栓塞,研究组38例患者在此基础上联合卡培他滨片口服,比较两组近、远期疗效。结果:研究组RR率为50.00%,对照组RR率为26.32%(P＜0.05)。研究组血流信号改善情况优于对照组(P＜0.05)。研究组患者12个月、24个月、48个月生存率分别为86.84%、65.79%、34.21%,均高于对照组(P＜0.05)。根据富血供与乏血供分为4个亚组,富血供患者疗效明显好于乏血供患者,富血供患者生存期明显长于乏血供患者(P＜0.05)。两组均无严重不良反应病例(P＞0.05)。结论:介入治疗联合卡培他滨片口服可显著延长大肠癌肝转移患者的生存期,值得临床采用。     

RAGE、sRAGE在结肠癌中的表达及其临床意义

目的：本研究通过检测结肠癌组织晚期糖基化终末产物受体（ the receptor for advanced glycation end product,RAGE）和血清可溶性RAGE（ soluble RAGE,sRAGE）的表达水平,探讨其在结肠癌发生、发展过程中的作用及其临床意义。方法：本研究纳入49例非糖尿病结肠癌（ I、II、III期）患者,以免疫组织化学评分法（ immunohistochemical score,IHS）评价结肠癌RAGE表达水平,以ELISA法检测患者外周血血清sRAGE浓度。结果：结肠癌组织RAGE表达水平（IHS为1.24±0.48）显著高于癌周组织（IHS为0.50±0.25）（P﹤0.05）, III期（IHS为1.42±0.51）显著性高于I期（IHS为1.01±0.35）（P﹤0.05）和II期（IHS为1.11±0.42）（P﹤0.05）;低分化结肠癌组织（IHS为1.58±0.49）显著高于中分化（IHS为1.08±0.45）（P﹤0.05）和高分化结肠癌组织（IHS为1.02±0.27）（P﹤0.05）。结肠癌患者血清sRAGE表达水平术前、术后分别为（344.77±68.06）ng/L、（265.43±76.85）ng/L,二者相比有显著性差异（ P ﹤0.05）,而且分别与健康志愿者 sRAGE （149.97±30.12）ng/L相比有显著性差异（ P﹤0.05）,但sRAGE表达水平与TNM分期、结肠癌组织分化程度没有关联。结肠癌组织RAGE高表达患者术前血清sRAGE浓度（415.02±85.08）ng/L高于中表达（334.26±44.56）ng/L和低表达（335.95±78.48）ng/L患者（P﹤0.05）。结论：结肠癌组织RAGE表达水平与分化程度呈负相关趋势,与TNM分期呈正相关趋势,作为术后诊断指标有一定的临床意义。血清sRAGE浓度与结肠癌组织RAGE表达水平呈正相关趋势,但与TNM分期和结肠癌组织分化程度没有关联,作为术前诊断指标仅能代表结肠癌风险,但无法考量结肠癌的恶性程度和进展情况。     

Promoting activities of the C-9 oxidized metabolites of N-2-fluorenylacetamide in liver carcinogenesis in the rat

The promoting potential of the C-9 oxidized metabolites of N-2-fluorenylacetamide (2-FAA) was examined and compared to that of the promoter 2-FAA in the two-stage hepatocarcinogensis system Male Sprague Dawley rats were initiated with a single intraperitoneal injection of diethylnitrosamine at 200 mg/kg of body weight (b.wt.). One, 2 and 5 weeks thereafter rats were given by gavage 5 doses per week of 2-FAA (0.05 mmol/kg of b.wt.), 9-hydroxy- or 9-oxo-2-FAA (0.05 and 0.1 mmol/kg of b.wt.) or the vehicle polyethylene glycol 400 (2 ml/kg of b.wt.). All rats underwent partial (70%) hepatectomy on day 3 after 5 initial doses. Following treatment with 2FAA, 9-hydroxy-or 9-oxo-2-FAA, the levels of gamma-glutamyl-transpeptidase activity and placental glutathione S-transferase (GST-P) were increased in the livers proportional to the increased numbers of foci of GST-P positive hepatocytes. The activities of the C-9 oxidized compounds were dose-dependent since treatment at 0.1 mmol/kg of b.wt. resulted in -2-fold greater effects than that at 0.05 mmol/kg of b.wt. The incidences of hepatocellular carcinoma increased with time after treatment and the relative order of potency was: 2-FAA>9-oxo-2-FAA>9-hydroxy-2-FAA. These data combined with our earlier evidence for preferential oxidations of 2-FAA at C-9 by hepatic microsomes of male Sprague Dawley rats support the significance of these metabolites as a promoting stimuli in liver carcinogenesis by 2-FAA.     

Inhibition of N-Methyl N'-nitro-N-nitrosoguanidine (MNNG) Induced Gastric Carcinogenesis by Phyllanthus amarus Extract

Chemopreventive activity of Phyllanthus amarus Schum & Thonn (Euphorbiaceae) extract was studied withregard to N-methyl N’-nitro-N-nitrosoguanidine (MNNG) induced stomach cancer in Wistar rats. Administration ofthe extract with MNNG significantly reduced the incidence of gastric neoplasms in rats (44%) as well as their numbers.Moreover, elevated levels of enzymes in the stomach were found to be reduced by P. amarus administration. Forexample, γ-glutamyl transpeptidase activity was decreased from 20.3 ± 6.7 mmol/min/mg protein to almost normallevels (2.8 ± 0.9) by 750mg/kg body weight of the extract. Similarly glutathione S-transferase activity (1317.6 ± 211 nmol/min/mg protein) and glutathione reductase (368 ± 66) levels in the MNNG treated group were found to belowered to 494.8 ± 76 and 192 ± 45, respectively, while reduced glutathione (GSH) was increased from 4.6 ± 0.9 to 8.5±1.4 n mol/min/mg protein. AgNOR dots and clusters, indicators of cellular proliferation, which were increased byMNNG treatment, became near to normal in P. amarus treated animals.     

增殖诱导配体小干扰RNA对人结直肠癌裸鼠移植瘤细胞增殖和凋亡的影响

目的 研究增殖诱导配体(APRIL)小干扰RNA(siRNA)对人结直肠癌裸鼠移植瘤细胞增殖和凋亡的抑制作用.方法 建立人结直肠癌裸鼠移植瘤模型,将裸鼠分为3组,瘤块内分别注射APRIL siRNA、空载体和PBS液,每2天注射1次,共2周.采用实时荧光定量PCR法和免疫组化SP法检测APRIL siRNA对APRILmRNA和蛋白表达的影响,采用酶联免疫吸附(ELISA)法检测肿瘤内增殖细胞核抗原(PCNA)含量的变化,采用免疫组化SP法检测肿瘤内bcl-2和bcl-xl蛋白的表达,采用原位末端标记(TUNEL)法检测肿瘤细胞的凋亡.结果 APRIL siRNA能敲低APRIL基因的表达水平,APRIL siRNA组移植瘤内APRIL mRNA的相对表达量为(0.13 ±0.05)×10-3,明显低于空载体组[(0.95 ±0.04)× 10-3]和空白对照组[(0.96±0.05)×10-3,P＜0.05].APRIL siRNA组APRIL蛋白的表达比空载体组和空白对照组降低了(87.5%±5.0%,P＜0.05).APRILsiRNA组裸鼠移植瘤生长缓慢,瘤块重量较空载体组和PBS对照组明显降低(P＜0.05).APRIL siRNA组移植瘤内PCNA的含量为(176.8±18.1)ng/ml,明显低于空载体组[(330.0±20.5)ng/ml]和空白对照组[(328.4±22.8)ng/ml,P＜0.05];bcl-2和bcl-xl蛋白的表达量也较空载体组和空白对照组明显降低(P＜0.05).APRIL siRNA组移植瘤内凋亡细胞的数量增多,凋亡率为40.1%±2.5%,明显高于空载体组(2.5%±0.1%)和空白对照组(2.5%±0.2%,P＜0.05).结论 APRIL siRNA能在裸鼠体内抑制人结直肠癌细胞的增殖,并促进细胞凋亡,达到显著的抑瘤效果.APRIL基因可能成为人结直肠癌基因靶向沉默治疗的重要候选基因之一.     

Dietary whey protein protects against azoxymethane-induced colon tumors in male rats.

Epidemiological studies have suggested a relationship between diet and colon cancer incidence. Results from animal studies suggest that whey protein, but not casein protein, may provide protective effects against experimentally induced breast cancer in animals. In the current study, we investigated the effects of casein and whey diets on chemically induced colon cancer in male rats. Pregnant female Sprague Dawley rats (days 3-4 of gestation) were maintained on modified AIN-93G diets formulated with a single protein source of either casein or whey. Life-time exposure to these diets was studied in the F1 generation (experiment A) or the F2 generation (experiment B). Male offspring were weaned to the same diets as the dams and were maintained on these diets throughout the study. At age 90 days, all rats received azoxymethane once a week for 2 weeks (s.c., 15 mg/kg). Forty weeks after the last azoxymethane injection, all rats were euthanized, the colon was examined visually for tumors, and each tumor was histologically evaluated. The weights and distribution of all of the tumors were recorded. In experiment A, rats fed the casein diet had a 56% incidence of colon tumors compared with 30% of the rats on whey-based diets (P < 0.05). In experiment B, rats fed the casein diet had 50% incidence of colon tumors compared with 29% in the whey group (P < 0.05). There were no significant effects of diet on tumor multiplicity or mass. These results suggest that consumption of whey protein-containing diets may reduce the risk of developing colon tumors.     

Runx3和CHD5基因在结直肠癌中的表达及临床意义

目的 探讨Runt相关转录因子3(Runx3)和染色质解旋酶DNA结合蛋白5(CHD5)基因在结直肠癌组织中的表达及临床意义.方法 采用实时荧光定量聚合酶链反应(qRT-PCR)和Western blotting方法检测96例新鲜结直肠癌组织、对应的癌旁10 cm正常组织以及72例腺瘤中的Runx3和CHD5基因的表达,分析其与结直肠癌临床病理特征的关系以及诊断价值和预后关系.结果Runx3和CHD5 mRNA、蛋白在结直肠癌组织中的相对表达量为0.35 ±0.00、0.28 ±0.02和0.26±0.02、0.31±0.01,显著低于癌旁正常组织中的相对表达量0.95 ±0.02、0.92 ±0.02和0.89 ±0.03、0.93 ±0.02(t=2.36,P＜0.05;t 1.25,P＜0.05;t=1.37,P＜0.05;t =1.13,P＜0.05)以及腺瘤中的相对表达量0.89±0.02、0.90 ±0.02和0.85±0.02、0.87 ±0.04(t =2.27,P ＜0.05;t =2.16,P＜0.05;t=1.25,P ＜0.05;t =2.65,P＜0.05).Runx3和CHD5的表达与结直肠癌TNM分期(x2=4.65,P=0.031;x2=7.89,P=0.005)、浸润深度(x2=4.17,P=0.041;x2 =4.86,P =0.028)、淋巴结转移(x2=4.20,P=0.040;x2=7.02,P=0.008)以及分化程度(x2=7.31,P=0.036;x2=9.54,P=0.023)呈负相关.线性相关性分析显示两基因表达呈正相关(r =0.572,P=0.001).受试者特征(ROC)曲线进行评价表明Runx3、CHD5在结直肠癌中具有诊断价值(曲线下面积分别为0.712、0.745,敏感性和特异性分别是45.9％、52.5％和83.6％、81.4％).Runx3和CHD5均低表达组与其他患者组在总生存时间(x2=8.156,P＜0.05)及无进展生存时间(x2=6.325,P＜0.05)上的差异具有统计学意义.结论 Runx3和CHD5在结直肠癌中低表达,两基因共同检测可以作为结直肠癌诊断靶标以及预后指标.     

阿霉素大鼠肾病模型足细胞线粒体体视学分析

目的探讨大鼠肾脏中足细胞线粒体形态和数目与阿霉素大鼠肾病模型蛋白尿的关系。方法清洁级雄性SD30只大鼠尾静脉分别注射阿霉素0．7mg／100g体重和生理盐水，分别在注射后2W（对照组3只，阿霉素组3只）、4w（对照组3只，阿霉素组6只）和6W（对照组8只，阿霉素组7只）处死大鼠，取肾皮质标本进行透射电镜观察，对线粒体形态和密度进行体视学分析。结果阿霉素组大鼠4W出现蛋白尿，持续至6W。对照组大鼠肾组织足细胞内线粒体多呈椭圆形，阿霉素组大鼠足细胞线粒体形态多样，大小不一。统计分析未发现阿霉素组和对照组大鼠足细胞线粒体面积、周长、形状因子和最大长宽比的差异。注射2W后蛋白尿出现前，阿霉素组大鼠肾组织足细胞内线粒体面数密度明显增多，与对照组比较差异具有统计学意义（0．17±0．00VS．0．14±0．01，t=6．173，P〈0．01），同时线粒体相对于细胞体的表面积密度有增加趋势（0．78±O．03VS．0．71±0．04，t=-2．526，P=0．065）。注射6w后，阿霉素组大鼠足细胞线粒体面数密度无显著变化，线粒体相对于足细胞胞浆的表面积密度显著减少（0．71±0．11VS．0．87±0．12，P=0．02）。结论线粒体多形性改变参与阿霉素大鼠肾病模型蛋白尿的发生，足细胞线粒体数量增多是阿霉素大鼠肾病模型蛋白尿发生过程中的早发事件，线粒体膜表面积密度下降参与足细胞损伤及阿霉素大鼠肾病的进展。     

The Effect of (E)-1-(4’-aminophenyl)-3-phenylprop-2-en-1-one on MicroRNA-18a,Dicer1, and MMP-9 Expressions against DMBA-Induced Breast Cancer

Background: Most of breast cancer patients are estrogen receptor alpha-positive and have high resistance and side effect of chemotherapeutic drug. Therefore, discovering an effective anticancer agent is needed. This research explored the effect of (E)-1-(4’-aminophenyl)-3-phenylprop-2-en-1-one (APE) on miR-18a, Dicer1, and MMP-9 expressions. Methods: Twenty four female Sprague-Dawley rats were invetigated in this study. The rats were divided into 6 groups of 4. G1 was considered as normal rat. G2, G3, T1, T2, and T3 were given DMBA 20 mg/kgBW twice a week for 5 weeks to induce mammary cancer. After being affiliated with cancer, G2 was given vehicle and G3 was treated with tamoxifen. T1, T2, and T3 were treated with APE intraperitoneally everyday for 21 days at doses of 5, 15, and 45 mg/kgBW/day, respectively. Blood plasma was collected to measure miR-18a expression using qRT-PCR. Mammary tissues were also collected to determine Dicer1 and MMP-9 expressions by using  immunohistochemistry. Results: The results showed significant down-regulation of miR-18a relative expression and up-regulation of Dicer1 expression in G3 and T1 compared to G2 (P<0.05). MMP-9 expression has significant decrease in T1 compared to G2 (P<0.05). Conclusion: APE can decrease miR-18a and MMP-9 expressions and increase Dicer1 expression in rat mammary cancer. Therefore, this compound could be a candidate of novel anticancer.     

围手术期营养支持对结直肠癌患者肿瘤细胞cyclin D1表达及复发转移的影响

目的 探讨结直肠癌患者围术期营养支持后对肿瘤细胞cyclinD1表达及复发转移的影响效果.方法 选择148例结直肠癌患者,按照患者是否进行围术期营养支持的意愿,将患者分为营养支持观察组和非营养支持对照组,观察组78例,对照组70例.围术期营养支持为我院配制的营养液,与术前一周及术后5~14 d对观察组患者进行全胃肠外营养,对照组患者进行常规的护理.采用SP免疫组织化学法检测cyclin D1的表达,用TUNEL方法检测细胞的凋亡,细胞核抗原免疫组织化学染色检测细胞增殖,对两组患者进行对比分析.结果 经围术期营养支持的患者肿瘤细胞cyclinD1的阳性表达率为(35.12±5.09)%,凋亡指数为(52.67±5.43)%,细胞增殖指数为(7.35±2.87)%;对照组的阳性表达率为(37.50±6.12)%,凋亡指数为(56.17±6.38)%,细胞增殖指数为(8.42±3.67)%;两组比较无统计学差异(P＞0.05).两组患者术后肿瘤的复发率分别为15.38%(12/78)、15.71%(11/70),两组比较无显著性差异(P＞0.05).结论 围术期是否对患者进行营养支持对结直肠癌患者肿瘤细胞cyclinD1的表达无显著的影响,同时不会促进肿瘤的复发和转移.     

应用MADB-106乳腺癌细胞构建大鼠转移性骨癌痛模型

目的应用MADB-106大鼠乳腺癌细胞构建转移性骨癌痛模型,通过对该模型痛行为学、影像学、组织学观察等检测进行治疗研究。方法将MADB-106大鼠乳腺癌细胞接种到SD大鼠胫骨上段骨髓腔内,构建骨癌痛动物模型。以大鼠自发性缩足反射次数评估大鼠自发性痛情况,以自由活动时下肢跛行程度评分观察大鼠行走诱发痛,以热痛觉阈值观察大鼠热痛觉过敏情况,以X线、骨组织病理学检测观察癌细胞对骨结构的破坏程度。结果造模组大鼠术后第15天自发缩足反射次数为（14．42±1．24）次,第22天为（18．33±1．37）次,第25天为（21．25±1．54）次,与术前比较差异有统计学意义（均P〈0．001）;术后第15天热刺激痛觉阈值为（11．86±1．63）S,第22天为（8．38±1．05）S,第25天为（7．47±1．25）s（P〈0．001）;术后第15天自由行走痛评分为（1．25±0．62）分,第22天为（2．00±0．95）分,第25天为（2．33±1．07）分。术后第15天开始出现明显的自发性痛、热痛觉过敏和行走诱发痛,胫骨X线片显示明显的骨破坏,组织学研究显示骨髓腔内肿瘤生长,向外侵蚀破坏骨皮质。结论成功应用MADB-106乳腺癌细胞构建大鼠骨癌痛模型,造模2周后可以用于癌痛治疗的评价。     

肠癌小鼠高血糖下的血清胰岛素样生长因子-1变化及瘤体血管内皮生长因子表达

 目的　观察小鼠肠癌移植瘤在高血糖下的生长并检测瘤体血管内皮生长因子（VEGF）表达以及血清胰岛素样生长因子-1（IGF-1）的变化情况,探讨2型糖尿病（T2DM）是否为促进结直肠癌的发生与进展的危险因素。方法　建立肠癌移植瘤合并T2DM的小鼠模型,观察移植瘤体积大小变化,5周末处死小鼠检测血清IGF-1及瘤体组织VEGF表达。结果　肠癌糖尿病组小鼠瘤体体积[（1628.5±882）mm3]大于肠癌组小鼠瘤体体积[（1950.2±726）mm3]（P＜0.05）,其血清IGF-1[（105.33±32.32）ng／ml]高于正常组[（69.83±25.57）ng／ml]及肠癌组小鼠[（70.17±25.27）ng／ml]（P＜0.05）,瘤体组织VEGF表达[（70.0±11.5）%]强于肠癌组[（42.9±7.5）%]（P＜0.05）。结论　T2DM可能为促进肠癌生长的原因之一,其可能机制与血液中升高的IGF-1作用有关,并通过诱导VEGF基因转录,上调VEGF的表达,促使肿瘤组织血管生成,从而导致肿瘤的发生和转移有关。     

Prognostic value of tumor-infiltrating dendritic cells in colorectal cancer: role of maturation status and intratumoral localization.

The clinical significance of tumor-infiltrating dendritic cells has been reported in a variety of human solid tumors as shown by the correlations found between the presence of tumor-infiltrating dendritic cells and clinical prognosis. In this study, we evaluated whether there is an association between the presence and maturation status of tumor-infiltrating dendritic cells, T lymphocytes, and clinical course in 104 primary tumor samples of patients with colorectal cancer. Dendritic cells were identified with four different markers (S-100, HLA class II, CD208, and CD1a) in double immunohistochemistry, with laminin as second marker to support the exact localization. Tumor-infiltrating dendritic cells showed a distinct infiltration pattern based on their maturation status. CD1a-positive dendritic cells resided in the advancing tumor margins in relatively high numbers, whereas mature CD208-positive dendritic cells were sparsely present in the tumor epithelium but mainly distributed in the tumor stroma and advancing tumor margin. Furthermore, high infiltration of CD1a-positive dendritic cells in the tumor epithelium was significantly correlated to the infiltration of CD4 lymphocytes (P = 0.006). Patients with relatively high numbers of mature CD208-positive infiltrating dendritic cells in the tumor epithelium had a shorter overall survival (P = 0.004). In addition, patients with relatively high numbers of CD1a-positive dendritic cells in the advancing margin of the tumor had a shorter disease-free survival (P = 0.03). We found that tumor-infiltrating dendritic cells had preferential infiltration sites within a tumor, affected local tumor cell-immune cell interactions, and correlated to the clinical prognosis of colorectal cancer patients.     

PEG-5-FU-MAMS靶向治疗裸鼠大肠癌的实验研究

目的 研究聚乙二醇(polyethylene glycol,PEG)修饰的5-氟尿嘧啶磁性白蛋白微球(PEG-modified 5-FU magnetic albumin microsphere,PEG-5-FU-MAMS)和5-氟尿嘧啶磁性白蛋白微球(5-FU-MAMS)对大肠癌组织的被动靶向性,为实现大肠癌的主动靶向治疗,减少化疗药物的不良反应寻找新的途径.方法 30只人大肠癌裸鼠随机分为3组:(1)游离5-FU组;(2) 5-FU-MAMS组;(3) PEG-5-FU-MAMS组.每组10只,分别将3种不同的制剂(按5-FU 8mg/kg)经尾静脉给药,30分钟后,经眼眶采血,处死裸鼠,用HPLC法测定血清、肿瘤、肾脏、肝脏组织5-FU的浓度.结果 (1)PEG-5-FU-MAMS组、5-FU-MAMS组和游离5-FU组肿瘤组织中的药物浓度分别为(51.2±2.1) mg/L、(33.1 ±8.2)mg/L和(20.3±1.9) mg/L,PEG-5-FU-MAMS组肿瘤组织中药物浓度明显高于5-FU-MAMS和游离5-FU组(P＜0.01);而在血清中药物浓度相反,PEG-5-FU-MAMS组[(1.7±0.5)mg/L]明显低于5-FU-MAMS[(6.8±0.2)mg/L]和游离5-FU组[(7.1±0.8)mg/L] (P ＜0.01);PEG-5-FU-MAMS组肝、肾脏中药物浓度[(22.7±2.4) mg/L和(21.1±2.3)mg/L]明显低于5-FU-MAMS[(44.3±6.7)mg/L和(38.2±4.9) mg/L]和游离5-FU组[(45.9 ±7.8)mg/L和(39.7±3.2)mg/L] (P ＜0.05).(2)PEG-5-FU-MAMS对裸鼠大肠癌的抑瘤率明显高于游离5-FU和5-FU-MAMS(45.3％ vs 15.0％、30.1％,P＜O.05).结论 PEG-5-FU-MAMS对大肠癌组织的靶向作用明显强于5-FU-MAMS和游离5-FU,但肝、肾脏的被动靶向作用明显减弱,有效地减轻药物对肝、肾脏的不良反应.PEG-5-FU-MAMS的抗结直肠癌作用明显强于游离5-FU和5-FU-MAMS.PEG-5-FU-MAMS有望成为肿瘤主动靶向治疗的有效药物.     

外周血黏附分子CD24在结直肠癌筛查中的作用

目的:探讨外周血CD24在结直肠癌及大肠息肉筛查中的作用。方法:应用流式细胞仪分别检测新发结直肠癌患者、大肠息肉患者及健康对照者外周血白细胞中的CD24表达情况。进一步根据病理类型将大肠息肉患者分为炎症性息肉、增生性息肉、腺瘤性息肉及腺瘤性息肉伴不典型增生四组,比较组间差异。结果:结直肠癌患者（15.56±5.28）、大肠息肉患者（12.05±3.43）外周血白细胞中CD24的荧光强度显著高于健康对照（9.42±1.48）（P＜0.01）。炎症性息肉（10.75±1.80）、增生性息肉（12.14±3.11）、腺瘤性息肉患者（10.77±2.67）外周血白细胞中CD24的荧光强度无统计学差异,腺瘤性息肉伴不典型增生组患者（17.08±5.12）外周血白细胞中CD24的荧光强度显著高于前三组（P＜0.01）。结论:非侵入性的外周血CD24表达水平的检测对于结直肠癌的筛查可能有一定的意义。