金雀异黄素对不同淋巴转移能力的小鼠肝癌细胞株的抑制作用

MTT法检测大豆提取物金雀异黄素（genistein,Gen）对不同淋巴转移能力的腹水型小鼠肝癌细胞株HepA-H和HepA-L的生长抑制作用。应用电镜观察细胞形态和流式细胞术检测细胞凋亡。将HepA-H和HepA-L接种NIH小鼠,制备荷瘤动物模型,Gen腹腔给药后第14天取淋巴结观察,并计算转移率。TUNEL法检测淋巴结凋亡细胞,并计算凋亡指数。实验发现：（1）Gen对两株细胞均具有良好的生长抑制作用,且对HepA-H的抑制优于HepA-L;（2）Gen在体外诱导两株细胞发生凋亡,且HepA-H的凋亡率高于HepA-L,并呈量效关系;（3）Gen抑制荷瘤动物肿瘤生长和肿瘤淋巴转移,且对HepA-H的抑制强于HepA-L,实验组H的凋亡指数（3．87％）也显著高于实验组L（1．69％）。故研究认为Gen显著抑制HepA-H和HepA-L肝癌细胞淋巴转移的作用机制,可能与诱导细胞凋亡有关,对高淋巴转移细胞的抑制作用强于低淋巴转移细胞,提示Gen抗肿瘤作用具有一定的选择性。     

中国人胃癌染色体17q21区域基因遗传不稳定性研究

To investigate microsatellite instability (MSI) and loss of heterozygosity (LOH) of locus D17S396, D17S579 and D17S855, and their effect on the expression of nm23H1 and BRCA1 of gastric cancer, which would provide experimental basis for clinical treatment and prognosis analysis of gastric cancer. DNA was extracted from paraffin-embedded materials. Polymerase chain reaction-single strand conformation polymorphism (PCR-SSCP) was used to analyze MSI and LOH. Expression of nm23H, and BRCA1 was detected by Envision immuno-histochemistry and Leica-Qwin computer imaging techniques. In the forty cases of gastric cancer, the frequency of MSI, LOH and nm23H1 protein were 20.00%, 17.50% and 55.00% respectively at locus D17S396, while at locus D17S579, the frequency of MSI, LOH and BRCA1 protein were 22.50%, 15.00% and 37.50% respectively; at locus D17S855, the frequency of MSI, LOH and BRCA1 of thirty-seven cases were 18.92%, 18.92%, 37.84% respectively. In tumor node metastasis (TNM) staging, at locus D17S396, D17S579 and D17S855, MSI in stages I + II appeared more frequently than that in stages III + IV, while LOH appeared the contrary tendency. In the group of metastasis of gastric cancer, MSI had a less frequency (5.00%) than that with no metastasis (35.00%, P < 0.05) at locus D17S396, but LOH appeared more frequently (30.00%) than that with no metastasis (5.00%, P < 0.05). At locus D17S579, MSI had an increasing tendency with the degree of tumor differentiation (50.00% in high differentiation cases, 20.00% in middle differentiation cases, and 0% in low differentiation cases, P < 0.05). The frequency of nm23H1 and BRCA1 protein in stages TNM I + II was higher than that in stages TNM III + IV; and that in higher differentiation cases was higher than in poor differentiation cases. The frequency of nm23H1 protein in the group of metastasis (30.00%) was less than that with no metastasis significantly (80.00%, P<0.01). The frequency of nm23H1 protein in the group positive to MSI (87.50%) was higher than that in the group negative to MSI (46.88%, P < 0.05). However, nm23H1 protein in group positive to LOH (14.29%) was lower than that in the group negative to LOH (63.64%, P < 0.05). The frequency of BRCA1 protein in the group positive to MSI (66.67%) was more than that in the group negative to MSI (29.03%, P < 0.05). The results of experiments indicate that MSI and LOH may separately control the development of sporadic colon cancer with different pathways. MSI may be an early period molecule marker for sporadic colon cancer, enhanced expression of nm23H1 protein can effectively inhibit colon cancer metastasis and improve prognosis of sporadic colon cancer patients. By comparison, LOH mostly arises in the late period of sporadic colon cancer and endows a high aggressive and poor prognostic phenotype. nm23H1 protein could effectively restrain gastric cancer metastasis and development; and BRCA1 protein could restain tumor from becoming lower differentiation.     

自身抗原，动力之源

众所周知,辅助T细胞在淋巴结中识别树突细胞或B细胞表面的MHCⅡ分子呈递的抗原而被激活,从而启动免疫应答的后续步骤。但是MHCⅡ分子所呈递的绝大部分是自身抗原,大多数时候辅助T细胞只能遇到这样的MHCⅡ分子而不能激活。那么自身抗原在免疫应答中有什么意义呢？最近的一项研究中,Fischer及其同事首先报道了自身抗原在免疫应答中的作用。     

Ⅰ期肠切除吻合术治疗急性梗阻性结肠癌的临床效果分析

目的:探讨Ⅰ期肠切除吻合术处理急性梗阻性结肠癌的临床效果,为普外科手术提供参考。方法:对我院2011年4月-2013年11月实施Ⅰ期肠切除吻合术的59例急性梗阻性结肠癌患者的临床资料进行分析,并与择期接受结肠癌根治术的42例患者对比。观察两组患者的平均术中出血量、手术时间、切口长度、切除肿瘤大小、清扫淋巴结数、术后肛门排气、并发症发生率、住院时间及肿瘤复发率等。结果:与根治切除术组比较,Ⅰ期肠切除吻合术组患者的平均手术时间及术中出血量无显著性差异(P0.05),但肿瘤切除率及淋巴结清扫情况优于根治切除术组,且手术切口小,组间比较差异具有统计学意义(P0.05)。Ⅰ期肠切除吻合术组患者术后肛门排气时间和下床活动时间早于根治切除术组患者(P0.05);Ⅰ期肠切除吻合术组患者术后并发症的发生率、肿瘤复发转移率均明显低于根治切除术组患者,差异具有统计学意义(P0.05);两组患者术后两年的生存率无显著差异(P0.05)。结论:Ⅰ期肠切除吻合术用于急诊处理急性梗阻性结肠癌具有良好的手术效果,并发症的发生率低,有利于改善患者术后的生存质量。     

氨肽酶N在结直肠癌中的表达及其临床意义

目的:检测结直肠癌中氨肽酶N(APN)的表达,并通过分析其与血管内皮生长因子(VEGF)和碱性成纤维细胞生长因子(bFGF)的关系,探讨APN在结直肠癌发生、发展过程中的作用及其临床意义。方法:应用免疫组织化学技术,检测40例结直肠癌组织及40例癌旁正常直肠组织中APN、VEGF和bFGF的表达,并结合临床病例特征进行分析。结果:40例结肠癌组织中APN的阳性率达52.5%(21/40),15例转移淋巴组织中达66.67%(10/15),在结直肠癌组织和转移淋巴结组织中的阳性表达率均明显高于癌旁正常直肠组织,差异有统计学意义(P0.05)。APN、VEGF与bFGF在结直肠癌中的阳性表达均与患者的性别、年龄、肿瘤大小、肿瘤位置、分化程度无关(P0.05),而与肿瘤的分期和有无转移显著相关(P0.05)。结直肠癌组织中APN与VEGF、bFGF的表达间均呈显著正相关性(P0.05)。结论:APN在结直肠癌组织中呈高表达,与VEGF、bFGF可能具有协同作用,共同参与调节结直肠癌的发展,对于评估结直肠癌的生物学行为和预测患者的预后可能具有重要价值。     

N-三甲基壳聚糖包裹喜树碱治疗卵巢癌淋巴结转移的研究

目的:采用N-三甲基壳聚糖包裹喜树碱(CPT-TMC)以增加其水溶性,并检测其抑制卵巢癌细胞淋巴结转移的效应。方法:体外实验,PI染色检测CPT-TMC促肿瘤细胞凋亡作用,体内实验,荷瘤裸鼠随机分为4组(5只/组):NS组、TMC组、CPT组及CPT-TMC组,通过尾静脉给药,每周2次,持续3周,Evan's Blue Dye染色,显影腹腔肿大转移的淋巴结。结果:PI染色提示CPT-TMC显著促进肿瘤细胞凋亡(P0.05);Evan's Blue Dye显影显示CPT-TMC显著抑制肿瘤细胞淋巴结转移(P0.05)。结论:CPT-TMC具有明显抑制卵巢癌细胞淋巴结转移效应,它可能成为一种新的有效的卵巢癌治疗药物。     

LLNA：BrdU-ELISA 改良法的建立及其在化妆品安全性评价中的应用

目的：建立可快速检测化学物致敏性和刺激性的局部淋巴结试验（ LLNA：BrdU-ELISA）改良法,并对化妆品产品进行评价。方法3种化学物（2,4二硝基氯苯（ DNCB）、丁子香酚、己基肉桂醛）和3种化妆品产品作为受试物,雌性BALB／c小鼠连续染毒3天,测量小鼠耳缘厚度,第5天腹腔注射BrdU,第6天称耳廓重,分离颌下淋巴结称重并制取单细胞悬液,用ELISA试剂盒检测淋巴细胞增殖。结果 DNCB（1.0％）、己基肉桂醛（25％、50％）和3号粉底霜引起耳肿胀和耳廓重显著增加（P ＜0.05或P ＜0.01）,可能为刺激物,其他均无刺激性;3种化学物和3号粉底霜致敏检测阳性,其他均为阴性。结论结合小鼠耳肿胀和耳廓重的LLNA：BrdU-ELISA改良法可较好地评价化学物和化妆品产品的致敏性和刺激性,有望在化妆品安全性评价中发挥重要作用。     

人EGFR显性负性突变体对胃癌细胞体内成瘤及淋巴结转移的影响

为了研究人表皮生长因子显性负性突变体(dominant negative mutant of EGFR, dnEGFR)对胃癌细胞体内成瘤及淋巴结转移的影响,用目的质粒pEGFPN1 dnEGFR,空质粒载体pEGFP N1分别转染胃癌细胞NCI-N87.筛选出稳定转染株,实验共分3组：NCI-N87细胞未转染组（untreated NCI-N87,UN组）,NCI-N87细胞pEGFP-N1转染组（EN组）;NCI-N87细胞pEGFPN1-dnEGFR转染组（DN组）.将3组细胞接种于裸鼠右后足垫,6周后测量移植瘤大小及对应腹股沟转移淋巴结数目,HE染色验证,real-time PCR和Western 印迹检测3组细胞中AKT1、MAPK3 mRNA和蛋白的表达改变.结果发现,DN组移植瘤较UN、EN组明显缩小（P<0.05）,且右腹股沟转移淋巴结数目较UN、EN组减少（P<0.05）.DN组细胞中,AKT1、MAPK3 mRNA和蛋白水平较UN、EN组降低（P<0.05）.提示pEGFPN1-dnEGFR可抑制裸鼠体内胃癌细胞成瘤及淋巴结转移,AKT1及MAPK3信号通路可能参与其中.     

细胞外基质和细胞粘附分子表达与细支气管肺泡癌淋巴结转移的关系

用免疫组化方法检测50例细支气管肺泡癌（BAC）（其中18例伴有淋巴结转移）及其三个亚型粘液型、非粘液型、硬化型中基底膜细胞外基质（ECM）层粘连蛋白（LN）、Ⅳ型胶原（CollⅣ）、纤粘连蛋白（FN）分布和细胞粘附分子A－CAM（又称N－钙粘素）表达情况。结果发现粘液型、非粘液型、硬化型BAC的淋巴结转移率分别为57％（8／14）、32％（7／22）和21％（3／14）。在BAC及BAC各亚型中,三种ECM在基底膜水平的分布状态与淋巴结转移无关（P均＞0.05）、A－CAM在伴或不伴淋巴结转移的BAC原发瘤中的表达率分别为56％（10／18）和53％（17／32）（P＞0.05）。提示BAC三个亚型中粘液型淋巴结转移率有高于其它两亚型的倾向。基底膜ECM（LN、CollⅣ、FN）分布完整与否及细胞粘附分子A－CAM表达高低与BAC淋巴结转移无关。