热疗与化疗对放射存活曲线影响的实验研究

目的：通过细胞学实验方法，观察加温与羟基喜树碱(HCPT)对人肺腺癌Anip973细胞放射存活曲线的影响，以期为提高肿瘤治疗效果提供更科学的方法。方法：采用细胞集落形成方法，得出加温与羟基喜树碱(浓度为0．5μg/mL)单独或联合与放射作用下的细胞存活曲线，并通过对各组曲线特征参数的分析来观察放射敏感性的变化。结果：单放、加温与放射、HCPT与放射、加温与HCPT并用放射各组的Do、Dq、N、SF2、β值逐渐减小，K、α、α/β值逐渐增大。结论：加温与羟基喜树碱均能增加Anip973细胞的放射敏感性，使细胞修复亚致死性损伤的能力减弱，直接杀伤效应增加，且HCPT作用强于加温，而二者联合则效果更佳。     

磁靶向热疗及其在肿瘤治疗中的应用

磁靶向热疗(MTH)是当前治疗恶性肿瘤的一种新手段,通过磁性材料与生物材料的耦联以及测温和控温技术实现了磁性材料靶向性治疗肿瘤的策略,主要包括动脉栓塞热疗、直接注射热疗和细胞内热疗等.MTH已在多种肿瘤动物模型中进行了抗肿瘤的实验治疗,在临床试验中也取得了一定疗效.     

原发性肝癌肿瘤血管密度及其表达的临床病理意义

目的研究肿瘤微血管密度（ＭＶＤ）和肿瘤血管生长因子（ＶＥＧＦ）的表达水平在原发性肝癌中的临床病理意义。方法对６３例手术切除的小肝癌（直径＜５ｃｍ）患者的临床病理资料进行了回顾性的研究。其中２年内转移复发组２９例,２年内无转移复发组３４例。每例取３张连续的切片分别进行ＨＥ染色、ＶＥＧＦ和生物素标记的荆豆凝集素Ｉ（Ｂｉｏ－ＵＥＡ－Ｉ）的免疫组化染色。根据Ｂｉｏ－ＵＥＡ－Ｉ染色的血管内皮细胞计数来测定ＭＶＤ。结果转移复发组和无转移复发组的ＭＶＤ分别为４９．６±２９．７、２２．７±２８．２（Ｐ＜０．０１）;转移复发组和无转移复发组的ＶＥＧＦ阳性率分别为８６．２％（２５／２９）、４７．１％（１６／３４）,两者差异有显著性（Ｐ＜０．０１）。两组在肿瘤分期、有无卫星灶、有无门脉癌栓３方面差异也具有显著性（Ｐ＜０．０５,Ｐ＜０．０５,Ｐ＜０．０１）。结论除肿瘤分期、卫星灶、门脉癌栓具有预后意义外,肝癌组织中ＭＶＤ和ＶＥＧＦ的表达也具有预后价值     

乳腺癌组织淋巴管内皮细胞生长因子表达及其意义

目的：研究人乳腺癌组织中淋巴管内皮细胞生长因子的表达及临床意义。方法：乳腺癌患者手术标本86例，采用蛋白质印迹法检测VEGF-B、VEGF-C、VEGF-D和flt-4表达含量。结果：86例乳腺癌患者中VEGF-B高表达44例(51．2％)，低表达42例(48．8％)；VEGF-C高表达46例(53．5％)，低表达40例(46．5％)；VEGF-D高表达43例(50．0％)，低表达43例(50．0%)；flt-4高表达48例(55．8％)，低表达38例(44．2％)。乳腺癌组织中VEGF-C、VEGF-D与flt-4蛋白的表达呈正相关。VEGF-C、VEGF-D、flt-4蛋白的表达与乳腺癌的淋巴结转移有关，但与乳腺癌组织病理学分型、肿瘤大小和分级无关。VEGF-C、VEGF-D高表达组总生存率较差。结论：VEGF-C、VEGF-D和flt-4蛋白的表达与乳腺癌淋巴转移有关，VEGF-C、VEGF-D可作为提示临床预后较差的指标。     

血管内皮生长因子在肿瘤中的表达及其临床意义

肿瘤的生长,浸润和转移依赖于肿瘤血管生成,血管内皮生长因子（VEGF）是最重要的一种血管生成刺激因子,VEGF特异性作用于血管内皮细胞,通过促进内皮细胞增殖,增加血管通透性,以诱导肿瘤血管生成,有肿瘤的发生发展中起关键作用。因而EGF在肿瘤的恶性程度和预后烦闷以及治疗方面有重要的临床应用价值。     

肿瘤多次热疗临床参数与疗效的相关性研究

单次热疗对肿瘤有一定疗效,但是容易肿瘤复发,多次热疗可以很好地解决这一问题.多次热疗不仅通过热来杀火肿瘤细胞,而且还可诱发机体的抗肿瘤免疫从而进一步杀灭肿瘤细胞.临床上大多数学者把热疗的间隔时间定在每周1-2次来解决多次热疗的热耐受问题.     

Artemin在软骨肉瘤中的表达及其对内皮细胞增殖和迁移的影响

目的：探讨Artemin在软骨肉瘤中的表达水平及其对内皮细胞增殖与迁移的作用及其机制。方法：收集2015年5月至2019年4月南京中医药大学连云港附属医院手术切除的40例软骨肉瘤患者的肿瘤组织标本,分为低（Ⅰ级）、高级别（Ⅱ、Ⅲ级）软骨肉瘤各20例;对照组为20例因车祸等意外伤截肢患者的正常软骨组织标本。用免疫组化法检测肿瘤组织中Artemin、血管内皮生细胞长因子（vascular endothelial growth factor,VEGF）、Ki-67表达水平和CD31+血管密度。用10 ng/ml的Artemin处理SW1353细胞后,用ELISA实验检测细胞培养上清中VEGF、基质细胞衍生因子-1（stromal cell derived factor-1,SDF-1）、基质金属蛋白酶2（matric metalloproteinase 2,MMP2）和MMP9的含量变化,用Transwell迁移实验和MTT细胞增殖实验分别检测Artemin处理的软骨肉瘤细胞对ECV304细胞增殖和迁移的影响。结果：低级别组软骨肉瘤组织中Artemin和Ki-67表达水平均显著高于对照组（均P<0.01）,高级别组软骨肉瘤组织中Artemin表达水平和Ki-67表达率显著高于低级别组（均P<0.01）。Artemin表达与软骨肉瘤组织中VEGF水平和血管密度呈正相关（均P<0.01）;Artemin促进软骨肉瘤细胞分泌VEGF,而对SDF-1、MMP2和MMP9分泌无显著的影响;Artemin通过促进软骨肉瘤细胞分泌VEGF,诱导ECV304细胞增殖和迁移（均P<0.01）。结论：Artemin在软骨肉瘤组织中高表达且与VEGF水平和血管密度呈正相关,Artemin能够增强软骨肉瘤细胞诱导血管生成功能。     

热疗与化疗对放射存活曲线影响的实验研究

目的 :通过细胞学实验方法 ,观察加温与羟基喜树碱 (HCPT)对人肺腺癌Anip973细胞放射存活曲线的影响 ,以期为提高肿瘤治疗效果提供更科学的方法。方法 :采用细胞集落形成方法 ,得出加温与羟基喜树碱 (浓度为 0 5 μg mL)单独或联合与放射作用下的细胞存活曲线 ,并通过对各组曲线特征参数的分析来观察放射敏感性的变化。结果 :单放、加温与放射、HCPT与放射、加温与HCPT并用放射各组的Do、Dq、N、SF2 、β值逐渐减小 ,K、α、α β值逐渐增大。结论 :加温与羟基喜树碱均能增加Anip973细胞的放射敏感性 ,使细胞修复亚致死性损伤的能力减弱 ,直接杀伤效应增加 ,且HCPT作用强于加温 ,而二者联合则效果更佳     

Caveolin-1对内皮细胞增殖的抑制作用研究

目的：研究Caveolin-1蛋白在内皮细胞增殖中的作用,其与Ras-p42/44MAPK细胞信号通路的关系.方法：以腺病毒介导在内皮细胞中表达外源Caveolin-1蛋白,分析其对内皮细胞增殖和p42/44MAPK分子磷酸化水平的影响.结果：感染腺病毒表达外源Caveolin-1蛋白的内皮细胞中,血管内皮生长因子（VEGF）诱导的3H-胸腺嘧啶掺入量降低了35%（n=3,P=0.009）,p42/44MAPK分子磷酸化水平降低了47.6%.结论：Caveolin-1蛋白抑制p42/44MAPK分子磷酸化水平,抑制内皮细胞增殖.     

血管内皮生长因子受体在骨巨细胞瘤中的表达及其与肿瘤增殖和血管生成关系的研究

目的 探讨血管内皮生长因子受体(KDR)在骨巨细胞瘤中的表达情况,及其与肿瘤血管生成和增殖状态的关系,并分析各种因素在骨GCT预后中的作用。方法 对58例患者之石蜡切片采用SP法检测VEGF/KDR、CD34及MIB-1的表达,同时收集临床及随访资料。分析VEGF、KDR与微血管密度(MVD)、增殖指数(PI)、临床因素及预后的关系。结果(1)骨GCT中新生血管主要位于肿瘤浸润边缘。VEGF阳性率为53.6％,KDR阳性率高达96.4％。(2)VEGF阳性组和KDR高表达组MVD均值分别高于VEGF阴性组和KDR低表达组(P<0.05)。VEGF/KDR与MVD呈正相关。KDR高表达组PI值高于KDR低表达组(P<0.05),二者呈正相关。VEGF与PI无显著相关性。(3)VEGF/KDR表达及MVD与PI与各临床因素无关(P>0.05)。(4)多因素logistic回归分析表明MVD、KDR以及手术方式与预后有关。KDR、MVD是影响其复发的危险因素。结论 VEGF及其受体与骨GCT的增殖和血管生成密切相关。VEGF能通过KDR以旁分泌或自分泌形式作用,间接或直接刺激肿瘤细胞增殖。MVD、KDR以及手术方式与预后有关,可作为判断患者预后的指标,并为骨GCT的抗血管生成治疗提供了实验依据。     

Crosstalk between H1975 tumor cells and platelets to induce the proliferation,migration and tube formation of vascular endothelial cells

Activated platelets (PLTs) participate in the regulation of tumor angiogenesis, and tumors can activate PLTs. Whether co-culture of lung carcinoma with PLTs improves the function of human umbilical vein endothelial cells (HUVECs) requires further investigation. The present study aimed to investigate the impact of H1975 cell crosstalk with PLTs on the proliferation, migration and tube formation of HUVECs. Following generation of cell-derived supernatants and construction of the co-culture system, Cell Counting Kit-8, flow cytometry, transmission electron microscopy and a meter for epithelial measurement were performed to detect the proliferative ability of HUVECs. Furthermore, the wound healing and Transwell migration assays were performed to detect the migratory ability of HUVECs. A tube formation assay was performed to assess angiogenesis, ELISA was applied to detect the content of vascular endothelial growth factor (VEGF) and western blotting was carried out to measure the expression levels of VEGF receptor 2 (VEGFR2) in HUVECs. Compared with single-cultured HUVECs (control), co-culture with H1975 cells and PLTs (Exp_HP) improved cell proliferation, increased the proportion of cells in the S-phase, destroyed the cell ultrastructure and decreased transepithelial electrical resistance in HUVECs. In addition, a higher relative migration rate, greater number of migrated cells, stronger tube-forming ability and increased expression of VEGF and VEGFR2 were detected in the Exp_HP group compared with the control group. The properties of HUVECs in Exp_H (co-cultured with H1975 cells) were similar to those in Exp_HP, but significantly weaker. Taken together, the results of the present study suggest that tumor cells interacting with PLTs may play an important role in tumor angiogenesis by affecting or mediating changes in the properties of vascular endothelial cells (VECs).     

Effect of heat treatment on tumor cells and antitumor effector cells

We examined the effects of heat treatment on tumor cells and antitumor effector cells in order to investigate the combined effects of hyperthermia and immunotherapy including adoptive immunotherapy. Plasmacytoma MOPC104E syngeneic to BALB/c mice was used as the tumor cell line, while fresh spleen cells immunized to MOPC104E (IM-FSC) and interleukin-2-cultured lymphocytes induced in vitro from IM-FSC (CL) were used as the effector cells. Tumor cells or effector cells were heat-treated in a water bath at 42 degrees C for 30 or 60 min. Tumor cells heat-treated at 42 degrees C for 30 min grew temporarily and then regressed in the tumor transfer test, whereas untreated tumor cells showed no regression under any conditions. Furthermore, fresh spleen cells of mice inoculated with heat-treated tumor cells from regressed tumors showed marked tumor-neutralizing activity. The antitumor effects of CL were markedly inhibited by heat treatment according to the results of the tumor-neutralizing test and the 51Cr release assay, whereas heat treatment had little influence on the antitumor activity of IM-FSC. However, the neutralizing activity of effectors and the killing activity of CL against heat-treated tumor cells were both markedly augmented, since the susceptibility of the tumor cells to the antitumor effector cells was augmented by heat treatment. These results suggest that heat treatment of tumor cells augments the antitumor effects of IM-FSC and CL, hence we speculate that hyperthermia augments the effects of immunotherapy including adoptive immunotherapy.     

姜黄素抑制HeLa细胞诱导的血管内皮细胞增殖作用及其机制研究

目的:研究姜黄素对HeLa细胞诱导的血管内皮细胞增殖的抑制作用及其机制.方法:采用台盼蓝排染法计数人脐静脉内皮细胞系ECV304细胞,MTT法测定不同浓度姜黄素对ECV304细胞和HeLa细胞诱导的ECV304细胞增殖的抑制,采用RT-PCR和Western blot检测40 μmol/L姜黄素作用于HeLa细胞诱导的血管内皮细胞4、8、16和24 h后,血管内皮生长因子(VEGF)及其受体VEGFR2(KDR)基因及蛋白的表达.结果:姜黄素直接作用于ECV304细胞,其细胞计数较对照组显著减少(P<0.05),并呈浓度依赖性.MTT法测得不同浓度姜黄素作用后,可产生显著的细胞增殖抑制作用,48 h体外半数抑制浓度(IC50)值为(38.4±0.031) μmol/L.随着姜黄素浓度的增加,其时ECV304细胞和HeLa细胞诱导的ECV304细胞的增殖有明显的抑制作用,P值分别为0.000 0和0.000 1.姜黄素作用于HeLa细胞诱导的血管内皮细胞4、8、16和24 h后,能明显下调VEGF与KDR的mRNA和蛋白表达.结论:姜黄素能直接抑制血管内皮细胞的增殖,对HeLa细胞诱导的血管内皮细胞的增殖也有抑制作用.此作用可能是通过押制VEGF及其受体KDR的mRNA和蛋白表达来实现的.     

Effect of hyperthermia, radiation and adriamycin combinations on tumor vascular function

Pathophysiologic studies of tumor vascular responses to hyperthermia, radiation or adriamycin given alone or in specific combinations have been made in the cervical carcinoma grown in the transparent cheek pouch chamber of the Syrian hamster. A specially designed chamber containing a compartment for flowing water enabled controlled heating of the tumor and pouch to within 0.2 degrees C; the desired temperatures were achieved within one minute. Heating at 42 degrees C for 30 minutes was followed, at 1, 5 or 24 hours, by a second heating for 30 minutes at 42 degrees C. In addition, the same period of heating was preceded or followed, at 1, 5 or 24 hour intervals, by a single exposure to 2000R or a single intravenous injection of adriamycin given at a rate of 0.45mg/100gm body weight. Of the three modalities, heat appeared to have the greatest acute effect on the tumor vascular system. A single dose of heat produced a rapid but transient constriction followed by a prominent dilation of vessels. Two heating periods given at a 1 hour interval caused persistent stasis in the tumor which progressed to coagulation necrosis. Although heating prior to irradiation or adriamycin, in general, increased the vascular responses to these agents, this sequence gave no tumor control. Radiation or adriamycin given prior to heating had relatively little effect on the vascular response to heating and produced no tumor control except when heat was applied shortly after irradiation. These studies indicate that changes in the microvasculature and perfusion in tumors, in response to hyperthermia alone or combined in specific sequences with radiation, can alter the internal environment of the tumor to produce a greater degree of tumor control than can be attributed to direct cell killing by these agents.     

抗KDR胞外Ⅲ区单抗抑制血管内皮细胞增殖的研究

目的研制能够封闭血管内皮生长因子(VEGF)受体KDR的单克隆抗体,探讨其体外抑制VEGF165诱导的生物学活性。方法以原核表达的KDR胞外Ⅲ区融合蛋白免疫Balb／c小鼠,采用传统杂交瘤技术制备抗KDR胞外Ⅲ区单克隆抗体,采用ELISA和FACS方法鉴定其抗原结合特异性,采用免疫沉淀和[^3H]-TdR掺入的方法分析该单克隆抗体阻断VEGF165刺激内皮细胞表面KDR酪氨酸激酶受体磷酸化,抑制VEGF165岱诱导内皮细胞增殖的活性。结果抗KDR胞外Ⅲ区单抗Ycom1D3不仅能与可溶性KDR结合,亦可与细胞表面表达的KDR结合,并可竞争性抑制VEGF165与可溶性KDR的相互作用,阻断VEGF165刺激内皮细胞表面KDR酪氨酸激酶受体磷酸化,显著抑制由VEGF165诱导脐静脉内皮细胞的增殖。结论抗KDR胞外Ⅲ区单抗Ycom1D3可通过封闭KDR而抑制VEGF活性,在肿瘤及其他血管新生疾病治疗中具有潜在的应用前景。     

氧化苦参碱对肿瘤诱导血管内皮细胞增殖的抑制作用

目的 探讨氧化苦参碱（Ｏｘｙ）对肺癌和胃癌细胞诱导血管内皮细胞（ＶＥＣ）增殖的抑制作用。方法 用ＭＴＴ法检测不同浓度Ｏｘｙ对ＶＥＣ、人肺腺癌ＳＰＣ－Ａ－１细胞、人低分化胃癌ＭＫＮ－４５细胞增殖及ＳＰＣ－Ａ－１和ＭＫＮ－４５细胞诱导ＶＥＣ增殖的抑制。结果 Ｏｘｙ浓度为２．５～１０ｍｇ／ｍｌ时,对ＶＥＣ增殖的抑制率为４．６％～３６．４％;Ｏｘｙ浓度为０．１５６～１０ｍｇ／ｍｌ时,对肺癌ＳＰＣ－Ａ－１和     

Inhibition of human vascular endothelial cells proliferation by terbinafine

We have demonstrated previously that terbinafine (TB), an oral antifungal agent used in the treatment of superficial mycosis, suppresses proliferation of various cultured human cancer cells in vitro and in vivo by inhibiting DNA synthesis and activating apoptosis. In our study, we further demonstrated that TB at a range of concentrations (0-120 microM) dose-dependently decreased cell number in cultured human umbilical vascular endothelial cells (HUVEC). Terbinafine was not cytotoxic at a concentration of 120 microM, indicating that it may have an inhibitory effect on the cell proliferation in HUVEC. The TB-induced inhibition of cell growth rate is reversible. [(3)H]thymidine incorporation revealed that TB reduced the [(3)H]thymidine incorporation into HUVEC during the S-phase of the cell-cycle. Western blot analysis demonstrated that the protein levels of cyclin A, but not cyclins B, D1, D3, E, CDK2 and CDK4, decreased after TB treatment. The TB-induced cell-cycle arrest in HUVEC occurred when the cyclin-dependent kinase 2 (CDK2) activity was inhibited just as the protein level of p21 was increased and cyclin A was decreased. Pretreatment of HUVEC with a p21 specific antisense oligonucleotide reversed the TB-induced inhibition of [(3)H]thymidine incorporation. Taken together, these results suggest an involvement of the p21-associated signaling pathway in the TB-induced antiproliferation in HUVEC. Capillary-like tube formation and chick embryo chorioallantoic membrane (CAM) assays further demonstrated the anti-angiogenic effect of TB. These findings demonstrate for the first time that TB can inhibit the angiogenesis.     

髓母细胞瘤的CT诊断(附50例分析)

 目前CT在诊断髓母细胞瘤方面虽有经验，但也经常遇到一些非典型CT表现的特殊病例误诊为其他颅内肿瘤。为进一步提高髓母细胞瘤的CT诊断水平，对经CT检查，并经手术病理证实的50例髓母细胞瘤总结如下。     

Rab31对神经胶质瘤细胞增殖迁移侵袭的影响及机制研究

目的:研究Rab31对人神经胶质瘤细胞增殖迁移侵袭的影响及相关分子机制.方法:Western blot检测正常人胶质细胞NHA和3株人神经胶质瘤细胞SHG-44、TJ905和LN229中Rab31表达;将3种Rab31沉默siRNA及其对照分别转染至神经胶质瘤SHG-44细胞中,分别记为si-Rab31-1组、si-R...