参麦注射液联合环磷酰胺抑制血管生成作用的实验研究

目的:探讨参麦注射液联合应用环磷酰胺对体内外血管生成的抑制作用。方法:以人肝癌细胞HepGⅡ作为对照,分别采用MTT法、三维血管模型、鸡胚尿囊膜(CAM)模型观察参麦注射液联合环磷酰胺对体外人微血管内皮细胞(HMVEC)增殖、迁移和体内血管生成的影响。结果:40μl/ml参麦注射液和0.4mg/ml环磷酰胺对血管内皮细胞增殖的抑制率分别为18%、11.2%,联合应用为39.1%;而对HepG II增殖的抑制率仅分别为1.2%、3.6%,联合应用为11.8%;对血管内皮细胞有抑制出芽的作用,联合应用具有显著的协同效应。对鸡胚尿囊膜新生血管生成具有协同抑制作用。结论:小剂量参麦注射液(40μl/ml)、环磷酰胺(0.4mg/ml):在体内外具有抑制血管生成作用,而对肝癌细胞无明显抑制作用;在体外具有抑制血管增殖和迁移的协同作用和体内具有抑制血管生成协同作用。     

参麦注射液联合热疗抑制血管生成作用的体内外实验研究

目的探讨参麦注射液联合应用热疗对体内外血管生成的抑制作用。方法采用MTT法观察参麦注射液联合热疗对人微血管内皮细胞(HMVEC)、人肝癌细胞HepGⅡ增殖的影响。利用三维血管模型观察参麦注射液联合热疗对HMVEC迁移的影响。利用鸡胚绒毛尿囊膜(CAM)模型,观察参麦注射液对鸡胚绒毛尿囊膜血管生成的影响。结果参麦注射液在40μl/ml~160μl/ml浓度范围内,具有抑制血管内皮细胞增殖和迁移作用(内皮细胞存活率在82.0%~36.8%),均与浓度呈负相关(相关系数分别为-0.949、-0.535,P值分别为0.0012、0.0043);具有抑制鸡胚绒毛尿囊膜血管生成的作用;而在40μl/ml~80μl/ml浓度范围内时,对人肝癌HepGⅡ细胞表现出亚细胞毒作用(细胞存活率在98.8%~76.2%之间);当浓度≥160μl/ml时对人肝癌HepGⅡ细胞发挥细胞毒作用,(细胞存活率低于50%)。40μl/ml~160μl/ml的参麦注射液与热疗联合应用在体外具有抗血管生成的协同效应。结论小剂量参麦注射液(40μl/ml~160μl/ml)联合热疗在体内外均具有抑制血管增殖和迁移协同作用。     

羟喜树碱联合热疗对体内外血管生成的抑制作用

目的:探讨羟喜树碱联合应用热疗(thermotherapy)对体内外血管生成的抑制作用。方法:采用MTT法观察羟喜树碱联合热疗对人微血管内皮细胞(HMVEC)、人肝癌细胞HepGⅡ增殖的影响。利用三维血管模型观察羟基喜树碱联合热疗对HMVEC迁移的影响。利用鸡胚绒毛尿囊膜(CAM)模型,观察羟喜树碱对鸡胚绒毛尿囊膜血管生成的影响。结果:羟喜树碱在20~40ng/ml、5~80ng/ml浓度范围内,具有抑制血管内皮细胞增殖和迁移作用,内皮细胞存活率(68.2%~44.7%),均与浓度呈负相关(相关系数r分别为-0.906、-0.469,P值分别为0.00003、0.0051);具有抑制鸡胚绒毛尿囊膜血管生成的作用;而在5~80ng/ml浓度范围内,对人肝癌细胞无细胞毒作用(细胞存活率:100%~90%)。20ng/ml的羟喜树碱与热疗联合应用在体外具有抗血管生成的协同效应;40ng/ml的羟喜树碱与热疗联合应用在体外具有抗血管生成的次加效应。结论:小剂量羟喜树碱(20~40ng/ml)联合热疗在体内外均具有抑制血管增殖和迁移协同或次加作用。     

参麦注射液联合轻基喜树碱抗肿瘤血管生成的实验研究

目的 观察小剂量经基喜树碱联合参麦注射液对小鼠H22 细胞移植瘤的生长抑制作用及对肿瘤组织中微血管密度（MVD ）和血管内皮生长因子（VEGF）表达的影响。方法 建立小鼠皮下肝癌H22 模型,随机分为5组：生理盐水注射液20ml/kg 阴性对照组、环磷酞胺（CTX) 20mg/kg 阳性对照组、经基喜树碱0. 2mg/kg组、参麦注射液20ml/kg组,参麦注射液20ml/kg 联合经基喜树碱0 . 2mg/kg 组,每组10 只小鼠。连续腹腔给予参麦注射液、经基喜树碱以及两药联合治疗10 天,观察用药后肿瘤生长情况,计算抑瘤率,并用免疫组化方法检测肿瘤组织中VEGF 和MVD 的表达。结果 参麦注射液、经基喜树碱具有显著抑制H22 移植瘤的作用,且两药联合具有协同作用;免疫组化结果显示治疗组H22 瘤体内MVD、VEGF 表达均明显低于对照组。结论 小剂量经基喜树碱对H22 荷瘤小鼠移植瘤具有一定的生长抑制作用,联合参麦注射液具有协同作用。同时,两药联合可明显抑制肿瘤组织VEGF、CD34 的表达,推测其血管生成抑制作用亦参与抗肿瘤。     

奥沙利铂联合复方苦参注射液抗血管生成作用的实验研究

背景与目的:肿瘤的生长、转移、复发和预后均与肿瘤血管生成密切相关.以肿瘤血管生成为靶点,开发血管生成抑制剂在抗肿瘤研究中是一个十分活跃的研究领域.近年来,研究发现某些化疗药物,在小剂量、高频率的给药方式下可以选择性地显著抑制肿瘤组织内新生血管内皮细胞的生长,使肿瘤细胞因为血供不足而生长受阻.这称为小剂量化疗或抗血管生成化疗.中医药治疗肿瘤具有独到的疗效,近年来发现许多中药制剂及中药来源的药物有抗血管新生作用.本研究旨在在国内外现有的研究基础上探讨奥沙利铂联合复方苦参注射液对体内外血管生成的抑制作用.方法:采用MTT法观察奥沙利铂(L-OHP)联合复方苦参注射液对人脐静脉内皮细胞(HUVEC)、人结肠癌LoVo细胞增殖的影响;采用transwell板,观察L-OHP联合复方苦参注射液对HUVEC迁移的影响;采用鸡胚绒毛尿囊膜(CAM)模型,观察L-OHP联合复方苦参注射液对鸡胚绒毛尿囊膜新生血管的抑制作用.结果:HUVEC和LoVo细胞对不同浓度两种药物进行联合的敏感性不同.2 μg/ml奥沙利铂和25 μl/ml复方苦参注射液对HUVEC增殖的抑制率分别为31.6%、32.1%,联合应用后的抑制率为54.4%,具有协同作用;0.5 μg/ml奥沙利铂和6.25 μl/ml复方苦参注射液具有抑制HUVEC迁移的作用,联合应用具有协同效应.奥沙利铂联合应用复方苦参注射液具有抑制鸡胚尿囊膜血管生成作用.结论:小剂量L-OHP和复方苦参注射液在体外有抑制细胞生长,在体内具有抑制血管生成作用,两者联合具有协同效应.     

TRAIL与羟基喜树碱联用协同杀伤人肺腺癌A549细胞

目的：探讨羟基喜树碱与肿瘤坏死因子相关凋亡诱导配体（tumor necrosis factor-related apoptosis inducing ligand，TRAIL）联用对肺腺癌A549细胞是否具有协同杀伤作用，并对其机制进行初步研究。方法：MTT法检测细胞毒性作用，流式细胞仪检测细胞凋亡和线粒体膜电位，Western blot分析bcl-2和bcl-xl蛋白水平变化。结果：单用100ng／mL TRAIL，4、40和400μg／mL羟基喜树碱对A549细胞杀伤率分别为13．9％、3．0％、23．4％和76．7％，联合用药的抑制率分别为43．6％、52．5％和83．1%。100ng／mL TRAIL和4μg/mL羟基喜树碱有协同作用，可使细胞线粒体膜电住降低，但不影响bcl-2和bcl-xl蛋白表达。结论：TRAIL与羟基喜树碱联用对肺腺癌A549细胞有明显的协同杀伤作用，其机制可能与羟基喜树碱降低A549细胞线粒体膜电位有关。     

重楼醇提物体外抑制血管生成作用研究

目的：探讨重楼醇提物对体外血管生成的抑制作用及其可能的机制。方法：采用MTT法观察重楼醇提物对人脐静脉血管内皮细胞（HUVEC）、人肠癌LOVO细胞增殖的影响。利用transwell小室趋化实验、体外管腔形成实验观察重楼醇提物对HUVEC迁移、成血管能力的影响。结果：重楼醇提物在（15—60）μg／ml浓度范围内，内皮细胞抑制率在24．2％-82．1％之间，具有抑制血管内皮细胞增殖作用；体外管腔形成实验发现，（15—60）μg／ml的重楼醇提物管腔形成数目减少，且管腔不完整，与对照组相比有显著性差异（P分别为0．0000011、0．0000417、0．0008752）。经（15—60）μg／m1的重楼醇提物处理后内皮细胞迁移数明显少于对照组（P分别为0．0000005、0．0000033、0．009993）。而在该浓度范围内，对人肠癌LOVO细胞无明显细胞毒作用（细胞抑制率在12．1％-13．9％之间）。（15—60）μg／ml的重楼醇提物可诱导内皮细胞凋亡，抑制内皮细胞DNA的合成，且呈剂量依赖性（相关系数分别为0．929、-0．922，P为0．036、0．039）。结论：重楼醇提物在体外能有效抑制血管生成。其机制可能与抑制内皮细胞增生、迁移和管腔形成，诱导内皮细胞凋亡，抑制内皮细胞DNA的合成有关。     

和厚朴酚抗血管生成作用的实验研究

目的：本研究通过检测和厚朴酚在体内对鸡胚尿囊膜（chicken chorilallantoic membranes，CAM）血管生成的抑制作用及体外对肿瘤血管生成的影响，探讨和厚朴酚的抗血管生成作用及其初步机制。方法：（1）用四甲基偶氮唑盐（MTT）法检测和厚朴酚对人脐静脉内皮细胞（human umbilical vein endothelial cell，HUVEC）、人成纤维细胞及人结肠癌RKO细胞增殖的抑制作用；（2）采用鸡胚绒毛尿囊膜模型，观测和厚朴酚对CAM新生血管生成的抑制作用；（3）逆转录聚合酶链反应（RT-RCR）方法检测和厚朴酚对RKO细胞血管内皮生长因子A（VEGF-A）mRNA表达及细胞培养上清液VEGF蛋白分泌的影响。结果：和厚朴酚对血管内皮细胞具有优先抑制作用，对HUVEC的半数抑制剂量（inhibitory concentration 50％，IC50）为14．5μmol/L，而对原代培养的人成纤维细胞的IC50高达150μmol/L，对人结肠癌RKO细胞的IC50为42μmol/L；和厚朴酚0．1μg／只和0．2μg／只剂量时显著抑制CAM新生血管形成，抑制率分别为58％和86％；和厚朴酚在10μmol/L和20μmol/L剂量时显著抑制RKO细胞的VEGF-A mRNA表达及细胞培养上清液中VEGF蛋白的分泌，具有显著性差异。结论：和厚朴酚在非凋亡剂量时具有抗血管形成作用，其机制与抑制血管内皮细胞增殖以及抑制肿瘤细胞表达VEGF有关。     

草分枝杆菌联合羟基喜树碱对SPCA／Ⅰ人肺腺癌细胞抗增殖效应的研究

目的：探讨草分枝杆菌（mycobacteriumphlei)对人肺腺癌细胞系SPCA／Ⅰ的抗增殖效应，以及不同浓度草分枝杆菌与羟基喜树碱联合应用时的协同效应。方法：应用细胞培养和MTT比色法，检测不同浓度组的草分支杆菌，以及草分枝杆菌与羟基鼓地碱联合应用对SPCA／Ⅰ人肺腺癌细胞生长的抑制作用，波长定于570nm，测定光密度（A值）并计算抑制率。结果：不同浓度的草分枝杆菌，在体外对SPCA／Ⅰ人肺腺朱均 直接的抑制作用，其抑制率与所用药物浓度呈正相关。1．720μg/ml及0．860μg/ml浓度的草分枝杆菌与羟基喜树碱（1．0PPC、0．5PPC）联用，在体外对人肺腺癌细胞的抑制率显著高于单用羟基喜树碱组。结：草分枝杆菌在体外对SPCA／Ⅰ人肺腺癌细胞有直接抑制作用，该药与羟基对碱联合应用有显著的协同抗肿瘤效应。     

长春瑞滨及联合热疗抗血管生成作用的实验

目的探讨长春瑞滨及联合热疗对血管生成的抑制作用。方法以人肺癌A549细胞为对照,采用MTT法观察长春瑞滨及联合热疗对人脐静脉内皮细胞（HUVEC）增殖的影响;通过Transwell趋化实验、小管形成实验及流式细胞术观察长春瑞滨对HUVEC迁移、小管形成及细胞凋亡的影响;利用鸡胚绒毛尿囊膜（CAM）模型,观察长春瑞滨对体内CAM新生血管的影响。结果小剂量（0.1～1 ng/ml）长春瑞滨对HUVEC和A549增殖抑制具有差异细胞毒性（P=0.000）。0.1～1 ng/ml 长春瑞滨呈剂量依赖性抑制HUVEC增殖（r=0.993,P=0.000）,联合热疗具有抑制HUVEC增殖的叠加或协同效应。小剂量长春瑞滨能够抑制HUVEC迁移和小管形成,诱导HUVEC凋亡,遏制CAM新生血管形成。结论小剂量长春瑞滨具有抗血管生成作用,联合热疗具有协同或叠加效应。     

Oncogénétique et psycho-oncologie, une collaboration recommandée...

Résumé: La possibilité depuis 1994, de connaître la probabilité individuelle de développer certains cancers a permis de proposer de nouvelles modalités de prévention, de traitements et contribué au développement actuel de loncogénétique. Une meilleure connaissance des répercussions psychologiques tant pour les patients que pour les apparentés est désormais possible et limplication des psycho-oncologues dans ce cadre de la réalisation des tests prédictifs, recommandée. La mission de «messager» qui incombe au «cas-index» doit faire lobjet dune attention particulière. La complexité de linformation et la dimension paradoxale que peut avoir parfois la communication à propos des choix, rend difficile lévaluation de la qualité du consentement. La situation particulièrement délicate dune aide à la décision à légard de la chirurgie prophylactique, exige une collaboration étroite des généticiens et des psycho-oncologues.Les soins de support en oncologie     

The role of papillomaviruses in human cancers

This review comprehensively evaluates the influence of gene-gene, gene-environment and multiple interactions on the risk of colorectal cancer (CRC). Methods of studying these interactions and their limitations have been discussed herein. There is a need to develop biomarkers of exposure and of risk that are sensitive, specific, present in the pathway of the disease, and that have been clinically tested for routine use. The influence of inherited variation (polymorphism) in several genes has been discussed in this review; however, due to study limitations and confounders, it is difficult to conclude which ones are associated with the highest risk (either individually or in combination with environmental factors) to CRC. The majority of the sporadic cancer is believed to be due to modification of mutation risk by other genetic and/or environmental factors. Micronutrient deficiency may explain the association between low consumption of fruit/vegetables and CRC in human studies. Mitochondrial modulation by dietary factors influences the balance between cell renewal and death critical in colon mucosal homeostasis. Both genetic and epigenetic interactions are intricately dependent on each other, and collectively influence the process of colorectal tumorigenesis. The genetic and environmental interactions present a good prospect and a challenge for prevention strategies for CRC because they support the view that this highly prevalent cancer is preventable.     

Antifungal combination therapy in localized candidosis

A mouse model of localized candidosis in air-filled subcutaneous cysts imitating thrush has been developed. We have now tested various antifungal combinations in this animal model. Flucytosine (5-FC) + amphotericin B (Amph B) showed the highest efficacy, a clear additive or even synergistic effect was seen. The combination of 5-FC + imidazole or triazole derivative was less efficacious, an additive effect was rare. The combination of 5-FC + Amph B was also tested against Candida albicans strains showing various degrees of 5-FC-resistance. A significant reduction in 5-FC-resistant mutants was seen after the treatment with the combination.     

Les génériques en cancérologie: à molécule identique, médicaments identiques? Les biosimilaires, des super-génériques?

Résumé: Les biosimilaires vont bientôt voir leur apparition en Europe. Comment un laboratoire peut-il aborder le développement de son dossier dAMM? Quelles sont les bases légales et les recommandations officielles? Comment la similarité et/ou le caractère générique peuvent-ils être démontrés? Les règles sont-elles identiques à celles des produits chimiques conventionnels pour lesquels, notamment en cancérologie, il existe des médicaments génériques? Comment faire pour que la sécurité et lefficacité des médicaments biosimilaires soient assurées pour les patients?     

Cancer immunotherapy

Unprecedented progress has seen made in the last decade in the field of cancer immunotherapy. The recent approval of nivolumab （Opdivo）, the first anti-programmed cell death-1 （PD-1） antibody, for metastatic melanoma in Japan, marked a milestone in the rapidly advancing field of cancer immunotherapy. Nivolumab together with ipilimumab （Yervoy）, the anti-cytotoxic T-lymphocyte-associated antigen-4 （CTLA-4） antibody, are the first 2 drugs in the class of ＂immune checkpoint inhibitors＂ that have delivered impressive responses in patients with metastatic melanoma and renal cell cancer （RCC） as well as a variety of solid tumors.     

Effect of tumor therapeutic irradiation on the mechanical properties of teeth tissue

Tumor irradiation of the head-neck area is accompanied by the development of a so-called radiation caries in the treated patients. In spite of conservative therapeutic measures, the process results in tooth destruction. The present study investigated the effects of irradiation on the demineralization and remineralization of the dental tissue. For this purpose, retained third molars were prepared and assigned either to a test group, which was exposed to fractional irradiation up to 60 Gy, or to a non-irradiated control group. Irradiated and non-irradiated teeth were then demineralized using acidic hydroxyl-cellulose gel; afterwards the teeth were remineralized using either Bifluorid12 or elmex gelee. The nanoindentation technique was used to measure the mechanical properties, hardness and elasticity, of the teeth in each of the conditions. The values were compared to the non-irradiated control group. Irradiation decreased dramatically the mechanical parameters of enamel and dentine. In nonirradiated teeth, demineralization had nearly the same effects of irradiation on the mechanical properties. In irradiated teeth, the effects of demineralization were negligible in comparison to non-irradiated teeth. Remineralization with Bifluorid12 or elmex gelee led to a partial improvement of the mechanical properties of the teeth. The enamel was more positively affected by remineralization than the dentine.     

Consultation multidisciplinaire pour les patients atteints d’une pathologie tumorale (carcinome infiltrant ou lésion précancéreuse) liée à HPV : la consultation multidisciplinaire papillomavirus

Given the recent increase in the number of human papillomavirus (HPV)-induced cancers in other locations than gynaecological, the number of patients with two cancers at distinct sites, and because of the lack of exhaustive data, we decided to create a multidisciplinary network around an HPV consultation at the Georges-Pompidou European Hospital (HEGP). This network aims to set up the best tools for detecting HPV-associated “multisite” precancerous lesions in order to determine the possible impact of dedicated care for this at-risk population. This monthly consultation was created at the HEGP in June 2014. It is currently organized around five consultations: gynaecological, ENT, urological, digestive and immunological. Every patient who has been diagnosed with HPV-related cancer and whose care is provided at the HEGP is offered this particular follow-up: systematically, once the initial lesion has been treated, the patient is convened annually for a day during which it benefits from the consultations mentioned above. A consultation with a psychologist is systematically proposed. Local samples are taken at each site: a cytological examination, the analysis of known predictive and prognostic virological markers are carried out. This study fits more broadly in a theme of clinical and fundamental research around cancers related to HPV.     

Differentiation state and invasiveness of human breast cancer cell lines

Summary Eighteen breast cancer cell lines were examined for expression of markers of epithelial and fibroblastic differentiation: E-cadherin, desmoplakins, ZO-1, vimentin, keratin and ₁ and ₄ integrins. The cell lines were distributed along a spectrum of differentiation from epithelial to fibroblastic phenotypes. The most well-differentiated, epithelioid cell lines contained proteins characteristic of desmosomal, adherens and tight junctions, were adherent to one another on plastic and in the basement membrane matrix Matrigel and were keratin-positive and vimentin-negative. These cell lines were all weakly invasive in anin vitro chemoinvasion assay. The most poorly-differentiated, fibroblastic cell lines were E-cadherin-, desmoplakin- and ZO-1-negative and formed branching structures in Matrigel. They were vimentin-positive, contained only low levels of keratins and were highly invasive in thein vitro chemoinvasion assay. Of all of the markers analyzed, vimentin expression correlated best within vitro invasive ability and fibroblastic differentiation. In a cell line with unstable expression of vimentin, T47D_CO, the cells that were invasive were of the fibroblastic type. The differentiation markers described here may be useful for analysis of clinical specimens and could potentially provide a more precise measure of differentiation grade yielding more power for predicting prognosis.