胎肝细胞因子的分离提取及其抗肿瘤生物效应的实验研究

目的：从胎肝细胞裂解液或上清液中分离提取中分子量的细胞因子和低分子量的细胞因子，并观察其对H7402肝癌细胞增殖和分化的影响。方法：从胎牛肝细胞分离细胞因子，采用^3H-TdR掺入法和MTT比色法。结果：将胎肝细胞因子分成分子量小于10KD的低分子细胞因子和分子量为10－100KD的中分子细胞因子。前者对H7402肝癌细胞的生长，脱氢酶活力以及DNA合成均有明显的抑制作用，半数抑制浓度（IC50）为136mg/L，而后者的作用则相反。结论：低分子胎肝细胞因子具有抑制肿瘤细胞生长的作用，而中分子因子则是促生长和分化作用。     

胎肝天然低分子抑瘤物对肿瘤细胞增殖和分化的影响

目的：观察胎肝天然低分子抑瘤物（LMW－NTS）对不同组织来源的肿瘤细胞增殖和分化的作用。方法：采用MTT比色法和^3H－TdR掺入技术,测定培养中的肿瘤细胞DNA合成和线粒体琥珀酸脱酶活力,间接反映肿瘤细胞的增殖抑制和诱导分化情况。结果：LMW－NTS对受试的4种肿瘤细胞都有生长抑制作用,但作用强度有所不同,对来源于造血系统的肿瘤细胞作用最为显著。不同浓度的LMW－NTS对肿瘤细胞线粒体琥酸脱氢酶活力的影响不同,低浓度时表现为促进作用,而高浓度时表现为轻度抑制作用。结论：胎肝低分子抑瘤物能够抑制不同各类的肿瘤细胞的增殖,并诱导其分化。     

人胎肌来源的生物因子抗肿瘤活性实验研究

研究人胚胎横纹肌条件培养液产生的生物因子（ＡＴＰ－ＦＭＣＭ）抗瘤活性。观察发现ＡＴＦ对８种肿瘤细胞株体外增殖有抑制作用。对ＢＧＣ－８２３等６种瘤细胞的抑制率均有５２％￣９４％之间。其中对ＢＧＣ－８２３、Ｖ－７２２１和Ｈｅｌａ３株瘤细胞的抑制率最高，分别为９４％、９３％和８５％。整体抑瘤实验证实，ＡＴＦ能明显减轻Ｓ－１８０荷瘤小鼠腹水的严重程度，生存时间延长，并能有效抑制ＬＡ－７９５肺腺癌荷瘤小鼠的     

胎肝天然低分子抑瘤物对肿瘤细胞增殖和分化的影响

目的:观察胎肝天然低分子抑瘤物(LMW-NTS)对不同组织来源的肿瘤细胞增殖和分化的作用.方法:采用MTT比色法和3H-TdR掺入技术,测定培养中的肿瘤细胞DNA合成和线粒体琥珀酸脱酶活力,间接反映肿瘤细胞的增殖抑制和诱导分化情况.结果:LMW-NTS对受试的4种肿瘤细胞都有生长抑制作用,但作用强度有所不同,对来源于造血系统的肿瘤细胞作用最为显著.不同浓度的LMW-NTS对肿瘤细胞线粒体琥珀酸脱氢酶活力的影响不同,低浓度时表现为促进作用,而高浓度时表现为轻度抑制作用.结论:胎肝低分子抑瘤物能够抑制不同种类的肿瘤细胞的增殖,并诱导其分化.     

参桃软肝丸抗肿瘤实验及其诱导肿瘤细胞凋亡的研究

 目的　探讨参桃软肝丸对肝癌的抑制作用及诱导细胞凋亡作用。方法　分别将参桃软肝丸以30 g/kg、2 0 g/kg和 10 g/kg、FT 2 0 70 .2 g/kg、生理盐水 0 .2ml/10 g ,对腹水型肝癌小鼠灌胃给药。 结果　各实验组与生理盐水对照组相比较有明显抑瘤作用 ,有诱导肝癌细胞凋亡。结论　参桃软肝丸可能是通过抑制瘤细胞增殖及诱导肿瘤细胞凋亡 ,从而达到抗肿瘤作用。     

胎牛肝细胞低分子天然抑瘤物的制备及其抗肿瘤作用

目的：探讨胎牛肝细胞低分子天然抑瘤物（LMW－NTS）的理化性质,生物学特性,急性与长期毒性作用。方法：从胎牛肝脏中分离提出取LMW－NTS,用高压液相色谱仪测定其分子量,紫外分光光度法测定OD260及OD280值,采用体外双层软琼脂细胞培养及腹腔注射LMW－NTS治疗荷瘤小鼠,观察共体内外抗肿瘤作用、急性与长期毒性作用。结果：胎牛肝细胞LMW－NTS的分子量为7000－8000,属核酸和多肽类物质,性质稳定。体外培养条件下胎牛肝细胞LMW－NTS对小鼠S－180细胞较对正常小鼠骨髓粒－巨噬系祖细胞有更强的抑制增殖及集落生成作用;腹腔注射胎牛肝细胞LMW－NTS治疗荷瘤小鼠能显著延长小鼠寿命（P＜0．001）。毒性试验证实胎牛肝细胞LMW－NTS无毒副作用,长期应用安全。结论：胎牛肝细胞LMW－NTS是一类分子量小、在体内外均有明显抗肿瘤作用、应用安全可靠的抑瘤物。     

基因枪介导的neu基因DNA疫苗抗肿瘤作用的实验研究

目的 探讨DNA疫苗pWRG-neu的皮内免疫,对高表达neu基因的小鼠移植瘤生长和转移的抑制作用。方法 向小鼠黑色素瘤B16F10细胞系转染pcDNA-neu,用有限稀释法筛选一株高表达neu基因的细胞株B16F10-neu。在基因枪介导下,向C57BL／6小鼠导入DNA疫苗pWRG-neu,通过观察免疫动物的生存期,评价DNA疫苗的抗肿瘤作用。分离免疫动物脾细胞,经自体淋巴细胞混合培养实验,分析DNA疫苗体内免疫后机体的CTL应答。结果 筛选到一株高表达neu基因的B16F10-neu细胞株,转基因过程和外源基因的表达没有改变细胞系的增殖特性。用基因枪轰击,进行DNA疫苗pWRG-neu皮内免疫,对小鼠黑色素瘤B16F10-neu进行预防、治疗和抗转移的实验研究,结果表明,DNA疫苗的免疫能够明显推迟移植瘤的生长,延长小鼠生存期,获得明显的抗肿瘤效果。DNA疫苗免疫后可诱导小鼠脾淋巴细胞CTL活性。结论 基因枪介导的DNA疫苗pWRG-neu经皮内免疫,能够有效的诱导机体的细胞免疫应答,预防和治疗小鼠移植瘤的发生,并有一定的预防肿瘤肺转移的作用。     

肿瘤坏死因子的抗肿瘤作用及其机理的研究

在体内肿瘤坏死因子Tumor Necrosis Factor,TNF)的抗肿瘤效果是剂量依赖的。我们研究发现,移植了S_(180)肉瘤的小鼠从尾静脉输入0.15μg天然的兔TNF即可致使移植瘤明显出血坏死,肿瘤生长抑制,动物存活时间延长,但未见完全治愈。0.3μg兔TNF可使多数动物完全治愈。这种治疗效果与出血坏死程度基本平行。细菌脂多糖(Lipopoly saccharid.LPS)在大于5μg时也能引起肿瘤明显出血坏死,但静脉输入1μgLPS不足以引起肿瘤的出血坏死。这个剂量的LPS与0.15μg TNF合用后协同增强了抗肿瘤作用,部分动物完全治愈。将用LPS和/或TNF完全治愈的小鼠睥细胞(免疫睥细胞)在S_(180)移植同时静脉输入到小鼠体内,结果有6/14的小鼠未长出肿瘤,而输注同样数量正常鼠脾细胞的动物中仅有1/14的小鼠未长出肿瘤,与接种失败率相当。输入免疫脾细胞后,其余长出的肿瘤在前10天生长缓慢,但10天后又恢复到对照水平,这些结果提示TNF的抗肿瘤作用还依赖于内体免疫机能的状态。     

蜂胶体外抗肿瘤实验及其诱发肿瘤细胞凋亡的研究

蜂胶是蜜蜂从植物的幼芽及树木的枝条上采集树脂类物质 ,与其唾液混合加工成一种芳香性胶状固体物 ,含有多种黄酮、阿巍酸、槲皮素、肉桂酸衍生物、多糖、氨基酸等有药理活性的化合物。它不仅具有抗病原微生物、抗氧化、抗炎、护肝等作用外 ,还具有抗肿瘤作用。我们通过对不同蜂胶提取物体外抗肿瘤实验研究 ,探讨其抗肿瘤发生的机制。一、材料与方法1 蜂胶提取物 :取本地产蜂胶 ,乙醚浸泡 2 4ｈ ,上清上硅胶柱 ,分别用乙醚、氯仿、乙酸乙酯、丙酮、乙醇、甲醇洗脱 ,并收集洗脱峰组分 ,然后浓缩、抽干 ,用二甲基亚砜 (ＤＭＳＯ)溶解成 2ｍｇ/…     

丹参酮IIA联合还原型谷胱甘肽治疗抗肿瘤药致急性肝损伤的效果

目的:探讨丹参酮IIA联合还原型谷胱甘肽治疗抗肿瘤药致急性肝损伤的临床效果。方法:选择抗肿瘤药致急性肝损伤患者70例,随机分为2组,对照组（n=35）行还原型谷胱甘肽保肝治疗,观察组（n=35）应用丹参酮IIA联合还原型谷胱甘肽保肝治疗,记录两组患者治疗前、后肝功能、临床疗效、外周血T淋巴细胞及不良反应情况。结果:治疗后,两组患者肝功能均优于治疗前（P<0.05）,且观察组AST、ALT、TBIL、ALP水平均显著低于对照组（P<0.05）;观察组治疗总有效率91.4%显著高于对照组68.6%,经比较差异有统计学意义（P<0.05）;观察组外周血CD3+、CD4+、CD8+ T淋巴细胞比率及CD4+/CD8+比值均较治疗前明显改善,差异有统计学意义（P<0.05）,且观察组同期CD3+、CD4+、CD8+ T淋巴细胞比率及CD4+/CD8+比值优于对照组,差异有统计学意义（P<0.05）;两组患者均未见严重不良反应,观察组不良反应发生率为8.57%,对照组为5.71%,差异无统计学意义（P>0.05）。结论:丹参酮IIA联合还原型谷胱甘肽治疗抗肿瘤药致急性肝损伤的治疗效果较还原型谷胱甘肽好,能更有效恢复患者的肝功能,用药安全,其作用机制可能与调控细胞免疫功能相关。     

EFFECT OF HEPATIC STIMULATOR SUBSTANCE (HSS) EXTRACTED FROM FETAL LIVER ON THE PROLIFERATION OF HUMAN ALEXENDER HEPATOMA CELLS

It's reported that hepatic stimulator substance (HSS) was extracte from the fetal liver of 4 - 6 months of fetus, and that the effect of HSS on the proliferation of human Alexender hepatoma cells was studied in this paper. The results showed that proliferation of Alexender cells varied with the amount of HSS in the culture medium, and the former was positively correlated with the latter significantly (P<0. 01). The study indicated that HSS from the fetal liver can stimulate the proliferation of human Alexender hepatoma cells.     

Th9细胞抗肿瘤免疫作用及机制研究进展

Th9细胞作为新命名的辅助T细胞亚群,在抗肿瘤免疫治疗中起着重要作用,由转化生长因子-β（transforming growth factor-β,TGF-β）和白细胞介素-4（interleukin,IL-4）共同诱导初始CD4+T细胞分化而来,也可在特定的条件下由其他T细胞转化而来,表现出一定的可塑性。动物实验表明Th9可以抑制肿瘤生长,主要通过分泌IL-9等细胞因子以及其他方式发挥抗肿瘤免疫作用,细胞因子等分子可通过不同的信号途径调控Th9细胞分化、发育。本文旨在对Th9细胞的来源、抗肿瘤免疫作用及机制、相关信号通路等进行阐述,为抗肿瘤治疗提供新的视野和思路。      

胎肝骨髓胸腺细胞悬液输注治疗22例骨髓增生异常综合征

胎儿肝脏、骨髓、胸腺细胞混悬液输注治疗22例骨髓增生异常综合征(MDS),包括11例难治性贫血(RA)、4例铁粒幼细胞贫血(RAS)、4例原始细胞增多难治性贫血(RAEB)和3例原始细胞增多难治性贫血转化型(RAEBT)。每例平均接受4.63个胎儿的这种细胞悬液。结果表明,本组病人治疗后第四周血红蛋白和网织红细胞显著增加(P 值分别<0.01;<0.001)。与治疗前比较,血红蛋白、粒细胞和血小板计数在治疗三个月后有效率为54.5%,6月后为50.0%。     

独活醇提物及其单体蛇床子素体外抗肿瘤活性的实验研究

目的：探讨独活醇提物及其单体蛇床子素在体外对人胃癌细胞株MKN-45、BGC-823、人肺腺癌细胞株A549、人乳腺癌细胞株MCF-7及人结肠癌细胞株LOVO生长的影响.方法：将不同浓度中药独活醇提物及其单体蛇床子素作用于人胃癌细胞株MKN-45、BGC-823、人肺腺癌细胞株A549、人乳腺癌细胞株MCF-7及人结肠癌细胞株LOVO,倒置显微镜下观察细胞形态变化,MTT法测定独活醇提物及其单体蛇床子素对5株肿瘤细胞的增殖抑制率.结果：蛇床子素浓度在3.75μg/ml-120 μg/ml之间对MKN-45、BGC-823、A549、MCF-7及LOVO均有明显的抑制作用,且呈剂量-效应关系,IC50值分别为43.299μg/ml、56.790μg/ml、37.444 μg/ml、38.397μg/ml、36.231 μg/ml;相同条件下,独活醇提物对5株肿瘤细胞的抑制作用都较弱.结论：蛇床子素在体外能够抑制MKN-45、BGC-823、A549、MCF-7和LOVO的增殖,且LOVO细胞对蛇床子素最敏感,蛇床子素对5株肿瘤细胞的抑制作用均强于独活醇提物,说明蛇床子素有可能是独活醇提物中抗肿瘤作用的主要成分.     

Increased rejection of primary tumors in mice lacking B cells: inhibition of anti-tumor CTL and TH1 cytokine responses by B cells

We investigated the role of B cells in tumor immunity by studying immune responses of mice genetically lacking B cells to primary tumors. IgM(-/-) B cell-deficient mice (BCDM) exhibited enhanced resistance to 3 histologically diverse syngeneic tumors as compared to the wild-type (WT) mice. EL4 thymoma and MC38 colon carcinoma grew progressively in WT mice, but regressed spontaneously in BCDM whereas growth of B16 melanoma was slowed significantly in BCDM as compared to the WT mice. BCDM exhibited increased T cell infiltration of tumors, higher T(H)1 cytokine response and, in the case of MC38, a higher anti-tumor CTL response. The increased tumor resistance of BCDM did not seem to result from intrinsic changes in their non-B immunocytes because adoptive transfer of WT splenic B cells to BCDM abrogated tumor rejection and resulted in diminished anti-tumor T(H)1 cytokine and CTL responses. Studies involving BCR-transgenic mice indicated that B cells may inhibit anti-tumor T cell responses by antigen-nonspecific mechanisms since neither tumor-specific antibodies nor cognate T:B interactions were necessary for inhibition of tumor immunity by B cells. IFN-gamma secretion in splenocyte:tumor co-cultures of tumor-challenged BCDM was inhibited by WT but not CD40(-/-) B cells indicating that B cells may inhibit anti-tumor T(H)1 cytokine responses in a CD40-dependent manner. Adoptive transfer of CD40(-/-) B cells into BCDM resulted in restored growth of MC38 suggesting additional factors other than CD40 are involved in dampening anti-tumor responses. The effects of B cells on anti-tumor response warrant further study.     

CpG-island methylation study of liver fluke-related cholangiocarcinoma

Background:

Genetic changes have been widely reported in association with cholangiocarcinoma (CCA), while epigenetic changes are poorly characterised. We aimed to further evaluate CpG-island hypermethylation in CCA at candidate loci, which may have potential as diagnostic or prognostic biomarkers.

Methods:

We analysed methylation of 26 CpG-islands in 102 liver fluke related-CCA and 29 adjacent normal samples using methylation-specific PCR (MSP). Methylation of interest loci was confirmed using pyrosequencing and/or combined bisulfite restriction analysis, and protein expression by immunohistochemistry.

Results:

A number of CpG-islands (OPCML, SFRP1, HIC1, PTEN and DcR1) showed frequency of hypermethylation in >28% of CCA, but not adjacent normal tissues. The results showed that 91% of CCA were methylated in at least one CpG-island. The OPCML was the most frequently methylated locus (72.5%) and was more frequently methylated in less differentiated CCA. Patients with methylated DcR1 had significantly longer overall survival (Median; 41.7 vs 21.7 weeks, P=0.027). Low-protein expression was found in >70% of CCA with methylation of OPCML or DcR1.

Conclusion:

Aberrant hypermethylation of certain loci is a common event in liver fluke-related CCA and may potentially contribute to cholangiocarcinogenesis. The OPCML and DcR1 might serve as methylation biomarkers in CCA that can be readily examined by MSP.     

中药独活的化学成分及其抗肿瘤活性的研究进展

独活在临床治疗中运用广泛。现代研究表明独活含有多种抗肿瘤活性成分,包括甲氧基欧芹素、补骨脂素、花椒毒素、香柑内酯、伞形花内酯、异欧前胡素。本文就近年来对独活的化学成分及其抗肿瘤作用研究进展情况进行综述。     

Impact of liver cirrhosis and portal hypertension on minimally invasive limited liver resection for primary liver malignancies in the posterosuperior segments: An international multicenter study

IntroductionTo assess the impact of cirrhosis and portal hypertension (PHT) on technical difficulty and outcomes of minimally invasive liver resection (MILR) in the posterosuperior segments.MethodsThis is a post-hoc analysis of patients with primary malignancy who underwent laparoscopic and robotic wedge resection and segmentectomy in the posterosuperior segments between 2004 and 2019 in 60 centers. Surrogates of difficulty (i.e, open conversion rate, operation time, blood loss, blood transfusion, and use of the Pringle maneuver) and outcomes were compared before and after propensity-score matching (PSM) and coarsened exact matching (CEM).ResultsOf the 1954 patients studied, 1290 (66%) had cirrhosis. Among the cirrhotic patients, 310 (24%) had PHT. After PSM, patients with cirrhosis had higher intraoperative blood transfusion (14% vs. 9.3%; p = 0.027) and overall morbidity rates (20% vs. 14.5%; p = 0.023) than those without cirrhosis. After coarsened exact matching (CEM), patients with cirrhosis tended to have higher intraoperative blood transfusion rate (12.1% vs. 6.7%; p = 0.059) and have higher overall morbidity rate (22.8% vs. 12.5%; p = 0.007) than those without cirrhosis. After PSM, Pringle maneuver was more frequently applied in cirrhotic patients with PHT (62.2% vs. 52.4%; p = 0.045) than those without PHT.ConclusionMILR in the posterosuperior segments in cirrhotic patients is associated with higher intraoperative blood transfusion and postoperative morbidity. This parameter should be utilized in the difficulty assessment of MILR.