IL-2联合银耳多糖激活的同种脾细胞对肝癌实验性治疗的研究

目的研究淋巴因子IL-2联合银耳多糖(Tremella Polysaccharide TP)共同激活的小鼠脾细胞对体外肿瘤细胞的杀伤作用及小鼠移植性肝癌的治疗作用.方法将IL-2激活的小鼠脾细胞(即LAK细胞)和IL-2与银耳多糖协同激活的小鼠脾细胞(即TP-LAK细胞)分别作用于体外培养的3H-TdR标记的肿瘤细胞P815,H22和B16,2h后用γ-闪烁计数仪进行检测,并计算杀伤程度.将上述激活的小鼠脾细胞在小鼠荷肝癌局部皮下注射,隔日1次,共5次.检测肿瘤质量、体积、组织学改变和主要脏器的形态学改变.结果 IL-2与银耳多糖激活的小鼠脾细胞对体外培养的肿瘤细胞有明显的杀伤作用,2种效应细胞对B16杀伤作用最强,H22次之,P815最弱,TP-LAK细胞的作用较之LAK细胞更强,P<0.05.治疗组肝肿瘤质量、体积均低于对照组.组织学上肿瘤坏死程度重且范围广,淋巴细胞、单核细胞浸润明显,纤维组织增生明显.银耳多糖协同IL-2实验组较单纯的IL-2实验组表现得更为明显.除脾脏外,其他主要脏器无明显形态学变化.结论 IL-2激活脾LAK细胞是有效的抗肝癌细胞,IL-2 与银耳多糖在活化脾细胞提高抗肿瘤方面具有协同作用.其对体内主要脏器无明显影响.     

银耳多糖在肝癌治疗中的作用及相关机制的实验研究

目的研究银耳多糖(Tremella Polysaccharide TP)对荷HAC小鼠T细胞表面分化抗原、细胞因子、体外肿瘤细胞的杀伤作用及小鼠移植性肝癌的影响.方法用环磷酰胺和银耳多糖分别作用于荷HAC肝癌小鼠,采用免疫荧光法、流式细胞仪及酶标仪观察各组免疫功能的变化及细胞因子的表达,并分离其脾细胞,将诱导的细胞毒性T细胞(CTL细胞)分别作用于体外培养的3H-TdR标记的肿瘤细胞P815,HAC和B16,4 h后用γ-闪烁记数仪进行检测,并计算杀伤程度.检测各组肿瘤质量、体积、组织学改变和主要脏器的形态学改变.结果银耳多糖实验组总T细胞、T杀伤细胞及肿瘤坏死因子(TNF)高于对照组,其中对总T细胞及TNF上调明显.在对HAC,P815,B16肿瘤细胞的杀伤作用中,银耳多糖实验组的杀伤活性明显高于对照组,分别为23.85%,20.81%,25.43%,P<0.01,其中对B16杀伤作用最强,HAC次之,P815最弱.环磷酰胺 银耳多糖实验组上述指标与银耳多糖实验组相比有所下降,但与对照组相比则明显增高(P<0.05).治疗组肝肿瘤质量、体积均低于模型组.组织学上肿瘤坏死程度重且范围广,淋巴细胞、巨噬细胞浸润明显,纤维组织增生显著.环磷酰胺组较实验组表现的更为明显.银耳多糖实验组除脾脏外,其他主要脏器无明显形态学变化.结论银耳多糖具有激活和提高特异性和非特异性杀伤细胞的抗肿瘤作用,是一种能调节机体免疫功能、抑制肿瘤细胞增殖、分化的抗癌中药制剂.     

人LAK细胞的诱导及其体外抗肿瘤作用

采用IL－2或IL－2＋PHA－P对人外周血单核细胞进行体外淋巴因子激活的杀伤细胞（LAK、LI－LAK细胞）的诱导及对其体外抗肿瘤细胞活性进行研究，结果显示：300～500IU／mL．IL－2＋5μg／mLPHA－P，孵育时间8～10d为最适的诱导条件，最高可诱导细胞增殖100倍以上。诱导出的LAK及LI－LAK细胞对数种自然杀伤细胞（NK）敏感或不敏感的瘤细胞均有明显杀伤作用。实验还显示，PHA－P的主要作用可能是促进细胞扩增，而非肿瘤杀伤活性的诱导。     

LAK细胞和HSV1-TKc/GCV基因治疗系统对卵巢癌细胞的体外作用

[目的]对LAK细胞和.HSV1-TKc/GCV基因治疗系统对卵巢癌细胞的体外作用进行研究分析.[方法]分别将不同比例的LAK细胞和AO细胞混合培养6 h(4:1,20:1,100:1,500:1).采用LDH释放法测定LAK的杀伤活性;HSV1-TKc+及HSV1-TKc-AO细胞按照不同比例培养60 h(15:85,35:65,50:50,75:25,65:35,85:15),后更换15μg/ml GCV培养液,6 d后采用LDH释放法测定其杀伤活性.对两组实验的效靶比与杀伤活性进行相关性检验,比较两组的杀伤活性.[结果]LAK细胞与靶细胞的比值与杀伤活性呈线性相关性(r=0.996,P＜0.01).其杀伤活性随着效靶比的增大而增加;AO/HSVI-TKc+与AO/HSV1-TKc-的细胞比例与HSV-1TKc/GCV的杀伤活性成线性相关(r=0.984,P＜0.01),其杀伤活性随着比例的增大而增加,LAK与HSV1-TKc/GCV两组比较,差异无统计学意义(P＞0.05).[结论]LAK细胞与HSV1-TKc/GCV基因治疗系统对卵巢癌细胞的体外具有一定的杀伤作用,但两者比较差异无统计学意义.     

抗CD3×CD87单链双特异抗体介导的单个核细胞对前列腺癌细胞杀伤作用的体外研究

本研究尝试探讨抗CD3×CD87单链双特异抗体介导的单核细胞对CD87+前列腺癌细胞的杀伤作用.首先,利用基因工程手段,构建了pET24a/抗CD3×CD87单链双特异抗体原核表达载体;将重组载体转化BL-21(DE3)感受态细胞,用IPTG诱导蛋白表达;获得的工程菌经超声破碎,先在变性条件下亲和纯化目的蛋白,再采用稀释复性方法对scBsAb进行复性.其次,利用流式细胞术检测表达纯化的scBsAb与两种相应抗原的结合情况.最后,通过CCK-8法检测外周血单个核细胞(PBMCs)对CD87阳性前列腺癌细胞PC-3的杀伤效应.结果表明,利用基因工程手段获得了较高纯度的scBsAb;scBsAb能与PC-3细胞表面CD87特异性结合,并且能与CD3特异性结合;scBsAb的浓度及效靶比影响scBsAb介导的杀伤作用.在效靶比为10∶1,scBsAb浓度为100μg/mL时最大杀伤率达到40.86%;在scBsAb浓度为100μg/mL,效靶比为40∶1时达到最大杀伤率60.9%.此外,ELISA检测表明,在scBsAb介导的PBMCs靶向CD87阳性肿瘤细胞杀伤过程中IFN-γ的水平显著增高.结果证明,scBsAb能够在体外有效介导效应细胞杀伤CD87阳性肿瘤细胞.     

IL-18的毕赤酵母表达及其激活NF-κB和p38途径促进BRL-3A细胞增殖

目的:探讨白细胞介素18(IL-18)在毕赤酵母中的表达条件及其对BRL-3A细胞的增殖作用和作用机制.方法:构建表达载体p PIC9K-IL-18,电转化至毕赤酵母GS115,PCR、Western blotting和SDS-PAGE方法鉴定表达产物的正确性,并对发酵条件进行优化,再经双水相萃取偶联DEAE离子交换层析分离纯化表达产物.然后,MTT法检测重组大鼠IL-18(rr IL-18)对BRL-3A细胞的增殖作用,RT-PCR和Western blotting方法检测IL-18信号通路相关基因的表达变化.结果:在3 L发酵罐规模下600的菌体密度、p H=6.0、诱导温度23℃、体积分数分别为20%DO和0.25%甲醇浓度条件下表达质量浓度最高约为280 mg/L,经分离纯化后纯度可达95%.此外,质量浓度为15~20 ng/m L rr IL-18孵育48 h,能明显促进BRL-3A细胞增殖(P0.5),且IL18R、My D88、NF-κB及其下游靶蛋白cyclin B1和cyclin B2的表达量均显著高于对照组(P0.5).同时,p38途径的ATF2及其下游靶蛋白cyclin A2和Bcl-2的表达量也随之升高(P0.5).结论:成功构建大鼠IL-18表达载体,优化在毕赤酵母中的发酵条件和纯化工艺,并首次证实IL-18能通过NF-κB和p38/ATF2途径激活细胞增殖相关靶蛋白cyclin B1、cyclin B2、cyclin A2和Bcl-2,进而促进BRL-3A大鼠肝细胞增殖.     

熊果酸对H22荷瘤小鼠抗肿瘤及免疫调节作用

目的 探讨熊果酸（UA）对H22荷瘤小鼠抗肿瘤作用及免疫功能的影响.方法 皮下移植建立H22荷瘤小鼠模型,腹腔注射不同剂量UA,检测抑瘤率和免疫器官指数,MTT法检测脾脏T、B淋巴细胞增殖能力,流式细胞术检测CD4＋、CD8＋T细胞亚群含量及比例,ELISA法检测血清细胞因子IL-2、IL-4和TNF-α的表达量.结果 UA对小鼠皮下移植性肿瘤H22有显著的抑制作用,可降低免疫器官中异常增大的脾指数,增强脾脏中T、B淋巴细胞增殖能力,提高淋巴细胞亚群CD4＋T细胞表达及CD4＋/CD8＋T细胞亚群比例,促进血清IL-2、TNF-α表达,降低IL-4表达.结论 UA可抑制小鼠肝癌H22肿瘤生长,体内可以提高荷瘤小鼠的免疫能力,其抗肿瘤作用可能与机体的免疫调节作用相关.     

pEgr1-Trail重组质粒联合电离辐射对MCF-7细胞杀伤效应的实验研究

目的研究p Egr1-Trail重组质粒联合电离辐射对MCF-7细胞的杀伤效应,为增强肿瘤放疗效果提供实验证据.方法应用Annexin V-EGFP/PI双染法检测细胞凋亡;应用酶标仪检测细胞增殖;应用流式细胞仪检测细胞周期.结果不同处理组作用MCF-7细胞后24 h、不同剂量X射线照射后24 h后,各处理组MCF-7细胞的生长抑制率呈上升态势,生长抑制由弱到强的顺序为:2.0 Gy,p Egr1-Trail+0.5 Gy,5.0 Gy,p Egr1-Trail+1.0 Gy,p Egr1-Trail+2.0 Gy,Egr1-Trail+5.0 Gy.各处理组联合2.0 Gy X射线照射后6 h,细胞凋亡率开始上升,48 h达高峰,其中p Egr1-Trail+2.0 Gy组与其他各处理组比较差异具有统计学意义,且细胞凋亡最为显著(P0.05).在细胞周期方面,2.0 Gy X射线照后6 h各处理组S期细胞百分数呈现上升趋势,24 h达高峰;照后24 h,G_2+M期细胞百分数开始上升,且各处理组比较差异均具有统计学意义(P0.05);照后48 h,G_2+M期细胞百分数达到最高,依旧是p Egr1-Trail+2.0 Gy组上升最显著.结论 p Egr1-Trail重组质粒联合电离辐射作用于肿瘤细胞表现出协同杀伤效应,促凋亡作用强于单独X射照射组或p Egr1-Trail基因干预组.     

海带硫酸多糖对小鼠腹腔巨噬细胞激活及细胞毒作用的影响

目的:探讨海带硫酸多糖(LPS)对小鼠腹腔巨噬细胞MΦ的激活及细胞毒作用,为研究LPS作用机体抑制S-180细胞生长的作用提供实验依据。方法:小鼠随机分为5组,生理盐水NC为对照组,Ⅰ、Ⅱ、Ⅲ、Ⅳ为实验组,分别腹腔注射不同浓度的LPS 20、40、80和120mg/kg·d,连续注射2 d,然后制备腹腔MΦ效应细胞和用传代8 d龄腹水型S-180瘤细胞制备靶细胞,使效:靶之比为5:1,在96孔培养板上进行MΦ细胞毒活性测定。结论:海带硫酸多糖能激活小鼠腹腔巨噬细胞,对S-180肿瘤细胞具有较好的杀伤作用。     

拟黑多刺蚁体外抑瘤作用研究

目的　通过拟黑多刺蚁对体外S180 ,H2 2 ,YAC 1肿瘤细胞DNA合成影响及细胞毒性作用 ,研究该蚁抑瘤效应及机制 .方法　采用3H TdR掺入法和12 5I UdR释放法 .结果　拟黑多刺蚁对S180 ,H2 2 ,YAC 1肿瘤细胞DNA合成均有明显抑制作用 (P <0 .0 5 ) ,其抑制作用随剂量增加而增强 .以对S180 肿瘤细胞抑制作用最显著 .同时证实 ,该蚁对YAC 1,S180 肿瘤细胞均有毒性作用 .结论　拟黑多刺蚁对肿瘤细胞具有直接杀伤和抑制生长的作用 .     

Markedness and its Interpretation in Language Use

Language markedness is a common phenomenon in languages, and is reflected from hearing, vision and sense, i.e. the variation in the three aspects such as phonology, morphology and semantics. This paper focuses on the interpretation of markedness in language use following the three perspectives, i.e. pragmatic interpretation, psychological interpretation and cognitive interpretation, with an aim to define the function of markedness.     

Features of Women's language

Language is a means of verbal communication. People use language to communicate with each other. In the society, no two speakers are exactly alike in the way of speaking. Some differences are due to age, gender, statue and personality. Above all, gender is one of the obvious reasons. The writer of this paper tries to describe the features of women＇s language from these perspectives： pronunciation, intonation, diction, subjects, grammar and discourse. From the discussion of the features of women＇s language, more attention should be paid to language use in social context. What＇s more, the linguistic phenomena in a speaking community can be understood more thoroughly.     

An Analysis of Catherine＇s Character——Wuthering Heights

Wuthering Heights, Emily Bronte＇s only novel, was published in December of 1847 under the pseudonym Ellis Bell. The book did not gain immediate success, but it is now thought one of the finest novels in the English language. Catherine is the key character of this masterpiece, because everybody and everything center on her though she had a short life. We can understand this masterpiece better if we know Catherine well.     

Lower Paleozoic oil relationships within Williston Basin, Canada

The Williston Basin is a significant petroleum province, containing oil production zones that include the Middle Cambrian to Lower Ordovician, Upper Ordovician, Middle Devonian, Upper Devonian and Mississippian and within the Jurassic and Cretaceous. The oils of the Williston Basin exhibit a wide range of geochemical characteristics defined as "oil families", although the geochemical signature of the Cambrian Deadwood Formation and Lower Ordovician Winnipeg reservoired oils does not match any "oil family". Despite their close stratigraphic proximity, it is evident that the oils of the Lower Palaeozoic within the Williston Basin are distinct. This suggests the presence of a new "oil family" within the Williston Basin. Diagnostic geochemical signatures occur in the gasoline range chromatograms, within saturate fraction gas chromatograms and biomarker fingerprints. However, some of the established criteria and cross-plots that are currently used to segregate oils into distinct genetic families within the basin do not always meet with success, particularly when applied to the Lower Palaeozoic oils of the Deadwood and Winnipeg Formation.     

低渗透非均质砂岩油藏启动压力梯度研究

理论推导与室内实验相结合,建立了低渗透非均质砂岩油藏启动压力梯度确定方法。首先借助油藏流场与电场相似的原理,推导了非均质砂岩油藏启动压力梯度计算公式。其次基于稳定流实验方法,建立了非均质砂岩油藏启动压力梯度测试方法。结果表明:低渗透非均质砂岩油藏的启动压力梯度确定遵循两个等效原则。平面非均质油藏的启动压力梯度等于各级渗透率段的启动压力梯度关于长度的加权平均;纵向非均质油藏的启动压力梯度等于各渗透率层的启动压力梯度关于渗透率与渗流面积乘积的加权平均。研究成果可用于有效指导低渗透非均质砂岩油藏的合理井距确定,促进该类油藏的高效开发。     

On Tragic Consciousness In The Works By Ernest Hemingway

As an American modern novelist who were famous in the literary world, Hemingway was not a person who always followed the trend but a sharp observer. At the same time, he was a tragedy maestro, he paid great attention on existence, fate and end-result. The dramatis personae's tragedy of his works was an extreme limit by all means tragedy on the meaning of fearless challenge that failed. The beauty of tragedy was not produced on the destruction of life, but now this kind of value was in the impact activity. They performed for the reader about the tragedy on challenging for the limit and the death.     

Design and analysis of generic LBS application developing platform

Location based services is promising due to its novel working style and contents.A software platform is proposed to provide application programs of typical location based services and support new applications developing efficiently. The analysis shows that this scheme is easy implemented, low cost and adapt to all kinds of mobile nework system.     

Achieving Unusual States of Matter Under High Pressure

正The periodicity of the elements and the non-reactivity of the inner-shell electrons are two related principles of chemistry,rooted in the atomic shell structure.Within compounds,Group I elements,for example,invariably assume the+1 oxidation state,and their chemical properties differ completely from those of the p-block elements.These general rules govern our understanding of chemical structures and reactions.Using first principles calcula-     

Exchange-Correlation and Electronic Excitation Energies from Pairing Matrix Fluctuations

We have developed an adiabatic connection to formulate the ground-state exchange-correlation energy in terms of pairing matrix linear fluctuations.This formulation of the exchange-correlation energy opens a new channel for density functional approximations based on the many-body perturbation theory.We illustrate the potential of such approaches with an approximation based on the particle-particle Random Phase Approximation(pp-RPA).This re-     

Density Functional Theory for the Response of Periodic Systems to Electric Fields Based on the Vector Potential Approach

正The electronic and nuclear(structural/vibrational)response of 1D-3D nanoscale systems to electric fields gives rise to a host of optical,mechanical,spectral,etc.properties that are of high theoretical and applied interest.Due to the computational difficulty of treating such large systems it is convenient to model them as infinite and periodic(at least,in first approximation).The fundamental theoretical/computational problem in doing so is that