重组人内皮抑素联合TP方案对MCF-7裸鼠移植瘤的影响

目的:探讨重组人内皮抑素联合TP方案对MCF-7裸鼠移植瘤模型血管生成及肿瘤生长的作用.方法:将36只荷瘤鼠随机分为A、B两大组,A、B两大组内均设有单一化疗组、联合用药组及对照组;单一化疗组给予紫杉醇 顺铂(TP方案).联合用药组给予重组人内皮抑素 TP,对照组等体积生理盐水;比较A大组内各组裸鼠的抑瘤率,肿瘤生长曲线.ELISA方法检测血清血管内皮生长因子(VEGF),取肿瘤组织病理切片观察微血管密度(MVD)、凋亡指数,并观察B组所有动物生存期.结果:联合用药组VEGF,MVD,较单一化疗组及对照组低,凋亡指数比单一化疗组高,有统计学差异(P＜0.05),联合用药组与化疗组生存时间的差异无统计学意义(P＞0.05),与对照组比较有统计学差异,生存时间较对照组长(P＜0.05).结论:重组人内皮抑素联合TP方案对MCF-7裸鼠移植瘤对肿瘤生长及肿瘤血管抑制优于单一化疗,但1个周期联合化疗用药不能延长生存期.     

人参皂苷Rg3联合苏拉明通过下调TGF-β1表达抑制小鼠肺癌生长转移

目的:肺癌是严重危害人类健康的恶性肿瘤之一,中晚期肺癌以放化疗为主,毒副作用大,观察低毒副作用的人参皂苷Rg3和苏拉明联合对小鼠Lewis肺癌生长转移的抑制作用及机制,为临床安全高效治疗肺癌提供实验资料.方法:将40只接种好Lewis肺癌细胞的C57BL6小鼠随机分为5组,每组8只:A组:对照组、B组:顺铂组、C组:人参皂苷Rg3组、D组:苏拉明组、E组:人参皂苷Rg3联合苏拉明组.接种第4天后开始药物干预,第20天处死,取肺组织,计肺转移结节数,剥离.移植瘤,计瘤重,计算抑瘤率,通过RT-PCR及Western-blot检测移植瘤TGF-β1的mRNA及蛋白的表达.结果:各药物干预组的肺转移结节数及瘤重明显低于对照组(P＜0.05,P＜0.01),人参皂苷Rg3联合苏拉明组最明显;BCDE各组抑瘤率分别为:(19.69±5.61)％、(35.37±8.97)％、(35.85±7.55)％和(49.39±6.99)％,ABCDE各组的肺转移率分别为100％、100％、100％、87.5％和62.5％;各药物干预组的TGF-β1的mRNA及蛋白的表达均低于对照组(P＜0.01),且人参皂苷Rg3联合苏拉明组TGF-β1的表达明显低于人参皂苷Rg3组和苏拉明组(P＜0.05).结论:人参皂苷Rg3联合苏拉明具有抑制小鼠Lewis肺癌的生长转移作用,对晚期肺癌的抑制可能与下调TGF-β1的表达有关.     

抗肝癌hdsFv-hEDN重组免疫毒素对荷人肝癌裸鼠皮下移植瘤生长的抑制作用

目的:研究抗肝癌hdsFv-hEDN重组免疫毒素对荷人肝癌裸鼠皮下移植瘤生长的抑制作用,评价其作用为导向治疗药物的临床应用价值.方法:将体外培养的人肝癌细胞系SMMC-7721细胞接种于裸鼠皮下,建立荷人肝癌裸鼠皮下移植瘤动物模型,随机分为hdsFv-hEDN治疗组和对照组,分别给予尾静脉注射hdsFv-hEDN和生理盐水,1次/日,共2周.比较各组裸鼠皮下移植瘤生长速度、肿瘤体积和重量,并计算肿瘤的抑制率.取各组裸鼠肿瘤组织,心,肺,肝,肾组织HE染色,光学显微镜下观察.结果:抗肝癌hdsFv-hEDN治疗组裸鼠皮下移植瘤生长抑制作用显著,肿瘤生长速度减慢,肿瘤体积从42.62±0.57 mm3增加到74.28±2.59 mm3、瘤重为155.82±14.43 mg,而对照组肿瘤体积从41.94±0.91 mm3增加到127.42±4.81 mm3、瘤重为283.28±15.21 mg,两组比较差异非常显著.肿瘤体积抑瘤率和瘤重抑瘤率分别达到41.59±0.02%和45.51±0.09%.组织学观察抗肝癌hdsFv-hEDN组肿瘤组织出现大片坏死,凋亡明显增加,心、肝、肺、肾等重要器官未见明显异常.结论:抗肝癌hdsFv-hEDN重组免疫毒素对荷人肝癌裸鼠皮下移植瘤生长具有良好的抑制作用.     

人EGFR显性负性突变体抑制胃癌细胞促血管形成能力

目的探讨人表皮生长因子显性负性突变体(dominant negative epidermal growth factor receptor,DNEGFR))对胃癌细胞促血管形成能力的影响及其分子机制,并检测其对裸鼠皮下移植瘤生长的影响。方法选用2株人胃癌细胞,分为如下6组:SGC-7901及NCI-N87细胞未转染组(US组,UN组),SGC-7901及NCI-N87细胞pEGFP-N1质粒转染组(ES组,EN组),SGC-7901及NCI-N87细胞pEGFPN1-DNEGFR质粒转染组(DS组,DN组)。采用人脐静脉内皮细胞(humanumbilical vein endothelial cell,HUVEC)管腔结构形成实验检测体外促血管形成能力,采用酶联免疫吸附测定法(enzyme-linked immunosorbent assay,ELISA)测定细胞培养液中血管内皮生长因子(vascular endothelial growth factor,VEGF)的水平,建立人胃癌细胞裸鼠移植瘤模型,标本微血管密度(microvessel density,MVD)检测体内促血管形成能力,标本体积检测其对裸鼠皮下移植瘤生长的影响。结果转染pEGFPN1-DNEGFR质粒的人胃癌细胞株出现HUVEC管腔结构形成抑制,培养液中VEGF水平降低,MVD计数降低,裸鼠皮下移植瘤体积变小。结论 DNEGFR可能通过下调VEGF分泌抑制胃癌细胞体外及裸鼠体内促血管形成能力,最终抑制裸鼠皮下移植瘤生长。     

顺铂联合鸦胆子油乳对人卵巢癌裸鼠原位移植瘤生长的影响

目的:探讨鸦胆子油乳以及联合顺铂对人卵巢癌裸鼠原位移植瘤生长的抑制作用。方法:构建人卵巢癌裸鼠原位移植瘤模型,随机分为对照组、鸦胆子油乳组、顺铂组和鸦胆子油乳顺铂联合组等4组,每组8只,观察各组裸鼠移植瘤体积、重量、抑瘤率及相对肿瘤增值率(T/C)的变化,检测裸鼠外周血白细胞、血小板数目及血清谷丙转氨酶(ALT)、乳酸脱氢酶(LDH)、血肌酐(CR)的变化。结果:鸦胆子油乳组、顺铂组、联合组均可有效遏制肿瘤细胞生长,各组的抑瘤率分别为23.56%±3.67%、42.57%±3.45%和48.36%±3.08%;各组的肿瘤体积和T/C明显下降,T/C均低于60%,且与对照组比较,差异有统计学意义(P0.05);与对照组比较,鸦胆子油乳组治疗后裸鼠LDH、CR值差异显著(P0.01),顺铂组与联合组ALT、LDH、CR值差异有统计学意义(P0.01);与对照组比较,鸦胆子油乳组白细胞数差异显著(P0.05),而顺铂组、联合组血小板、白细胞数量均明显减少,且差异均具有统计学意义(P0.01)。结论:顺铂联合鸦胆子油乳可以抑制人卵巢癌裸鼠移植瘤的生长,鸦胆子油乳相对于顺铂对人卵巢癌SKOV3细胞裸鼠白细胞及血小板、肝肾功能影响较小,具有低毒作用。     

塞来昔布联合survivin反义寡核苷酸对胰腺癌荷瘤裸鼠的治疗作用

目的:探讨环氧化酶-2(COX-2)抑制剂塞来昔布和survivin反义寡核苷酸(ASODN)联合作用对胰腺癌荷瘤裸鼠的治疗作用。方法:构建胰腺癌荷瘤裸鼠模型,将成瘤裸鼠随机分为4组:对照组、塞来昔布组(给予1000ppm的塞来昔布饮水)、survivin ASODN组(瘤内注射40μg/200μL的survivin ASODN)、联合组(给予1000ppm的塞来昔布饮水的同时瘤内注射40μg/200μL的survivin ASODN);观测裸鼠肿瘤生长情况并测量不同时间的体积变化,于接种肿瘤的35天处死裸鼠,测瘤体质量,应用cas- pase-3试剂盒检测caspase-3活性,免疫组织化学法检测肿瘤增殖指数(PI)和微血管密度(MVD)。结果:治疗组平均体积均明显低于对照组(491.97±4.62mm~3,427.34±14.62mm~3,300.39±6.59mm~3 vs 703.56±12.51 mm~3,P<0.01),其中联合治疗组体积明显低于塞来昔布组或survivin ASODN组(P<0.01);平均抑瘤率分别为32.26%、50.86%、62.07%。治疗组caspase-3相对活性明显高于对照组(0.026±0.003、0.040±0.018、0.059±0.005 vs 0.006±0.001,P<0.01),其中联合治疗组caspase-3相对活性明显高于塞来昔布组(P<0.01)或survivin ASODN组(P<0.05);治疗组平均PI和MVD均明显低于对照组(P<0.01,或P<0.05),其中联合治疗组平均PI和MVD明显低于塞来昔布组或survivin ASODN组(P<0.01,或P<0.05)。结论:COX-2抑制剂塞来昔布和survivin ASODN联合应用可显著抑制荷胰腺癌裸鼠的肿瘤生长,其作用机制可能是通过共同提高caspase-3活性来诱导细胞凋亡,通过抑制肿瘤细胞的增殖和新生血管的形成来发挥抗肿瘤效应,为胰腺癌的治疗提供了新的思路。     

格列卫与多西紫杉醇联合对人乳腺癌细胞株MCF-7及其裸鼠移植瘤的作用

目的：探讨分子靶向药物格列卫与多西紫杉醇联合对人乳腺癌细胞株MCF-7的凋亡及其裸鼠皮下移植瘤生长的影响。方法：采用流式细胞检测仪检测MCF-7细胞在格列卫与多西紫杉醇单独处理及共同处理条件下的凋亡率;建立人乳腺癌细胞株MCF-7裸鼠皮下移植瘤模型,观察格列卫与多西紫杉醇单独治疗组及联合治疗组移植瘤的生长,计算抑瘤率。结果：MCF-7细胞在格列卫与多西紫杉醇共同处理条件下的凋亡率（50.86%）远高于多西紫杉醇单独处理（22.06%）及同剂量格列卫组单独处理（8.13%）条件下的凋亡率;高剂量多西紫杉醇与格列卫联合组肿瘤质量与单独多西紫杉醇组比较,差异虽然没有显著性,但联合组抑瘤率高达99.55%,高于多西紫杉醇组（97.43%）,q值为1.014;中剂量多西紫杉醇与格列卫联合组肿瘤质量与单独多西紫杉醇组比较,差异有高度显著性,联合组抑瘤率达96.53%,高于多西紫杉醇组（92.01%）,q值为1.02;低剂量多西紫杉醇与格列卫联合组肿瘤质量与单独多西紫杉醇组比较,差异有高度显著性,联合组抑瘤率达68.20%,高于多西紫杉醇组（58.40%）,q值为1.004。结论：格列卫与凋亡诱导剂多西紫杉醇共同处理MCF—7细胞能达到协同诱导凋亡的效果;高、中、低剂量的多西紫杉醇与格列卫联合对人乳腺癌细胞株MCF-7裸鼠移植瘤增殖的抑制具有相加作用。     

沙利度胺联合放疗对宫颈癌裸鼠移植瘤的VEGF、bFGF、TNF-α表达的影响

目的:分析沙利度胺联合放疗对宫颈癌裸鼠移植瘤的VEGF、bFGF、TNF-α表达的影响,探究其增敏作用的机制。方法:选择24只裸鼠,均在左侧腋部皮下注射0.2 m L Hela细胞悬浊液进行造模。将造模成功的裸鼠随机分成对照组(Control group,CG)、沙利度胺组(Thalidomide group,TG)、放疗组(Radiotherapy group,RG)和联合治疗组(TG+RG),每组6只,分别给予羧甲基纤维素钠、沙利度胺溶液、放疗、沙利度胺溶液+放疗处理。治疗14 d后,通过检测裸鼠肿瘤体积、肿瘤抑制率、肿瘤延迟生长时间(Delayed tumor growth time,TGD)、放射增敏比(Radiosensitization ratio,SER)、肿瘤细胞的坏死程度、微血管密度、血清中血管内皮生长因子(VEGF)、碱性成纤维细胞生长因子(bFGF)及肿瘤组织中VEGF、bFGF、肿瘤坏死因子-α(TNF-α)分析沙利度胺联合放疗对宫颈癌裸鼠移植瘤的放疗增敏作用及其机制。结果:治疗14 d后,与对照组相比,沙利度胺组、放疗组和联合治疗组裸鼠肿瘤体积均明显变慢,肿瘤抑制率均显著增高,TGD均显著均明显延长,坏死细胞个数均明显增多,微血管密度、血清和肿瘤组织中VEGF、bFGF、TNF-α表达量均明显降低(P0.05);与沙利度胺组和放疗组相比,联合治疗组裸鼠瘤重均明显变慢,肿瘤抑制率均显著增高,TGD均显著均明显延长,坏死细胞个数均明显增多,微血管密度、血清和肿瘤组中VEGF、bFGF、TNF-α表达量均明显降低(均P0.05);放射增敏比(SER)大于1。结论:沙利度胺可显著增强放疗对宫颈癌裸鼠移植瘤的效果,可能与其显著降低VEGF、bFGF、TNF-α的表达有关。     

Bevacizumab 联合吉西他滨对荷肝癌裸鼠移植瘤生长的抑制作用

目的:探讨抗血管生成药物Bevacizumab联合吉西他滨对人肝癌裸鼠皮下移植瘤生长的抑制作用。方法:构建人肝癌细胞HepG2裸鼠皮下移植瘤模型,随机分为空白对照组、Bevacizumab组、吉西他滨组和联合用药组。观察用药前后肿瘤体积,绘制肿瘤生长曲线;应用免疫组化检测肿瘤微血管密度(MVD);Western Blot检测Bcl-2蛋白的表达。结果:Bevacizumab和吉西他滨单药均能抑制肿瘤生长,两药联合疗效明显增强(P=0.000)。与对照组和吉西他滨组相比,Bevacizumab组和联合用药组能明显抑制肿瘤血管生成,MVD值均明显降低,以联合用药组最为明显(P均0.000)。Bevacizumab和吉西他滨单药均能下调Bcl-2的表达,两药联合下调作用明显增强。结论:Bevacizumab联合吉西他滨能增强对人肝癌裸鼠移植瘤的生长及微血管生成的抑制作用,其机制可能与调控Bcl-2的表达有关。     

存活素反义寡核苷酸在人甲状腺髓样癌裸鼠模型中的抑瘤作用

目的探讨存活素(survivin)反义寡核苷酸(antisense oligodeoxyribonucleotide,ASODN)在人甲状腺髓样癌裸鼠模型中的抑瘤作用。方法构建12只人甲状腺髓样癌裸鼠模型,随机分配到正常组,SODN(sense oligodeoxyribonucleotide,正义寡核苷酸)组和ASODN组,每组4只。各组每日一次瘤内注射相应处理液,连续7d。治疗结束后1周,杀死全部瘤鼠,切取瘤体,测算瘤重和抑瘤率以评价肿瘤生长情况,瘤组织石腊切片分别行HE、免疫组化(IHC)和DNA末端原位标记法(TUNEL法)染色以评价组织病理构造、survivin与VEGF蛋白表达和细胞凋亡的变化。结果HE染色显示ASODN组具有明显减低的血管新生特点。此外,相对于正常组和SODN组,ASODN组具有显著的survivin和VEGF低表达、高凋亡、低瘤重和高抑瘤率特点(各P<0.01),而正常组与SODN组间未见上述各特点的统计学差异。结论本研究证实针对人甲状腺髓样癌survivin基因沉默治疗能够达到明显抑瘤效果,此效果至少部分通过促进肿瘤调亡和抑制肿瘤血管新生而实现。这提示survivin基因在人甲状腺髓样癌中具有重要意义和其可成为人甲状腺髓样癌基因靶向沉默治疗的重要候选基因之一。     

Recent advances in the study of Kaposi's sarcoma-associated herpesvirus replication and pathogenesis

It has now been over twenty years since a novel herpesviral genome was identified in Kaposi's sarcoma biopsies. Since then, the cumulative research effort by molecular biologists, virologists, clinicians, and epidemiologists alike has led to the extensive characterization of this tumor virus, Kaposi's sarcoma-associated herpesvirus(KSHV; also known as human herpesvirus 8(HHV-8)), and its associated diseases. Here we review the current knowledge of KSHV biology and pathogenesis, with a particular emphasis on new and exciting advances in the field of epigenetics. We also discuss the development and practicality of various cell culture and animal model systems to study KSHV replication and pathogenesis.     

The structure, reproduction and taxonomy of Vidalia and Osmundaria (Rhodophyta, Rhodomelaceae)

Comprises species occurring mostly in subtidal habitats in tropical, subtropical and warm-temperate areas of the world. An analysis of the type species, V. spiralis (Sonder) Lamouroux ex J. Agardh, a species from Australia, establishes basic characters for distinguishing species in the genus. These characters are (1) branching patterns of thalli, (2) flat blades that may be spiralled on their axis, (3) width of the blade, (4) primary or secondary derivation of sterile and fertile branchlets and (5) position of sterile and fertile branchlets on the thalli. Application of the latter two characters provides an important basic method for separation of species into three major groups. Osmundaria , a genus known only in southern Australia, was studied in relation to Vidalia , and its separation from the Vidalia assemblage is not accepted. Species of Vidalia therefore are transferred to the older genus name, Osmundaria. Two new species, Osmundaria papenfussii and Osmundaria oliveae are described from Natal. Confusion in the usage of the epithet, Vidalia fimbriala Brown ex Turner has been clarified, and Vidalia gregaria Falkenberg, described as an epiphyte on Osmundaria pro/ifera Lamouroux, is revealed to be young branches of the host, Osmundaria prolifera.     

New or rare chromosome counts in Artemisia L. (Asteraceae, Anthemideae) and related genera from Kazakhstan

Fifteen chromosome counts of six Artemisia taxa and one species of each of the genera Brachanthemum, Hippolytia, Kaschgaria, Lepidolopsis and Turaniphytum are reported from Kazakhstan. Three of them are new reports, two are not consistent with previous counts and the remainder are confirmations of very scarce (one to four) earlier records. All the populations studied have the same basic chromosome number, x = 9, with ploidy levels ranging from 2x to 6x. Some correlations between ploidy level, morphological characters and distribution are noted.     

肝癌中HBV和HCV基因和抗原的分布及意义

采用原位分子杂交方法检测HCV RNA及HBV X基因;采用免疫组织化学方法研究HCV核心抗原,非结构区C33c抗原及HBxAg在肝细胞肝癌中的定位及分布.结果表明(1)HCV RNA、HBV X基因在肝细胞肝癌组织检出率分别为40%(55/136)和82%(112/136).HCV RNA定位于癌细胞的胞浆内,阳性细胞呈散在、灶状及弥漫分布三种形式;HBV X基因在肝癌细胞中的分布呈胞浆型、核型及核浆型,阳性细胞也呈上述三种分布形式;(2)HCV C33c抗原、核心抗原在肝细胞肝癌中的阳性率为81%(133/164)及86%(141/164).C33c抗原定位于癌细胞及肝细胞的胞浆内;核心抗原既定位于癌细胞核中,又可定位于胞浆中.C33c抗原阳性细胞以灶状分布为主;而核心抗原阳性细     

Selection with two alleles and complete dominance

For a plant selection model with frequency-independent viabilities, fertilities and selfing rates, it is shown that apart from global fixation, for certain parameter combinations a protected polymorphism and facultative fixation (either allele may become fixed according to initial frequencies) may both occur. Facultative fixation requires different selling rates for the dominant and recessive type. Protection of the polymorphism requires resource allocation for male and female function. In this connection the problem of purely genetically caused population extinction is discussed.
For general frequency dependence and regular segregation, the chances for establishment of a completely recessive gene are compared to those of a completely dominant gene. It is proven that the process of establishment of the recessive gene, despite a fitness advantage, may be considerably endangered by drift effects if random mating prevails. The recessive gene may reach the same effectivity in establishment as a dominant gene, only if the recessive homozygote mates exclusively with its own type during the period of establishment.