抗凋亡基因bcl—2在急性白血病骨髓与细胞株中的转录水平表达

采用原位杂交方法检测了急性白血病骨髓活检标本及细胞株中ｂｃｌ－２基因表达水平。结果２２例白血病患骨髓中２１例存在ｂｃｌ－２ｍＲＮＡ高表达；６种白血病细胞株中５种ｂｃｌ－２ｍＲＮＡ阳性（Ｍｏｌｔ－４例外），它们分别为ＣＥＭ、Ｒａｊｉ、ＨＬ－６０、Ｕ９３７及Ｋ５６２。表明造血系统肿瘤中广泛存在ｂｃｌ－２基因转录水平激活，其基因产物可能通过抑制细胞凋亡过程而影响急性白血病细胞的生物学特性。     

bcl—x和bcl—2基因在急性白血病患者的表达及其临床意义

观察凋亡调控基因ｂｃｌ－ｘ和ｂｃｌ－２在急性白血病患中的表达,探索ＡＬ的病理和化疗反应的分子机制。方法应用半定量逆转录－聚合酶链反应技术检测ｂｃｌ－ｘ和ｂｃｌ－２在３４例ＡＬ患中ｍＲＮＡ水平的表达。结果抑凋亡基因ｂｃｌ－ｘＬ和ｂｃｌ－２在ＡＬ细胞中的表达比在完全缓解和正常对照组骨髓细胞中的表达明显增高（Ｐ〈０．０１）,同时在复发患ＡＬ细胞中的表达水平分别是初治患的１．８倍和１．９倍（Ｐ〈０     

bcl-2基因在急性白血病细胞中的表达及其意义

目的 :探讨急性白血病 (AL)患者骨髓细胞中bcl- 2基因的表达及其与AL的分型、临床特征、疗效及预后因素的关系。方法 :采用免疫组化SP法检测 5 4例初治AL骨髓细胞的bcl- 2基因的表达情况 ,分析其与FAB分型、临床特征、疗效及预后因素的关系。结果 :(1)AL骨髓细胞中bcl- 2基因的表达显著高于正常对照组 (P <0 .0 5 ) ,bcl- 2基因表达在急性髓系白血病 (AML)与急性淋巴细胞白血病 (ALL)无显著差异 ;(2 )按FAB分型AML各亚型比较 :bcl- 2在M 4及M5中的表达高于M1、M2和M3(P <0 .0 5 ) ;bcl- 2的表达与初诊时的白细胞数呈正相关 (R =0 .4 4 ,P <0 .0 5 ) ,与疗效呈负相关 (R =- 0 .4 0 ,P<0 .0 5 )。结论 :bcl- 2基因在AL骨髓细胞中呈高表达 ,可作为AL疗效观察及预后判断的指标之一。     

细胞凋亡与bcl—2在皮肤鳞形细胞癌中的表达及意义

目的：研究人类皮肤肿瘤中细胞凋亡相关基因bcl-2的表达,探讨癌组织中凋亡和bcl-2的关系。方法：应用三羟基末端标记法和免疫组化（SABC）法原位观察了40例皮肤癌中细胞凋亡与bcl-2蛋白的表达,以10例正常组织作对照。结果：细胞凋亡广泛存在于皮肤鳞吕中和正常组织中,皮肤鳞癌中细胞凋亡明显高于正常组织（P＜0．05）;皮肤鳞癌中细胞凋亡与bcl-2阳性表达有显著差异（P＜0．01）。结论：皮肤癌组织中bcl-2与细胞凋亡关系密切,bcl-2有抑制细胞凋亡的作用。,     

bcl—2和bax基因在乳腺癌中的表达及其病理特征

目的探讨bcl-2和bax基因在乳腺癌中的表达及其病理特征.方法应用免疫组织化学方法检测54例乳腺癌及31例癌旁组织和8例良性乳腺病变组织标本的bcl-2和bax基因表达情况.结果bcl-2蛋白在乳腺癌中的阳性表达率为57.4%.bax蛋白的阳性表达率为79.6%.Ⅰ级乳腺癌的bcl-2蛋白阳性表达率为72.2%,Ⅱ级为61.5%,Ⅲ级为30%.bcl-2蛋白各级别表达程度间无显著性差别(χ2=5.839,ν=4,P=0.2496).Ⅰ级乳腺癌的bax蛋白阳性表达率为100%,Ⅱ级为88.5%,Ⅲ级为20%.bax蛋白的表达程度间有显著性差异(χ2=27.889,v=4,P＜0.0001).bcl-2蛋白在良性乳腺病变,癌旁组织和原位及浸润型乳腺癌4组病例中的表达阳性率之间有显著性差异(χ2=23.694,v=3,P＜0.0001);bax蛋白在4组间的阳性表达率亦有差异性(χ2=10.772,v=3,P=0.013).结论bcl-2和bax基因在乳腺癌的发生,发展过程中起着重要的作用,并与乳腺癌细胞凋亡的调节过程有关.     

bcl—2基因与淋巴瘤和白血病

ｂｃｌ－２基因为一原癌基因，它的重排与淋巴瘤和白血病的关键密切，是一抑制程序性细胞死亡基因，可作为判断预后的因素之一，本文对此做一综述。     

急性白血病患者bcl—2和bax基因表达的临床意义及其与mdr—1基因表达的关系

细胞凋亡受抑作为肿瘤细胞的耐药机制,是急性白血病(AL)预后不良的原因之一.bcl-2家族是目前最受重视的调控细胞凋亡的基因家族,本研究为探讨bcl-2和bax基因在急性白血病表达的意义,应用逆转录-聚合酶链反应(RT-PCR)方法测定70例初治AL患者及20例正常人的bcl-2,bax,mdr-1基因mRNA水平的表达,用流式细胞术检测其蛋白表达.结果表明,bcl-2和bax基因在AL患者广泛表达,bcl-2 mRNA平均表达水平明显高于正常对照(1.46 vs 0.71,P＜0.05).bcl-2和bax基因表达及bax/bcl-2比值与AL患者的年龄、性别、血小板计数、血红蛋白水平、骨髓原始细胞百分率、FAB分型和S+G2M%均未发现相关.Bcl-2蛋白表达(34.6%vs 69.2%,P＜0.05),bax/bcl-2 mRNA比值(37.1%vs 82.9%,P＜0.01)决定AL对化疗的敏感性,与AL CR率密切相关.bax/bcl-2 mRNA比值还是影响总生存期的因素.bcl-2与bax基因表达和mdr-1表达两者无相关关系.     

bcl—2基因与头颈部肿瘤

抗凋零基因ｂｃ12(ＢＣｅｌｌＬｅｕｋａｅｍｉａ/Ｌｙｍｐｈｏｍａ2)是一种原癌基因,最早是从Ｂ细胞淋巴瘤染色体14和18位断裂点ｔ(14∶18)(ｑ31∶ｑ21)中得到,免疫电镜检测ｂｃ12基因表达蛋白定位于细胞内氧自由基产生的位置。如线粒体、核周膜和内质网〔1〕。ｂｃ12是已明确的抗凋亡基因。研究表明ｂｃ12基因与许多肿瘤发生、发展密切相关,可作为评估肿瘤预后的一个参数,是细胞凋亡的调控因子。高等动物中第一个被确认的存活基因就是ｂｃ12基因,它能阻遏程序性细胞凋亡(Ａｐｏｐｔｏｓｉｓ)和延长细胞生命,但不促进细胞增殖〔2〕。关于ｂ…     

肠MALT淋巴瘤细胞凋亡与p53和bcl—2的表达

目的：探讨肠黏膜相关淋巴组织（MALT）淋巴瘤细胞增殖和凋亡的特征以及调亡相关调控基因的表达。方法：应用TUNEL技术检测21例肠淋巴瘤的凋亡指数（apoptosis index,AI),免疫组化S－P法检测PCNA增殖指数（proliferative index,PI）及bcl-2和p5e蛋白的表达。结果：随着肠MALT淋巴瘤恶性度的增高,AI和PI显著增加,并且二者呈显著正相关。低度恶性、高度恶性伴低度恶性以及高度恶性肿瘤组中,bcl-2阳性率分别为79．4％、57．1％、40．9％。三组bcl-2呈阳性率为AI均差异显著（P＜0．05）。bcl-2与AI呈显著负相关（P＜0．05）。p53阳性率为31．4％。高度恶性组p53阳性率显著高于其余两组。p53与bcl-2表达呈负相关（P＜0．05）。结论：凋亡和增殖在肿瘤的发生、发展、转化中起着重要作用,检测AI、PI可能是诊断恶性淋巴瘤、评价其生物学行为的可靠指标。在MALT的恶性度从低到高的转化中,p53和bcl-2基因可能起着重要作用。     

经体外高热联合液体培养净化的自体骨髓移植治疗成人急性白血病

用体外高热联合液体培养净化骨髓移植物后作自体骨髓移植（ＡＢＭＴ），治疗７例成人急性白血病和１例骨髓受侵的霍奇金病，移植后除１例早期死于败血症，其余皆造血重建。６例持续完全缓解（ＣＣＲ），中位缓解１８８天，１例ＣＣＲ４个月后死于非病意外，作者认为本法近期疗效较好，远期疗效尚需更多病例的长期随访证实。     

骨髓移植物体外液体培养净化处理在白血病自体骨髓移植中的应用

应用骨髓液体培养的方法对白血病患者的骨髓移植物进行大规模体外培养净化处理并进行自体骨髓移植。实验证明骨髓移植物体外液体培养净化处理后，骨髓有核细胞和ＣＦＵ－ＧＭ的回收率分别可达到６８．６±２０．０％和７２．１±２７．４％。骨髓移植后患者中性粒细胞数量恢复到＞０．５×１０￣９／Ｌ和血小板恢复到＞５０×１０￣９／Ｌ的平均时间分别为２４．７±７．２天和５１．０±１１．１天。临床研究证明，骨髓移植物体外液体培养净化处理后的自体骨髓移植，在患者体内成功的获得了造血重建。     

PNH患者骨髓红系祖细胞体外培养的研究

采用造血祖细胞体外培养技术研究了阵发性睡眠性血红蛋白尿症（ＰＮＨ）患者骨髓红系祖细胞（ＢＦＵ－Ｅ和ＣＦＵ－Ｅ）的增殖能力；骨髓细胞经新鲜酸化ＡＢ型血清处理后培养的ＢＦＵ－Ｅ和ＣＦＵ－Ｅ的增殖能力；以及ＢＦＵ－Ｅ和ＣＦＵ－Ｅ对红细胞生成素（Ｅｐｏ）的反应能力。发现ＰＮＨ患者骨髓ＢＦＵ－Ｅ和ＣＦＵ－Ｅ集落数明显低于正常；骨髓细胞经新鲜酸化ＡＢ型血清处理后培养的ＢＦＵ－Ｅ和ＣＦＵ－Ｅ集落数明显低于经热灭活酸化ＡＢ型血清处理后培养的集落数；ＢＦＵ－Ｅ和ＣＦＵ－Ｅ对Ｅｐｏ的反应回归直线呈低平状态。认为ＰＮＨ患者骨髓红系祖细胞膜缺陷可导致其在酸性条件下对补体的敏感性增高和导致其对Ｅｐｏ的敏感性降低。     

低剂量辐射联合环磷酰胺对肿瘤细胞凋亡、细胞周期及骨髓增殖影响

目的探讨低剂量辐射联合环磷酰胺对小鼠移植肿瘤细胞的凋亡、细胞周期以及骨髓增殖的影响。方法昆明种雄性小鼠60只随机分为空白对照组、荷瘤对照组（假照组）、低剂量照射组（LDR组）、CTX化疗组（CTX组）和低剂量照射联合化疗组（LDR＋CTX组）,小鼠左后肢腹股沟皮下接种S180肉瘤细胞（空白组除外）,接种后第5、8天LDR组和LDR＋CTX组小鼠给予γ射线全身照射75 mGy,照射36 h后CTX组和LDR＋CTX组小鼠采用环磷酰胺（CTX）化疗,测量肿瘤大小变化,并取肿瘤组织采用流式细胞仪分析凋亡、细胞周期,取冲洗骨髓组织分析骨髓增殖情况。结果与假照组相比,各处理组肿瘤生长缓慢;LDR组肿瘤细胞凋亡增加;CTX组、LDR＋CTX组G1期细胞比例明显增加,S期细胞比例明显减少,后者较前者为著（t=2.52,P〈0.05）。与空白对照组相比,假照组、LDR组骨髓细胞浓度未见明显变化,化疗组（CTX组、LDR＋CTX组）明显减少,后者较前者有明显提高（t=4.71,P〈0.05）;CTX组、LDR＋CTX组的增殖指数（PI）明显增加,LDR＋CTX组较CTX组进一步升高（t=3.41,P〈0.05）。结论低剂量辐射联合环磷酰胺可使机体肿瘤细胞G1期阻滞加剧,对肿瘤增殖细胞杀伤作用增强,明显提高机体抗肿瘤的作用,同时保护机体的骨髓造血机能,具有辅助肿瘤化疗的实际临床意义。     

THE PROLIFERATION OF PLASMA CELLS FROM MOUSE BONE MARROW IN VITRO : I. THE ROLE OF THYMUS

An in vitro proliferative system for immunoglobulin-G-forming plasma cells from the bone marrow of mice was established by the addition of antigenic protein and thymic cells or their homogenate to bone marrow cultures. The promoting activity of the thymus on plasma cells was independent from mouse strain, but it differed in strength with the variations of donor strains. Synthesis of immunoglobulin-G in proliferating plasma cells and its antibody reactivity against the administered antigen were demonstrated by immunocytological analyses.     

多元耐药相关蛋白P_(170)糖蛋白在白血病骨髓细胞中的表达

本文应用P_(170)糖蛋白的单克隆抗体MRK_(16),采用流式细胞仪法对10例正常骨髓细胞和21例白血病骨髓细胞P_(170)糖蛋白表达进行了测定。结果表明P_(170)糖蛋白仅出现于白血病骨髓细胞,其中10例难治性白血病有7例阳性表达,11例初治性白血病仅3例阳性表达。这表明 P_(170)糖蛋白与白血病相关,但 P_(170)糖蛋白与白血病疗效的关系则尚待更大宗病例的进一步研究。     

CELL TO CELL INTERACTION IN THE IMMUNE RESPONSE : II. THE SOURCE OF HEMOLYSIN-FORMING CELLS IN IRRADIATED MICE GIVEN BONE MARROW AND THYMUS OR THORACIC DUCT LYMPHOCYTES

The number of discrete hemolytic foci and of hemolysin-forming cells arising in the spleens of heavily irradiated mice given sheep erythrocytes and either syngeneic thymus or bone marrow was not significantly greater than that detected in controls given antigen alone. Thoracic duct cells injected with sheep erythrocytes significantly increased the number of hemolytic foci and 10 million cells gave rise to over 1000 hemolysin-forming cells per spleen. A synergistic effect was observed when syngeneic thoracic duct cells were mixed with syngeneic marrow cells: the number of hemolysin-forming cells produced in this case was far greater than could be accounted for by summating the activities of either cell population given alone. The number of hemolytic foci produced by the mixed population was not however greater than that produced by an equivalent number of thoracic duct cells given without bone marrow. Thymus cells given together with syngeneic bone marrow enabled irradiated mice to produce hemolysin-forming cells but were much less effective than the same number of thoracic duct cells. Likewise syngeneic thymus cells were not as effective as thoracic duct cells in enabling thymectomized irradiated bone marrow-protected hosts to produce hemolysin-forming cells in response to sheep erythrocytes. Irradiated recipients of semiallogeneic thoracic duct cells produced hemolysin-forming cells of donor-type as shown by the use of anti-H2 sera. The identity of the hemolysin-forming cells in the spleens of irradiated mice receiving a mixed inoculum of semiallogeneic thoracic duct cells and syngeneic marrow was not determined because no synergistic effect was obtained in these recipients in contrast to the results in the syngeneic situation. Thymectomized irradiated mice protected with bone marrow for a period of 2 wk and injected with semiallogeneic thoracic duct cells together with sheep erythrocytes did however produce a far greater number of hemolysin-forming cells than irradiated mice receiving the same number of thoracic duct cells without bone marrow. Anti-H2 sera revealed that the antibody-forming cells arising in the spleens of these thymectomized irradiated hosts were derived, not from the injected thoracic duct cells, but from bone marrow. It is concluded that thoracic duct lymph contains a mixture of cell types: some are hemolysin-forming cell precursors and others are antigen-reactive cells which can interact with antigen and initiate the differentiation of hemolysin-forming cell precursors to antibody-forming cells. Bone marrow contains only precursors of hemolysin-forming cells and thymus contains only antigen-reactive cells but in a proportion that is far less than in thoracic duct lymph.     

微波体外净化自体骨髓移植治疗急性白血病35例分析

用微波加温４２℃持续１小时净化３５例急性白血病患者缓解期骨髓后进行自体骨髓移植。移植时中位年龄２７岁，３５例均为初次缓解。移植后中位无病生存期２２个月，４例移植后４～８个月复发死亡，２例分别于移植后，１２，１５个月出现中枢神经系统白血病，治疗后仍存活。全组患者５年无病生存率及复发率分别为６５．３％及１８．３％。     

CONTRIBUTION OF A THYMIC HUMORAL FACTOR TO THE DEVELOPMENT OF AN IMMUNOLOGICALLY COMPETENT POPULATION FROM CELLS OF MOUSE BONE MARROW

The hypothesis that cells located in mouse bone marrow can acquire immunological competence by a process that involves interaction with a noncellular component of the thymus was tested using an in vitro assay of graft-versus-host reactivity as a criterion of cell competence. When suspensions of C57BL bone marrow cells were incubated in thymus extract and injected into mice incapable of inducing a response in the graft-versus-host assay as a result of neonatal thymectomy, or adult thymectomy plus irradiation, or because of genetic similarity with the (C3H x C57BL)F₁ tissue used for challenge in the assay, competent cells were recovered from the spleens of the injected mice. The reactive cells were shown to be of bone marrow origin since immune reactivity was related to the genetic makeup of the bone marrow cells rather than that of the intermediate recipients. A thymic factor was involved in the process leading to immune reactivity by these cells, as bone marrow cells incubated in xenogeneic or syngeneic thymic extracts induced a graft-versus-host response after passage through nonresponsive mice, whereas incubation of bone marrow cells in xenogeneic lymph node or spleen extracts or in culture medium only did not lead to subsequent reactivity. Participation of peripheral lymphoid tissue seemed essential in this process since bone marrow cells tested directly after exposure to thymic extract failed to induce a graft-versus-host response. C57BL bone marrow cells exposed to thymus extract and cultured together with fragments of (C3H x C57BL)F₁ spleen tissue in vitro were competent to induce a graft-versus-host response; thus, these components would seem to be sufficient as well as necessary for the immunodifferentiation process leading to graft-versus-host activity. It is concluded that one step in the process by which bone marrow cells acquire competence vis-a-vis the graft-versus-host response depends upon a thymic agent that is noncellular and extractable, and that another stage in this process is under the influence of components found within the peripheral lymphoid tissue environment. It is suggested that differentiation of precursor cells to competence could occur by progressive development of the cells in separate compartments of the lymphoid system.     

干细胞复合纳米晶羟基磷灰石胶原修复骨缺损的研究

目的　研究纳米材料作为组织工程骨基质材料的特点。方法　分离培养人骨髓间充质干细胞 ,与纳米晶羟基磷灰石胶原材料 (nHAC/PLA)于体外联合培养 ;通过大体观察、组织学分析及电镜观察了解成骨情况 ,进一步临床应用于修复骨缺损。结果　人骨髓间充质干细胞在体外可以大量扩增 ,复合细胞的材料植入骨缺损处后 ,X光摄片动态观察可见骨缺损处连接良好。结论　骨髓间充质干细胞具有成骨细胞作用 ,纳米晶羟基磷灰石胶原材料是一种很好的构建组织工程骨的支架材料。