急性髓细胞白血病骨髓基质细胞对其白血病祖细胞体外生长的影响及其与疗效关系的初探

用cactromalaspina法对11例急性髓细胞白血病骨髓基质细胞进行体外培养,观察其对CFU-L(液相-半固相集落培养法)体外生长的影响及疗效的关系.发现基质细胞数与CFU-L高峰值呈正相关.随基质细胞产率的增加其悬浮细胞及CFU-L液相集落也增加.基质细胞与CFU-L50%衰减时间越长,首次诱导缓解效果越差.基质细胞是具有多分化、多功能的细胞,参与骨髓造血微环境组成,具有支持及调节造血的作用.白血病诱导缓解的治疗效果除了与CFU-L集落形成能力有关外,也与CFU-L及基质细胞由高峰值衰减50%的时间密切相关,时间越长,治疗效果越差,可见白血病的治疗不但要消灭白血病细胞,也要摧毁白血病细胞赖以生存增殖的异常微环境.     

急性白血病病人细胞体外药敏试验与临床的联系

应用克隆培养方法,以及Ara-C,6-TG,MTX,VCR,Adr,HHT六种药物。对51例初治急非淋白血病(ANLL)病人细胞进行体外药敏试验,20例同时有体内治疗结果资料。体内外结果比较表明,病人使用的治疗药物体外敏感,临床72.7%缓解,其中4例同时有二种治疗药物体外敏感,临床均获得缓解;只有一种治疗药物体外敏感。7例中4例缓解。9例病人所有治疗药物体外试验均不敏感,8例不缓解。因此本组病人体外药敏测定能够联系临床结果,并提示如体外选择一或一种以上敏感药物治疗,有可能显著提高白血病人的治疗缓解率。     

钙调素拮抗剂与三尖杉酯碱合用增强对HL—60细胞的抗癌活性

三尖杉酯碱(harringlonine Har)对治疗急性非淋巴细胞白血病有良好疗效,是一种较好的抗癌药物.但由于Har对心脏和造血组织的毒性作用而限制了用药剂量和疗效的提高.用钙调素拮抗剂处理白血病、肝癌等肿瘤细胞,可抑制其增殖.钙调素拮抗剂W_7对HL—60细胞有明显抑制作用.但W_7与Har合用,是否能增强抑制作用,尚未见报道.本实验表明W_7与Har单用和联合用,对体外HL—60细胞抑制作用的对比研究,初步结果报导如下:     

体外短期药敏试验预测急性白血病化疗反应

作者用体外短期药敏试验测定50例急性白血病对药物的敏感性。病人的白血病细胞分别与HH、Adr、Ara-C、VCR和Pred等培养2小时,接着加入~3H-UR继续培养1小时。药物的抑制作用以~3H-UR的掺入抑制百分率SI(％)来表示。对38例在体外和体内均用过HH的病人作了回顾性——相关性研究,发现如HH的SI>15％时,在体内敏感,<15％则抗药,体外与体内的相符率为87％(33/38).我们认为使用~3H-UR掺八的体外短期药敏试验比使用~3H-TdR更为可靠.在急性白血病化疗前可采用此法预测疗效.     

肾上腺素影响小鼠粒巨噬系祖细胞生长的实验研究

目的了解肾上腺素(Adrenaline,Adr)对小鼠粒巨噬系祖细胞的影响.方法在体外半固体培养的粒-巨噬系集落形成单位(Colony Form Unit-Granulocyte and Macrophage,CFU-GM)中加入不同浓度的Adr后,计数集落数,并与阴性对照组比较.结果Adr浓度在10-6～10-10mol/ml范围内,CFU-GM集落数与对照组相比,P＞0.05;Adr浓度为10-5mol/ml时,P＜0.05;Adr浓度为10-4mol/ml时,培养体系中没有CFU-CM形成.结论高浓度Adr可抑制CFU-GM在体外的生长,儿茶酚胺类物质能参与血发生的调节.     

白血病细胞体外培养~3H-TdR掺入法药敏试验临床观察

目前，化疗是治疗白血病的主要方法。化疗方案的选择应遵循个体化的原则，白血病细胞对化疗药物的敏感性是影响疗效的关键因素之一，应用体外药敏试验能合理选用化疗药物。笔者选择阿糖胞着（Ara－C）、三尖杉酯碱（Har）、长春新碱（VCR）和阿克拉霉素（ACR）4种常用化疗药物，用核酸前体物质’H－TdR（氖一胸腺呼唤核昔）掺入法做体外微量短期细胞培养，用掺入抑制率表示对某药物的敏感程度，以指导临床医生选择化疗方案。1材料和方法1．l病例选择25例白血病中男性13例，女性12例。初治12例，复治13例，平均年龄45．5岁。急淋L。型3…     

氮酮促三尖杉酯碱透皮吸收作用的研究

本文研究比较了三种不同浓度的氮酮（Azone）对三尖杉酯碱（Harringtonine,Har）的体外透皮促进作用。实验结果表明，1％、3％和5％Azone对Har均有促透皮吸收作用，而以含3％Azone的Har溶液透皮吸收率最高，这为设计制剂处方时选择最佳Azone量提供了理论依据。     

环孢素A治疗再生障碍性贫血体外药敏试验的初步研究

目的：为预测环孢素A（CSA）治疗再生障碍性贫血（AA）疗效而建立体外药敏试验。方法：采用微量甲基纤维素半固体培养粒一单核细胞集落（GM－CFU），并用CSA干预。结果：体外／体外（S：敏感，R耐药）：S／S11例，R／R10例，R／S：20例，与临床总符合率51．2％，阳性符合率100％，阴性符合率33．3％，试验敏感性35．35％，特异性100％，结论：CSA体外敏感试验与临床疗效有一定相关性，如体外敏感，则体内100％敏感，但存在一定假阴性。     

阿霉素在耐药株LoVo/Adr细胞内的摄入及分布特点

目的观察阿霉素在耐药株LoVo/Adr细胞内的摄入及分布特点,探讨其耐药机制。方法采用流式细胞术检测阿霉素在细胞中的摄入;利用荧光显微镜观察阿霉素在细胞内的分布;采用免疫组化法检测P-糖蛋白(P-gp)的表达。结果LoVo/Adr细胞内阿霉素含量显著低于LoVo细胞,且阿霉素在核区分布明显减少,相对在胞质中增多;而在敏感LoVo细胞中,阿霉素集中分布在核区,胞质中很少。加入维拉帕米后,不仅LoVo/Adr细胞对阿霉素的蓄积能力显著增强,而且可使阿霉素在LoVo/Adr细胞中的分布恢复到敏感状态。P-gp染色在LoVo细胞株均为阴性,而耐药株LoVo/Adr细胞膜呈强阳性。结论阿霉素在耐药细胞中不仅摄入减少,而且分布异常,主要与P-gp有关,P-gp是药物耐药的机制之一。     

乙酰阿地咪逆转肿瘤细胞多药耐药及其机制的研究

目的 探讨乙酰阿地咪体外逆转多药耐药细胞株人乳腺癌耐阿霉素细胞（MCF-7/Adr）以及人非小细胞肺癌耐阿霉素细胞（A549/Adr）的耐药作用及其作用机理。方法 采用乳酸脱氢酶释放法检测乙酰阿地咪与阿霉素共处理MCF-7、A549、MCF-7/Adr以及A549/Adr细胞的细胞毒作用,以观察增敏和逆转耐药作用;用全功能酶标仪测定乙酰阿地咪与阿霉素共同处理MCF-7/Adr和A549/Adr细胞后细胞内积累阿霉素的荧光强度,并采用Western blot检测细胞膜上多药耐药蛋白P-糖蛋白（P-gp）表达的变化。结果 乙酰阿地咪能降低MCF-7/Adr和A549/Adr对阿霉素的耐药性,10 μmol/L的乙酰阿地咪对MCF-7/Adr和A549/Adr逆转倍数分别为10.8、20.1,其逆转作用与乙酰阿地咪呈剂量依赖关系,且对MCF-7和A549细胞也具有增敏作用;随着乙酰阿地咪浓度的增加,MCF-7/Adr和A549/Adr细胞内积累阿霉素的荧光强度增加率增加,呈剂量依赖性;经乙酰阿地咪处理的MCF-7/Adr和A549/Adr细胞P-gp蛋白表达水平降低,且降低水平随乙酰阿地咪浓度增加而更明显。结论 乙酰阿地咪可显著逆转肿瘤细胞的多药耐药性,其机理与抑制多药耐药蛋白P-gp的表达有关,提示乙酰阿地咪具有潜在的克服肿瘤化疗中多药耐药现象的应用价值。     

G-CSF和GM-CSF对髓系白血病细胞增殖的影响

目的 了解粒细胞集落刺激因子（Ｇ－ＣＳＦ）和粒细胞－巨噬细胞集落刺激因子（ＧＭ－ＣＳＦ）对髓系白血病细胞增殖的影响。方法 采用甲基纤维素半固体培养法培养白血病细胞集落（ＣＦＵ－Ｌ）,观察Ｇ－ＣＳＦ和ＧＭ－ＣＳＦ对１１例ＡＭＬ和ＣＭＬ患者外周血ＣＦＵ－Ｌ形成的影响。结果 ＣＭ－ＣＳＦ对ＣＦＵ－Ｌ的促进作用明显高于Ｇ－ＣＳＦ（Ｐ＝０．０１２）,而Ｇ－ＣＳＦ对ＣＦＵ－Ｌ形成影响与对照组没有显著性差别（Ｐ＝０．４９５）。结论 Ｇ－ＣＳＦ对髓系白血病细胞体外增殖无明显影响,作为髓系白血病大剂量化疗的支持治疗可能更为安全。     

益气养阴方对小鼠白血病祖细胞及化疗药物敏感性的影响

通过测定 L_(7212)小鼠白血病祖细胞(CFU—L)集落形成率及在化疗药体外综合药敏实验基础上加用一定量中药,发现益气养阴方能降低 CFU—L 集落形成率和提高体外化疗药物的敏感性。提示此方能抑制白血病细胞和协同化疗药杀伤白血病细胞。清热解毒方也有抑制白血病细胞的作用。     

重组人肿瘤坏死因子对白血病细胞生长抑制的研究

应用ＭＴＴ比色法研究了重组人肿瘤坏死因子（ｒｈＴＮＦα）对白血病细胞株ＨＬ６０和Ｋ５６２生长的抑制作用。实验结果表明，ｒｈＴＮＦα对白血病细胞生长的抑制作用有时相和剂量依赖关系。ｒｈＴＮＦα与ＨＬ６０和Ｋ５６２细胞分别培养１２ｈ，抑制率仅有６．２％和４．３％。当作用时间延长到２４ｈ时，抑制度分别达到５５．４％和４７．１％。ｒｈＴＮＦα的用量由０．０４μｇ／２×１０５细胞增加到５μｇ／２×１０５细胞时，对ＨＬ６０和Ｋ５６２细胞的生长抑制率也分别由１６．８％和６．４％增长到７７．１％和５９．３％。上述两种依赖关系的实验数据经单因素方差处理，有显著性差异。ｒｈＴＮＰα同抗肿瘤药Ｖｐ１６合用对白血病细胞生长的抑制有叠加作用。ｒｈＴＮＦα加入急性粒细胞白血病患者骨髓细胞培养体系后，对白血病细胞集落形成单位（ＣＦＵ－Ｌ）的生长有一定抑制作用。当ｒｈＴＮＦα为１μｇ／５×１０５白血病细胞时，对ＣＦＵ－Ｌ的抑制率可达４３．４％。     

The Com prehensive Evaluation on Four Indices of Drug Re-sistance in Acute Myeloid Leukem ia

Ｔｈｅｄｒｕｇｒｅｓｉｓｔａｎｃｅｏｆａｃｕｔｅｍｙｅｌｏｇｅ－ｎｏｕｓｌｅｕｋｅｍｉｃｃｅｌｌｓ（ＡＭＬ）ｉｓａｍａｉｎｃａｕｓｅｏｆｔｒｅａｔｍｅｎｔｆａｉｌｕｒｅ．Ｔｈｅａｆｆｅｃｔｉｎｇｆａｃｔｏｒｓｉｎｖｏｌｖｅｄｉｎｔｒａｃｅｌｌｕｌａｒｄｒｕｇｒｅｓｉｓｔａｎｃｅ，ｋｉ－ｎｅｔｉｃｓｏｆａｎｔｉ－ｃａｎｃｅｒｄｒｕｇ，ａｂｉｌｉｔｙｏｆｄｒｕｇｄｉｆｆｕｓｉｏｎａｎｄｃａｐａｃｉｔｙｏｆｌｅｕｋｅｍｉｃｃｅｌｌｇｒｏｗｔｈ，ｅｔｃ．［１］．Ｔｈｅｄｅｔｅｃｔｉｏｎｏｆｄｒｕｇｒｅ－…     

NEOPLASTIC TRANSFORMATION AND CYTOGENETIC ABERRATIONS OF RAT TRACHEAL EPITHELIAL CELLS INDUCED BY a-PARTICLE IRRADIATION

Primary rat tracheal epithelium (RTE) cells can be transformed in vitro by α-particles irradiation. One out of 3 isolated morphologically transformed colonies gave rise to an immortal cell line and ultimately became neoplastic by successive passaglng. Cytogenetic analysis was performed on the cell line in order to understand how the specific chromosome alterations participate in the process of neoplastic transformation of RTE cells. The 5th, 18th, 39th passage cells and_ the colony cells isolated from soft agar (SA) culturing of the 40th passage cells were analyzed. The results showed that the 20th passage cells were nontumorigenlc,but the 40th passage cells were tumorigenic. The increase of the length of the long arm of chromosome 15(15q^ ) was frequent at the 5th passage (67. 8% ), especially in SA cells in which it occurred in a clonal fashion (100%). Therefore, 15q might be the critical gene lesion after α-particles irradiation. Monosomy of chromosome 8 that occurred frequently at passage 5 (54%) and 40 (53%) might be correlated with earlier transformation of RTE cells. In addition, three marker chromosomes (MI, M2 and M3)showed a passage-dependent increase. In particular M1 and M2 that were highly frequent at passage 39 (90%) suggested that they might play an important role in the process of cell transformation.     

组胺受体激动剂和拮抗剂对粒单系祖细胞增殖的影响

用体外琼脂培养法研究了2-噻唑乙胺、dimaprit及甲氰咪胍对小鼠CFU—GM产率的影响。结果表明:10~(-8)M～10~(-4)M的2-噻唑乙胺不影响CFU—GM产率,而10~(-8)M的dimaprit即可明显增加CFU—GM产率。此作用随剂量的递增而加强,至10~(-5)M时,增至最大值,此后不再增高。单用甲氰咪胍不影响CFU—GM产率,但伍用dimaprit时,甲氰咪胍阻断dimaprit的上述作用;二者对CFU—GM产率的影响呈现竞争现象。分析了该结果的理论意义和临床意义。     

苦参抗白血病作用及其机制

By using CFU-GM/CFU-L colony assay, NBT/MTT reductant test and DNA fragmentation analysis, we studied the effects of Sophora flavescens (SF) on CFU-GM proliferative ratio in human normal bone marrow/umbilical cord blood and on proliferation, differentiation and apoptosis in human acute myelogenous leukemia HL-60 cells. The results showed that 5, 10, 15, 20 micrograms.microliter-1 of SF significantly inhibited proliferation and induced apoptosis in the HL-60 cells in a dose-dependent fashion. Fifteen micrograms.microliter-1 of SF also induced differentiation in HL-60 cells. Furthermore, cytotoxic activity of SF(5-15 micrograms.microliter-1) was not apparent on human normal hematopoietic progenitors(CFU-GM). The results indicate that an appropriate concentration of SF has a selective antileukemic effect. Thus, these are important impetuses for further research of SF as an anticancer agent.     

Study on the relationship between the Bcl-2/Bax ratio and the growth types of leukemic cells and drug resistance in acute myelogenous leukemia

Summary The ratio of Bcl-2 to Bax (Bcl-2/Bax) in 40 patients with acute myelogenous leukemia (AMD were determined. At the same time, the CFU-L of AML patients and drug resistance were detected by cell culture and MTT assay. The results showed that Bcl-2/Bax in AML was significantly higher than that in normal control (P<0.001). Bcl-2/Bax ratio in colony growth group was higher than that in no-growth group (P <0. 05). Between drug resistance group and drug sensitivity group there was a significant difference in Bcl-2/Bax ratio (P <0. 05). There were more cases of drug resistance in high ratio group (H) than in low ratio group (L) (P <0.05). Meanwhile, Complete Remission (CR) rate in group H was obviously lower than that in group L (P <0.05) and patients in group H tended to have poor response. The above results indicated that the alteration of Bcl-2 and Bax is an important mechanism in the pathogenesis, progress and the development of drug resistance in AML. The determination of Bcl-2/Bax ratio has far-reaching implication in the treatment, choice of chemotherapeutic agents, and prediction of prognosis.     

组胺和甲氰咪胍对粒-单系祖细胞的影响

用体外琼脂培养法研究组胺和甲氰咪胍对小鼠粒-单系祖细胞的影响,结果表明,10~(-9)M～10~(-5)M 组胺和10~(-8)M～10~(-4)M 甲氰咪胍分别作用时,对粒-单系祖细胞的产率没有影响,用甲氰咪胍阻断 H_2-受体后,组胺可增加粒-单系祖细胞的产率和自杀率。对此结果的理论意义和临床意义进行了讨论。     

The effects of total saponins of panax ginseng on hematopoietic progenitor cells in healthy humans and aplastic anemia patients

Ginseng is said to have beneficial effects on anemia. The proliferation effects of total saponins of Panax ginseng (TSPG) on hematopoietic progenitor cell in healthy individuals and 29 patients with aplastic anemia (AA) were observed through bone marrow cultures of burst forming unit-erythroid (BFU-E), colony forming unit-erythroid (CFU-E) and colony forming unit-granulocyte/macrophage (CFU-GM) in vitro compared with methyltestosterone (MT). The results suggest TSPG might prompt the proliferation of normal progenitor cells at a concentration of 20 μg/ml. The numbers of BFU-E,CFU-E and CFU-GM increased by 37.8±2.9%, 31.4±2.9% and 33.3±4.0% respectively overthe controls; furthermore TSPG was still useful to BFU-E, CFU-E growth without Epo in vitro, although the colony numbers were much lower. Otherwise MT was useless to CFU-GM. Of the 29 patients with AA, 14 who responded to MT showed sensitivity to TSPG in marrow culture (the rising rate of colony formation exceeded 30%), but immune-mediated AA (patient’s peripheral blood mononucleated cell suppressed normal hematopoiesis) and stem cell decreased AA (few of colonies were formed) showed almost no expression for TSPG activity because of the immunological suppression system and the absence of progenitors.