人参总皂甙对人血细胞生成的研究

从人参中提取有效成份人参总皂甙（ＴＳＰＧ），测定它对人骨髓粒单系祖细胞（ＣＦＵ－ＧＭ），红系祖细胞（ＣＦＵ－Ｅ，ＢＦＵ－Ｅ）和多向祖细胞（ＣＦＵ－ＧＥＭＭ）的刺激增殖作用。结果显示：在培养中加入不同浓度的ＴＳＰＧ５．２５，５０，７５，１００ｍｇ／Ｌ，可促进造血细胞增殖，在体外形成集落。在ＴＳＰＧ５－７５ｍｇ／Ｌ时，ＣＦＵ－ＧＭ集落产率均明显高于不加ＴＳＰＧ的对照组浓度的ＴＳＰＧ使集落产率明显高于对     

IFN—α及其联合GM—CSF对CML患者骨髓细胞凋亡相关基因？…

目的：探讨ＩＦＮ－α及ＩＦＮ－α联合ＧＭ－ＣＳＦ调节慢性期慢性粒细胞白血病（ＣＭＬ－ＣＰ）患者骨髓单个核细胞（ＭＮＣｓ）ｂｃｒ－ａｂｌ,ｂｃｌ－２和ｃ－ｍｙｃ基因表达的影响。方法：淋巴细胞分离液离心富集１４例ＣＭＬ－ＣＰ患者骨髓ＭＮＣｓ,经干扰素－α（ＩＦＮ－α）及ＩＦＮ－α联合粒细胞－巨噬细胞集落刺激因子（ＧＭ－ＣＳＦ）培养２４ｈ后,采用相对定量逆转录－聚合酶链反应（ＲＴ－ＰＣＲ）及扩增片段光密     

抗CD3McAb和rhGM—CSF对再障患者CFU—GM生长的体外影响的研究

用ＣＦＵ－ＧＭ单固体琼脂培养技术研究了抗ＣＤ３单克隆抗体（ＭｃＡｂ）和重组人集落刺激因子对再生障碍性贫血（ＡＡ）患者骨髓ＣＦＵ－ＧＭ体外生长进行研究。结果表明：１０μｇ／ｍｌ的抗ＣＤ３ＭｃＡｂ和０．１μｇ／ｍｌ的ｒｈＧＭ－ＣＳＦ均可增加ＡＡ患者ＣＦＵ－ＧＭ的集落数,抗ＣＤ３ＭｃＡｂ作用较ｒｈＧＭ＝ＣＳＦ显著,且不同患者对抗ＣＤ３ＭｃＡｂ和ｒｈＧＭ－ＣＳＦ的敏感性不同,对ＡＡ患者,临床治疗时,应先行     

3种细胞因子在体外对阿糖胞苷细胞毒性的影响

采用对碘硝基四唑（ＩＮＴ）比色法,观察了１７例急性髓性白血病（ＡＭＬ）患者和８例正常人的骨髓,在体外对阿糖胞苷的敏感性,同时分别观察了粒细胞集落刺激因子（Ｇ－ＣＳＦ）,粒－巨噬细胞集落刺激因子（ＧＭ－ＣＳＦ）,白细胞介素－３（ＩＬ－３）３种细胞因子对其敏感性影响。结果显示ＡＭＬ细胞与正常细胞对Ａｒａ－Ｃ敏感性存在显著差异;Ｇ－ＣＳＦ,ＧＭ－ＣＳＦ,ＩＬ－３此３种因子均能增加Ａｒａ－Ｃ对ＡＭＬ细胞的     

脐血血浆,rhGM—CSF增强急性髓性白血病细胞对Ara—C敏感性的…

研究了脐血血浆、基因重组人粒－单细胞集落刺激因子对２０例急性髓性白血病细胞体外对糖胞苷敏感性的影响。采用ＭＴＴ法测定Ａｒａ－Ｃ的细胞毒作用发现，ＣＢＰ及ｒｈＧＭ－ＣＳＦ可显著增强Ａｒａ－Ｃ对耐药ＡＭＬ细胞的毒性作用，与对照组相比，Ｐ〈０．０５；     

重组人粒细胞集落刺激因子在治疗急性髓细胞白血病中的应用

目的：报告重组人粒细胞集落刺激因子（ｒｈＧ－ＣＳＦ）在急性细胞白血病（ＡＭＬ）治疗中的应用及其疗效观察：方法应用中等剂量、短疗程（３－５天）ｒｈＧ－ＣＳＦ治疗ＡＭＬ２８例,并与同期未用ｒｈＧ－ＣＳＦ治疗的ＡＭＬ２８例进行比较。结果：ｒｈＧ－ＣＳＦ能显著缩短化疗后中性粒细胞计数（ＡＮＣ）的低下期及感染的持续时间,同时能明显减少化疗后的输血用量。     

外周血干细胞的变化与化疗因素的影响

为探讨外周血干细胞移植（ＰＢＳＣＴ）时采取干细胞的最适条件，以粒一巨噬细胞集落形成单位（ＣＦＵ－ＧＭ）为指标，对１２例造血系恶性肿瘤化疗后ＰＢＳＣ的变化进行了观察。结果表明：化疗结束约２周后，白细胞数２×１０￣９／Ｌ、中性粒细胞数０．８×１０￣９／Ｌ、血小板数１１０×１０￣９／Ｌ左右时采取于细胞是最适宜的；化疗强度使外周血白细胞数抑制得越低，血象恢复期出现的ＣＦＵ－ＧＭ就越多，外周血象最低时的白细胞数与ＣＦＵ－ＧＭ数之间呈负相关（Ｐ＜０．０５）；随化疗次数的增多，ＣＦＵ－ＧＭ数逐渐减少；两者之间呈负相关（Ｐ＜０．０５）；粒细胞集落刺激因子（Ｇ－ＣＳＦ）可增加ＣＦＵ－ＧＭ数量；化疗后血象恢复期出现的ＣＤ３４阳性细胞与ＤＦＵ－ＧＭ之间呈大致相同的变化趋势。ＣＤ３４阳性细胞与ＣＦＵ－ＧＭ之间的确切关系有待于进一步探讨。     

用WEHI_3－条件培养液代替肺－条件培养液培养CFU－GM

用ＷＥＨＩ＿３－条件培养液（ＷＥＨＩ＿３－ＣＭ）代替个条件培养液（Ｌ－ＣＭ）对小鼠粒单系集落形成单位（ＣＦＵ－ＧＭ）生长的影响进行研究。结果表明，ＷＥＨＩ＿３－ＣＭ对体外小鼠ＣＦＵ－ＧＭ的形成具有明显的刺激作用；ＷＥＨＩ＿３－ＣＭ与Ｌ－ＣＭ均主要刺激混合型集落生长。提示ＷＥＨＩ＿３－ＣＭ内可能含粒单集落刺激因子（ＧＭ－ＣＳＦ）活性。     

用WEHI3—条件培养液代替肺—条件培养液培养CFU—GM

用ＷＥＨＩ３－条件培养液（ＷＥＨＩ３－ＣＭ）代替肺－条件培养液（Ｌ－ＣＭ）对小鼠粒单系集落形成单位（ＣＦＵ－ＧＭ）生长的影响进行研究，结果表明，ＷＥＨＩ３－ＣＭ对体外小鼠ＣＦＵ－ＧＭ的形成具有明显的刺激作用，ＷＥＨＩ３－ＣＭ与Ｌ－ＣＭ均订刺激混合型集落生长，提示ＷＥＨＩ３－ＣＭ内可能含粒单集落刺激因子（ＧＭ－ＣＳＦ）活性。     

粒细胞集落刺激因子在急性髓细胞白血病化疗中的作用

①目的　探讨粒细胞集落刺激因子（Ｇ－ＣＳＦ）在急性髓细胞白血病（ＡＭＬ）化疗中对骨髓造血细胞的作用。②方法　随机选择ＡＭＬ化疗病人５１ 例,其中化疗后应用Ｇ－ＣＳＦ治疗的２７ 例为治疗组,未应用Ｇ－ＣＳＦ治疗的２４ 例为对照组,动态观察骨髓抑制期和恢复期的血细胞变化。③结果　治疗组骨髓抑制期短于对照组,差异有显著意义（ｔ＝ ２．３２,Ｐ＜ ０．０５）。两组缓解率和复发率比较,差异无显著意义（χ２＝ ２．０１０,２．０３１,Ｐ＞ ０．０５）。④结论Ｇ－ＣＳＦ对化疗后骨髓的恢复作用明显。     

Recombinant human G-CSF and retinoic acid in synergistically inducing granulocyte differentiation of human promyelocytic leukemic cells.

The effects of recombinant human G-CSF (rhG-CSF) and retinoic acid (RA) were studied on the proliferation and differentiation of HL-60 cells and human acute myeloid leukemic cells. Synergistic effect on granulocyte differentiation was observed when HL-60 cells and primary acute promyelocytic leukemic cells were cocultured with RA plus rhG-CSF. rhG-CSF combined with RA increased more significantly the percentage of mature cells than RA alone and greatly increased NBT reduction activity (P < 0.001). These results suggested that proliferated effect of rhG-CSF on leukemic cells may be important for inducing differentiation of myeloid leukemic cells. But this effect might expose the patients to the risk of acute myeloblastic leukemia if G-CSF was used alone. However, RA could not only rule out the latter situation but retain former merit as well. The authors suggest that the combined use of G-CSF with RA is probably a new approach to the treatment of leukemia.

     

Granulocyte differentiation of human promyelocytic leukemic cells induced by coordinate action of granulocyte colony stimulating factor and retinoic acid]

We analysed the effects of recombinant human G-CSF (rhG-CSF) and retinoic acid (RA) on proliferation and differentiation of HL-60 cells and human acute myeloid leukemic (AML) cells. A synergistic effect on granulocyte differentiation was observed when HL-60 cells and primary cultured acute promyelocyte leukemic cells were cocultured with 10(-8)mol/L RA plus 1:2000 or 1:1000 rhG-CSF. The rhG-CSF plus RA treated cells demonstrated significant increase in the percentage of mature cells. Morphological changes and nitroblue tetrazolium (NBT) reduction activity evidenced more increase than RA treatment alone (P less than 0.001). The results suggest that RA not only inhibits the proliferative action of G-CSF, but also retains and enhances the action of G-CSF to induce differentiation. Therefore, we believe that the combined use of G-CSF with RA may improve the treatment of leukemia.     

针灸对环磷酰胺荷瘤小鼠血清细胞集落刺激因子含量的影响

目的观察针刺和艾灸前后环磷酰胺(CTX)模型荷瘤小鼠血清中粒-巨噬细胞集落刺激因子(GM-CSF)和粒细胞集落刺激因子(G-CSF)含量的变化,探讨针灸改善荷瘤小鼠骨髓抑制、升高白细胞数的作用机制。方法选用清洁级昆明种雄性小鼠64只,植种S180肉瘤造成移植瘤模型,按体重随机分为空白组、模型组、针刺组、艾灸组,每组16只。其中模型组、针刺组、艾灸组腹腔注射CTX,制备荷瘤小鼠骨髓抑制模型,空白组腹腔注射等量生理盐水。针刺组、艾灸组均选取大椎、膈俞、肾俞、足三里,每天固定时间分别进行针刺、艾灸治疗,空白组、模型组每日陪同抓取、固定,不做任何治疗。均每日1次,连续5 d。各组小鼠分别采尾部静脉血,人工镜检外周血白细胞计数。并分别于治疗3 d和治疗5 d后的第2天,每组各处死8只小鼠,摘取眼球,取其血清,运用酶联免疫吸附试验(ELISA)观察血清中GM-CSF和G-CSF的含量。结果注射CTX的各组荷瘤小鼠白细胞均降低,且第4天降至最低,血清中GM-CSF和G-CSF含量明显下降,与空白组比较有统计学意义(P0.05)。第5天,针刺组与艾灸组白细胞开始回升,血清中的GM-CSF和G-CSF含量升高,与模型组比较有统计学意义(P0.05)。第6天,针刺组、艾灸组白细胞数均已接近基础白细胞水平,血清GM-CSF和G-CSF含量明显升高,与模型组比较有统计学意义(P0.05)。结论CTX造成荷瘤小鼠骨髓造血细胞损伤,导致外周血白细胞减少,且血清GM-CSF和G-CSF的含量降低。针刺和艾灸可以提高CTX荷瘤小鼠血清中GM-CSF和G-CSF的含量,诱导粒细胞前体和巨噬细胞前体细胞呈集落生长,刺激造血细胞分化,以利于髓系造血细胞的增值、分化和成熟,从而缓解化疗后的骨髓抑制,提升外周血白细胞。     

联用重组的IL-3、GM-CSF对阿糖胞苷诱导的HL-60细胞凋亡的影响

为研究在阿糖胞苷（Ara－C）的作用下，基因重组的人白细胞介素3（rh－IL－3）、粒单祖细胞集落刺激因子（rh－GM－CSF）对白血病细胞凋亡的影响，选用了HL－60白血病细胞系，采用DNA电泳、流式细胞仪检测、单克隆抗体C－myc、bcl－2抗原表达的分析以及白血病细胞集落的培养观察方法，观察了0～36h8个不同时相凋亡率与其他指标的变化，表明Ara－C凋亡的作用随药物孵育时间的延长而逐渐增强，凋亡率从0h的1．5％升至36h后的36．4％，而IL－3和GM－CSF能使这种作用明显提高，使凋亡率从7．6％升至19．6％，并能增强Ara－C对白血病细胞的杀灭，引起HL－60细胞C－myc抗原表达下降，而bcl－2抗原表达变化不大，同时发现这两种细胞因子对正常造血细胞影响较小，这为临床上白血病诱导缓解期联用rh－IL－3、rh－GM－CSF和细胞毒药物，提高缓解率，提供了理论依据。     

Effect of Concurrent Use of rh-IL-3 and rh-GM-CSFon Apoptosis of HL-60 Cells Induced by Ara-C

Accordingtotherecentstudy,thechemotherapeuticagentswhichhaveeffectsondifferentlevels0fnucleicacidmetabolismcaninduceapopt0siswhichoc-curredinacuteleukemiccells.Thefinalef-fectofchemotherapeutantdepends0ntwofactors,thegenetoxicity0fdrugsandin-duction0fap0pt0sis.Ara-Cisachem0thera-peutantwh1chaffectstheSphaseincellcy-c1e.ItsuppressesDNAp0lymeraseandnu-cleoside-diph0phate-reductase.Italsoper-meatesam0nggen0t0xicityofDNAandkillstheleukemicceIls.Thehemopoieticgrowthfactors,namelyrh-IL-3andrh-…     

体外造血微环境对L_(801)白血病细胞增殖和分化的影响

本文观察到同系正常小鼠骨髓基质细胞层对L_(801)白血病细胞有明显抑制增殖和促进分化的作用,而基质细胞条件培养液只能抑制L_(801)细胞增殖,不能诱导其分化。提示基质层能产生抑白血病细胞增殖的抑制因子,基质细胞的直接接触或短距离因子的作用与L_(801)的细胞分化有关。这种抑制增殖和促进分化的作用可能不通过GM-CSF。     

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％。     

In vitro proliferation, expansion, and differentiation of a CD34+ cell-enriched hematopoietic cell population from human umbilical cord blood in response to recombinant cytokines

BACKGROUND: The conditions and mechanisms that control the in vitro growth of hematopoietic stem/progenitor cells (contained within the population of CD34+ cells) are still not completely understood. METHODS: By using an immunomagnetic system, we have enriched for umbilical cord blood (UCB)-derived CD34+ cells (55% of total cells recovered vs. 0.8% of total cells prior to the enrichment procedure) and analyzed their in vitro growth (proliferation, expansion, and differentiation) in a liquid culture system in the absence or presence of different recombinant cytokine combinations. RESULTS: When the selected cells were cultured in the absence of recombinant cytokines, no proliferation or expansion was observed. In the presence of steel factor (SF) and interleukin-6 (IL-6), total cell number was increased nearly fourfold; however, no progenitor cell expansion took place. When cultures were supplemented with SF and IL-6 together with IL-3 and erythropoietin (EPO), a rapid proliferation of the CD34+ -enriched cell population was observed with a selective stimulation of erythropoiesis. However, this stimulation was only transient, suggesting that there was a rapid exhaustion of erythroid progenitor cells within the first 10 days. Significantly higher levels of proliferation and expansion of progenitor cells were observed in the presence of SF, IL-6, GM-CSF, and G-CSF with preferential stimulation of myelopoiesis. Interestingly, such stimulation of myelopoiesis was sustained for the entire culture period (>30 days). The highest levels of proliferation and expansion were observed in the presence of all six cytokines. Under these conditions, erythropoiesis was also sustained only transiently (10 days), whereas myelopoiesis was sustained for >30 days. CONCLUSIONS: This study indicates that significant proliferation and expansion of hematopoietic progenitors can be achieved in vitro when culturing a cell population in which CD34+ cells comprise only >50% of the total cells. Our results also suggest that myeloid progenitors (those responding to GM-CSF and G-CSF) possess higher expansion potentials in vitro than their erythroid counterparts. The methods described here for the enrichment and culture of CD34+ cells may be relevant in the development of protocols for the ex vivo proliferation and expansion of hematopoietic progenitors for transplantation.     

基质细胞衍生因子1对人牙髓干细胞增殖、迁移和成牙本质能力的影响

目的：对比基质细胞衍生因子1（stromal cell-derived factor-1,SDF-1）和粒细胞集落刺激因子（granulocyte colony-stimulating factor,G-CSF）对人牙髓干细胞（dental pulp stem cell, DPSC）的体外增殖、迁移和成牙本质能力的影响。方法：分别在培养基中加入100 μg/L SDF-1或100 μg/L G-CSF,采用细胞计数试剂盒（cell counting kit-8,CCK-8)和集落形成实验（colony-forming unit,CFU）检测SDF-1和G-CSF对DPSC增殖的影响;采用划痕实验和Transwell迁移实验检测两者对DPSC迁移能力的影响;对DPSC进行成牙本质诱导,通过碱性磷酸酶（alkaline phosphatase,ALP）染色、测定ALP活性、茜素红染色和real-time RT-PCR检测成牙本质相关基因的表达,以检测两者对DPSC体外成牙本质能力的影响。结果：SDF-1和G-CSF能够轻度提高DPSC的增殖及集落形成能力,但差异无统计学意义。加入SDF-1或G-CSF的实验组划痕汇合速率明显高于对照组（P<0.01）,但两种因子间差异无统计学意义。Transwell迁移实验中,对照组每视野的迁移细胞数量为(5.0±1.4)个,SDF-1组每视野的迁移细胞数量为(24.3±6.8)个,G-CSF组每视野的迁移细胞数量为(11.8±3.3)个,各组间差异有统计学意义（P<0.05）。经成牙本质诱导后,实验组细胞ALP染色加深,ALP活性上升,矿化结节形成数量增加,成牙本质相关基因的表达均显著高于对照组。结论：SDF 1对DPSC的增殖能力影响不显著,但能明显提高DPSC的迁移能力和成牙本质分化能力,效果优于G-CSF。     

抑癌基因p14~（ARF）对慢性粒细胞白血病细胞增殖、细胞周期及凋亡的影响机制研究

目的探讨抑癌基因p14ARF对慢性粒细胞白血病细胞的增殖、细胞周期及凋亡的的影响机制研究。方法应用慢病毒载体系统,将抑癌基因p14ARF导入慢性粒细胞白血病细胞系及患者来源的慢性粒细胞白血病细胞中,应用WST-8法检测细胞生长存活率,应用流式细胞术检测细胞周期和凋亡情况。结果应用表达p14ARF的VSV-G假构型慢病毒载体可明显抑制慢性粒细胞白血病细胞增殖,诱导细胞凋亡,但对细胞周期没有明显的影响。结论利用p14ARF基因治疗可明显抑制慢性粒细胞白血病细胞的增殖。