氨磷汀对急性放射病小鼠早期骨髓造血功能的防护作用

本研究旨在探究氨磷汀（WR2721,Amifosfine）对60coY射线6．5Gy单次全身照射（TBI）所致急性放射病小鼠早期骨髓造血的防护作用。60只C57／BL6J小鼠随机分成正常对照（normal）、照射对照（IR）和WR2721预防给药（WR2721）3组。观察WR-2721照射前30min给药对6．5Cy60C01射线照射小鼠照射后60d内小鼠外周血白细胞（WBC）、中性粒细胞（Neut）、血小板（Pit）和红细胞（RBC）数的影响;照射后2h和24h计数骨髓有核细胞数（BMNC）、流式细胞术分析造血干／祖细胞（LSK／LK）数量和观察多系骨髓细胞集落形成能力。结果显示,WR2721组的WBC和Neut在照射后4—18d、Pit数7—18d、RBC数10—30d均明显高于瓜组（P〈0．05）;照射后24hBMNC、LSK和LK数均明显增加（P〈0．05）;2h和24h髓系祖细胞集落形成单位（CFU—GEMM）、粒系-巨噬系集落形成单位（CFU—GM）、巨核系集落形成单位（CFU—MK）、红系暴式集落形成单位（BFU-E）和红系集落形成单位（CFU-E）集落数均明显增加（P〈0．01）。结论：WR2721能有效减轻急性放射病小鼠造血系统早期损伤,促进外周血细胞更早更快恢复。     

化疗对造血微环境的影响及化疗后回输自体骨髓基质细胞对造血功能恢复的作用

目的观察化疗对造血祖细胞、造血微环境的影响,化疗后回输体外扩增的自体骨髓基质细胞(ABMSC)对造血功能恢复的作用.方法对化疗患者进行骨髓CFU-GM、CFU-E、BFU-E及基质细胞集落(CFU-F)培养、长期骨髓培养,观察基质层融合情况,计算基质层覆盖率及完全融合时间.长期化疗的10例患者进行自身前后对照,在同一化疗方案下,单纯化疗与化疗后回输体外扩增的ABMSC[(1.1～8.7)×103/次]作对比,观察两者造血恢复情况.结果①长期化疗组的CFU-GM、BFU-E、CFU-E、CFU-F显著低于正常对照组及短期化疗组,后两组差异无显著性;②三组基质层覆盖率及完全融合时间差异无显著性;③长期化疗组ABMSC输注后的CFU-GM、CFU-E、BFU-E、CFU-F显著高于未输注组;④输注ABMSC后白细胞计数及血小板计数降至最低点的值显著高于未输注组,前者白细胞计数及血小板计数恢复正常时间较后者明显缩短;⑤基质细胞回输过程及输后临床观察无不良反应.结论长期化疗明显损伤造血祖细胞、基质祖细胞,但对骨髓基质细胞融合功能无明显影响,化疗后给予ABMSC可加速造血功能的恢复.     

骨髓基质细胞系QXMSC_1体外促进造血及其作用机理的研究

本实验研究了骨髓基质细胞系QXMSC_1体外对CFU-F,CFU-GM,BFU-E及CFU-E集落形成的影响,并通过建立QXMSC_1基质细胞与骨髓造血细胞的长期共培养体系,研究了QXMSC-1在体外促进造血的机理。结果表明,QXMSC_1基质细胞条件培养上清液可促进小鼠CFU-F,CFU-GM,BFU-E及CFU-E集落的生长。在无GM-CSF和Epo存在的情况下,QXMSC_1细胞能维持CFU-GM,BFU-E和CFU-E集落生长。表明QXMSC_1细胞本身可分泌GM-CSF和Epo。QXMSC_1细胞与骨髓造血细胞共培养时,可明显提高CFU-GM集落数。共培养体系中,用微孔滤膜隔断QXMSC_1与造血细胞的直接接触,也能促进CFU-GM集落生成,但比混合培养时低,说明QXMSC_1通过直接接触和分泌细胞因子促进造血的过程。     

川芎嗪促进同基因骨髓移植小鼠骨髓造血重建机制的探讨

本研究探讨川芎嗪对骨髓移植（BMT）后小鼠骨髓中干细胞因子（stem cell factor，SCF）mRNA表达水平的影响及其促进骨髓造血重建的机制。BMT后统计小鼠存活率和计数脾集落形成单位（CFU—S），并采用RT—PCR方法从mRNA水平动态检测骨髓基质细胞中SCF的表达。结果表明：川芎嗪组小鼠在BMT后第10天CFU—S计数、第7、14、21天存活率及川芎嗪组干细胞因子的表达水平均显著高于对照组（p〈0．01或p〈0．05）。结论：）川芎嗪能够促进骨髓造血细胞的恢复，改善骨髓微环境，促进骨髓造血重建。     

不同剂量TPO对小鼠骨髓MSC增殖的影响

为了探讨不同剂量血小板生成素（TPO）对小鼠骨髓间充质干细胞（MSC）增殖的影响，将20只昆明小鼠（35±5g）随机分为低、中、高3个剂量实验组与对照组：给实验组分别腹腔注射TPO25、50和100μg／kg，而给对照组腹腔注射生理盐水0．1ml／g，每日1次，连用5日：每组分别于最后1次注射后12小时收集小鼠骨髓，计数骨髓有核细胞数（BMNC），以10^6／cm^2接种、培养并计数原代成纤维样细胞集落形成单位（CFU—F），同时对其进行成骨、成脂肪诱导分化，用流式细胞术检测BMNC中CD90^＋、CD105^＋、CD34^＋细胞比例并鉴定CFU—F的表型。结果显示：与对照组相比，实验组所获得的BMNC、CD90^＋、CD105^＋、CD34^＋细胞比例和CFU—F集落数明显增加（P〈0．05）。3个剂量中以50μg/kgTPO组增加最明显，但50μg/kg组的CFU—F集落数与100μg/kgTPO组的CFU—F集落数相比，差异无统计学意义（P〉0．05）。CFU—F样MSC具有成骨、成脂肪分化的能力。结论：TPO促进BMNC数、CD90^＋、CD105^＋细胞数和CFU—F集落数增多，即促进骨髓MSC的增殖，但是TPO促进骨髓MSC增殖的作用不随剂号的增加而增加．     

中药对骨髓造血功能重建作用机理的研究现状

骨髓是出生后的主要造血器官。骨髓造血既需要一定数量和功能正常的适血干细胞(HSC)，又需要有利于其生长发育的造血微环境。HSC具有分化和增殖成粒-单系集落形成单位(CFu—GM)、红系爆式集落形成单位(BFU—E)、红系集落形成单位(CFU-E)、混合集落形成单位(CFU—GEMM)等多系祖细胞的功能，并与所处的微环境有密切的关系。骨髓造血微环境由基质细胞、细胞     

IL-6基因转染的骨髓基质细胞系对骨髓移植小鼠造血功能恢复的促进作用

目的:探讨白细胞介素6(IL-6)基因转染的骨位基质细胞系QXMSC, IL6对骨髓移植后造血功能的重建作用。方法:将骨髓造血细胞和骨髓基质细胞系一起经尾静脉注射给同系小鼠,建立骨髓移植(BMT)模型。小鼠的造血功能用脾结节(CFU-S)、粒-单系祖细胞(CFU-GM)、红系祖细胞(CFU-E、BFU-E)测定及外周血各项血液学指标来确定。结果:QXMSC_1 IL-6转基因骨髓基质细胞可明显增强BMT小鼠形成的CFU-S、CFU-GM、CFU-E和BFU-E数,促进外周血象的恢复。结论:QXMSC_1 IL-6细胞可明显促进小鼠骨髓移植后早期造血功能重建。     

去卵巢大鼠骨密度变化与骨髓造血功能改变的关系

本研究探讨去卵巢大鼠不同时段骨密度的改变与骨髓造血功能改变的关系。探讨骨质疏松的发生与造血功能的关联。50只3月龄SD雌性大鼠随机分为去卵巢（OVX）组和假手术（Sham）组,分别在4周、8周、12周、16周和20周处死。取左侧股骨测量骨密度。右侧远端股骨干骺端松质骨甲醛固定,常规制片观察骨髓组织病理改变,集落形成实验观察造血干细胞集落形成能力,酶联免疫吸附试验（ELISA）检测血清造血生成因子粒-巨噬细胞集落刺激因子（GM—CSF）的浓度。结果表明,大鼠去卵巢4周还未出现明显的骨量丢失时,其骨髓组织即出现明显的脂肪组织容量增加,大鼠去卵巢8周、12周、16周和20周,与同时段假手术组比较股骨骨密度显著减低（P〈0．05）,同时伴有骨髓造血组织容量减少,脂肪组织容量增高,巨核细胞数目减少,破骨细胞数目和肥大细胞数目显著增加,与假手术组比较均有显著性差异（P〈0．05）。大鼠去卵巢16周后,造血干细胞形成粒-巨噬细胞集落能力明显低于假手术组。大鼠去卵巢12周后,GM—CSF水平明显低于假手术各组（P〈0．05）。结论：去卵巢大鼠骨密度减低的同时存在骨髓造血功能的减低,3月龄SD雌性大鼠去卵巢可以作为造血功能减低的动物模型,有助于临床上诸多造血功能减低的研究。     

骨髓基质细胞促进γ射线照射小鼠的造血恢复

用体外骨髓贴壁细胞长期传代法,建立一株骨髓基质细胞QXMSC1,经鉴定可能为巨噬细胞来源。用5Gyγ射线照射BALB/c小鼠建立造血功能缺损动物模型,将QXMSC1细胞输入到照射小鼠体内,用半固体琼脂法测定粒-巨噬系祖细胞,甲基纤维素法测定红系祖细胞,血细胞自动计数仪测定外周血各项血液学指标。结果表明,在照射后第10天,骨髓基质细胞可增加辐射损伤小鼠骨髓有核细胞数,增加CFU-GM,CFU-E和mBFU-E集落数。在照射后第20天,可促进外周血WBC,RBC,Hct和Hb的恢复。说明骨髓基质细胞QXMSC1可促进辐射损伤小鼠造血功能恢复。此外,QXMSC1细胞条件培养上清液具有刺激体外CFU-GM集落生长的作用。     

骨髓基质细胞移植加快化疗小鼠造血与免疫功能恢复的实验研究

目的 探讨骨髓基质细胞移植对化疗小鼠造血和免疫功能的影响。方法 BABL／C小鼠骨髓基质细胞体外扩增培养，然后移植化疗后的同系小鼠，观察不同时点外周血象、骨髓集落形成单位、骨髓细胞增殖指数和脾细胞增殖反应的变化。结果 输注扩增培养的骨髓基质细胞无毒副反应，移植了骨髓基质细胞的小鼠各项指标到第7天就已恢复至正常水平，而对照组则需14d，两组间差异有统计学意义（P〈O．05）。结论 骨髓基质细胞移植不仅能够加快化疗小鼠造血功能的重建，而且有助于免疫功能的恢复。     

G－CSF对化疗后外周血干细胞动员作用的影响

通过对１１例急性白血病患者单独化疗与化疗后加用粒细胞集落刺激因子（Ｇ－ＣＳＦ）的对比，动态观察了Ｇ－ＣＳＦ对外周血造血干细胞（ＰＢＳＣ）的动员作用。发现化疗后加用Ｇ－ＣＳＦ比单用化疗的粒－巨噬细胞集落形成单位（ＣＦＵ－ＧＭ）增加５．１倍，红系爆式集落形成单位（ＢＦＵ－Ｅ）增加４．５倍。Ｇ－ＣＳＦ还可使ＣＦＵ－ＧＭ＞１００／ｍｌ和ＢＦＵ－Ｅ＞２００／ｍｌ的持续时间延长；化疗后ＣＦＵ－ＧＭ的最高值提早出现，而不影响ＢＦＵ－Ｅ／ＣＦＵ－ＧＭ比值。结果表明，化疗后加用Ｇ－ＣＳＦ可明显提高ＰＢＳＣ的收集效率，Ｇ－ＣＳＦ是一种有效的ＰＢＳＣ动员剂。     

严重型再生障碍性贫血患者骨髓和外周血HLA—DR＋T细胞的变化…

目的：进一步阐明免疫紊乱在严重型再生障碍性贫血（ＳＡＡ）发病中的作用。方法：对１３例初诊ＳＡＡ、７例抗淋巴细胞球蛋白（ＡＬＧ）治疗后恢复期ＳＡＡ（ｒＳＡＡ）患者骨髓和外周血及对照组（包抱９名骨髓对照和１１名外周血对照）的ＨＬＡ＝ＤＲＴ细胞进行工对部分患者和对照组刺激外周务去单核细胞的单个核培养上清液（ＰＨＡ－ＬＹＣＭdisplay status     

人脐血源基质细胞与骨髓基质细胞对造血功能损伤修复作用的比较研究

目的比较人脐血源基质细胞(hUCBDSCs)与骨髓基质细胞(hBMSCs)对造血功能损伤的修复作用。方法取63只BALB/c小鼠均给予6.0 Gy60Co射线全身照射,随机分为hUCBDSCs组(n=21)、hBMSCs组(n=21)和对照组(生理盐水组)(n=21),分别于移植后1、3、5、7、10、14、21 d检测1次外周血细胞,于照射后7和21 d分别制备股骨骨髓涂片观察骨髓细胞形态学改变,收集小鼠骨髓细胞并作集落培养。结果 3组小鼠移植后白细胞、血小板、红细胞在3～5 d降至最低,hUCBDSCs组白细胞和血小板均从7 d开始回升,10 d后回升速度与程度提高,21 d时回升接近未照射水平,hBMSCs组从7 d开始回升,回升趋势与hUCBDSCs组相似,21 d后回升程度低于未照射水平,对照组从7 d后开始回升,回升较慢,21 d回升程度低于实验组(P<0.05);hUCBDSCs组,hBMSCs组及对照组红细胞均在3 d时降至最低,21 d后恢复,但实验组较对照组回升更快,(P<0.05),hUCBDSCs组与hBM-SCs无甚差异(P>0.05)。集落培养CFU-E、BFU-E、CFU-GM、CFU-GMEM,hUCBDSCs组21 d为(113±6)、(167±6)、(131±6)、(14±4)个,hBMSCs组(46±6)、(141±11)、(118±18)、(10±1)个,对照组为(87±3)、(122±4)、(84±7)、(5±1)个(P<0.05)。hUCBDSCs组所有小鼠均存活。结论 hUCBDSCs较hBMSCs修复造血功能损伤和血小板恢复的作用更佳,hUCBDSCs移植是1种安全有效的促进造血损伤修复的方法。     

Hemopoietic colony growth-promoting activities in the plasma of bone marrow transplant recipients.

Plasma samples were obtained from 34 bone marrow transplant (BMT) recipients before and after administration of the preparative regimen and tested for their ability to promote and/or support growth of hemopoietic colonies. The ability of plasma samples to promote colony formation on their own was tested on normal nonadherent target cells without addition of exogenous growth factors. The growth-supporting activity was examined in the presence of medium conditioned by phytohemagglutinin-stimulated leukocytes (PHA-LCM) and/or erythropoietin (EPO). A series of kinetic changes was routinely observed. Pretransplant samples rarely gave rise to colonies without addition of exogenous growth factors. Plasma samples obtained after completion of the preparative regimen demonstrated increments of growth-promoting activities for megakaryocyte and granulocyte-macrophage progenitors (CFU-Meg and CFU-GM), respectively, that peaked between 7 and 21 d after transplantation. By day 30, activity levels of some patients had returned to pretransplant values, whereas in other patients, activities remained elevated. Persisting activity levels were associated with delayed engraftment. In contrast, activities for progenitors committed to erythropoiesis (BFU-E) and pluripotent precursors (CFU-GEMM) were only rarely observed. The activities were independent of febrile episodes. Their growth-promoting influence on CFU-GM could be neutralized completely by anti-granulocyte-macrophage colony-stimulating factor (GM-CSF) antibodies. These data suggest that at least some of the observed activities in post-BMT plasma are related to GM-CSF. The growth-supporting activities of pretransplant plasma samples are lower than normal plasma when tested on CFU-Meg and CFU-GM. The growth-supporting activities improved transiently within the first month after BMT. A decline during the second and third month was followed by a gradual return to activity levels that were comparable to normal plasma. The effects of these plasma samples on BFU-E and CFU-GEMM were assessed with PHA-LCM and EPO. Similar to CFU-Meg- and CFU-GM-supporting capabilities, they improved transiently after BMT with a return of normal support function after 5-6 mo. The observed endogenous production of growth-promoting and growth-supporting activities for hemopoietic progenitors may serve as a background to design clinical trials for the timely administration of recombinant hemopoietic growth factors to BMT recipients.     

短程大剂量粒细胞集落刺激因子动员外周血造血干细胞的研究

研究短程大剂量粒细胞集落刺激因子对外周血造血干细胞的动员作用。方法采用短程大剂量Ｇ－ＣＳＦ对１１例患者进行外周血造血干细胞动员，Ｇ－ＣＳＦ５μｇ／ｋｇ皮下注射，每日２次，共３天，动员当天及第４天，分别取骨髓及外周血增生明显活跃，外周血白细胞计数明显升高。     

Effect of corticosteroid treatment on hemopoiesis in vivo and in vitro in a patient with Felty's syndrome.

We have studied the CFU-GM and BFU-E in vitro growth in a neutropenic and anemic patient with Felty's syndrome, either before or one and three months after steroid therapy when neutrophils and erythrocytes returned to normal. Both CFU-GM growth and CSA production were found to be low before therapy, and prednisone was shown to raise them to normal levels. The in vitro growth of BFU-E and the production of BPA by T lymphocytes of the patient were significantly lower than normal when studied before therapy. However, the T lymphocytes incubated in vitro with hydrocortisone regained their ability to stimulate the BFU-E growth. After prednisone therapy both BFU-E growth and BPA production by T lymphocytes returned to normal. Possible pathogenetic mechanisms of impaired granulo- and erythropoiesis in Felty's syndrome are discussed. The in vitro study with hydrocortisone can help to identify steroid-sensitive patients.     

骨髓增生异常综合征患者间充质干细胞的生物学特性及其支持造血的体外研究

为了解骨髓增生异常综合征（MDS）患者间充质干细胞（MSC）的生物学特性,探讨其对体外造血的支持能力,采集MDS的骨髓标本,分离、培养和扩增MSC,进行细胞形态观察和免疫表型、成骨分化及增殖能力检测,并对骨髓细胞进行成纤维细胞集落（CFU-F）形成分析.对MDS患者的MSC进行贴壁培养,接种脐血单个核细胞（MNC）,以观察细胞数和CFU-GM的变化,作为MDS患者MSC支持造血体外实验.结果表明：来自MDS患者的MSC呈典型的成纤维样细胞形态,表型鉴定CD34,CD45阴性,SH2（CD105）,SH3（CD73）,Thy-1（CD90）阳性,体外诱导可向成骨细胞分化,其增殖能力和CFU-F形成能力与源于正常供者的MSC相当.支持造血的体外实验显示,实验组培养上清中的细胞总数和CFU-GM计数在第2周后均显著低于对照组（P＜0.05）.结论：来源于MDS患者骨髓的间充质干细胞的生物学特性与正常供者的MSCs无差异,但其体外支持造血的功能较后者显著减弱.     

Variation of erythroid and myeloid precursors in the marrow and peripheral blood of volunteer subjects infected with human parvovirus (B19).

Infection of normal individuals with human parvovirus (B19) results in a mild disease (erythema infectiosum) but gives rise to aplastic crises in patients with chronic hemolytic anemias. The effects of this disease on hemopoiesis were investigated following intranasal inoculation of the virus into three volunteers. A typical disease ensued with a viremia peaking at 9 d. Marrow morphology 6 d after inoculation appeared normal but at 10 d there was a severe loss of erythroid precursors followed by a 1-2-g drop in hemoglobin, and an increase in serum immunoreactive erythropoietin. Erythroid burst-forming units (BFU-E) from the peripheral blood were considerably reduced, starting at the time of viremia and persisting for 4-8 d depending on the individual. Granulocyte-macrophage colony-forming units (CFU-GM) were also affected but the loss started 2 d later. Both CFU-GM and BFU-E showed a sharp overshoot at recovery. In the marrow, BFU-E and CFU-E were reduced at 6 and 10 d in the individual having the longest period of peripheral progenitor loss. In contrast, there was an increase in BFU-E and CFU-E in the subject with least change in peripheral progenitors. In the third subject, with an intermediate picture, there was a loss at 6 d but an increase at 10 d of erythroid progenitors. It is suggested that the architecture of the marrow might partially isolate progenitors from high titers of virus in the serum and individual variation in this respect might give the results observed.