Therapeutic effects of combined cytokines on hematopoietic injuries induced by 4.5 Gy γ-rays irradiation in beagles

Objectivc To observe the therapeutic effects of combined cytokines on hematopoietic injuries induced by 4.5 Gy60 Co γ-rays irradiation in beagles,and to provide experimental evidences for the clinical treatment of extremely severe myeloid acute radiation sickness(ARS).Methods 16 beagles were given 4.5 Gy60 Co γ-rays total body irradiation,and then randomly assigned into irradiation control group,supportive care group and cytokines group.In addition to supportive care,recombinant human granulocyte colony-stimulating factor (rhG-CSF),recombinant human interleukin-11(rhIL-11)and recombinant human interleukin-2(rhIL-2)were administered subcutaneouly to dogs in cytokines group.Peripheral blood hemogram was examined once every two days.Bone marrow and peripheral blood were collected to proceed colony cultivation 4 d pre-irradiation and 1 and 45 d post-irradiation.Conventional histopathological sections of sternum were prepared to observe the histomorphology changes. Results After irradiation,the population of all kinds of cells in peripheral blood declined sharply.WBC nadir Was elevated(1.04×109/L,but 0.28×109/L and 0.68×109/L for the irradiation control group and the supportive care group separately),the duration of thrombocytopenia was shortened (24 days,but 33 days for the supportive care groug) and red blood cell counts were maintained in the range of normal values after cytokincs treatment in combination.The colony forming efficiency of haemopoietic stem cells(HSCs)in bone marrow and peripheral blood decreased obviously 1 d post irradiation,but recovered to the level of that before irradiation 45 d post irradiation after supportive care and cytokines treatment.Hematopoietic cells disappeared in bone marrow of animals in irradiation control group,but hematopoietic functions were recovered after cytokines were administrated.Conclusions RhG-CSF.rhIL-11 and rhIL-2 used in combination could elevate WBC nadir,accelerate the recovery of leukocytes,platelets and red blood cells and promote the proliferation,differentiation and maturity of HSPCs left in the body after 4.5 Gy γ-rays total body irradiation,eventually restore the hematopoietic function.Hence,combination of rhG-CSF,rhIL-11 and rhIL-2 could serve as better therapeutic strategy to treat extremely severe myeloid ARS.     

细胞因子组合治疗急性放射损伤犬的组织病理学观察

急性放射性损伤的主要病理变化不仅是骨髓造血功能受到严重抑制,外周血白细胞和血小板极度缺乏,由此引起感染和出血,同时,放射性损伤还累及消化、呼吸、泌尿和生殖等多个系统,导致体内多个系统器官的损伤"[1-2].研究发现,动物放射性损伤后体内多种细胞因子、化学增活素和细胞外基质表达水平增高,以保护受损脏器的损伤"[3-4].而采用细胞因子治疗急性放射性损伤能促进造血功能的恢复[5].     

PEG IFN α-1b重复给药对恒河猴的毒性实验

目的评价PEG IFN α-1b对恒河猴多次皮下注射给药的毒性作用,提供PEG IFN α-1b对恒河猴毒性反应的靶器官及其损害的性质,确定PEG IFN α-1b的无毒反应剂量,为制定临床用药方案提供参考资料。方法24只成年健康恒河猴(雌雄各半)分为溶剂对照、PEG IFN α-1b600、100和15μg/kg4组,每组雌、雄动物各3只。各组动物隔天一次分别皮下注射溶剂或不同浓度的PEG IFNα-1b各2ml,90d共注射45次。PEG IFN α-1b给药量分别相当于成人一次剂量的230、30和6倍。结果给药期间所有动物反应无异常,无死亡。动物的体重、体温、血压、呼吸、尿常规及凝血指标无异常。30项外周血象结果表明,给药期间各组动物外周血白细胞及中性粒细胞数减少,以大剂量组尤为明显,减少幅度约50%;单核细胞有升高,血小板数有降低的趋势,但各剂量组间无明显的量效关系。给药期间所有3个剂量PEG IFN α-1b组动物的16项血浆生化指标与同期溶剂对照组比较,差异无统计学意义。各组动物血清免疫球蛋白IgA、IgG、IgM和IgE检查结果与药前值比较,差异均无统计学意义。3个不同剂量PEG IFN α-1b给药后20d,血清中结合抗体滴度均开始明显升高,以大剂量组尤为明显。此后仍呈波动性升高的趋势,至停药后30d仍处峰值。PEG IFN α-1b皮下注射90d和停药后30d3个给药组均未查到中和抗体的产生。组织病理学检查结果显示大剂量PEG IFN α-1b给药组动物给药90d处死猴肾小球毛细血管减少,基底膜增厚,间质细胞增生;胸腺小叶蜕化,细胞数减少,胸腺脂肪化,停药后30d,上述病理变化消失,未见异常改变;肝中央静脉旁可见淋巴小结形成,肝细胞出现散在大片状水肿,胞体肿胀,但未见到肝细胞坏死现象,停药30d时可见个别动物仍有肝细胞广泛性水肿;骨髓细胞明显疏松,数量明显减少,可见多发性细胞坏死小灶,伴有明显地髓腔内出血。PEG IFN α-1b中、小剂量及溶剂对照组未见上述异常病理改变。结论PEG IFN α-1b100和15μg/kg隔天一次、连续90d皮下注射恒河猴给药期间除外周血白细胞、血小板数降低外,其他各项指标包括组织病理学检查均无明显异常改变。PEG IFN α-1b600μg/kg组动物给药期间外周血白细胞和血小板数降低至药前值的52%和80%,肾小球毛细血管减少,间质细胞增生;肝细胞胞体肿胀;骨髓细胞疏松,数量减少,可见有细胞坏死小灶。因此,在临床应用中,尤其是在大剂量、长期给药时,应密切观察肝、肾功能指标及外周血象。     

Studies on mechanism of treatment of granulocyte colony-stimulating factor,recombinant human interleukin-11 and recombinant human interleukin-2 on hematopoietic injuries induced by 4.5 Gy γ-rays irradiation in beagles

Objective To investigate the mechanism of treatment of granulocyte colony-stimulating factor(rhG-CSF),recombinant human interleukin-11(rhIL-11)and recombinant human interleukin-2 (rhIL-2)on hematopoietic injuries induced by 4.5 Gy60 Coγ-ray irradiation in beagles,and to provide experimental evidence for the clinical treatment of extremely severe myeloid acute radiation sickness (ARS).Methods Sixteen beagle dogs were given 4.5 Gy60 Co γ-ray total body irradiation(TBI),then randomly assigned into irradiation control group,supportive care group or cytokines+supportive care (abbreviated as cytokines)group.In addition to supportive care,rhG-CSF,rhlL-11 and rhIL-2 were administered subcutaneously to treat dogs in cytokines group.The percentage of CD34+cells,cell cycle and apoptosis of nucleated cells in peripheral blood were examined by Flow cytometry.Results After 4.5 Gy 60 Co γ-ray irradiation,the CD34+cells in peripheral blood declined obviously(61.3%and 52.1% of baseline for irradiation control and supportive care group separately).The cell proportion of nucleated cells in Go/G1 phase was increased notably(99.27% and 99.49% respectively).The rate of apoptosis(26.93% and 21.29% separately)and necrosis(3.27% and 4.14%,respectively)of nucleated cells were elevated significantly when compared with values before irradiation(P<0.05) 1 d post irradiation.When beagles were treated with cytokines and supportive care,the CD34+cells in peripheral blood were markedly increased(135.6% of baseline).The effect of G0/G1 phase blockage of nucleated cells became more serious(99.71%).The rate of apoptosis(5.66%)and necrosis(1.60%)of nucleated cells were significantly lower than that of irradiation control and supportive care groups 1 d after exposure.Conclusions Cytokines maybe mobilize CD34+cells in bone marrow to peripheral blood,indce cell cycle block at G0/G1 phase and reduce apoptosis,and eventually cure hematopoieticinjuries induced by irradiation.     

蓝莓花青素调控Notch1/Hes-1通路对糖尿病大鼠视网膜神经节细胞氧化损伤的影响及机制研究

目的 探讨蓝莓花青素(BA)对糖尿病大鼠视网膜神经节细胞(RGC)氧化损伤的影响及可能的作用机制。方法将50只雄性SD大鼠随机分为正常组(NC组,n=10)和造模组,造模组一次性腹腔注射链脲佐菌素(STZ)50 mg/kg,构建糖尿病模型,造模成功后再随机分为糖尿病模型组(DM组)、蓝莓花青素组(BA组)、DAPT组(Notch1通路抑制剂)和蓝莓花青素+DAPT组(BA+DAPT组),每组10只。BA组每天给予20 mg/kg的BA灌胃,DAPT组给予10μmol/L的DAPT干预,BA+DAPT组给予20 mg/kg的BA灌胃+10μmol/L的DAPT干预,NC组和DM组用等量0.9%氯化钠溶液(生理盐水)灌胃,连续8周。于末次给药24 h后,摘取大鼠眼球,用酶联免疫吸附分析(ELISA)试剂盒检测各组大鼠视网膜组织中活性氧(ROS)、过氧化氢酶(CAT)、超氧化物歧化酶(SOD)和丙二醛(MDA)含量;苏木精-伊红(HE)染色观察大鼠视网膜组织病理变化,并进行RGC计数;用荧光定量聚合酶链式反应(qRT-PCR)测量HO-1和iNOS mRNA表达;Western blot法检...     

不同剂量HS6101对环磷酰胺损伤小鼠造血功能的影响

目的观察不同剂量HS6101对环磷酰胺（cyclophosphamide，CTX）损伤小鼠造血功能恢复的影响。方法正常ICR小鼠连续3 d腹腔注射CTX 100 mg/（kg·d）制备化疗药损伤模型，3组动物于首次CTX前1 d，每只分别皮下注射HS61010、9和27μg，每组动物数分别为20只，观察各组小鼠外周血细胞计数、第4和9天骨髓造血祖细胞集落数和骨髓病理组织学变化。结果不同剂量HS6101均可明显升高CTX化疗小鼠外周血白细胞和中性粒细胞数（P<0.05），增加骨髓各系造血祖细胞集落生成，刺激化疗损伤后骨髓细胞增殖。其中HS610127μg给药组效果更佳。结论 HS610127μg可明显促进CTX化疗ICR小鼠造血功能的恢复。     

重组人粒细胞集落刺激因子融合蛋白对急性放射病小鼠的治疗作用

目的研究重组人血清白蛋白-粒细胞集落刺激因子融合蛋白（GW003）对急性辐射损伤小鼠的治疗作用。方法 80只BALB/c小鼠用60Coγ射线一次全身照射5.0 Gy后随机分为照射对照,GW003 1、3和9 mg/kg 4组,照射后1 h和7 d分别皮下注射生理盐水,GW003 1、3和9 mg/kg。照前1 d和照后30 d内隔日一次检测外周血细胞,照射后30 d处死小鼠取胸骨行组织病理学观察。结果 60Coγ射线5.0 Gy照射后小鼠外周血白细胞、中性粒细胞、淋巴细胞和单核细胞数急剧下降,其中1和3 mg/kg GW003组白细胞和中性粒细胞减少持续时间较照射对照组显著缩短,开始恢复时间提前,白细胞最低值明显升高。GW003 9 mg/kg组上述指标与照射对照组相比有所改善,但无显著差异。3 mg/kg GW003组小鼠单核细胞数于照射后9～13 d明显高于照射对照组。各组间外周血淋巴细胞、血小板及红细胞数无明显差异。胸骨病理组织切片显示照射后30 d各组活存小鼠骨髓造血均十分活跃。结论 GW003对60Coγ射线5.0 Gy照射所致急性放射病小鼠有明显的治疗作用。     

2D-STI检测急性心肌梗死患者经皮冠状动脉介入治疗前后跨壁力学的改变

目的使用超声二维斑点追踪显像技术(two-dimensional speckle tracking imaging,2D-STI)观察左室前壁急性心肌梗死患者(acute myocardial infarction,AMI)经皮冠状动脉介入(PCI)治疗前和成功PCI治疗后心内膜与心外膜下心肌跨壁力学的变化。方法左室前壁AMI患者38例,应用2D-STI获取左心室心内膜、心外膜下心肌的应变-时间曲线,测量梗死及非梗死节段心内膜、心外膜下心肌应变参数:纵向峰值应变(LSpeak)、峰值应变延迟时间(TPSLS),与正常对照组39例进行比较分析。结果 (1)梗死组梗死节段心内膜下心肌及心外膜下心肌LSpeak与梗死组非梗死节段及正常对照组对应节段比较明显减低(P<0.01),TPSLS明显延长,随PCI术后时间的延长,梗死节段心内膜下心肌及心外膜下心肌LSpeak逐渐恢复,TPSLS明显缩短;至成功PCI介入治疗后3个月,心内膜下心肌仍未恢复至正常水平,而心外膜下心肌可恢复至正常水平,与梗死组非梗死节段及正常对照组比较,成功PCI介入治疗后3～7 d和3个月组间差异均有统计学意义(P<0.05)。(2)PCI治疗前、成功PCI介入治疗后3～7 d和3个月:梗死组梗死节段心外膜下心肌LSpeak均高于同期心内膜下心肌LSpeak(P<0.05),梗死节段心外膜下心肌TPSLS均低于同期心内膜下心肌TPSLS(P<0.05)。结论 AMI患者PCI介入治疗前及成功PCI介入治疗后,梗死节段心内膜下心肌与心外膜下心肌间存在跨壁力学改变,2D-STI技术通过测定LSpeak及TPSLS可以发现这一变化。     

肿瘤治疗中的一个稳定靶标

由于肿瘤细胞的遗传不稳定性,肿瘤抗原并非癌症治疗的理想靶标。相反,与肿瘤有关的成纤维细胞因其遗传稳定性而成为倍受关注的靶点。Re-isfeld等用一种DNA疫苗拮抗成纤维细胞激活蛋白(FAP),结果显示,此疫苗单独使用或联合化疗均可抑制肿瘤细胞的生长。对pFAP DNA疫苗免疫的小鼠     

补肾壮骨丸抗骨质疏松的药效学作用及机制探讨

目的：探讨北京积水潭医院院内制剂补肾壮骨丸抗骨质疏松的药效学作用及其机制。方法6个月龄雌性SD大鼠60只,随机分为正常对照组、模型对照组、补肾壮骨丸低(1.2 g/kg)、中(2.4 g/kg)、高(4.8 g/kg)剂量组以及阳性药雷洛昔芬组,每组10只。双侧卵巢摘除术建立大鼠骨质疏松模型,去势手术1周后,补肾壮骨丸低、中、高剂量组及阳性药组分别灌胃给予不同剂量补肾壮骨丸混悬液及雷洛昔芬混悬液治疗12周,正常及模型对照组灌服等体积的生理盐水。分别检测血清碱性磷酸酶(ALP)、抗酒石酸酸性磷酸酶(TRAP)、甲状旁腺激素(PTH)、促甲状腺激素(CT)以及垂体-性腺轴相关激素雌二醇(E2)、睾酮(T)、黄体生成素(LH)、卵泡刺激素(FSH)含量;测定骨密度、骨生物力学性能。结果去势后,模型对照组血清ALP、TRAP、PTH活性显著升高(P<0.05或P<0.01),含量分别为(16.347±2.957)U/100 mL、(27.691±7.042)U/L、(45.829±17.007)pg/mL,CT水平显著下降,含量为(20.469±7.343)pg/mL,全股骨和股骨头骨密度为(0.1859±0.0137)、(0.1871±0.0193)g/cm2,股骨最大载荷为(63.448±17.540)N,最大挠度为(0.5848±0.1595)mm,均较正常对照组显著降低(P<0.05或P<0.01);性激素E2和T含量显著降低(P<0.01),分别为(5.935±1.621)pg/mL、(3.382±1.405)ng/mL,促性腺激素LH和FSH水平明显上调(P<0.05),含量分别为(12.549±2.376)、(15.841±1.896)mU/mL;与模型对照组比较,补肾壮骨丸中、高剂量组ALP含量为(13.619±2.881)、(12.720±3.044)U/100 mL,TRAP含量为(20.904±6.454)、(19.933±5.988)U/L,PTH含量为(26.753±13.013)、(25.630±15.618)pg/mL,三者活性均显著下调(P<0.05或P<0.01);高剂量组CT含量显著升高,为(28.350±7.827)pg/mL,中、高剂量组全股骨密度及高剂量组股骨头密度显著升高,分别为(0.2081±0.0113)、(0.2135±0.0139)、(0.2304±0.0290)g/cm2,高剂量组股骨最大载荷及挠度分别为(81.314±16.413)N、(0.7358±0.1488)mm,较模型组显著升高(P<0.05);高剂量组E2、T显著升高(P<0.05或P<0.01),含量为(8.799±2.204)pg/mL、(5.086±1.668)ng/mL;中、高LH含量分别为(10.740±1.652)、(10.575±0.886)mU/mL,FSH含量分别为(13.244±2.089)、(13.348±2.170)mU/mL,二者水平均显著下调(均P<0.05)。结论补肾壮骨丸可有效纠正骨代谢异常,增加骨密度,增强生物力学性能,其发挥抗骨质疏松作用的部分机制与其调节垂体-性腺轴相关激素水平有关。