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Objective To observe the early changes of related indexes after high dose of 60Co γ-ray irradiation on rhesus monkey hematopoietic system.Methods A total of 33 rhesus monkeys were randomly divided into normal control and different irradiation control group,rhesus monkeys in irradiation control group were given different doses(4,8,12 Gy) irradiation to establish acute radiation sickness(ARS) models.XE-2100 automatic blood cell analyzer detected the peripheral blood before and after the irradiation of 3,6,9,12,24,48,80 h.The rhesus monkeys were sacrificed to have a observation of sternum pathological changes at 6,48 and 80 h after 4,8,12 Gy 60Co γ-ray irradiation.Results The number of white blood cell in peripheral blood of the rhesus monkeys after 4 and 8 Gy 60Co γ-ray irradiation were lower than that before irradiation at 3 h after irradiation,as was significant increased at 6 h after irradiation,the highest values were 136.04%.and 221.38% after 9 h(with before irradiation values was 100.00%,the same below),become obviously drooped from 12 h after irradiation,show clearly temporary peak.But the number of white blood cell after 12 Gy 60Co γ-ray irradiation was significant increased at 6 h after irradiation,at the highest of 9 h,become obviously drooped from 12 h after irradiation.Peripheral blood neutrophile count was significant increased at 6 h after irradiation,at the highest of 9 h,become obviously drooped from 12 h after irradiation.Peripheral blood lymphocyte count fell sharply after irradiation,3 h detection value was only 12.02%-25.04% of before irradiation.Sternal bone marrow nucleated cell number decreased sharply after irradiation,the more irradiation dose,the less residual hematopoietic cells.Conclusion In the early stage of BM-ARS,temporary peaktime node of the white blood cell and neutrophil count could be regarded as the best delivery time of hematopoietic cytokine therapy.
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Objective To study the radioprotective potential of CBLB502 protein against hemopoiesis injury by irradiat-ion. Methods C57BL/6J mice were assigned to normal irradiation, and CBLB502 0.2mg/kg 30min pre-irradiation group at random before 6.5 Gy 60Co-γray total body irradiation (TBI). The peripheral blood was obtained to assay hemogram pre and post-irradiation. The bone marrow nucleated cell counts were evaluated in mice at 2 h and 24 h after irradiation. Colony-forming unit (CFU) assay was used to analyze the alteration of hemopoietic progenitor cells in bone marrow. Competitive repopulation assay was conducted to observe the implantation ability of bone marrow cells. The percentage of each lineage hemopoietic cells was measured by flow cytometry analysis. Results CBLB502 significantly alleviated the sharp decrease of peripheral blood counts including leukocyte (WBC), erythrocyte (RBC), and platelet, and accelerated recovery. Counts of various hematopoietic progenitor cell colonies of mouse bone marrow in CBLB502 group were significant higher than those of irradiation group (P<0.01). CBLB502 significantly improved the implantation ability of bone marrow cells. Conclusion CBLB502 showed obvious radioprotective effects on haemopoiesis injury by irradiation.
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Objective To observe the effect of different doses of HS6101 on the recovery of hematopoietic injury in ICR mice treated with cyclophosphamide (CTX). Methods Normal ICR mice were intraperitoneally injected with CTX at 100 mg/kg once a day for 3 consecutive days,and the mouse model of chemotherapy-induced hematopoietic injury was established. Three groups of mice (with 20 per group),were respectively injected with HS6101 at 0,9 or 27 μg subcutaneously at one hour before the first administration of CTX. The peripheral blood cell counts of the mice were observed once every 2 days. Hematopoietic progenitor cell colony counting and histopathological assessment of bone marrow cells were evaluated at 4 d and 9 d after the first administration of CTX. Results In ICR mice after chemotherapy with CTX,all doses of HS6101 significantly increased peripheral leukocytes and neutrophils (P<0.05),elevated the number of multilineage hematopoietic progenitor cell colonies of bone marrow,and stimulated the proliferation of bone marrow cells after CTX injury. Mice receiving 27 μg HS6101 were better than those of the other two groups. Conclusion HS6101 at 27 μg could significantly promote the recovery of hematopoiesis in ICR mice treated with CTX chemotherapy.
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Objective To evaluate the effects of combined administration of recombinant human interleukin-11(rhIL-11),recombinant human G-CSF(rhG-CSF)and recombinant human interleukin-2 (rhIL-2)on acute radiation sickness(ARS)beagles.Methods Sixteen beagle were irradiated with 4.5 Gy60 Co γ-rays to establish ARS models,and were divided into irradiation control group,supportive care group and combined cytokines treatment group.After irradiation irradiation control group was given no treatment,the dogs in supportive care group received purely symptomatic treatment,while combined cytokines treatment group received rhIL-11 50μg/(kg·d)and rbG-CSF 10μg/(kg·d)subcutaneously(0-14 d)and rhIL-2 1×1 06 U/d(29-43 d)besides symptomatic treatment.Manifestation and characteristics of ARS beagles were observed,and the survival time were recorded.At last,post-mortem examination and histological examination were performed.Results All animals underwent nausea,diarrhea and fever.After irradiation,all animals in irradiation control group died in two weeks,and the mean survival time was 12.7 d,while only one died at 33 d in supportive care group.All dogs in combined cytokine group survived at 45 day after exposure,and their haematopoiesis and gastrointestinal tract were recovered.Conclusions Combination of rhIL-11 + rhG-CSF + rhIL-2 treatment could be significantly effective on ARS beagles irradiated by 4.5 Gy60 Co γ-rays,which could accelerate injured haemotopoiesis and intestinal tract recovery,increase the survival rate and improve the life quality of animals.
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Objective To explore the mechanisms of cytokines on acute radiation disease in irradiated beagles.Methods The sera of beagles irradiated with 4.5 Gy γ-rays with cytokines treatment was collected at different time points post irradiation.The two-dimensional gel electrophoresis(2-DE)was used to isolate and compare the differentially expressed proteins in sera.HD-MS was used to analyze the differentially expressed proteins with significance,and the amino acid sequences should be determined. Results High resolution 2-DE gel map was obtained.There were six differentially expressed proteins in sera of irradiated beagles at different time points.Four protein spots were successfully identified by MS.A significant spot was identified as serum amyloid A(SAA)by HD-MS,with relative molecular mass of 13 077 and isoelectfie point of 6.26.Expression of SAA was not found 1 d pre-irradiation and 36 d postirradiation,but increased slightly 1 d(0.2166)and significantly 14 d post-irradiation(0.4577). Conclusions The expression of serum amyloid A was consistent with the process of acute radiation injury,which might indicate the turnover of the disease.
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Objective To explore the clotting mechanism in beagles irradiated by 4.5 Gy γ-rays after treatment with supportive care,or supportive care and combined cytokines.Methods Sixteen beagles were divided into irradiation control group,Supportive care group and combined cytokines treatment group.Platelet aggregation test,thrombelagtography (TEG) and the time measurement were analyzed in vitro.Results In irradiation group and supportive care group,the platelet aggregation rates in beagles were decreased markedly and the k value of TEG was increased 7 d post-irradiation,while those indexes in combined cytokines treatment group changed little.At 14 d post-irradiation,each parameter of TEG in irradiated group changed obviously.The values of r,k,r+k and M were elevated significantly,clotting time and the maximum coagulation time of thrombus delayed,the Ma value was decreased markedly,and the maximum elasticity amplitude of thrombus was diminished.All parameters in combined cytokines treatment group were better than those in supportive care group.The thrombin time was prolonged obviously in irradiated group 14 d post-irradiation,while the thrombin time was the longest at 2-3 weeks post irradiation in supportive care group and combined cytokines treatment group(P>0.05).Conclusions Cytokines could improve the platelet aggregation and the blood clotting functions of beagles suffering from acute radiation sickness.
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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.
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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.
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To evaluate the effects of recombinant human stem cell factor on mobilization of peripheral blood stem cells, 30 monkeys were divided into control, rhG CSF 10?g?kg -1 ?d -1 (?KD), rhSCF 50?KD, rhG CSF 10?KD+rhSCF 20?KD, rhG CSF 10?KD+rhSCF 50?KD and rhG CSF 10?KD+rhSCF 125?KD groups. The monkeys were given subcutaneous injections of rhSCF for 8 days and/or rhG CSF for 5 days. The highest value of leukocyte in rhG CSF and rhSCF groups was 206% and 200% respectively, and that of rhG CSF+rhSCF groups was 280% to 310%. CFU GM of peripheral blood was up to 1 1, 1 3 and 5 8～7 9 times respectively after the mobilization. CD34 positive cell number of joint mobilization group was 20, 24 and 35 times respectively of that before mobilization. rhSCF can mobilize the effect of peripheral blood stem cells, and the effect of joint rhSCF+rhG CSF is better.
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Objective To reproduce a simple and practicable animal model of rheumatoid arthritis(RA)for evaluating the therapeutic effects of recombinant human interleukin-1 receptor antagonist(rhIL-1Ra).Methods The animal model of RA was reproduced in Wistar rats by intradermic injection of an admixture of bovine type Ⅱcollagen and Freund's complete adjuvant on the foot pad,tail,and back of the rats and the injections were repeated 7 days later.Results Two weeks after the second injection,all the extremities were markedly swollen.The diameter of ankle joints,the level of TNF-? in serum,and the size of the paws were increased obviously.There was destruction and absorption of the bone trabeculae of distal end of the tibia.There was also necrosis and fibrosis of cartilage of the joints.The articular space was narrowed.Thus RA model was successfully reproduced in rat.Conclusion Bovine type Ⅱcollagen and Freund's complete adjuvant were a favorable agent for the reproduction of RA model in rats.
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Objective To evaluate the therapeutic effects of combined recombinant human IL-11(rhIL-11) and recombinant human G-CSF (rhG-CSF) on the neutron-irradiated dogs. Methods 18 male beagle dogs were divided into radiation control group (C, n=7), symptomatic treatment group (S, n=4), and combination therapy group (T, n=7). All dogs were exposed to total body 2Gy 90% neutron radiation. From the first day after radiation, the animals of group T received rhG-CSF 10?g/(kg?d) and rhIL-11 50?g/(kg?d) subeutaneously for 14d and 21d respectively. The cell counts of peripheral blood and CFU-GM of bone marrow were carried out. Results All animals of group S and T survived, however, the survival rate of group C was only 57%. The cellcounts of T group peripheral blood cells (white blood cell at any time point , the platelet and red blood cell of recovery phase) were higher than that of C or S group. The count of T group bone marrow CFU-GM was 6 fold higher than that of group C or S on day 1, and still 1.75, 1.46 fold higher than that group C or S, respectively, on day 33. Conclusion the combination therapy of rhIL-11 and rhG-CSF significantly raised the white cell and platelet counts in ARS dogs induced by neutron irradiation by accelerating the recovery of bone marrow hematopoietic function.
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Objective To observe the curative effect of rhG-CSF or rhG-CSF+rhSCF mobilized peripheral blood stem cells (PBSC) on rhesus monkeys after irradiation of 7.0Gy ? ray.MethodsFifteen healthy adult rhesus monkeys were divided into placebo control, rhG-CSF, and rhG-CSF+rhSCF mobilized groups. rhSCF 20?g?kg -1 ?d -1 (?KD) was administered for 8 consecutive days. Then from the 4th day to the 8th day, animals of both rhG-CSF and rhG-CSF+rhSCF mobilized groups were also given rhG-CSF 10?KD. The placebo control animals were given 0.9% sodium chloride of the same volume per kilogram. On day 0 of the irradiation, PBSCs were collected from all subjects. 3～4 hrs after irradiation, all animals received autologous PBSC infusion. ResultsFrom day 21 to day 29 after TBI, rhG-CSF+rhSCF mobilized group showed higher WBC counts compared to placebo control and rhG-CSF mobilized group. Platelet counts of both mobilized groups recovered more quickly than those of placebo control. Bone marrow nucleated cells culture demonstrated that rhG-CSF and rhG-CSF+rhSCF mobilized PBSC had stimulated bone marrow nucleated cells to form more CFUs. Histopathological evaluation showed the number of hematopoietic cells in the bone marrow of rhG-CSF mobilized group was greater than those in placebo control but less than those in rhG-CSF+rhSCF mobilized group. ConclusionThe infusion of rhG-CSF or rhG-CSF+rhSCF mobilized PBSC can improve the hematopoietic recovery in rhesus monkeys after 7.0Gy ? ray irradiation. The combined use of two mobilizing factors gives better result than using single factor.