HPLC-ELSD法同时测定复方丹参缓释片中丹参酮ⅡA和人参皂苷Rb1含量

目的建立同时测定复方丹参缓释片中丹参酮ⅡA和人参皂苷Rb1含量的方法。方法采用高效液相色谱-蒸发光散射检测法。色谱柱为Agilent C18(250mm×4.6mm,5μm),乙腈为流动相A,0.02%磷酸溶液为流动相B,梯度洗脱(0~5min,20%~25%流动相A;5~30min,25%~45%流动相A),流速:1.0mL.min-1,蒸发光散射检测器,氮气流量1.6L.min-1,漂移管温度70℃。结果丹参酮ⅡA和人参皂苷Rb1分别在7.295~116.7μg.mL-1和4.063~65.00μg.mL-1的范围内呈良好线性关系,平均回收率分别为96.70%和98.92%,RSD分别为1.46%和0.93%。结论该方法操作简单、快速、准确,重复性好,可用于复方丹参缓释片的质量控制。     

保守性手术治疗输卵管妊娠的疗效及生育结局

目的探讨药物治疗、保守性手术、根治性手术三种方式治疗输卵管妊娠的疗效及生育结局。方法该院2008年1月—2010年1月共接诊的242例输卵管妊娠患者,分别予保守性治疗(药物治疗及保守性手术)及根治性输卵管切除术治疗,比较不同治疗方案的治愈率、住院时间、血β-HCG降至正常时间、输卵管通畅率、宫内妊娠率及再次异位妊娠率。结果药物治疗的治愈率最低,而住院时间、血β-HCG降至正常时间最长,输卵管通畅率最高(P0.05);保守性手术的宫内妊娠率最高(P0.05);再次异位妊娠率三者无统计学差异,均在10%左右。结论保守性手术可保留输卵管的完整性,同时提高宫内妊娠率,生育结局最为乐观。对于有生育要求的输卵管患者而言,在患者自身条件允许的情况下,保守性手术是最佳治疗方法。     

15例Gastaut型特发性儿童枕叶癫痫的临床特征及预后

目的：探讨Gastaut型特发性儿童枕叶癫痫的临床特征及预后情况。方法：对确诊为迟发型枕叶癫痫的15例患儿应用抗癫痫药物进行治疗,随诊并分析其临床特点、脑电图特征及预后情况。结果：发病平均年龄为9．1岁,男9例,女6例,临床特征为全部以全面性强直阵挛发作来就诊,追问病史后,常在全面性强直阵挛发作前1～2年有视幻觉、头眼偏斜及偏头痛等症状。简单视幻觉11例（73％）,发作性视盲或视力模糊8例（5a％）,头痛7例（47％）,眼球偏转10例（67％）。发作间期脑电图示枕部导联为主的后头部棘波放电,单侧或双侧枕部导联出现,左右同步或不同步,常为闭眼诱发,睁眼抑制,伴同侧后颞导联棘波活动11例（73％）,弥漫性棘波放电3例（20％）。大部分单药治疗有效,部分需要联合用药。13例（87％）服药期间未再出现临床发作,2例服药后发作减少,但不能完全控制发作,其中有4例患儿服药后超过3年以上未再出现临床发作,并有停药后再次复发病史。结论：Gastaut型枕叶癫痫起病较晚,其临床表现较有特征,日间视觉症状突出,大多发作频繁,脑电图以枕导棘波放电为特点,抗癫痫药物控制效果较好,但停药后易复发,其归类为良性儿童癫痫综合征存在一定争议。     

细胞因子联合对4.5Gyγ射线照射比格犬造血损伤的治疗作用

目的 观察细胞因子组合对4.5 Gy γ射线照射比格犬造血系统损伤的治疗效果,为极重度骨髓型急性放射病的临床救治提供实验依据。方法 16只比格犬均给予4.5 Gy ⁶⁰Co γ射线全身照射,随机分为照射对照、综合对症和细胞因子3组。细胞因子组动物在综合对症支持治疗的基础上应用rhG-CSF、rhIL-11和rhIL-2联合治疗。2 d检测1次外周血象,分别于照射前4 d,照射后1和45 d收集骨髓和外周血进行造血细胞集落培养,制备胸骨病理切片观察组织形态学改变。结果 照射后各组动物外周血各类细胞数急剧下降,细胞因子联合治疗可提高白细胞最低值(1.04×10⁹/L,而照射对照组和综合对症组分别为0.28×10⁹/L和0.68×10⁹/L),缩短血小板减少持续时间(细胞因子组24 d,综合对症组33 d),使红细胞维持在正常值范围;照射后1 d骨髓及外周血中造血干细胞集落形成率明显下降,照射后45 d 2个治疗组造血干细胞集落数均恢复为照射前水平;照射对照组动物骨髓造血细胞完全消失,细胞因子治疗使得骨髓造血功能完全恢复,与照射前水平比较差异无统计学意义。结论 rhG-CSF、rhIL-11和rhIL-2联合应用可提高极期时外周血白细胞最低值,加速白细胞、血小板和红细胞恢复,促进4.5 Gy γ射线照射犬体内残留造血干/祖细胞的增殖、分化和成熟,从而加速造血功能的重建。rhG-CSF、 rhIL-11和rhIL-2不失为治疗极重度骨髓型急性放射病的有效措施。     

Experimental therapeutic effect of combined cytokines on beagle dogs exposed to 4.5 Gy γ-rays

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.     

Initial research of screening for the differentially expressed proteins in beagles irradiated with 4.5 Gy60 CO γ-rays by two-dimensional gel electrophoresis and mass spectrometry

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.     

Clotting mechanism in beagles irradiated by 4.5 Gy γ-rays

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.     

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].