Objective To explore the effects of artesunate on the proliferation,cell cycle,apoptosis and apoptosis-related protein of bortezomib-resistant multiple myeloma (MM) cells.Methods Human MM cell line NCI-H929 was treated with bortezomib in a dose-dependent manner to establish a bortezomib-resistant cell line NCI-H929BR.The logarithmic growth phase NCI-H929BR cells were divided into control group(blank culture medium)and two concentration experimental groups (artesunate,25,50 μg · mL-1)and treated for 48 h.The inhibitory role of artesunate on NCI-H929BR proliferation was measured by MTT assay.Cell apoptosis and cell cycle were determined by flow cytometry.The Bcl-2 family proteins (Bcl-2,Bcl-Xl,Bax) were detected by Western blot.Results Bortezomib resistance index of NCI-H929BR is 20.12 times than NCI -H929.After administration artesunate for 48 h,the proliferation rate of NCI-H929BR in the low and high concentration experimental groups were 62.46％,33.48％;there was significant difference compared with control group (all P＜0.05).The artesunate dramatically decreases NCI-H929BR proliferation in a dose-dependent manner (P ＜ 0.05).After treatment for 48 h with artesunate,the proportion of G1 phase cell cycle in high concentration experimental group was increased compared with control group [(51.63 ± 4.02) ％ vs (34.72 ± 2.43) ％,P ＜ 0.05].The proportion of apoptosis cell was increased,early apoptosis in high concentration experimental group and control group were (13.10 ± 1.53)％,(2.32 ± 1.02)％;and late apoptosis in the two groups were (10.26 ±2.42)％,(1.93 ± 0.69) ％;comparison between two groups,the difference had significantly (all P ＜ 0.05).In addition,artesunate significant decreased of Bcl-2 and Bcl-Xl protein expression and increased of Bax protein expression in a dose -dependent manner.Conclusion Artesunate could inhibit NCI-H929BR proliferation,increase apoptosis,cause G0/G1-arrest and down-regulate Bcl-2 and Bcl-Xl expression,while up-regulate Bax expression.

Objective:To investigate the effects of Artesunate on suppressing the proliferation of Bortezomib-resistant multiple myeloma (MM) cell line and to explore its molecular mechanism on reversal of drug resistance.Method:Human MM cell line NCI-H929 was treated with Bortezomib in a dose-dependent manner to establish Bortezomib-resistant cell line NCI-H929BR.The inhibitory role of Artesunate on NCI-H929BR and its reversal effect against Bortezomib-resistance were determined by methylthiazolyldiphenyl-tetrazolium bromide (MTT) assay.Cell apoptosis was determined by flow cytometry,and the protein expression levels of nuclear factor-κB p65 (NF-κB p65),phospho-nuclear factor-κB p65 (NF-κB p-p65),P-glycoprotein (P-gp),B-cell lymphoma/leukemia-2 (Bcl-2) protein and Bcl-2 associated X protein (Bax) were detected by Western blot assay.Result:Bortezomib-resistance index of NCI-H929BR was 20.2 times.Artesunate treatment had significant inhibitory effect on the proliferation of NCI-H929BR and the inhibitory effect was in a concentration-dependent manner.Bortezomib (50 nmol·L-1) alone had less effect on NCI-H929BR proliferation,while the inhibition rate of artesunate (12.5 mg·L-1) alone was (23.53 ± 2.21)％ (P ＜0.05);Bortezomib combined with artesunate treatment had greater inhibitory effect (60.71 ± 3.43) ％ (P ＜ 0.01).Artesunate treatment increased NCI-H929BR apoptosis,down-regulated NF-κB p65,NF-κB p-p65,P-gp and Bcl-2 expression levels,and upregulated Bax expression level in a concentration-dependent manner.Conclusion:Artesunate could inhibit NCI-H929BR proliferation,promote apoptosis and reverse Bortezomib-resistance,and its mechanism may be associated with down-regulating expression levels of NF-κB p65,NF-κB p-p65,P-gp,Bcl-2 and up-regulating expression level of Bax.

Objective To investigate the effect of imatinib on coagulation function ,platelet parameters and fibrinolytic indexes in patients with chronic myelocytic leukemia (CML) in chronic phase.Methods A total of 112 patients with CML admitted to our hospital between February 2006 and August 2016 were selected as the study group ,and another 112 healthy people who were admitted to our hospital for physical examination were selected as the normal control group at the same time .The study group were treated with imatinib .The platelet parameters ,coagulation function and fibrinolytic indexes were detected before treatment and 3 months after treatment.The adverse reactions were observed and recorded .Results Before treatment,the PLT level was significantly higher in the study group than the control group (P<0.05),and after treatment,PLT levels were significantly decreased (P<0.05).There was no significant difference in PCT ,MPV and PDW between the two groups before and after treatment ( P>0.05 ) .The levels of Fbg and D-D were significantly higher in the study group than the control group before treatment ( P<0.05 ) .After treatment ,the levels of Fbg and D-D decreased significantly ( P<0.05 ) .There was no significant difference in PT , TT and aPTT between the two groups before and after treatment ( P>0.05 ) .The non-hematological adverse reactions were mild , and there were hematological adverse reactions ,such as neutropenia in 57 cases ( 50.89%) and thrombocytopenia in 26 cases(23.21%).After timely decreasing the dosage of imatinib and treatment with G-CSF,the symptoms were relieved .Conclusion The treatment of CML with imatinib can significantly reduce the platelet count and has little effect on platelet morphology .It can restore the levels of Fbg and D-D to normal levels and regulate between coagulation and fibrinolytic system .

目的 探讨青蒿琥酯对耐地塞米松人多发性骨髓瘤(MM)细胞株MM.1R增殖的影响,并探讨其可能作用机制.方法 取对数生长期MM.1S、对塞米松敏感的MM细胞株MM.1R,各分为空白对照组、地塞米松组、青蒿琥酯组、地塞米松+青蒿琥酯组,分别加入等量培养液、20 ng/mL的地塞米松、25μg/mL的青蒿琥酯、20 ng/mL的地塞米松+ 25 μg/mL的青蒿琥酯,观测各组细胞增殖情况,计算细胞增殖抑制率.取MM.1R细胞加入终浓度为25 μg/mL的青蒿琥酯,分别于给药前、给药24 h、给药48 h检测细胞核转录因子κB (NF-κB) p65蛋白、P糖蛋白(P-gp)蛋白的表达.结果 给药24h时,MM.1S细胞地塞米松组、青蒿琥酯组、地塞米松+青蒿琥酯组的细胞增殖抑制率分别为20.41％±2.75％、29.07％ ±2.71％、68.92％ ±4.12％,组间两两比较,P均＜0.05;给药24 h,MM.1R细胞地塞米松组、青蒿琥酯组、地塞米松+青蒿琥酯组的细胞增殖抑制率分别对为4.13％±3.10％、27.43％ ±2.67％、60.14 ±3.24％,组间两两比较,P均＜0.05;青蒿琥酯给药前、给药24 h、给药48 h,MM.1R细胞NF-κB p65蛋白的相对表达量分别为0.89±0.21、0.42 ±0.13、0.23±0.08;P-gp蛋白的相对表达量分别为0.86±0.19、0.43 ±0.13、0.25 ±0.09,随干预时间增加,MM.1R细胞NF-κB p65、P-gp蛋白的相对表达量均降低(P均＜0.05).结论 青蒿琥酯可抑制MM.1S以及MM.1R细胞的增殖,与地塞米松联用有协同效应.青蒿琥酯抑制MM.1R细胞增殖的机制可能为下调NF-κB p65、P-gp蛋白的表达.

Objective To investigate the combination effect of artesunate and lenalidomide on the proliferation of bortezomib-resistant multiple myeloma cell line MM .1R, and its molecular mechanism .Methods MM.1R cells were cultured in RPMI 1640 and treated with DMSO (control group), lenalidomide (20 μmol/L), artesunate (25 μg/mL), or lenalidomide (20μmol/L) plus artesunate (25μg/mL), separately.After 24 or 48 h, the cell proliferation rates were assessed by MTT assay, and the NF-κB (P65), Bcl-xl, Mcl-1, p-Bad, and P-gp protein levels were measured by Western blot .Results The inhibitory rate of cell proliferation was 15.16%by lenalidomide for 24 h, 28.28%by lena-lidomide for 48 h, 45.98%by artesunate for 24 h, 17.19%by artesunate for 24 h, 34.53%by lenalidomide plus artesu-nate for 24 h, or 65.16%by lenalidomide plus artesunate for 48 h (P<0.05).NF-κB (P65), Bcl-xl, Mcl-1, p-Bad, and P-gp proteins were down -regulated by lenalidomide plus artesunate treatment in a time -dependent manner . Conclusion Both artesunate and lenalidomide inhibit MM .1R cell proliferation .Combination of artesunate with lenalido-mide can enlarge this effect through down -regulating NF-κB (P65), Bcl-xl, Mcl-1, p-Bad, and P-gp.

目的 探讨尼古丁对脐带间充质干细胞(MSCs)形态学、细胞周期、细胞增殖及凋亡的影响.方法 不同浓度尼古丁作用于MSCs,以相差显微镜及原子力显微镜(Atomic Force Microscope,AFM)观察形态学变化;MTT法分别于24h、48h、72h检测细胞增殖;流式细胞仪法于24 h检测细胞周期与细胞凋亡.结果 尼古丁作用于MSCs后,细胞固缩,细胞膜表面形成空洞及凹陷,微绒毛样突起多见;细胞周期改变,G0/G1期的细胞比例明显增加(P＜0.05),G2期及S期细胞比例则逐渐减少(P＜0.01),周期阻滞随着尼古丁浓度递增而增加;细胞增殖受抑,呈时间剂量依赖性,凋亡率增高,0.5、1、1.5mg/mL的尼古丁作用MSCs 24h后,细胞凋亡率均高于对照组.0.5mg/mL组凋亡率(4.867±0.404)％与对照组(3.300±0.400)％比较,差异无统计学意义(P＞0.05);1mg/mL组凋亡率(10.333±0.961)％、1.5mg/mL组凋亡率(44.367±3.612)％与对照组比较,差异均有统计学意义(P＜0.01).结论 尼古丁使MSCs形态及超微结构发生早期凋亡改变,阻滞细胞周期,抑制细胞增殖且促进其凋亡.

目的探讨急性髓系白血病患者化疗后应用重组人粒细胞集落刺激因子(rhG-CSF)治疗对中性粒细胞碱性磷酸酶活性(NAP)的变化及其临床意义。方法选择97例急性髓系白血病患者作为观察组,检测使用rhG-CSF治疗前后NAP的活性,另选29例急性髓系白血病患者作为对照组,单用常规化疗,化疗前后均未予以rhG-CSF治疗,检测化疗前后NAP的变化。结果观察组与对照组比较,应用rhG-CSF治疗后NAP的活性明显增高,两者比较有显著性差异(p<0.01)。结论 rhG-CSF能明显提高急性髓系白血病患者化疗后NAP的活性,从而提高机体抗感染能力。

Objective To investigate the expression level of regulatory T cells(Tregs) in aplastic anemia patients and its changing in the evolution of the disease,and to explore the rule of Tregs on the pathogenesis of apalastic anemia and the possibility of Tregs as the indicator of prognosis and the effect of treatment. Methods Tregs of peripheral blood mononuclear cells were analyzed before and after immunocompressive therapy in 21 patients with severe aplastic anemia by FACS,and,compared with normal control. Results The quantities of Tregs of severe aplastic anemia reduced dramatically compared with normal healthy,and recovered after effective therapy;non response patients remained the level of Tregs unchanged. Conclusion Tregs may play an important role in maintaining normal hematopoiesis,and the reduction of Tregs may be related to the pathogenesis of aplastic anemia.Meanwhile,Tregs could be looked as indicator to monitor the effectiveness of treatment.

Objective:To observe the effect of Chinese medicine of nourishing yin and clear heat for prevention of chronic graft-versus-host disease(cGVHD) in mice.Methods:Mice in the Chinese medicine groups were administrated with high-dose,middle-dose and low-dose yin-nourishing and heat-clearing Chinese medicine respectively from the day of model induction.Positive control group and negative control group were administrated with the same volume of normal saline.At the 2nd,5th,8th,12th week of model induction,the number of Y chromosome in bone marrow cells was detected for the analysis of to chimerism.The body weight,hair and incidence of cGVHD were observed every 3 days.One hundred days after model induction,the pathological changes of mice skin,liver and intestine were observed.Results:Stable chimerism formed in all Chinese medicine groups and positive control group.Incidence of cGVHD in high-dose and middle-dose Chinese medicine groups was lower than that in low-dose Chinese medicine group and positive control group.As for the symptomatic grading and pathological grading,statistical difference was found between high-dose Chinese medicine group and positive control group while not found between low-dose Chinese medicine group and positive control group.Conclusion:Appropriate dose of Chinese medicine of nourishing yin and clear heat can prevent cGVHD in mice with haploidentical bone marrow transplantation.

目的观察30例健康供者在应用rhG-CSF动员前后外周血中性粒细胞形态有无改变。方法注射rhG-CSF前后采集静脉血制备血涂片,pH6.4～6.8条件下瑞特-姬姆萨混合染色后在油镜下观察中性粒细胞形态,同时进行中性粒细胞碱性磷酸酶(NAP)染色、pH5.4条件下姬氏染色观察中毒颗粒。结果应用rhG-CSF前1 d中性粒细胞形态无明显异常,中毒颗粒、NAP阳性率及积分值均在正常范围内;应用rhG-CSF后中毒颗粒、NAP阳性率及积分明显增高,96%的中性粒细胞胞体偏大,胞浆内见大量"中毒"性颗粒、D hle小体、空泡数量增加。结论应用rhG-CSF可引起外周血中性粒细胞出现明显的形态变化。

例1,男,2岁1个月,1岁时出现丑陋面容,智力和体格发育迟缓,患儿头大呈舟状,脐疝,肝肋缘下5 cm,脾肋缘下3 cm.手足宽而短、肥厚.X线摄片显示肋骨胸骨端增宽,如飘带状;脊柱后突,胸腰椎呈鸟嘴状突出,手掌指骨粗短呈弹头样改变.尿甲苯胺蓝斑点试验阳性.

CD4+CD25+Treg细胞具有免疫失能活性,经体外非特异性和体内特异性抗原激活后,通过细胞接触依赖或细胞因子非依赖方式抑制自身反应性T细胞或效应性T细胞增殖/应答反应,发挥免疫抑制活性,进而控制过度的免疫应答、诱导自身免疫耐受、维持免疫稳态,具体的作用机制尚在进一步研究及探讨中.