Pathophysiology, clinical features and radiological findings of differentiation syndrome/all-trans-retinoic acid syndrome

In acute promyelocytic leukemia, differentiation thera-py based on all-trans-retinoic acid can be complicated by the development of a differentiation syndrome(DS). DS is a life-threatening complication, characterized by respiratory distress, unexplained fever, weight gain, interstitial lung infiltrates, pleural or pericardial effusions, hypotension and acute renal failure. The diagnosis of DS is made on clinical grounds and has proven to be difficult, because none of the symptoms is pathognomonic for the syndrome without any definitive diagnostic criteria. As DS can have subtle signs and symptoms at presentation but progress rapidly, end-stage DS clinical picture resembles the acute respiratory distress syndrome with extremely poor prognosis; so it is of absolute importance to be conscious of these complications and initiate therapy as soon as it was suspected. The radiologic appearance resembles the typical features of cardiogenic pulmonary edema. Diagnosis of DS remains a great skill for radiologists and haematologist but it is of an utmost importance the cooperation in suspect DS, detect the early signs of DS, examine the patients' behaviour and rapidly detect the complications.

Effects of All-trans Retinoic Acid (ATRA) against Bacterial Infection in Chickens

In order to investigate the effects of all-trans retinoic acid （ATRA） against bacterial infection in chickens, 35 3-day-old AA broiler chickens were fed adaptively for two days and randomly divided into five groups, including Escherichia coli experimental group （ group 1 ）, Escherichia coli control group （group 2）, blank control group （ group 3 ）, PasteureUa experimental group （ group 4）, and PasteureUa control group （ group 5 ）. At 5 days of age, the chickens in group 1 and group 4 were drenched with 5 p.mol/kg ATRA for seven consecutive days according to their weight; the chickens in group 2, group 3 and group 5 were drenched with an equal volume of dimethyl sulfoxlde （DMSO）. The clinical symptoms and weight changes in each group were observed and recorded. Seven days later, the chickens were euthanized and dissected to determine the immune organ indexes. The results showed that there were significant differences in body weight between ATRA-administrated chickens and non-administrated chickens after bacterial infection （P 〈 0.05 ） ; moreover, the immune organ indexes of ATRA-administrated chickens exhibited significant differences compared with control group （P 〈 0.05 ）, indicating that ATRA could promote the development of immune organs of poultry, thereby enhancing the body immunity against bacterial infection.

ATRA调节人异位子宫内膜间质细胞GJIC Connexin基因、蛋白的作用及意义

目的:探讨全反式维甲酸(ATRA)对人异位子宫内膜间质细胞间隙连接细胞间通讯(GJIC)、connexin基因、蛋白的调节作用及意义。方法:取24例子宫内膜异位症患者卵巢处异位内膜标本,分离纯化得到间质细胞,分别用0.1mmol/L、1mmol/L、10mmol/LATRA处理24h、48h、72h、96h、120h,同时设置空白对照组,采用荧光漂白后恢复技术检测异位内膜间质细胞的GJIC功能,并检测与GJIC功能密切相关的Cx43、Cx26、Cx32的基因及蛋白表达。结果:ATRA处理72h内,1mmol/L、10mmol/L组的异位内膜间质细胞荧光恢复功能明显增强;0.1mmol/LATRA组的异位内膜间质细胞荧光恢复功能无明显变化。未经ATRA处理的异位内膜间质细胞中无Cx43、Cx26、Cx32的mRNA及蛋白表达;经1mmol/LATRA处理后,异位内膜间质细胞中检测到Cx43mRNA和蛋白表达,但未见Cx26、Cx32的mRNA及蛋白表达。结论:ATRA能选择性地诱导Cx43基因及蛋白表达,从而有效上调异位内膜间质细胞的GJIC功能;ATRA可能是治疗子宫内膜异位症的潜在药物。

FAM及FAM辅加ATRA方案对胃癌术后患者疗效对比观察

目的对比观察FAM及FAM辅加ATRA方案对胃癌术后患者的疗效。方法将62例胃癌根治性手术(D2、D3)后患者随机分为FAM+ATRA组及FAM组。结果FAM+ATRA组术后第1、2、3年的肿瘤复发转移率分别为16.1%,38.7%和67.7%;FAM组分别为25.8%,54.8%和74.2%。术后FAM+ATRA组1、2、3年生存率分别为93.5%、74.2%和38.7%;FAM组分别为80.7%、61.3%和35.5%。2组术后第1、2年的肿瘤复发转移率和患者1、2年生存率比较均有显著性差异(P<0.05),2组术后第3年肿瘤复发转移率和患者3年生存率比较无显著性差异(P>0.05)。结论FAM辅加ATRA治疗能有效提高胃癌术后患者的近期疗效及1、2年生存率。

HSP90β在ATRA诱导的白血病细胞株HL60耐药中的作用研究

目的:通过检测HSP90β基因在白血病细胞株HL60及其耐药株HL60/ATRA中的表达差异,探讨HSP90β基因在ATRA相关多药耐药性发生中的作用。方法:采用ATRA浓度递增及间歇诱导法建立HL60/ATRA耐药细胞株;用MTT法和流式细胞检测技术(Flow cytometry,FCM),分别检测HL60和HL60/ATRA细胞的增殖及细胞周期;RT-PCR、Western blot法检测HSP90β基因在HL60及HL60/ATRA细胞中的表达;通过MTT法检测HL60和HL60/ATRA细胞对常用化疗药物(VCR、CTX、VP-16、ADM)敏感性。结果:成功构建了白血病细胞耐药株HL60/ATRA;在mRNA和蛋白两个水平,均检测到HSP90β在耐药株HL60/ATRA中的表达较HL60明显增高(P<0.01);药敏结果显示HL60/ATRA细胞对VCR、CTX、ADM、VP-16的耐药倍数分别为10.74、5.51、4.01、3.24。结论:ATRA能够诱导HL60细胞中HSP90β高表达,经ATRA诱导后的HL60/ATRA细胞对抗癌药物的敏感性显著降低、耐药性增高,这提示HSP90β可能在ATRA相关多药耐药性的发生过程中发挥着重要作用。

ATRA对雏鸡抗细菌感染作用的研究

为了研究全反式维甲酸(all-trans retinoic acid,ATRA)对雏鸡抗细菌感染的作用,试验选择3日龄AA肉雏鸡35只,适应性喂养2 d,随机分为5组,即大肠杆菌试验组(1组)、大肠杆菌对照组(2组)、空白对照组(3组)、巴氏杆菌试验组(4组)、巴氏杆菌对照组(5组),5日龄时1,4组雏鸡按体重分别灌服5μmol/kg ATRA,连续灌服7 d;同时2,3,5组雏鸡分别灌服等量的二甲基亚砜(DMSO),观察各组雏鸡临床症状并记录体重变化。7 d后剖检,测定免疫器官指数。结果表明:经细菌感染后,口服ATRA的雏鸡与未口服ATRA的雏鸡体重变化差异显著(P<0.05),而口服ATRA雏鸡的免疫器官指数与对照组相比差异显著(P<0.05)。说明ATRA能够促进家禽免疫器官的发育,进而提高机体免疫力,对抗细菌感染。

ATO,ATRA/G-CSF联合使用加速RAR α降解诱导NB4细胞末端分化

目的:探讨三氧化二砷(ATO)、全反式维甲酸(ATRA)及粒细胞集落刺激因子(G-CSF)(A/G)联合使用,诱导NB4细胞末端分化的分子机理。方法:台盼蓝染色记录细胞生长,吉姆萨染色观察细胞的形态学变化,流式细胞仪分析确定细胞分化分子标志及凋亡,Western blot观察RAR α蛋白的表达。结果:NB4细胞经过ATO+A/G处理,出现形态学末端分化特征和免疫表型的成熟。Western blot结果显示,RAR α基因是ATO特定的靶向目标。结论:ATO联合ATRA/G-CSF能够诱导NB4细胞末端分化,其机理与调节RAR α的表达有关。

8-MOP、ATRA以及二者联合应用对Mc3细胞EGFR表达的影响

目的 :研究 8 -甲氧基补骨脂素 (8-Methoxypsoralen ,8-MOP)、全反式维甲酸 (AllTransRetinoicAcid ,ATRA)以及两者联合应用对粘液表皮样癌高转移克隆细胞株 (The 3rdCloneofMucoepidermoidCarcinomaCellLine - 1,Mc3)细胞表皮生长因子受体 (EpidermalGrowthFactorReceptor,EGFR)表达的影响。 方法 :免疫组织化学染色方法。结果 :EGFR表达分别为 :对照组 (+ + ) ;8-MOP组 (+ ) ;ATRA(+ ) ;8-MOP与ATRA联合用药组 (+ )。结论

ATRA联合HSV1-TK/GCV系统治疗卵巢癌的实验研究

目的探讨全反式维甲酸(all-trans retinoic acid,ATRA)能否提高Ⅰ型单纯疱疹病毒胸苷激酶(herpes simplex virus thymidine kinase,HSV1-TK)/丙氧鸟苷(ganciclovir,GCV)系统对卵巢癌的体内治疗作用。方法用携带tk基因的重组逆转录病毒上清转染人卵巢癌细胞系SKOV-3,用含G418的培养液筛选获得抗性克隆(命名为SKOV-3/TK)。采用PCR方法检测HSV1-tk基因整合情况。分别用SKOV-3细胞建立皮下移植瘤裸鼠模型作为对照组和ATRA组,用SKOV-3细胞与SKOV-3/TK细胞按8∶2比例混合细胞建立的动物模型作为HSV1-tk/GCV组和HSV1-tk/GCV联合AT-RA组。于接种细胞后15天开始实验用药,从用药第1天开始每5天测量肿瘤体积1次,至用药结束后2周,绘制肿瘤生长曲线,计算抑瘤率,并取瘤组织做病理学检查。结果与对照组比较,ATRA组、HSV1-tk/GCV组抑瘤率分别为25.5%、38.8%,联合用药组抑瘤率高达67.8%,有统计学意义,P<0.05。病理结果显示实验组出现不同程度点、片状坏死,以联合用药组为重。结论HSV1-TK/GCV系统对卵巢癌有体内杀伤作用;ARTA可以提高HSV1-TK/GCV系统对卵巢癌的体内杀伤作用。

All-trans retinoic acid upregulates VEGF expression in glioma cells in vitro

All-trans retinoid acid （ATRA） is one of the most potent and most thoroughly studied differentiation inducers that induce the differentiation and apoptosis of glioma cells. However, the effect of ATRA on angiogenesis of glioma re- mains poorly understood. We examined the effect of ATRA on the expression of vascular endothelial growth fac- tor （VEGF） in different glioma cell lines and investigated the underlying mechanism, intending to partially reveal the effects of ATRA on angiogenesis of glioma. Glioma cells were treated by ATRA at 5 and 10 μmol/L. The VEGF mRNA transcript levels were determined by real-time RT-PCR and the protein levels of VEGF in glioma cells were evaluated by Western blotting assays. Moreover, hypoxia-inducible factor-1α （HIF-la） mRNA expression was analyzed by using real-time RT-PCR. After treatment with 5 and 10 μmol/L ATRA, the VEGF mRNA tran- script levels in glioma cells increased remarkably, compared with that in the control group, and the relative protein expression of VEGF was also up-regulated. Meanwhile, the HIF-la mRNA expression also increased. ATRA in- creases the expression of VEGF in glioma cells at both transcriptional and translational levels.

All-trans retinoic acid increases ARPE-19 cell apoptosis via activation of reactive oxygen species and endoplasmic reticulum stress pathways

AIM:To explore the apoptosis of ARPE-19 cells after the treatment with different doses of all-trans-retinoic acid(ATRA).METHODS:ARPE-19 cells were used in the in-vitro experiment.Flow cytometry assay was employed to evaluate the level of reactive oxygen species(ROS)and apoptosis.The effects of ATRA(concentrations from 2.5 to 20μmol/L)on the expression of endoplasmic reticulum stress(ERS)markers in vitro were evaluated by Western blot and realtime quantitative polymerase chain reaction(qRT-PCR)assays.The contribution of ROS and ERS-induced apoptosis in vitro was determined by using N-acetyl-L-cysteine(NAC)and Salubrinal,an antagonist of NAC and ERS,respectively.RESULTS:Flow cytometry showed that ATRA significantly increased ARPE-19 cell apoptosis and ROS levels in each group(F=86.39,P<0.001;F=116.839.P<0.001).Western blot and qRT-PCR revealed that levels of CHOP and BIP were elevated in a concentration-dependent pattern after the cells were incubated with ATRA(2.5-20μmol/L).The upregulation of VEGF-A and CHOP induced by ATRA could be inhibited by NAC(antioxidant)and Salubrinal(ERS inhibitor)in vitro.CONCLUSION:ATRA induces the apoptosis of ARPE-19 cells via activated ROS and ERS signaling pathways.

ATRA在卵巢癌细胞H08910增殖中的作用

目的：探讨全反式雏甲酸在人卵巢癌细胞生长和增殖中的作用机制。方法：选用人卵巢癌细胞株H08910在体外进行培养，培养时添加不同浓度的全反式维甲酸（all—tralns retinoie acid，ATRA），作用24h、48h、72h后，采用MTT比色法测定细胞生长抑制率；采用流式细胞术（flow cytometry，FCM）进行细胞周期分析；采用RT—PCR法检测维甲酸仅受体（retinoic acid receptorct，RARer）mRNA的表达。结果：不同浓度的全反式雏甲酸对H08910细胞生长均起到抑制作用（P〈0．05），并呈浓度和时间依赖关系；能够增加G1期细胞比例，同时降低S、G2／M期细胞比例（P〈0．05），细胞增殖指数PI（P〈0．05）降低；能够上调雏甲酸α受体mRNA的表达（P〈0．05），从而抑制卵巢癌细胞增殖。结论：说明全反式维甲酸对卵巢癌细胞H08910的增殖具有显著的抑制作用，其机制可能是通过上调维甲酸α受体mRNA的表达来实现的。

atRA对原代培养的猪甲状腺细胞生存率及细胞损伤的实验研究

目的 研究全反式维甲酸(atRA)对猪甲状腺细胞生长调节及细胞损伤的作用。方法 采用正常猪甲状腺组织,以细胞单层培养、噻唑蓝(MTT)法及乳酸脱氢酶(LDH)的活性测定,检测不同浓度的atRA (10 -8～10 - 5mol/L)对甲状腺细胞存活率及损伤的影响。结果 atRA呈剂量依赖性抑制甲状腺细胞的生长,亦呈剂量依赖性损伤细胞。结论 一定浓度的atRA对甲状腺细胞具有一定的细胞毒作用。

Apoptosis of Human Pancreatic Carcinoma Cells Induced By All-Trans Retinoic Acid and Interferon

Objective： To investigate the apoptosis of human pancreatic carcinoma PC3 cells induced by the combination of all-trans retinoic acid （ATRA） with interferon alpha （IFN-α）. Methods： PC3 cells were treated with ATRA and IFN-α. The inhibitory rate of PC3 cell proliferation was detected using MTT method. Cellular apoptosis was determined with flow cytometry. The percentage of PC3 cell apoptosis was assayed using TUNEL methods. Results： ATRA and IFN-α could inhibit cellular proliferation and induces cellular apoptosis of PC3 cells. The inhibitory effect was stronger when the ATRA and IFN-α were combined as a therapy. Conclusion： ATRA inhibits the proliferation of PC3 cells and induce the apoptosis of PC3 cells. The combination of IFN-α with ATRA may enhance these effects on PC3 cells.

ATO单用或联合ATRA治疗首发急性早幼粒细胞白血病临床对比研究

目的探讨ATO单用或联合ATRA治疗首发急性早幼粒细胞白血病临床疗效及安全性差异。方法选取收治的首发急性早幼粒细胞白血病患者150例,随机分为对照组和联合组,每组75例,分别采用ATO单用和在此基础上与ATRA联用治疗;比较2组患者完全缓解率,完全缓解所需时间,早期死亡率,随访生存率,毒副作用发生率,治疗后肝功能、心肌酶学及血脂指标水平等。结果 2组患者完全缓解率比较差异无统计学意义(P>0.05);联合组患者完全缓解所需时间显著短于对照组,差异有统计学意义(P<0.05);2组患者早期死亡率和随访1年生存率比较差异无统计学意义(P>0.05);联合组患者随访3年生存率显著高于对照组,差异有统计学意义(P<0.05);2组患者药物毒副作用发生率比较差异无统计学意义(P>0.05);联合组患者肝功能指标水平显著高于对照组,差异有统计学意义(P<0.05);联合组患者心肌酶学指标水平显著低于对照组,差异有统计学意义(P<0.05);2组患者治疗后血脂指标水平比较差异无统计学意义(P>0.05)。结论相较于ATO单用,ATO联合ATRA治疗首发急性早幼粒细胞白血病可有效缩短完全缓解所需时间,提高远期生存率,且未增加不良反应发生风险。

Study of Bcl-2 siRNA Enhancement of Sensitivity of HL-60 Cells to All Trans Retinoic Acid

OBJECTIVE To study whether siRNA targeting against the Bcl-2 gene can enhance sensitivity of HL-60 cells to all trans retinoic acid(ATRA). METHODS siRNA,which is a leading sequence selected by previous experiments,was transferred into HL-60 cells.At 6 h after transfection,the cells were cultured with ATRA.The cell growth of the HL-60 cells was measured by the MTT assay at 24, 48,72 h.The level of the Bcl-2 protein and ROS(reactive oxygen species)as well as membrane potential of the mitochondria were determined by flowcytometry. RESULTS siRNA significantly increased the inhibitory effect of ATRA on growth of the HL-60 cells.The combination of siRNA with ATRA resulted in a decrease in the Bcl-2 protein level and an increase in the ROS level as well as significantly lowering the mitochondrial membrane potential of the HL-60 cells(P<0.05). CONCLUSION Effective siRNA targeting of Bcl-2 increases the sensitivity of HL-60 leukemic cells to ATRA by inhibiting the expression of the Bcl-2 protein.

ATRA DAP方案治疗早幼粒细胞白血病的临床研究

应用ATRA和DAP方案治疗APL50例,ATRA组完全缓解(CR)率92.85％,DAP组CR率54.54％,两组疗效差异显著(P<0.05)。ATRA组CR后必须进行强化治疗,否则易复发。此外对应用ATRA后血细胞的动态变化和造血细胞CFU—GM、CFU—E治疗前后的改变进行了研究。

1,25(OH)_2D_3和ATRA对HO8910生长的协同抑制作用

目的:通过单独或联合使用1,25-二羟维生素D3[1,25-dihydroxy vitaminD3,1,25(OH)2D3]和全反式维甲酸(all trans rinoic acid,ATRA),研究其对人卵巢癌细胞HO8910生长、细胞周期和细胞凋亡的影响,以及维生素D受体(vitamin D recept VDR)及维甲酸α受体受体(retinoic acid receptorα,RARα)mRNA的表达变化,探讨其作用的分子机制。方法:选用人卵巢癌细株HO8910在体外进行培养,培养时分别添加1,25(OH)2D3、ATRA或二者联合,采用MTT比色法测定细胞生长抑制率,流式细胞术(flow cytometry,FCM)进行细胞周期和细胞凋亡分析,用RT-PCR检测VDR及RARαmRNA的表达。结果:结果是单独或联合使用1,25-二羟维生素D3和全反式维甲酸诱导后,HO8910细胞生长均受到不同程度的抑制(P<0.05),并呈时间依赖关系;能够引起细胞周期时相的改变,G1细胞增多(P<0.05);能够诱导细胞产生细胞凋亡(P<0.05);能够上调VDR及RARαmRNA的表达(P<0.05),进而抑卵巢癌细胞增殖,其中以联合用药组最为显著。结论:说明1,25-二羟维生素D3和全反式维甲酸能抑制人卵巢癌细胞株HO8910生长;诱导细胞周期发生G1期阻滞,同时产生细胞凋亡作用;通过上调VDR及RARαmRNA的表达,从而抑制癌细胞增当1,25-二羟维生素D3与全反式维甲酸联合用药时,能够对HO8910细胞生长产生协同抑制作用。

ATRA、DAP方案治疗早幼粒细胞白血病临床和实验研究

应用ＡＴＲＡ和ＤＡＰ方案治疗ＡＰＬ５０例，ＡＴＲＡ组ＣＲ率９２．８５％，ＤＡＰ组ＣＲ率５４．５４％，两组疗效差异显著（Ｐ＜０．０５）。ＡＴＲＡ组ＣＲ后必须进行强化治疗，否则易复发。此外对应用ＡＴＲＡ后血细胞的动态变化和造血干细胞ＧＭ－ＣＦＵ、ＣＦＵ－Ｅ治疗前后的改变进行了研究。

全反式视黄酸调控颌骨骨髓间充质干细胞成骨分化双向效应的体外研究

目的·探究不同浓度全反式视黄酸(all-trans retinoic acid,ATRA)对大鼠颌骨骨髓间充质干细胞(jaw bone marrow mesenchymal stem cells,jBMSCs)成骨分化的影响。方法·通过全骨髓贴壁法分离培养4周龄Sprague-Dawley(SD)大鼠jBMSCs。运用流式细胞术鉴定jBMSCs表面抗原。利用碱性磷酸酶(alkaline phosphatase,ALP)染色/茜素红染色、油红O染色及阿尔辛蓝染色分别对成骨诱导、成脂诱导及成软骨诱导后的jBMSCs进行多向分化潜能检测。分别使用ATRA浓度为0.01、0.1、1、5、10、20μmol/L的成骨诱导液对jBMSCs进行体外成骨诱导,并使用二甲基亚砜(dimethyl sulfoxide,DMSO)作为对照组,利用CCK8进行细胞活力检测。采用ALP染色和茜素红染色对各浓度组jBMSCs的成骨分化能力进行检测,并筛选后续实验浓度。利用实时荧光定量聚合酶链反应(quantitative real-time polymerase chain reaction,qPCR)、免疫荧光染色分析不同浓度ATRA下jBMSCs成骨相关基因mRNA与蛋白表达水平。结果·流式细胞术分析显示,98%以上的P1代jBMSCs表现为CD29+CD90+CD31−CD45−,与骨髓间充质干细胞表面抗原特点相符。ALP染色/茜素红染色、油红O染色及阿尔辛蓝染色结果证明P1代jBMSCs具有成骨、成脂、成软骨多向分化能力。ALP染色/茜素红染色结果显示,0.01、0.1和1μmol/L ATRA组jBMSCs成骨活性和矿化能力较对照组增强,而继续提升ATRA浓度则使成骨活性与矿化能力减弱,浓度高于5μmol/L时开始低于对照组水平(均P<0.05)。qPCR分析发现,0.1、1μmol/L ATRA组成骨相关基因如Alp、骨唾液酸蛋白(bone sialoprotein,Bsp)、Ⅰ型胶原蛋白α1(collagen typeⅠα1,Col1a1)、骨钙素(osteocalcin,Ocn)表达水平较对照组上升,而继续提升ATRA浓度则使基因表达水平下降,ATRA浓度高于5μmol/L时开始低于对照组水平(均P<0.05)。免疫荧光染色结果显示,0.1、1μmol/L ATRA组较对照组成骨相关蛋白成骨细胞特异性转录因子SP7、ALP及OCN表达增强,而继续提升ATRA浓度则使蛋白表达下降,浓度高于5μmol/L时开始低于对照组水平(均P<0.05)。结论·较低浓度(0.1、1μmol/L)ATRA可促进大鼠jBMSCs成骨分化能力,且该促进效应在0.1μmol/L浓度时达到峰值,进一步提升浓度可使该促进效应减弱。较高浓度(5、10、20μmol/L)ATRA对大鼠jBMSCs成骨分化呈现抑制效应。研究在体外证明ATRA对大鼠jBMSCs成骨分化具双向效应,并鉴定了0.1μmol/L ATRA为大鼠jBMSCs成骨分化的最适浓度,为体内研究的开展与全反式视黄酸的临床应用提供了参考依据。