The contribution of c-Jun N-terminal kinase activation and subsequent Bcl-2 phosphorylation to apoptosis induction in human B-cells is dependent on the mode of action of specific stresses

The c-Jun N-terminal kinase (JNK) pathway can play paradoxical roles as either a pro-survival or a pro-cell death pathway depending on type of stress and cell type. The goal of the present study was to determine the role of JNK pathway signaling for regulating B-cell apoptosis in two important but contrasting situations--global proteotoxic damage, induced by arsenite and hyperthermia, versus specific microtubule inhibition, induced by the anti-cancer drug vincristine, using the EW36 B-cell line. This cell line over-expresses the Bcl-2 protein and is a useful model to identify treatments that can overcome multi-drug resistance in lymphoid cells. Exposure of EW36 B-cells to arsenite or lethal hyperthermia resulted in activation of the JNK pathway and induction of apoptosis. However, pharmacological inhibition of the JNK pathway did not inhibit apoptosis, indicating that JNK pathway activation is not required for apoptosis induction by these treatments. In contrast, vincristine treatment of EW36 B-cells resulted in JNK activation and apoptosis that was suppressed by JNK inhibition. A critical difference between the two types of stress treatments was that only vincristine-induced JNK activation resulted in phosphorylation of Bcl-2 at threonine-56, a modification that can block its anti-apoptotic function. Importantly, Bcl-2 phosphorylation was attenuated by JNK inhibition implicating JNK as the upstream kinase. Furthermore, arsenite and hyperthermia treatments activated a p53/p21 pathway associated with apoptosis induction, whereas vincristine did not activate this pathway. These results reveal two stress-activated pathways, one JNK-dependent and another JNK-independent, either of which can bypass Bcl-2 mediated resistance, resulting in cell death.     

Results with Floxuridine,Actinomycin D,Etoposide, and Vincristine in Gestational Trophoblastic Neoplasias with International Federation of Gynecology and Obstetrics Scores ≥5

Background5‐fluorouracil‐based multiagent chemotherapy has been used as the primary treatment for high‐risk gestational trophoblastic neoplasia (GTN) in China for a few decades. This study aims to assess the efficacy and toxicity of floxuridine, actinomycin D, etoposide, and vincristine (FAEV) as a primary treatment for patients with GTN who had International Federation of Gynecology and Obstetrics (FIGO) scores ≥5.Materials and MethodsA total of 207 patients with GTN who had FIGO scores ≥5 were treated with FAEV as first‐line chemotherapy at Peking Union Medical College Hospital between January 2002 and December 2017. Complete remission (CR), resistance, survival, toxicity, and reproductive outcomes were analyzed.ResultsOf the 207 patients treated with FAEV, 9 (4.3%) required a change of chemotherapy owing to toxicity and 1 (0.5%) died of cerebral hernia 5 weeks after commencing treatment. The remaining 197 patients were assessable to determine the response to FAEV; among them, 168 (85.3%) achieved CR with FAEV and 29 (14.7%) developed resistance to FAEV. The 5‐year overall survival rate of the entire cohort was 97.4%. Grade 3–4 neutropenia, thrombocytopenia, and anemia occurred in 28.4%, 6.8%, and 6.2% of cycles, respectively. No acute toxicity‐related deaths occurred. Five patients developed acute myeloid leukemia 10–50 months after exposure to chemotherapy; another patient developed duodenal cancer 2 years after completing therapy. Sixty‐one patients who preserved fertility wanted to become pregnant; 56 of them conceived.ConclusionThe FAEV regimen is an effective primary treatment for patients with GTN who have FIGO scores ≥5 and has predictable and manageable toxicity.Implications for PracticeThe most commonly used multiagent chemotherapy for high‐risk gestational trophoblastic neoplasia (GTN) is etoposide, methotrexate and actinomycin D/cyclophosphamide and vincristine (EMA/CO) worldwide. However, 5‐fluorouracil‐based multiagent chemotherapy has been used as a primary treatment for high‐risk GTN in China for a few decades. This study evaluated the efficacy and toxicity of floxuridine, actinomycin D, etoposide, and vincristine (FAEV) as a primary treatment for patients with GTN who have International Federation of Gynecology and Obstetrics (FIGO) scores ≥5. The study''s data demonstrated that FAEV as a primary treatment achieved favorable outcomes for patients with FIGO scores ≥5. Toxicities that result from the FAEV regimen are predictable and manageable. The FAEV regimen may provide another option for the treatment of GTN.     

长春新碱联合糖皮质激素治疗11例婴儿期Kasabach-Merritt综合征

目的 探讨Kasabach-Merritt（K-M）综合征的有效治疗方案。方法 收集2015—2017年首都医科大学附属北京儿童医院皮肤科收治的11例K-M综合征患儿的临床资料,分析长春新碱联合糖皮质激素治疗的疗效。结果 11例患儿就诊年龄1～212（87.91 ± 72.01） d,男4例、女7例。血管瘤多呈紫红色斑块,质地偏硬,其中5例瘤体周围伴有皮肤紫癜。血小板计数（4 ~ 32） × 109/L。给予口服泼尼松（2 ~ 5） mg·kg-1·d-1与静脉注射长春新碱每周0.05 mg/kg联合治疗。10例患儿应用糖皮质激素联合长春新碱治疗（1.8 ± 1.23）周后血小板达到正常,治疗（3.6 ± 1.26）周后纤维蛋白原恢复正常,治疗（3.9 ± 0.74）周后血管瘤开始软化或缩小。1例治疗5周后,血小板计数仍没有恢复正常,联合使用静脉栓塞后,血小板恢复正常,并可维持。结论 长春新碱联合糖皮质激素治疗可控制K-M综合征患儿瘤体发展,促进血小板恢复。     

天麻素对化疗痛模型大鼠脊髓IL-1β和TNFα表达的影响

目的 观察天麻素对长春新碱致大鼠神经病理性疼痛的治疗作用,观察大鼠脊髓腰段IL-1β和TNF α表达的变化,探讨其在脊髓水平的作用机制。方法 用长春新碱隔日腹腔注射制作大鼠化疗痛模型,模型成功制备后腹腔注射天麻素治疗,用电子Von Frey测痛仪测定大鼠机械性痛阈,热辐射仪测定大鼠热刺激痛阈值;采用ELISA方法检测脊髓腰段IL-1β和TNFα表达情况。结果 实验第8天化疗痛大鼠模型成功建立,用不同剂量天麻素治疗模型大鼠1周后,治疗组大鼠的机械和热刺激痛阈值与模型组比较均有不同程度的提高 (P<0.01, P<0.05);与模型组比较,治疗组大鼠的IL-1β和TNFα的表达明显降低（P<0.01）。结论 天麻素可减轻长春新碱诱导的大鼠化疗痛反应,其机制可能与抑制化疗痛大鼠脊髓IL-1β和TNFα的表达有关。     

芳姜黄酮衍生物对A375人黑素瘤细胞增殖及凋亡作用的机制研究

目的：研究一种芳姜黄酮衍生物（ATD）对人皮肤黑色素瘤A375细胞增殖及凋亡的影响。方法不同浓度（5、10、20、40、80μmol/L）ATD、长春新碱及芳姜黄酮体外作用A375及人皮肤成纤维细胞（HSF）48 h。CCK?8法检测细胞增殖抑制率;吖啶橙/溴化乙锭（AO/EB）染色,倒置显微镜观察细胞凋亡形态;DNA片段化检测细胞凋亡;比色法检测半胱氨酸天冬氨酸蛋白酶3（caspase?3）活性;流式细胞仪检测细胞凋亡及周期。结果ATD、长春新碱及芳姜黄酮对A375细胞有抑制增殖作用,且呈剂量依赖性（ATD：R2=0.99,F=340.96;长春新碱：R2=0.99,F=349.19;芳姜黄酮：R2=0.89,F=25.41,均P<0.05）,三者IC50分别为（15.96±0.02）、（77.00±0.04）及（356.95±0.01）μmol/L。当药物浓度为5μmol/L及10μmol/L时,ATD对HSF增殖抑制率分别为（8±0.06）%和（25±0.02）%,长春新碱为（33±0.04）%和（29±0.08）%,芳姜黄酮为（49±0.09）%和（34±0.07）%;ATD对A375细胞抑制率分别为（26±0.06）%和（39±0.02）%,长春新碱为（8±0.04）%和（17±0.08）%,芳姜黄酮为（6±0.09）%和（10±0.07）%,与二甲基亚砜相比,差异均有统计学意义（P＜0.05）,且ATD对A375细胞增殖抑制活性强于长春新碱及芳姜黄酮（P＜0.05）,但对HSF细胞毒性却明显低于长春新碱及芳姜黄酮（P＜0.05）。ATD、长春新碱及芳姜黄酮均可诱导A375细胞凋亡,caspase?3活性随3种药物浓度增加而增强,且药效为ATD>长春新碱>芳姜黄酮。流式细胞仪检测证实,3种药物都能诱导细胞发生不同程度凋亡,同芳姜黄酮及长春新碱相比,ATD能显著诱导细胞凋亡,且以晚期凋亡为主。随药物浓度增加,ATD组G1期A375细胞逐渐增多,G2期及S期细胞数明显减少。结论 ATD对A375细胞有抑制增殖及促凋亡作用,该作用明显强于芳姜黄酮及长春新碱,其机制可能是激活caspase?3,使细胞周期阻滞在G1期,进而抑制肿瘤细胞分化与增殖。     

Impaired function and expression of P-glycoprotein in blood-brain barrier of streptozotocin-induced diabetic rats

The aim was to investigate the effect of diabetes mellitus (DM) on P-glycoprotein (P-GP) function and expression in rat blood-brain barrier (BBB). P-GP function in BBB was assessed by measuring the brain-to-plasma concentration ratios (Kp values) of rhodamine 123 (Rho123) and vincristine (VCR), two well-known P-GP substrates, in control rats and 5-week streptozotocin (STZ)-induced diabetic rats. Evans blue (EB) dye was used as a BBB integrity indicator for examining the extravasation from the blood into the brain. P-GP expression in the brain cortex was evaluated with Western blot. The uptakes of Rho123 and VCR by cultured rat brain microvessel endothelial cells (rBMECs) incubated in diabetic and control rat serum for 72 h were also used to examine P-GP function, respectively. It was found that the Kp value of Rho123 (0.022+/-0.005 vs. 0.016+/-0.002 ml/g brain, p=0.033) and VCR (0.072+/-0.028 vs. 0.023+/-0.006 ml/g brain, p=0.006) in diabetic rats was significantly higher than that in control rats. The uptakes of Rho123 and VCR by cultured rBMECs incubated in the diabetic rat serum were higher than that in the control rat serum, respectively. No significant difference of the EB concentration in the brain cortex was found between the diabetic rats and control rats. Electron microscope examination of the brain cortex did not show a clear damage to the endothelial cells of microvessel in diabetic rats. In addition, the protein level of P-GP in the brains of the diabetic rats examined was significantly lower than that of control rats. These results suggested that the function and expression of P-GP might be impaired in the BBB of STZ-induced diabetic rats.     

VAD方案治疗多发性骨髓瘤疗效观察

 目的 评估VAD方案治疗多发性骨髓瘤（MM）的疗效。方法 24例Ⅲ期MM,采用VAD方案,即长春新碱0.5 mg／d（或长春地辛1 ~ 2 mg／d）缓慢静脉注射,第1～4天;多柔比星（或表柔比星）10 ~ 20 mg／d缓慢静脉注射,第1～4天;地塞米松40 mg／d口服,第1～4天,第9～12天,第17～20天,28 d为1疗程。连续应用2疗程以上评估疗效。观察项目包括：血清M蛋白、肝肾功能、尿蛋白、骨髓穿刺、血象等,记录毒副反应。MM疗效标准,分为完全缓解（CR）、部分缓解（PR）、未缓解（NR）。结果 CR8例（33.3 %）,PR12例（50 %）,NR4例（16.7 %）,总有效率为83.3 %。结论 VAD方案治疗Ⅲ期MM的临床疗效显著,临床症状改善明显。     

长春新碱对口腔鳞癌中Stathmin调控作用的实验研究

目的: 先前的研究发现,微管解聚蛋白（Stathmin）是潜在的指导局部晚期口腔鳞癌诱导化疗的生物标志物,本研究旨在进一步探索长春新碱对Stathmin的调控作用以及其作为TPF化疗替代药物的潜能。方法: 构建Stathmin过表达及敲减稳转细胞系,通过细胞增殖试验、q-PCR、Western免疫印迹、皮下移植瘤等实验方法,采用SPSS 23.0软件包进行统计学差异分析。结果: 长春新碱可抑制口腔鳞癌细胞系Stathmin的表达。高表达Stathmin后,口腔鳞癌细胞系对长春新碱的敏感性提高。针对高表达Stathmin的口腔鳞癌细胞系移植瘤,长春新碱具有良好的抑瘤效果。结论: 对于高表达Stathmin的口腔鳞癌,长春新碱是潜在的替代化疗药物。