CFSE/PI双标法检测CIK细胞对顺铂预处理肿瘤细胞的杀伤活性

细胞因子诱导的杀伤细胞(cytokine-induced killer,CIK)是将人外周血单个核细胞在体外用多种细胞因子共同培养一段时间后获得的一群异质细胞,具有增殖速度快、杀瘤活性高、杀瘤谱广等优点.如何对CIK细胞的效应功能进行客观准确的评价,如何使CIK和化疗有机地结合在一起,对CIK的临床应用具有重要的意义.

局部晚期宫颈癌IB2至ⅡB期34例全身新辅助化疗的术前疗效观察

近年来,宫颈癌的发病率呈明显上升趋势,且发病年龄呈年轻化趋势.传统放疗对巨块型(直径≥4cm)及局部晚期宫颈癌的局部控制率较低,疗效较差.新辅助化疗(neoadj uvant chemotherapy,NAC,)作为一种新的治疗手段应用于局部晚期宫颈癌患者,能缩小局部病灶,降低肿瘤分期,为根治手术创造条件.本研究对2005年1月至2007年1月本院收治的34例ⅠB2～ⅡB期局部晚期宫颈癌患者用BIP方案[博来霉素(bleomycin,BLM)+异环磷酰胺(isofosfamide,IFO)+顺铂(cisplatin,DDf)]静脉全身化疗1～2个疗程,探讨其疗效,现报道如下.

Ototoxic drug-induced apoptosis of inner ear cel s has been shown to be associated with calpain expression. Cisplatin has severe ototoxicity, and can induce cochlear cel apoptosis. This study assumed that cisplatin activated calpain expression in apoptotic cochlear cel s. A mouse model of cisplatin-induced ototoxicity was established by intraperitoneal injection with cisplatin (2.5, 3.5, 4.5, 5.5 mg/kg). Immunofluorescence staining, image analysis and western blotting were used to detect the expression of calpain 1 and calpain 2 in the mouse cochlea. At the same time, the auditory brainstem response was measured to observe the change in hearing. Results revealed that after intraperitoneal injection with cisplatin for 5 days, the auditory brainstem response threshold shifts increased in mice. Calpain 1 and calpain 2 expression significantly increased in outer hair cel s, the spiral ganglion and stria vascularis. Calpain 2 protein expression markedly increased with an in-creased dose of cisplatin. Results suggested that calpain 1 and calpain 2 mediated cispla-tin-induced ototoxicity in BALB/c mice. During this process, calpain 2 plays a leading role.

顺铂致溶血性贫血1例

病例:患者,女,56岁,因卵巢癌复发于2008年5月13日入院.既往因绝经8年,阴道出血2个月伴腹胀,检查发现盆腔肿物,于2005年7月26日于外院行全子宫、双附件大网膜、阑尾切除术加盆腔淋巴结清扫术.

奈达铂治疗恶性肿瘤研究进展

铂类化合物是治疗实体肿瘤的主要药物,自从1967年人们发现顺铂( cisplatin,DDP)有抗肿瘤活性以来,铂类金属抗肿瘤药的应用和研究得到了迅速的发展.奈达铂( nedaplatin,NDP;顺甘醇酸二氨合铂)是日本盐野义制药公司开发的第2代铂类抗肿瘤药,1995年在日本首次获准上市,目前,NDP主要用于头颈部肿瘤、肺癌、子宫癌、食道癌、膀胱癌、睾丸癌等,单药及联合化疗、放疗显示出了良好的抗肿瘤活性,本文对近年NDP在治疗恶性肿瘤方面的研究做一综述.

Objective To explore the inhibitory effect of recombinant mutant human tumor necrosis factor-α(rmhTNF-α) in combination with cisplatin on human lung adenocarcinoma cell line A549.
Methods Human lung adenocarcinoma cell line A549 was treated with varying concentrations of rmhTNF-α (0.38, 0.75, 1.50, 6.00 and 12.00 IU/ml) or cisplatin (3.91, 7.81, 15.63, 31.25 and 62.50 μg/ml) for 24 hours. Viable cell number was analyzed by using crystal violet staining. The inhibitory rates of A549 cells growth by the two drugs were calculated. For analyzing whether there was a synergistic effect of rmhTNF-α with cisplatin, A549 cells were treated with 0.75 IU/ml rmhTNF-αand increased concentrations of cisplatin.
Results rmhTNF-αor cisplatin inhibited the growth of A549 cell lines in a dose-dependent manner. The inhibitory effect of rmhTNF-αcombined with cisplatin was significantly greater than cisplatin alone at the same concentration (all P<0.01).
Conclusion rmhTNF-αcombined with cisplatin might have synergistic inhibitory effect on human lung adenocarcinoma cell line A549.

1例重组人p53腺病毒注射液联合顺伯、替加氟治疗舌癌的护理

肿瘤是一个全身性疾病,综合治疗已成为共识.继手术、放疗、化疗等治疗方法之后,肿瘤的基因治疗逐步应用并且显示了一定的疗效.2007年3月,我院应用重组人p53腺病毒注射液联合顺伯、替加氟治疗舌癌1例,疗效显著.现报告如下.

顺铂与白藜芦醇联合对胃癌细胞影响的研究

胃癌是常见的消化道恶性肿瘤之一,并且容易发生转移,其中人胃癌MGC803 细胞对多种化疗药物敏感.虽然我国在胃癌综合治疗方面做了大量研究,并取得一定效果,但胃癌术后5年生存率仍不足40%[1-3].顺铂(Cisplatin,CDDP)具有独特的抗癌机理和神奇的抗癌功效,CDDP是近年来常用的抗肿瘤药物.本研究以人胃癌SGC7901细胞株为研究对象,观察了CDDP对胃癌细胞及正常人体细胞的作用,并进一步研究CDDP与常规化疗药物白藜芦醇(Resverat rol,RES)联合应用对胃癌细胞作用的影响.为指导临床治疗提出一些新的化疗方法.

Cisplatin is widely used in the treatment of many tumors,particularly in ovarian cancer.GST-π,metallothionein(MT), multidrug resistance associated proteins(MRPs), nucleotide excision repair(NER), mismatch repair(MMR) and oncogenes contribute to drug resistance of cisplatin.

Objective:The purpose of the study is to investigate the ef ects of up-regulation of Raf kinase inhibitor protein (RKlP) on the chemosensitivity of cervical cancer Hela cells. Methods:Eukaryotic expression plasmid pcDNA3.1(+)-ssRKIP containing human overal length RKIPcDNA was transfected into cervical cancer Hela cellby lipofectin assay, establishing a stable cellline containing a target gene by G418. Expression of RKIP in Hela cells was measured by Western blot analysis. After treatment with cisplatin of dif erent concentrations and intervals of time, the ef ect of RKIP on the proliferation of Hela cells was evaluated by MTT method. The flow cytometry was used to investigate whether the RKIP could inhibit apoptosis in Hela cells induced by cisplatin. Results:The expression of RKIP in Hela cells transfected with pcDNA3.1-ssRKIP was increased obviously. After dif erent concentrations of cisplatin treatment cells for 24, 48 and 72 h, the growth inhibition rate in Hela cells transfected with pcDNA3.1-ssRKIP was significantly higher than in control cells (P<0.05). With 5μg/mL cisplatin treatment for 24 h, pcDNA3.1-ssRKIP-transfected Hela cells had an obviously higher percentage of apoptosis (23.2 ± 0.24)%than non-transfected cells (12.4 ± 0.31)%and empty vector-transfected cells (13.4 ± 0.47)%. Without treatment of cisplatin, the percentage of apoptosis for Hela cells transfected with pcDNA3.1-ssRKIP was (5.7 ± 0.12)%, which was stil higher than those of the non-transfected cells (2.9 ± 0.21)%and empty vector-transfected cells (3 ± 0.08)%. Conclusion:Higher expres-sion of RKIP gene can improve chemosensitivitv of cervical cancer Hela cells to cisplatin.

Objective:To compare the ef icacy and safety of Lobaplatin plus Etoposide (EL) and Cisplatin plus Etoposide (EP) regimens in chemonaive with extensive-stage smal-cel lung cancer (SCLC). Methods:Between July 2010 and July 2011, a total of 62 patients with extensive-stage smal-cel lung cancer who received initial treatment in our hospital and 309 hospital of PLA. 31 patients were randomly assigned to the EL Group:Lobaplatin was given intravenously at a dose of 30 mg/m2 on day 1 and Etoposide 100 mg/m2 on days 1 to 3 of 21-day cycles for a maximum of six cycles. Another 31 patients were assigned to the EP Group:Cisplatin was given intravenously at a dose of 75 mg/m2 on day 1 and Etoposide 100 mg/m2 on days 1 to 3 of 21-day cycles for a maximum of six cycles. We evaluated the ef icacy, overal response rate (ORR), disease control rate (DCR), the progression-free survival (PFS) and toxicity between the patients of the two groups. Results:Al 62 patients were eligible. In the EL group, 2 (6.5%) patients had complete response, 20 (64.5%) patients had partial response, 5 (16.1%) patients had stable disease and 4 (12.9%) patients had progress disease. In the EP group, 2 (6.5%) patients had complete response, 22 (70.9%) patients had partial response, 4 (12.9%) patients had stable disease and 3 (9.7%) patients had progress disease. The ORR of EL and EP group were 70.9%and 77.4%, respectively, showing no significant dif erence (P=0.562). The DCR of both groups were 87%and 90%, respectively, showing no significant dif erence (P=0.688). Median PFS of patients with EL and EP regimens were 5.5 months and 5 months, respectively, showing no significant dif erence (P=0.637). Adverse events were observed in al 62 patients. Grade 1 to 4 anemia was higher in the EP group than in EL group, showing significant dif erence (P=0.02). Grade 3 and 4 thrombocytopenia was seen in 4 patients (12.9%) in EL group and 1 patient (3.2%) in EP group. Although one patient had platelet transfusion owing to Grade 4 thrombocytopenia in EL group, no significant dif erence (P=0.637) were shown. The incidence of nausea/vomiting was higher in the EP group than in the EL group (96.7%vs 51.6%, P=0.00). Conclusion:The EL regimen is an ef ective and low-toxicity chemotherapy and no inferior to EP regimen in treatment response, therefore, EL regimen maybe is a good choice for patients with extensive-stage SCLC.

Objective:This randomized control ed clinical study was to assess and compare the ef icacy and safety of two chemoradiotherapy regimens [cisplatin+5-fluorouracil+3 dimensional conformal radiation therapy (3DCRT) and cisplatin+weekly docetaxel+3DCRT] in patients with local y advanced esophageal squamous cel carcinoma. Methods:A total of seventy-four patients with clinical stages IIB to IIIB esophageal squamous cel carcinoma were enrol ed. Chemotherapy for PF group comprised 5-fluorouracil at days 1-5 (250 mg/m2/d) and cisplatin (20 mg/m2) at days 1-3 of every 28-day cycle;ful treatment course included 2 cycles. Chemotherapy for DP group comprised docetaxel (20 mg/m2) and cisplatin (20 mg/m2) at days 1, 8, 15, 22, 29, and 36. Both groups treated with concurrent 60 Gy 3DCRT at 200 cGy/d. Results:Seventy-four patients were enrol ed and 71 completed the planned treatment, with a fol ow-up rate of 95.94%. Short-term curative ef ect was not statistical y significant between the two groups (P=0.471). The 2-year survival rates were 65.7%and 61.1%, respectively (P=0.806), 5 years survival rates were 34.29%and 27.78%, respectively (P=0.221), and there was no significant dif erence by Fisher test (P=0.734). As common side ef ects, incidence rates of radioactive esophagitis and hematological toxicity were lower in DP group. Conclusion:For local y advanced esophageal cancer patients, current chemoradiotherapy with chemo-therapy regimen of weekly docetaxel plus cisplatin has equal curative ef ect with 5-fluorouracil plus cisplatin, but wel-tolerated by reducing side ef ects such as radioactive esophagitis and bone marrow suppression.

临床药师查房英语

Clinical pharmacist:Good morning,how are you feeling today?药师:早上好,今天感觉怎么样?Patient:Not so good,I've been feeling like nausea and vomit since yesterday,just after finished my chemotherapy.患者:不太好,从昨天化疗完成后就一直恶心、想吐.Clinical pharmacist:Ok,let me check your medical order.Yes,the GP(Gemcitabine+Cisplatin) regimen performed yesterday is very good for NSCLC(Non-Small Cell Lung Cancer),but the Cisplatin in the regimen can cause severe nausea and vomit.And I find the doctor has given antiemetic for you: Tropisetron with intravenous injection and Metoclopramide for oral administration three times a day.Did you follow the advice?药师:好吧,让我看看你的医嘱.是的,你用的GP化疗方案确实对非小细胞肺癌有效,但是其中的顺铂也很容易引起恶心呕吐.而且医嘱上也给你开了止吐药了,有静脉给的托烷司琼,还有口服的甲氧氯普胺需要一天吃三次,你是按这种方法用药的吗?

Neoadjuvant chemotherapy plus radiotherapy is the most common treatment regimen for advanced nasopharyngeal carcinoma (NPC). Whether chronomodulated infusion of chemotherapy can reduce its toxicity is unclear. This study aimed to evaluate the toxic and therapeutic effects of sinusoidal chronomodulated infusion versus flat intermittent infusion of cisplatin (DDP) and 5-fluorouracil (5-FU) followed by radiotherapy in patients with locoregionally advanced NPC. Patients with biopsy-diagnosed untreated stages III and IV NPC (according to the 2002 UICC staging system) were randomized to undergo 2 cycles of sinusoidal chronomodulated infusion (Arm A) or flat intermittent constant rate infusion (Arm B) of DDP and 5-FU fol owed by radical radiotherapy. Using a“MELODIE”multi-channel programmed pump, the patients were given 12-hour continuous infusions of DDP (20 mg/m2) and 5-FU (750 mg/m2) for 5 days, repeated every 3 weeks for 2 cycles. DDP was administered from 10:00 am to 10:00 pm, and 5-FU was administered from 10:00 pm to 10:00 am each day. Chronomodulated infusion was performed in Arm A, with the peak deliveries of 5-FU at 4:00 am and DDP at 4:00 pm. The patients in Arm B underwent a constant rate of infusion. Radiotherapy was initiated in the fifth week, and both arms were treated with the same radiotherapy techniques and dose fractions. Between June 2004 and June 2006, 125 patients were registered, and 124 were eligible for analysis of response and toxicity. The major toxicity observed during neoadjuvant chemotherapy was neutropenia. The incidence of acute toxicity was similar in both arms. During radiotherapy, the incidence of stomatitis was significantly lower in Arm A than in Arm B (38.1%vs. 59.0%, P = 0.020). No significant differences were observed for other toxicities. The 1-, 3-, and 5-year overal survival rates were 88.9%, 82.4%, and 74.8%for Arm A and 91.8%, 90.2%, and 82.1%for Arm B. The 1-, 3-, and 5-year progression-free survival rates were 91.7%, 88.1%, and 85.2%for Arm A and 100%, 94.5%, and 86.9% for Arm B. The 1-, 3-, and 5-year distant metastasis-free survival rates were 82.5%, 79.1%, and 79.1%for Arm A and 90.2%, 85.2%, and 81.7%for Arm B. Chronochemotherapy significantly reduced stomatitis but was not superior to standard chemotherapy in terms of hematologic toxicities and therapeutic response.

The prognosis of locally advanced or recurrent squamous cell carcinoma (SCC) of the penis after conventional treatment is dismal. This study aimed to evaluate the therapeutic effects of intraarterial chemotherapy with gemcitabine and cisplatin on local y advanced or recurrent SCC of the penis. Between April 1999 and May 2011, we treated 5 patients with locally advanced penile SCC and 7 patients with recurrent disease with intraarterial chemotherapy. The response rate and toxicity data were analyzed, and survival rates were calculated. After 2 to 6 cycles of intraarterial chemotherapy with gemcitabine and cisplatin, 1 patients with locoregional y advanced disease achieved a complete response, and 4 achieved partial response. Of the 7 patients with recurrent disease, 2 achieved complete response, 3 achieved partial response, 3 had stable disease, and 1 developed progressive disease. An objective tumor response was therefore achieved in 10 of the 12 patients. The median overal survival for the patients was 24 months (range, 10-50 months). Three out of 10 patients who responded were long-term survivors after intraarterial chemotherapy. Intraarterial chemotherapy with gemcitabine and cisplatin may be effective and potential y curative in locoregional y advanced or recurrent penile SCC. The contribution of this therapy in the primary management of advanced or recurrent penile SCC should be prospectively investigated.

E2F-1基因沉默可增强人胃癌耐药细胞株SGC-7901/DDP对顺铂的敏感性

目的：探讨E2F转录因子1（E2F transcription factor 1, E2F-1）基因沉默对人胃癌耐药细胞株SGC-7901/DDP对顺铂敏感性的影响及可能机制。方法将SGC-7901/DDP细胞接种于六孔板，分为3组：以E2F-1沉默重组慢病毒颗粒（Lv-shRNA-E2F-1）感染SGC-7901/DDP，作为Lv-shRNA-E2F-1组；以阴性对照慢病毒颗粒（Lv-shRNA-NC）感染SGC-7901/DDP，作为Lv-shRNA-NC组；空白对照组不做任何处理。以MTT法测定转染后各组细胞对DDP的半数抑制浓度（IC50），并以AxnnexinV-PE/7-AAD双染色流式细胞术检测各组细胞凋亡率及周期分布。应用Western blot和半定量RT-PCR技术分别对各组细胞中E2F-1、survivin、Bcl-2的mRNA及蛋白水平进行检测。结果与Lv-shRNA-NC组和空白对照组相比，Lv-shRNA-E2F-1组细胞对顺铂的IC50显著降低，细胞凋亡率显著提高，细胞周期阻滞于G0/G1期，差别均有统计学意义（P<0.05）；Lv-shRNA-E2F-1组E2F-1 mRNA水平较Lv-shRNA-NC组和空白对照组分别下降45.0%和41.3%，蛋白水平分别下降66.7%和70.5%（均P<0.01）；Lv-shRNA-E2F-1组survivin mRNA水平较Lv-shRNA-NC组和空白对照组分别下降30.3%和28.7%，蛋白水平分别下降56.5%和53.6%（均P<0.01）；Lv-shRNA-E2F-1组Bcl-2 mRNA水平较Lv-shRNA-NC组和空白对照组分别下降76.6%和76.8%，蛋白水平分别下降74.6%和79.9%（均P<0.01）；Lv-shRNA-NC组与空白对照组比较，上述指标差别均无统计学意义（均P>0.05）。结论：沉默E2F-1基因可有效提高人胃癌耐药细胞株SGC-7901/DDP对顺铂的敏感性，其机制可能与survivin及Bcl-2表达下调有关，提示E2F-1可能成为胃癌药物治疗的新靶点。

下调CDH17基因对cisplatin耐药性人胃癌BGC823细胞裸鼠移植瘤模型的影响

目的 研究下调 CDH17 基因对cisplatin耐药的人胃癌 BGC823 细胞裸鼠移植瘤模型的影响,从体内实验研究治疗胃癌新的治疗手段.方法 以人胃癌BGC823细胞为研究材料,建立人胃癌BGC823细胞裸鼠移植瘤模型.同时,构建干扰质粒从基因和蛋白水平对CDH17进行干扰,成功构建慢病毒载体.将细胞分为三组,分别是WT对照组,siNC阴性对照组,siCDH17干扰组.通过 TUNEL方法和Annexin V/PI双染色法检测各组细胞的凋亡水平.结果 Real-time PCR和蛋白免疫印迹实验证实成功构建了CDH17的干扰表达载体.TUNEL检测实验表明下调CDH17基因可以促进对cisplatin耐药的人胃癌BGC823细胞裸鼠移植瘤模型细胞凋亡.Annexin V/PI双染色检测siCDH17干扰组细胞早期凋亡率远高于WT对照组和siNC阴性对照组的细胞.结论 下调CDH17基因可以促进cisplatin耐药性人胃癌BGC823细胞凋亡,抑制肿瘤细胞增殖而达到治疗胃癌的目的.

姜黄素对乳腺癌细胞顺铂敏感性的影响

目的 探讨姜黄素(curcumin)对乳腺癌细胞中顺铂(Cisplatin,CDDP)治疗敏感性的影响及其可能机制.方法 CCK-8检测、Hoechst染色观察CDDP、姜黄素单独及联合用药48 h对MCF7细胞增殖抑制和细胞凋亡的影响;Western blot分析MCF7细胞在CDDP(0、1.25、2.5、5、10、20 μg/mL)、姜黄素(0、1、5、20、30、50、100 μmol/L)单独及联合处理后FEN1(flap endonuclease 1)表达水平的变化;CCK-8检测沉默FEN1对MCF7细胞中CDDP敏感性的影响.结果 CDDP、姜黄素均可以剂量依赖方式抑制MCF7细胞增殖,其IC50分别为5、30 μmol/L;与单独2μg/mLCDDP处理组比较,2μg/mL CDDP联合20-μmol/L姜黄素、30 μmol/L姜黄素处理组对细胞增殖的抑制效应明显增强[分别为(84.1±0.8)％、(51.1±0.5)％、(29.4±0.3)％,P＜0.05];与单独20μmol/L姜黄素处理组比较,20 μmol/L姜黄素联合2μg/mL CDDP、5μg/mL CDDP处理组对细胞增殖的抑制效应也明显增强[分别为(76.9±0.7)％、(42.4±0.3)％、(31.6±0.4)％,P＜0.05];2μg/mL CDDP联合20 μmol/L姜黄素组的细胞凋亡明显增强(P＜0.05);与Control siRNA组比较,FEN1 siRNA+CDDP组可显著增强CDDP抑制MCF7细胞增殖的敏感性[(25.4±0.3)％vs(18.7±0.2)％,P＜0.05];CDDP增殖抑制无效浓度或低浓度时,FEN1蛋白的表达随CDDP的处理剂量依赖性上调,而姜黄素可剂量依赖性下调FEN1蛋白表达;与单独CDDP和姜黄素处理组比较,2μg/mL CDDP联合20 μmol/L姜黄素组可显著降低FEN1蛋白表达(P＜0.05).结论 姜黄素可增强CDDP对乳腺癌细胞的敏感性,其机制与降低细胞FEN1的表达有关.

肺癌GP方案化疗后急性心梗死亡1例报告

1 临床资料患者,女,48岁,因咳嗽、咳痰半月入院.入院查体:神志清,浅表淋巴结未触及肿大,双肺呼吸动度均等,叩诊两侧清音,双肺呼吸音粗,可闻及少量干湿啰音,未闻及胸膜摩擦音.CT检查示:右肺门增大并软组织密度影,纵隔内多发淋巴结肿大.