鸦胆子油乳对膀胱癌影响的实验分析

目的通过实验分析鸦胆子油乳对膀胱癌的影响。方法在人体外培养人膀胱癌细胞(BIU-87),并将不同浓度的鸦胆子油乳加入其中,之后观察人膀胱癌细胞的生长、组织结构和细胞周期。对ICR小鼠进行亚硝胺的膀胱癌诱导,再给ICR小鼠膀胱灌注鸦胆子油乳,在光镜和电镜下观察膀胱癌的进展或抑制情况。结果鸦胆子油乳对人膀胱癌细胞产生了抑制作用,并破坏了膀胱癌细胞的微结构和超微结构,改变了膀胱癌细胞的性质并致其坏死,同时它还能够阻止人膀胱癌细胞由G0期向S期进展以及抑制DNA的合成。ICR小鼠实验结果表明,亚硝胺诱导的膀胱癌在膀胱灌注鸦胆子油乳之后得到有效的抑制。结论鸦胆子油乳能够抑制膀胱癌细胞的生长与繁殖,临床中可加以应用。     

鸦胆子油乳联合NP 方案治疗晚期非小细胞肺癌的临床疗效观察

目的:观察鸦胆子油乳联合NP方案化疗与单纯NP方案化疗对晚期肺小细胞癌患者的疗效、生活质量等因素的影响。方法:将154例患有晚期非小细胞肺癌患者随机分为两组:治疗组,在NP方案化疗基础上使用鸦胆子油乳治疗;对照组,单纯接受NP方案化疗。对154例患者追踪3个月,观察并比较患者的疗效、体质量、生活质量、毒副反应等方面的差异。结果:在NP方案化疗基础上使用鸦胆子油乳治疗在患者疗效评价方面与单纯NP方案化疗相比不具有统计学差异(P>0.05);在提高患者体质量、改善患者临床症状和生活质量等方面均优于单纯NP方案化疗(P<0.05);在化疗所引发的白细胞、血红蛋白和血小板降低等部分毒副反应发生率方面低于单纯NP方案化疗组(P<0.05)。结论:鸦胆子油乳联合NP方案化疗治疗晚期非小细胞肺癌不仅可以起到化疗增效的作用,而且能够明显提高患者生活质量,有效的改善临床症候,降低化疗引发的毒副反应,这一联合治疗方案可以在临床治疗中广泛推广。     

鸦胆子油乳对肺癌荷瘤鼠上呼吸道菌群定植的影响

目的探讨鸦胆子油乳治疗荷瘤肺癌小鼠对上呼吸道菌群定植的影响。方法建立A549细胞株肺癌荷瘤小鼠模型16只,随机分为2组:鸦胆子油乳治疗组和对照组,每组8只。用无菌咽拭子取材后进行测序分析上呼吸道菌群特征。结果鸦胆子油乳治疗后CB17-SCID肺癌荷瘤鼠上呼吸道菌群结构发生改变。鸦胆子油乳治疗荷瘤肺癌小鼠提高呼吸道菌群多样性,抑制条件致病菌的定植。结论鸦胆子油乳治疗影响荷瘤肺癌小鼠上呼吸道定植菌群,调节了机体的微生态平衡。     

顺铂联合鸦胆子油乳对人卵巢癌裸鼠原位移植瘤生长的影响

目的:探讨鸦胆子油乳以及联合顺铂对人卵巢癌裸鼠原位移植瘤生长的抑制作用。方法:构建人卵巢癌裸鼠原位移植瘤模型,随机分为对照组、鸦胆子油乳组、顺铂组和鸦胆子油乳顺铂联合组等4组,每组8只,观察各组裸鼠移植瘤体积、重量、抑瘤率及相对肿瘤增值率(T/C)的变化,检测裸鼠外周血白细胞、血小板数目及血清谷丙转氨酶(ALT)、乳酸脱氢酶(LDH)、血肌酐(CR)的变化。结果:鸦胆子油乳组、顺铂组、联合组均可有效遏制肿瘤细胞生长,各组的抑瘤率分别为23.56%±3.67%、42.57%±3.45%和48.36%±3.08%;各组的肿瘤体积和T/C明显下降,T/C均低于60%,且与对照组比较,差异有统计学意义(P0.05);与对照组比较,鸦胆子油乳组治疗后裸鼠LDH、CR值差异显著(P0.01),顺铂组与联合组ALT、LDH、CR值差异有统计学意义(P0.01);与对照组比较,鸦胆子油乳组白细胞数差异显著(P0.05),而顺铂组、联合组血小板、白细胞数量均明显减少,且差异均具有统计学意义(P0.01)。结论:顺铂联合鸦胆子油乳可以抑制人卵巢癌裸鼠移植瘤的生长,鸦胆子油乳相对于顺铂对人卵巢癌SKOV3细胞裸鼠白细胞及血小板、肝肾功能影响较小,具有低毒作用。     

鸦胆子油乳联合TP方案治疗晚期非小细胞肺癌近期临床观察

目的:观察鸦胆子油乳联合TP方案治疗晚期非小细胞肺癌(NSCLC)近期临床疗效。方法:选取2013年11月-2015年11月我院收治的80例晚期NSCLC患者进行分组研究,根据不同治疗方法分为参照组(n=40)和研究组(n=40),参照组给予TP化疗方案进行治疗,研究组给予鸦胆子油乳联合TP方案进行治疗,比较两组近期疗效及生活质量改善情况。结果:研究组治疗总有效率为67.5%,生活质量改善率为80.0%,参照组治疗总有效率为47.5%,生活质量改善率为67.5%,经统计比较,组间差异明显,具有统计学意义(P0.05)。结论:鸦胆子油乳联合TP方案治疗晚期NSCLC近期疗效优于单纯TP化疗,可显著改善患者生活质量,值得临床借鉴和进一步推广应用。     

运用HPLC-MS对鸦胆子油自微乳给药系统肠吸收的研究

目的:建立高效液相色谱-串联质谱法检测油酸和亚油酸含量的方法,从而对鸦胆子油自微乳给药系统中鸦胆子油的肠吸收进行研究.方法:以甲醇-水(95∶5 v/v)为流动相,流速为0.4 mL/min,柱温为35℃作为高效液相色谱的检测条件.利用大鼠小肠膜建立体外药物扩散体系研究鸦胆子油的肠吸收特性.结果:油酸和亚油酸的保留时间分别为10.46± 0.02和8.55±0.01 min,线性范围分别为0.50～50.0 ng/mL和5.06～101.2 ng/mL,平均绝对回收率分别为97.49±3.11％和105.76± 3.13％.日间和日内精密度都小于5％.在肠吸收实验中,鸦胆子油自微乳体系中测得油酸和亚油酸含量是单独给予鸦胆子油测得量的2.8和4.1倍.结论:该方法高效、灵敏、选择性高,可以用于鸦胆子油及鸦胆子油自微乳肠吸收中油酸和亚油酸的质量测定.     

鸦胆子油乳联合放疗治疗鼻咽癌的临床疗效观察

目的:探讨鸦胆子油乳联合放疗治疗鼻咽癌的临床疗效及安全性。方法:将在我科就诊的154例鼻咽癌患者随机分为对照组和治疗组,治疗组患者在常规放射治疗的基础上联合鸦胆子油乳治疗,而对照组患者单纯接受常规放射治疗。待治疗结束后,观察并比较两组患者的临床疗效、血液学指标及不良反应的发生情况。结果:对照组和治疗组的临床有效率分别为63.16%、82.05%,治疗组的临床有效率明显高于对照组,两组间比较有统计学差异(P0.05)。治疗后,治疗组患者的血液学指标如白细胞计数、红细胞计数、血红蛋白水平、CD4+/CD8+等均显著高于对照组,不良反应如恶心呕吐、张口困难、吞咽困难、声音嘶哑、口腔炎均明显低于对照组,差异具有统计学意义(P0.05)。结论:鸦胆子油乳联合放射治疗能够有效提高鼻咽癌患者的临床疗效,并且降低放疗所致的各种不良反应,值得临床推广应用。     

鸦胆子油乳、艾迪注射液对非小细胞肺癌化疗效果的影响

目的:分析鸦胆子油乳、艾迪注射液配合GP化疗治疗非小细胞肺癌的疗效及对免疫功能的影响。方法:以2010年1月至2016年3月宝鸡市解放军第三医院收治的非小细胞肺癌患者80例为研究对象,根据住院病历号顺序分为观察组和对照组,每组40例,对照组接受GP化疗方案,观察组接受GP化疗方案联合鸦胆子油乳、艾迪注射液治疗,两组均10天为一个周期,4个周期为一个疗程,均治疗1个疗程。比较两组的临床疗效、不良反应、T细胞亚群水平及生活质量评分。结果:观察组的总缓解率和疾病控制率分别为62.5%和92.5%,均显著高于对照组(47.5%和75.0%)(P0.05);治疗后,两组的CD3~+、CD4~+、CD8~+、CD4~+/CD8~+水平均较治疗前显著降低(P0.05),观察组治疗后的CD3~+、CD4~+、CD8~+水平均明显高于对照组(P0.05);两组治疗后的KPS评分均较治疗前升高,且观察组的KPS评分显著高于对照组(P0.05);治疗期间,观察组的恶心呕吐、腹泻、白细胞计数减少、血小板减少、肝功能损害和肾功能损害的发生率均显著低于对照组(P0.05);半年内,观察组的生存率(87.5%)显著高于对照组的生存率(77.5%)。结论:鸦胆子油乳、艾迪注射液配合GP化疗治疗非小细胞肺癌的疗效显著,不良反应发生率低,提高半年生存率,而且可降低化疗对T细胞亚群水平的影响,并降低患者血清VEGF水平。     

鸦胆子苦醇抑制人前列腺癌DU145细胞生长及作用机制

中药鸦胆子是一种常用的抗肿瘤中草药,鸦胆子苦醇是来源于鸦胆子的主要成分。该研究探讨了鸦胆子苦醇(brusatol)对人前列腺癌DU145细胞的生长抑制及其作用机制。采用四甲基偶氮唑盐(MTT)法检测鸦胆子苦醇对不同细胞株的生长抑制情况,以及不同浓度的鸦胆子苦醇对DU145细胞的增殖抑制率;应用Hoechst 33258染色法观察鸦胆子苦醇处理DU145细胞后所发生的形态学变化;分别采用PI单染及AnnexinV-FITC双染法流式细胞术分析细胞周期分布个凋亡率的变化;以Western blot测定鸦胆子苦醇对MAPK信号通路相关蛋白表达的影响。结果表明:鸦胆子苦醇对人前列腺癌DU145细胞的抑制作用更为显著,并且可以时间和剂量依赖性地抑制人前列腺癌DU145细胞的生长,其半数有效抑制浓度IC50为(0.27±0.04)μmol·L-1;鸦胆子处理DU145细胞后,Hoechst 33258染色可见到明显的凋亡特征;细胞周期图中可见明显的亚二倍体峰,且随着作用时间的延长凋亡比例增加,FCM检测鸦胆子苦醇作用24 h后凋亡图中,可见凋亡的发生;Western blot检测表明鸦胆子苦醇处理后可使磷酸化的p38和JNK表达增加,使磷酸化的ERK表达降低。鸦胆子苦醇能显著抑制DU145细胞增殖,诱导DU145细胞凋亡。磷酸化的P38和JNK的表达增加,但磷酸化的ERK表达下降,这表明MAPK途径的活化可能是鸦胆子苦醇对DU145细胞生长抑制的作用机制之一。因此,鸦胆子苦醇是潜在的抗前列腺癌药物,有必要进一步在动物水平阐明其抗前列腺癌活性。     

鸦胆子油乳注射液联合化疗对晚期胃癌的治疗效果及对血清CEA与CA199的影响

目的:研究鸦胆子油乳注射液联合化疗对晚期胃癌的治疗效果及安全性。方法:选取2014年8月至2015年8月我院接诊的80例晚期胃癌患者作为本次研究对象,按治疗方法分为观察组40例和对照组40例。对照组采用5-氟尿嘧啶及奥沙利铂化疗;观察组采用鸦胆子油乳注射液联合化疗。观察和比较两组患者的治疗疗效,血清癌胚抗原(CEA)、糖类抗原-199(CA199)、白细胞(WBC)、血红蛋白(HGB)、血小板(PLT)水平变化、生存质量改善情况及不良反应的发生情况。结果:治疗后,观察组总有效率为87.50%,显著高于对照组(65.00%,P0.05);生存质量改善率为90.00%,显著高于对照组(72.50%,P0.05);血清CEA、CA199水平、不良反应总发生率均显著低于对照组(P0.05);两组患者WBC、HGB、PLT水平比较差异无统计学意义(P0.05)。结论:鸦胆子油乳注射液联合化疗用于晚期胃癌的治疗效果显著,且安全性高。     

鸦胆子抗肿瘤活性成分的化学研究

从苦木科植物鸦胆子[Brucea javanica(L.)Merr]干躁果实的硅胶干柱柱层析所得的活性部位中经层析分离得到7个四环三萜苦木内酯成份(A,B,C,D,E,F,G),经UV,IR,~1H-NMR,~(13)C-NMR等方法鉴定分别为鸦胆苦醇(Brusatol A),双氢鸦胆苦醇(Dihydrobrusatol,B),鸦胆因B(Bruceine B,C),鸦胆因D(Bruceine D,D),鸦胆因H(Bruceine H,E),鸦胆子甙A(Bruceoside A,F)和双氢鸦胆子甙A(Yadanzioside A,G)。据报道,鸦胆苦醇和鸦胆子甙A具有较强的抗肿瘤活性。     

Mechanisms of the antimetastatic effect in the liver and of the hepatocyte injury induced by alpha-galactosylceramide in mice

The role of mouse liver NK1.1 Ag(+) T (NKT) cells in the antitumor effect of alpha-galactosylceramide (alpha-GalCer) has been unclear. We now show that, whereas alpha-GalCer increased the serum IFN-gamma concentration and alanine aminotransferase activity in NK cell-depleted C57BL/6 (B6) mice and B6-beige/beige mice similarly to its effects in control B6 mice, its enhancement of the antitumor cytotoxicity of liver mononuclear cells (MNCs) was abrogated. Depletion of both NK and NKT cells in B6 mice reduced all these effects of alpha-GALCER: Injection of Abs to IFN-gamma also inhibited the alpha-GalCer-induced increase in antitumor cytotoxicity of MNCS: alpha-GalCer induced the expression of Fas ligand on NKT cells in the liver of B6 mice. Whereas alpha-GalCer did not increase serum alanine aminotransferase activity in B6-lpr/lpr mice and B6-gld/gld mice, it increased the antitumor cytotoxicity of liver MNCS: The alpha-GalCer-induced increase in survival rate apparent in B6 mice injected intrasplenically with B16 tumor cells was abrogated in beige/beige mice, NK cell-depleted B6 mice, and B6 mice treated with Abs to IFN-gamma. Depletion of CD8(+) T cells did not affect the alpha-GalCer-induced antitumor cytotoxicity of liver MNCs but reduced the effect of alpha-GalCer on the survival of B6 mice. Thus, IFN-gamma produced by alpha-GalCer-activated NKT cells increases both the innate antitumor cytotoxicity of NK cells and the adaptive antitumor response of CD8(+) T cells, with consequent inhibition of tumor metastasis to the liver. Moreover, NKT cells mediate alpha-GalCer-induced hepatocyte injury through Fas-Fas ligand signaling.     

秋枫属的染色体数目及其进化意义

对秋枫属两个种Bischofia javanica和B·polycarpa的体细胞进行了染色体计数研究。结果表明这两个种在形态学上虽然存在差异,如秋枫是圆椎花序,重阳木是总状花序,但染色体数目均为2n=196。同时,结合细胞学、形态学和生态学特点探讨了秋枫属的染色体基数,多倍化的起源及其演化意义。     

荧光显微镜观察大蒜油对腹水癌细胞的影响

用吖啶橙染色,荧光显微镜观察大蒜油对—S180,昆明小鼠,U14 C57BL/6J小鼠和L1210DBA小鼠的癌细胞作用的影响,结果表明大蒜油有直接破坏癌细胞的DNA,RNA和分裂中期染色体的作用。给药后2—6小时作用最强,12小时后癌细胞数逐渐恢复,因此给药间隔不应超过12小时。我们研究证实大蒜油有明显的抗癌作用,为临床应用提供有力的依据。     

Antioxidant and antigenotoxic activities of Angelica keiskei, Oenanthe javanica and Brassica oleracea in the Salmonella mutagenicity assay and in HCT116 human colon cancer cells

Epidemiological studies indicate that consumption of green-yellow vegetables rich in chlorophyll, vitamin C, vitamin E, and carotenoids reduce the risk of cancer. We sought to examine the antigenotoxic and antioxidant properties of chlorophyll-rich methanol extracts of Angelica keiskei, Oenanthe javanica, and Brassica oleracea (kale). In the Salmonella mutagenicity assay, A. keiskei caused dose-dependent inhibition against three heterocyclic amine mutagens in the presence of S9, O. javanica was antimutagenic only at the highest concentration in the assay (2 mg/plate), and B. oleracea showed no consistent inhibitory activity at non-toxic levels. None of the extracts were effective against three direct-acting mutagens in the absence of S9. Extracts of A. keiskei and, to a lesser extent O. javanica, inhibited two of the major enzymes that play a role in the metabolic activation of heterocyclic amines, based on ethoxyresorufin-O-deethylase and methoxyresorufin-O-demethylase assays in vitro. All three plant extracts were highly effective in assays which measured ferric reducing/antioxidant power, oxygen radical absorbance capacity, and Fe2+/H2O2-mediated DNA nicking. Finally, using the 'comet' assay, all three plant extracts protected against H2O2-induced genotoxic damage in human HCT116 colon cancer cells. These findings provide support for the antigenotoxic and antioxidant properties of chlorophyll-rich extracts of A. keiskei, O. javanica, and B. oleracea, through mechanisms that include inhibition of carcinogen activation and scavenging of reactive oxygen species.     

Combined blockade of TIM-3 and TIM-4 augments cancer vaccine efficacy against established melanomas

Cancer vaccines have been developed to instruct the endogenous immune responses to autologous tumors and to generate durable clinical responses. However, the therapeutic benefits of cancer vaccines remain insufficient due to the multiple immunosuppressive signals delivered by tumors. Thus, to improve the clinical efficacy of cancer immunotherapy, it is important to develop new modalities to overcome immunosuppressive tumor microenvironments and elicit effective antitumor immune responses. In this study, we show that novel monoclonal antibodies (mAbs) specifically targeting either T cell immunoglobulin mucin protein-3 (TIM-3) or T cell immunoglobulin mucin protein-4 (TIM-4) enhance the therapeutic effects of vaccination against established B16 murine melanomas. This is true for vaccination with irradiated B16 melanoma cells engineered to express the flt3 ligand gene (FVAX). More importantly, combining anti-TIM-3 and anti-TIM-4 mAbs markedly increased vaccine-induced antitumor responses against established B16 melanoma. TIM-3 blockade mainly stimulated antitumor effector activities via natural killer cell-dependent mechanisms, while CD8⁺ T cells served as the main effectors induced by anti-TIM-4 mAb. Our findings reveal that therapeutic manipulation of TIM-3 and TIM-4 may provide a novel strategy for improving the clinical efficacy of cancer immunotherapy.     

Lysosomal membrane permeabilization contributes to elemene emulsion-induced apoptosis in A549 cells

Abstract

Elemene is a broad-spectrum antitumor agent. In the present study, lysosomal membrane permeabilization (LMP) was detected after short elemene emulsion - exposure (12 h) that preceded a decrease of the mitochondrial membrane potential and DNA damage (24 h) in A549 cells. At later time points (36 h) elemene emulsion caused the appearance of A549 cells with apoptotic features, including apoptotic morphology, phosphatidylserine exposure, and caspase-3 activation. A significant increase in protein expression for cathepsin D was also observed utilizing Western blot analysis after exposure to elemene emulsion for 12 h. The present study showed that elemene emulsion induced the increased levels of reactive oxygen species (ROS) and depletion of glutathione (GSH) in A549 cells. Cells treated with pepstatin A, an inhibitor for cathepsin D, showed a significant inhibition in DNA damage, mitochondrial membrane permeabilization, caspase-3 activation, and phosphatidylserine exposure. These results demonstrated that apoptosis induced by elemene emulsion in A549 cells is mediated in part through LMP and lysosomal protease cathepsin D.     

Lysosomal membrane permeabilization contributes to elemene emulsion-induced apoptosis in A549 cells

Elemene is a broad-spectrum antitumor agent. In the present study, lysosomal membrane permeabilization (LMP) was detected after short elemene emulsion--exposure (12 h) that preceded a decrease of the mitochondrial membrane potential and DNA damage (24 h) in A549 cells. At later time points (36 h) elemene emulsion caused the appearance of A549 cells with apoptotic features, including apoptotic morphology, phosphatidylserine exposure, and caspase-3 activation. A significant increase in protein expression for cathepsin D was also observed utilizing Western blot analysis after exposure to elemene emulsion for 12 h. The present study showed that elemene emulsion induced the increased levels of reactive oxygen species (ROS) and depletion of glutathione (GSH) in A549 cells. Cells treated with pepstatin A, an inhibitor for cathepsin D, showed a significant inhibition in DNA damage, mitochondrial membrane permeabilization, caspase-3 activation, and phosphatidylserine exposure. These results demonstrated that apoptosis induced by elemene emulsion in A549 cells is mediated in part through LMP and lysosomal protease cathepsin D.     

不同基因型水生植物对铵态氮和硝态氮吸收动力学特性研究

针对不同营养状况的富营养化水体修复而选择吸收养分效率较强的水生植物,采用改进常规耗竭法比较研究了6种不同基因型水生植物凤眼莲(Eichhornia crassipes Solms)、黄花水龙(Jussiaea stipulacea Ohwi)、再力花(Thalia dealbata Fraser)、美人蕉(Canna glauca L.)、水芹[Oenanthe javanica(Bl).DC]和豆瓣菜(Nasturtium officinaleR.Br.)对铵态氮和硝态氮吸收动力学特性。结果表明,不同基因型水生植物吸收铵态氮和硝态氮的动力学特性可用Michaelis-Menten方程来描述。在低浓度培养下,不同基因型水生植物对NH4+-N和NO3--N吸收的动力学参数Imax和Km差异较大,其吸收NH4+-N和NO3--N的Imax最大是水芹,其次是豆瓣菜;Km值最小的是水芹,其次是豆瓣菜;且水芹对NH4+-N和NO3--N不仅具有较强的亲和力,还具有较高的离子吸收速率。结果还表明,当介质中氮浓度较低时,水芹有优先吸收硝态氮的趋势,而豆瓣菜和再力花有优先吸收铵态氮的趋势。     

A comparison and critical analysis of preclinical anticancer vaccination strategies

Anticancer vaccines have been extensively studied in animal models and in clinical trials. While vaccination can lead to tumor protection in numerous murine models, objective tumor regressions after anticancer vaccination in clinical trials have been rare. B16 is a poorly immunogenic murine melanoma that has been extensively used in anticancer vaccination experiments. Because B16 has been widely used, different vaccination strategies can be compared. We reviewed the results obtained when B16 was treated with five common vaccine types: recombinant viral vaccines, DNA vaccines, dendritic cell vaccines, whole-tumor vaccines, and peptide vaccines. We also reviewed the results obtained when B16 was treated with vaccines combined with adoptive transfer of tumor antigen-specific T cells. We found several characteristics of vaccination regimens that were associated with antitumor efficacy. Many vaccines that incorporated xenogeneic antigens exhibited more potent anticancer activity than vaccines that were identical except that they incorporated the syngeneic version of the same antigen. Interleukin-2 enhanced the antitumor efficacy of several vaccines. Finally, several effective regimens generated large numbers of tumor antigen-specific CD8(+) T cells. Identification of vaccine characteristics that are associated with antitumor efficacy may aid in the development of more effective anticancer vaccination strategies.