紫草素逆转埃克替尼对肺癌EGFR-TKI 耐药及其机制的研究

目的:观察紫草素联合埃克替尼对肺腺癌耐药细胞H1975增殖的影响,探讨克服耐药可能的作用机制。方法:应用MTT法检测紫草素(1.25~20μmo/L)、埃克替尼(5~100μmol/L)及两药联合干预对H1975细胞生长的抑制作用;流式细胞术观察紫草素(1.25μmol/L)、埃克替尼(10μmol/L)及联合使用对H1975凋亡作用;Western blot检测不同干预对H1975细胞EGFR、p-EGFR、AKT、p-AKT、ERK、p-ERK和凋亡相关蛋白PARP表达水平的影响。结果:MTT检测结果显示,与单药组相比,联合用药组细胞H1975增殖能力明显减弱,差异有统计学意义(P0.05);流式细胞术结果显示,联合用药组细胞的凋亡率达到(52.45±3.04)%,较紫草素组细胞凋亡率(22±1.17)%和埃克替尼处理组细胞凋亡率(15.35±5.85)%明显提高,差异有统计学意义(P0.05)。Western blot结果显示,单药组下调了p-EGFR、p-Akt蛋白水平的表达,而联合用药组显著抑制了p-ERK、PARP蛋白水平的表达,差异有统计学意义(P0.05),EGFR、AKT、ERK蛋白表达无差异(P0.05)。结论:紫草素联合埃克替尼能明显抑制H1975细胞增殖,促进肿瘤细胞凋亡;抗肿瘤机制可能与调节EGFR信号通路相关蛋白表达有关。     

培美曲塞化疗对非小细胞肺癌患者T淋巴细胞亚群的影响

目的:探究培美曲塞化疗对非小细胞肺癌患者T淋巴细胞亚群的影响,评价其临床应用价值。方法:选取2016年3月到2017年3月我院肿瘤内科收治的100例非小细胞肺癌患者进行研究,其中,50例非小细胞肺癌患者采用培美曲塞化疗、50例采用多西他赛进行化疗。比较患者治疗前后T淋巴细胞亚群变化情况。结果:与化疗前比较,非小细胞肺癌患者化疗后CD_3~+、CD_4~+、CD_4~+/CD_8~+比值以及NK细胞显著上升,而CD_8~+显著下降(P0.05)。为了验证培美曲塞对非小细胞肺癌患者的安全性,本研究将部分患者采用多西他赛化疗,非小细胞肺癌患者培美曲塞和多西他赛化疗后CD_3~+、CD_4~+、CD_4~+/CD_8~+比值以及NK细胞相比差异不大(P0.05)。此外,两组不良反应如恶心、呕吐,细胞减少,血小板减少,肾功能障碍差异不大(P0.05)。结论:培美曲塞化疗可以显著改变非小细胞患者的T淋巴细胞亚群构成,提高患者免疫功能和治疗效果。     

培美曲塞联合顺铂治疗晚期非小细胞肺癌的临床疗效观察

目的:探讨培美曲塞联合顺铂治疗晚期非小细胞肺癌(NSCLC)的临床疗效。方法:随机选取我院肿瘤科晚期NSCLC患者177例,随机将其分为3组,培美曲塞联合顺铂治疗(PP组)72例,多西他赛联合顺铂治疗(DP组)53例,吉西他滨联合顺铂治疗(GP组)52例,比较三组治疗方法的临床疗效与不良反应之间的差异,根据临床疗效将PP组分为有效组与无效组,分析培美曲塞联合顺铂治疗晚期NSCLC的影响因素。结果:PP组疾病控制率(DCR)与客观有效率(ORR)均显著高于GP组(均P0.05);PP组与DP组近期疗效之间的比较无显著差异(均P0.05)。PP组的药物毒副作用均显著优于DP组与GP组(均P0.05)。PP组的中位生存期显著高于DP组与GP组(均P0.05),在无吸烟、腺癌与IV期晚期NSCLC患者中,培美曲塞联合顺铂治疗有效率更高。结论:培美曲塞治疗晚期NSCLC的疗效佳,与多西他赛相当并显著优于吉西他滨治疗,药物毒副作用小,且受吸烟状况、病理类型与临床分期影响。     

多西他赛单药与培美曲塞联合顺铂二线治疗老年晚期胃癌的疗效及对患者生活质量的影响

目的:探讨多西他赛单药与培美曲塞联合顺铂二线治疗老年晚期胃癌的疗效及对患者生活质量的影响,为临床用药提供参考。方法:选取我院2014年6月-2017年6月期间收治的120例一线化疗失败的老年晚期胃癌患者,按照随机数字表法将患者分为观察组和对照组各60例,观察组使用培美曲塞联合顺铂治疗,对照组单独使用多西他赛治疗,两组均治疗3个疗程。治疗3个疗程后,采用实体肿瘤的疗效评价标准(RECIST)对两组患者的临床疗效进行评价,参照抗癌药物常见毒副反应分级标准统计患者出现的不良反应,采用生活质量量表评价患者的生活质量,并对所有患者进行为期半年的随访,统计两组患者的生存率。结果:观察组有效率(RR)为30.00%,略高于对照组的25.00%,但两组比较差异无统计学意义(P>0.05)。两组患者不良反应发生率比较差异无统计学意义(P>0.05)。治疗后,观察组患者日常生活、社会活动、抑郁、焦虑得分均明显高于对照组(P<0.05)。观察组患者半年生存率为71.67%,明显高于对照组的48.33%(P<0.05)。结论:相比于多西他赛单药治疗,培美曲塞联合顺铂二线治疗老年晚期胃癌能够改善患者生活质量,延长其生存期,安全可靠。     

顺铂联合多西他赛、培美曲塞及吉西他滨治疗晚期肺腺癌的临床效果比较

目的:比较吉西他滨、培美曲塞、多西他赛联合顺铂三种化疗方案治疗晚期肺腺癌患者的近期疗效与安全性。方法:选择2014年7月至2015年8月在本院肿瘤科住院的经病理或细胞学证实为ⅢB~Ⅳ期肺腺癌的患者共140例,随机分为三组,分别采用多西他赛+顺铂(多西他赛组,n=38)、培美曲塞+顺铂(培美曲塞组,n=56)、吉西他滨+顺铂(吉西他滨组,n=46)三种化疗方案。对三组患者的近期疗效和Ⅲ、Ⅳ度毒性反应的发生情况进行比较。结果:吉西他滨组无完全缓解(CR)患者,部分缓解(PR)患者20例,稳定(SD)患者16例,进展(PD)患者10例,总有效率(RR)43.5%,疾病控制率(DCR)为78.3%;多西他赛组无CR患者,PR患者16例,SD患者12例,PD患者10例,RR42.1%,DCR73.7%;培美曲塞组无CR患者,PR患者28例,SD患者20例,PD患者8例,RR50.0%,DCR为85.7%。三组患者RR及DCR相比较差异无统计学意义(P0.05)。三组化疗方案的主要毒副反应为骨髓抑制,无Ⅲ~Ⅳ度皮疹和末梢神经炎等毒性反应发生。其中,培美曲塞组的严重骨髓抑制即Ⅲ度+Ⅳ度白细胞减少、中性粒细胞减少及血小板减少的发生率明显低于吉西他滨组和多西他赛组(P0.05)。三组化疗方案Ⅲ度+Ⅳ度血红蛋白下降、胃肠道反应、脱发、肝肾功能异常等毒性反应的发生率相比较差异均无显著性(P0.05)。结论:培美曲赛、多西他赛、吉西他滨联合顺铂方案治疗晚期肺腺癌的疗效相当,但培美曲塞组安全性更高。     

Mcl-1参与非小细胞肺癌对吉非替尼耐药的实验研究

目的:探究人髓细胞白血病基因-1(myeloid cell leukemia-1,Mcl-1)是否参与非小细胞肺癌对吉非替尼的耐药。方法:应用Western blot检测Mcl-1在吉非替尼敏感细胞PC-9和耐药细胞H1975表达差异;梯度浓度的吉非替尼作用于PC-9细胞后,Western blot实验检测B淋巴细胞瘤-2基因(B-cell lymphoma-2,Bcl-2)家族中抗凋亡蛋白的表达变化;应用Western blot实验检测Mcl-1在PC-9和H1975细胞内降解速度差异。结果:Mcl-1在PC-9细胞内的表达明显低于H1975细胞,差异有统计学意义(P0.05),并且随着吉非替尼作用浓度的升高,Mcl-1表达逐渐降低,而Bcl-2和Bcl-x L表达基本不变,并且PC-9细胞内Mcl-1降解更迅速,半衰期明显缩短。结论:非小细胞肺癌对吉非替尼耐药可能与Mcl-1的表达量上调,降解速度减慢,半衰期延长有关。     

埃克替尼治疗晚期非小细胞肺癌的疗效及对患者血清相关指标水平的影响

目的:分析埃克替尼对晚期非小细胞肺癌的治疗效果及对血清指标的影响。方法:将86例晚期非小细胞肺癌患者按抽签法分成对照组与观察组,各43例。对照组采用多西他赛治疗,观察组采用埃克替尼治疗。观察并比较两组患者治疗前后血清细胞角蛋白21-1(Cyfre21-1)、鳞状细胞癌抗原(SCCA)、血管内皮生长因子(VEDF)、自然杀伤(NK)细胞、CD4~+、CD8~+、CD4~+/CD8~+,白细胞介素-8(IL-8)、肿瘤坏死因子-α(TNF-α)、基质金属蛋白酶-2(MMP-2)及基质金属蛋白酶-9(MMP-9)水平、临床疗效以及安全性。结果:观察组疾病控制率高于对照组,差异具有统计学意义(P0.05)。治疗后,两组患者血清Cyfre21-1、SCCA、CD8~+、LI-8、TNF-α、MMP-2及MMP-9均低于治疗前,且观察组低于对照组,,差异具有统计学意义(P0.05);治疗后,两组患者血清VEGF水平均降低,且观察组低于对照组,差异具有统计学意义(P0.05);治疗后,两组患者血清NK、CD4~+及CD4~+/CD8~+均高于治疗前,且观察组高于对照组,差异具有统计学意义(P0.05);观察组不良反应发生率低于对照组,差异具有统计学意义(P0.05)。结论:埃克替尼对晚期非小细胞肺癌的临床效果肯定,可下调血清VEGF表达。     

多西他赛固体脂质纳米粒抗乳腺癌效果及机制研究

目的:探讨多西他赛固体脂质纳米粒抗乳腺癌效果及机制研究。方法:本实验采用MTT法考察了多西他赛固体脂质纳米粒对人乳腺癌MCF-7细胞增殖的抑制作用,采用流式细胞术检测多西他赛固体脂质纳米粒对MCF-7肿瘤细胞凋亡作用,并进一步应用Western-Blot印迹法观察多西他赛固体脂质纳米粒对MCF-7细胞中Src、E-cadherin、β-catenin蛋白表达的影响,探索了其抗乳腺肿瘤的作用机制。结果:多西他赛固体脂质纳米粒能够显著抑制人乳腺癌MCF-7肿瘤细胞的增殖,且浓度越高,抑制率越大(P0.05)。经25、50、100μg/m L多西他赛固体脂质纳米粒制剂作用24 h后,人乳腺癌细胞MCF-7的凋亡率分别为14.56%、21.21%、29.94%,细胞凋亡率随着药物浓度的增加而增加,且各实验组间比较有显著性差异(P0.05)。人乳腺癌MCF-7肿瘤细胞经不同浓度的多西他赛固体脂质纳米粒处理后,细胞中E-cadherin蛋白表达显著升高,Src、β-catenin蛋白表达显著降低,且呈现出明显的剂量依赖性。结论:多西他赛固体脂质纳米粒能够抑制人乳腺癌MCF-7细胞增殖,促进其凋亡,可能与下调β-catenin蛋白的表达,上调E-cadherin蛋白表达以及抑制Src激酶活性有关。     

培美曲塞单药或联合铂类治疗晚期复治非小细胞肺癌54例临床观察

目的:培美曲塞是一种多靶点抗叶酸化疗药,目前已成为晚期非小细胞肺癌二线治疗的标准药物.本研究回顾分析培美曲塞单药或联合铂类治疗晚期复治非小细胞肺癌的疗效及不良反应.方法:对既往至少接受过1个标准含铂方案化疗的54例晚期非小细胞肺癌怠者,分为单药治疗组21例,联合铂类治疗组33例.单药治疗组给予培美曲塞单药治疗,培美曲塞500mg/m2,第1天,21天为1个周期;联合铂类治疗组给予培美曲塞联合顺铂或卡铂,培美曲塞500mg/m2,第1天,顺铂75 mg/m2或卡铂AUC=5,第1天,21天为1个周期.评价疗效及不良反应.结果:54例患者均可评价疗效.单药治疗组PR 1例,RR4.8%,SD10例,疾病控制率(DCR)52.4%,PD10例(47.6%).中位无进展生存期3.8个月;联合治疗组PR4例,RR12.1%,SD20例,疾病控制率(DCR)72.7%,PD9例(27.3%).中位无进展生存期4.8个月.与药物相关的不良反应主要为:Ⅰ/Ⅱ度骨髓抑制、胃肠道反应.结论:培美曲塞或与铂类联合治疗晚期复治非小细胞肺癌有效,不良反应轻微、可耐受.     

依维莫司协同拉帕替尼降低HER2+乳腺癌对拉帕替尼耐药

研究依维莫司(Everolimus)联合拉帕替尼(Lapatinib)对HER2~+乳腺癌耐药细胞的影响。CCK-8法测定乳腺癌亲本细胞(skbr3)和耐药细胞(skbr3-LR)的细胞活性、半数抑制浓度(Half inhibitory concentration,IC_(50));CompuSyn软件计算联合作用指数(combined effect index, CI)值;流式细胞术检测对照组实验组skbr3-LR的周期;Western blotting方法检测各组AKT、m TOR、S6相关蛋白的表达量。依维莫司联合拉帕替尼使skbr3-LR对拉帕替尼的IC_(50)从(27.13±0.3)下降到(6.79±0.04);20 nmol/L依维莫司联合5μmol/L、10μmol/L、20μmol/L、40μmol/L拉帕替尼CI指数分别为0.243 67、0.278 61、0.282 19、0.366 79、0.416 67;拉帕替尼联合依维莫司组G1期细胞百分数(69.84±0.93)%显著大于对照组(52.91±1.64)%(p0.001),也显著高于拉帕替尼组(59.77±0.52)%(p0.001)和依维莫司组(58.73±0.64)%(p0.001);Western blotting结果显示skbr3-LR中p-mTOR比skbr3表达上调(p0.001),20 nmol/L依维莫司或10μmol/L拉帕替尼单独处理skbr3-LR,以及联合用药组与skbr3-LR组相比p-mTOR均有显著下调(p0.001, p0.01, p0.001)。依维莫司联合拉帕替尼可协同抑制skbr3-LR增殖,降低其拉帕替尼耐药性,其作用机制部分是通过下调AKT/mTOR/S6通路实现的。     

六种藓类植物茎的比较解剖学研究

通过对6种藓类植物,即褶叶青藓(Brachythecium salebrosum(Web.et Mohr.)B.S.G.)、湿地匐灯藓(Plagiomnium acutum(Lindb.)Kop.)、侧枝匐灯藓(Plagiomnium maximoviczii(Lindb.)Kop.)、大凤尾藓(Fissidensnobilis Griff.)、大羽藓(Thuidium cymbifolium(Doz.et Molk.)B.S.G.)和大灰藓(Hypnum plumaeforme Wils.)嫩茎和老茎的石蜡切片和显微观察发现,同一藓类植株的嫩茎和老茎,茎结构稳定,不同种藓类植物茎横切面具有不同特征.植物体茎横切面形状、表层细胞的层数、细胞大小和细胞壁厚薄、皮层细胞大小和形状、中轴的有无以及比例等特征可以作为藓类植物的分科分类依据之一.     

Photoregulation of seed germination of wild-type and of an aurea-mutant of tomato

Seed germination of an aurea mutant of tomato ( Lycopersicon esculentum Mill.) is promoted by continuous irradiation with red, far-red or long-wavelength far-red (758 nm) light as well as by cyclic irradiations (5 min red or 5 min far-red/25 min darkness). Far-red light applied immediately after each red does not change the germination behaviour. Seed germination of the isogenic wild-type, cv. UC-105, is promoted by continuous and cyclic red light while it is inhibited by continuous and cyclic far-red light and by continious 758 nm irradiation. Far-red irradiation reverses almost completely the promoting effect of red light. The promoting effect (in the aurea mutant) and the inhibitory effect (in the wild-type) of continuous far-red light do not show photon fluence rate dependency above 20 nmol m⁻² s⁻¹. It is concluded that phytochrome controls tomato seed germination throgh low energy responses in both the wild type and the au mutant. The promoting effect of continuous and cyclic far-red light in the au mutant can be attributed to a greater sensitivity to P_fr.     

Cause of Senescence of Nine Sorts of Flowers

The levels of endogenous phytohormones and respiratory rate in nine sorts of flowers such as Cymbidium faberi Rolfe, Nopalxochia ackermannii Kunth and others were investigated both at full bloom and senescence and meanwhile the effect of exogenous phytohormones on prolonging the blossoms and promoting ethylene production were tested. There is a high content of endogenous ethylene in all the long-lived flowere, about 3–16 folds higer than the short-lived ones. There is a high level of ABA at full blooming flowers of short-lived flowers, in which there is no or only some cytokinins in it, but the ratio of CTK (6BA+zeatin)/ABA is smaller(l.7). The endogenous ABA reached a much higher level at senescence in all nine sorts of flowers, so it is reasonable to consider that it is ABA which plays an important role of regulation in controlling flower's senescence. There is a much higher level of GA3 and zeatin in the long-lived flowers which is not demonstrated in the shortlived ones. The respiratory rate is one of the factors controtling the longevity of flowers, but it does not play a decided role. Application of 6BA and zeatin prolongs distinctly orchid’s longevity, however exogenous IAA through the promotive action on ethylene production, evidently extends the longevity of the flowers of the Nopalxochia ackermannii Kunth.     

龙胆科药用植物化学成分的研究现状

龙胆科植物在我国的分布范围很广,且多数为药用植物,其多数种属的药用植物,至今其化学成分尚未被系统研究。综述了目前龙胆科药用植物的化学成分的研究现状及一般提取方法,对近年来发现的环烯醚萜及裂环烯醚萜类化合物进行了总结,为本科药用植物的更深入研究提供了参考。     

Interconnection of parameters of regulation system of lipid peroxidation and of morphophysiological parameters of mouse liver

A complex analysis of seasonal fluctuations of the mean group parameters of the system of regulation of lipid peroxidation has been performed in liver of Balb/c mice. Association of lipid characteristics and morphophysiological parameters is studied in the Balb/c mouse liver. An inter-connection is revealed between the liver index and the amount of lysoforms of phospholipids, the scale and character of the interconnection differing essentially depending on proportion of phos-phatidylcholine in mouse liver phospholipids.     

真菌类遗传学分析的知识结构教学

本文以认知结构理论为指导,讨论了真菌类遗传分析与高等动植物遗传分析的内在联系,认为利用这种内在联系进行教学可收到好的效果并说明了作者的具体教学过程。 Abstract:In the paper, the relationship between genetic analysis of Fungi and genetic analysis of high animal and plant was discussed.A good results were obtained when we adopted this method in the teaching.     

Physiological characteristics of various species of strains of carboxydobacteria

Seven strains of aerobic carbon monoxide-oxidizing bacteria (carboxydebacteria) when growing on CO as sole source of carbon and energy had doubling times which ranged from 12–42 h. The activity profiles obtained after discontinuous sucrose density gradient centrifugation indicated that the CO-oxidizing enzymes are soluble and the hydrogenases are membrane-bound in all strains examined. The CO-oxidizing enzymes of Pseudomonas carboxydohydrogena, Pseudomonas carboxydoflava, Comamonas compransoris, and the so far unidentified strains OM2, OM3, and OM4 had a molecular weight of 230,000; that of Achromobacter carboxydus amounted to 170,000. The molecular weights of the CO-oxidizing and H₂-oxidizing enzymes turned out to be identical. The cell sonicates were shown to catalyze the oxidation of both CO and H₂ with methylene blue, thionine, phenazine methosulfate, toluylene blue, dichlorophenolindophenol, cytochrome c or ferricyanide as electron acceptors. Methyl viologen, benzyl viologen, FAD⁺, FMN⁺, and NAD(P)⁺ were not reduced. The spectrum of electron acceptors was identical for all strains tested. Neither free formate, hydrogen nor oxygen gas were involved in the CO-oxidation reaction. Methylene blue was reduced by CO at a 1:1 molar ratio. The results indicate that CO-oxidation by carboxydobacteria is catalyzed by identical or similar enzymes and that the reaction obeys the equation CO+H₂OCO₂+2H⁺+2e^- as previously shown for Pseudomonas carboxydovorans.Dedicated to Otto Kandler remembering almost three decades of enjoyable cooperation     

Modulation of allostery of pyruvate kinase by shifting of an ensemble of microstates

Since the introduction of the concepts of allostery about four decades ago, much advancement has been made in elucidating the structure-function correlation in allostery. However, there are still a number of issues that remain unresolved. In this review we used mammalian pyruvate kinase (PK) as a model system to understand the role of protein dynamics in modulating cooperativity. PK has a triosephosphate isomerase (TIM)(α/β)₈ barrel structural motif. PK is an ideal system to address basic questions regarding regulatory mechanisms about this common (α/β)₈ structural motif. The simplest model accounting for all of the solution thermodynamic and kinetic data on ligand-enzyme interactions involves two conformational states, inactive E^T and active E^R. These conformational states are represented by domain movements. Further studies provide the first evidence for a differential effect of ligand binding on the dynamics of the structural elements, not major secondary structural changes. These data are consistent with our model that allosteric regulation of PK is the consequence of perturbation of the distribution of an ensemble of states in which the inactive E^T and active E^R represent the two extreme end states. Sequence differences and ligands can modulate the distribution of states leading to alterations of functions. The future work includes: defining the network of functionally connected residues; elucidating the chemical principles governing the sequence differences which affect functions; and probing the nature of mutations on the stability of the secondary structural elements, which in turn modulate allostery.