砷剂对K_(562)细胞凋亡作用的基础研究

[目的]研究三氧化二砷对K562细胞凋亡的诱导.[方法]采用人红白血病细胞株K562细胞常规培养,给不同浓度的三氧化二砷,在不同的时间收获细胞,用台盼蓝排染法,DNA荧光染料Hoechst33342荧光染色法,及碘化丙啶(PI)与Hoechst33342共染计数坏死细胞的PI阳性率等方法,检测其对K562细胞的影响.[结果]三氧化二砷能够诱导K562细胞凋亡.并且呈现浓度依赖性和时间依赖性.[结论]三氧化二砷主要以诱导肿瘤细胞凋亡而表现其毒性作用.     

刺五加多糖对人白血病K562细胞作用的研究

目的:研究刺五加多糖对体外培养人白血病K562细胞有无增殖抑制和凋亡诱导作用。方法:取培养至对数生长期的K562细胞(密度为5×104/mL和1×106/mL)分别接种于96孔培养板(100μL/孔)及50mL培养瓶(1.5mL/瓶)中,加入不同浓度的刺五加多糖作用24h后,用CCK-8法检测刺五加多糖对K562细胞增殖抑制作用;荧光显微镜检测细胞凋亡。结果:不同浓度刺五加多糖(0.405、0.810、1.620、2.430、3.240mg/mL)作用K562细胞24 h,抑制率分别为16%、27%、48%、50%、55%;荧光显微镜下观察发现培养K562细胞中出现核固缩、凋亡小体。结论:刺五加多糖对体外培养K562细胞生长有明显的抑制作用,可诱导K562细胞凋亡。     

硫氧还蛋白还原酶抑制剂乙烷硒啉逆转顺铂耐药及其机制研究（英文）

目的:研究凋亡调控相关蛋白来了解顺铂耐药成因,同时考察乙烷硒啉(Ethaselen)在K562耐药细胞中逆转顺铂耐药的作用,并初步探讨其作用机制。创新点:首次研究乙烷硒啉在逆转顺铂耐药中的作用,且此作用与乙烷硒啉诱导细胞凋亡相关。方法:通过长时间脉冲诱导得到顺铂耐药K562细胞,并观察耐药细胞形态及倍增时间。采用MTT法考察乙烷硒啉、顺铂及其联用组在不同细胞株间的生长抑制作用。流式细胞术分析细胞凋亡情况以及细胞内活性氧(ROS)水平。最后,通过蛋白质免疫印迹(Western blot)考察凋亡调控相关蛋白水平的变化。结论:脉冲诱导得到的K562耐药细胞对顺铂的耐受性是原K562细胞的5.34倍。形态学观察发现,耐药细胞体积增大,粘附性进一步降低。乙烷硒啉与顺铂联用表现出协同效应。当加入少量的乙烷硒啉(顺铂与乙烷硒啉的摩尔比率为10:1),顺铂作用K562耐药细胞的半抑制浓度(IC50)值可以减少21倍。流式细胞术及Western blot表明,乙烷硒啉能够诱导耐药细胞凋亡。其逆转顺铂耐药主要是通过调控Bcl-2及Bax蛋白比例以及通过提高细胞内活性氧水平引起线粒体通透转运孔道(PTP)蛋白孔道的形成来促使释放细胞色素c,进而引起Caspase凋亡途径。     

黄芪成分F_3新制剂对胃癌细胞株的抑制作用

[目的]观察黄芪成分F3新制剂对胃癌(BGC-823)细胞株体外细胞生长、抑制作用.[方法]利用基因组DNA电泳,流式细胞仪检测细胞凋亡.[结果]基因组DNA电泳出现“梯形”条带;流式细胞仪分析出现低于G1期DNA含量的亚二倍体凋亡率.[结论]黄芪成分F3新制剂对胃癌细胞有明显的抑制作用.     

立体定向血肿抽吸对大鼠脑出血细胞凋亡的影响

[目的]探讨大鼠脑出血双侧纹状体区细胞凋亡的时程变化规律和早期立体定向血肿抽吸术干预治疗对细胞凋亡的影响.[方法]雄性Sprague-Dawley(SD)大鼠80只,随机分成正常对照组,假手术组,脑出血自然恢复组,6 h血肿抽吸组,12 h血肿抽吸组以及24 h血肿抽吸组.采用立体定向注入Ⅶ型胶原酶建立脑出血模型.采用原位末端标记法检测出血周边组织细胞的凋亡.[结果]①大鼠脑出血周边组织12 h出现凋亡细胞,3d凋亡细胞达峰值,11 d仍有凋亡细胞表达.②血肿抽吸术干预后出血周边组织凋亡细胞数与脑出血组对应时间点比较显著下降(P<0.05).③6 h血肿抽吸组与24 h血肿抽吸组比较细胞凋亡数明显低于24 h血肿抽吸组,差异有显著性(P<0.05).[结论]立体定向血肿抽吸术干预治疗大鼠脑出血可明显影响细胞凋亡的表达,脑出血各种临床治疗的“时间窗”应为出血后12 h之内.早期(6 h)立体定向血肿抽吸术治疗可以显著减少脑出血后细胞凋亡.     

高效液相色谱法测定K562细胞对顺铂的摄取

顺铂是目前治疗癌症最有效的药物之一,测定细胞内顺铂的含量对于了解细胞摄取顺铂和顺铂发挥药效有重要作用．本研究通过MTT法检测了顺铂杀伤K562细胞的有效浓度,建立并利用高效液相色谱法（HPLC）检测了K562细胞对顺铂的摄入．结果表明K562细胞经10μg/mL顺铂处理12h后,细胞活性显著下降;细胞外有36．75％顺铂进入胞内．实验数据显示,顺铂对K562细胞的杀伤与其在胞内的累积相关．     

针刺预处理对全脑缺血大鼠微血管损伤及脑水肿的保护作用

[目的]针刺预处理对全脑缺血大鼠微血管损伤及脑水肿的保护作用.[方法]采用四动脉阻断法制作大鼠全脑缺血模型,针刺预处理组:术前给予针刺,针刺7 d后给予全脑缺血10 min,干湿法测脑组织水含量,应用HE染色、TUNEL染色及透射电镜技术,观察大鼠脑缺血后,组织病理学及超微结构改变.[结果]与脑缺血组相比,针刺预处理组不引起明显的神经元损伤,全脑缺血10 m in后72 h,微血管损伤及脑水肿明显减轻,血管内皮细胞凋亡减少.[结论]针刺预处理能够通过减轻微血管损伤及脑水肿对脑缺血产生保护作用.     

抑制口腔白色念珠菌感染的实验研究

[目的]筛选对口腔白色念珠菌具有明显拮抗作用的菌株.[方法]通过纸片琼脂扩散法(K-B法)观察各菌株对白色念珠菌的拮抗作用;再利用试管法观察有拮抗作用菌株对白色念珠菌的抑菌效果.[结果]K-B法表明大肠杆菌、甲型链球菌、卡他球菌对白色念珠菌无抑菌作用,表皮葡萄球菌、微球菌有抑菌作用,但抑菌环小;枯草杆菌有明显抑菌作用.试管法表明表皮葡萄球菌和枯草杆菌对白色念珠菌均有生物拮抗作用,其抑菌率分别为98%和90%.[结论]枯草杆菌是白色念珠菌的理想拮抗菌株.     

β-连环素与p63在大鼠舌部鳞状细胞癌癌变过程中作用的初步研究

[目的]研究β-连环素与p63基因在大鼠舌部鳞状细胞癌中的表达,探讨它们在口腔鳞状细胞癌增殖中的作用.[方法]免疫组织化学方法检测48例不同分化程度大鼠舌部鳞状细胞癌中β-连环素与p63基因的表达.[结果]8例高分化口腔鳞状细胞癌中分别有7和6例高表达p63与β-连环素.β-连环素和p63表达与大鼠舌部鳞状细胞癌的组织学分级不具有显著性差异(P>0.05).[结论]β-连环素在大鼠舌鳞状细胞癌增殖中的作用与在人回顾性研究中的作用相似.     

砷剂对肿瘤多药耐药细胞株作用机理的研究

三氧化二砷（As2O3）是人们观念中的剧毒物砒霜中的有效成分．它既是一种致癌剂又有一定有益的生物学作用．本实验研究证实，砷剂在非细胞毒性剂量下能降低化疗药物阿霉素（ADM）对多耐药（multi drug resistance，MDR）肿瘤细胞株K562/ADM细胞的IC50（细胞抑制率达50％时化疗药物的浓度），表明三氧化二砷具有逆转人红白血病细胞株K562/ADM细胞MDR作用．     

Protective paracrine effect of mesenchymal stem cells on cardiomyocytes

Objective  The aim of this study was to test the protective effect of mesenchymal stem cells (MSCs) on cardiomyocytes in vitro and to investigate the anti-apoptotic signaling pathway. Methods  MSCs from Sprague-Dawley (SD) rats were separated and cultured. MSC medium was collected from MSCs cultured in serum-free Dulbecco’s modified eagle medium (DMEM) under hypoxia. Cultured cardiomyocytes from neonatal SD rats were exposed to hypoxia/reoxygenation (H/R) and treated with MSC medium. The apoptotic cardiomyocytes were stained with Annexin-V-fluorescein isothiocyanate (FITC), Hoechst 33342 and terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling (TUNEL). The mitochondrial transmembrane potential of cardiomyocytes was assessed using a fluorescence microscope. The expression of Bcl-2, Bax, cytochrome C, apoptosis-induced factor (AIF), and caspase-3 was tested by Western blot analysis. Results  Our data demonstrated that MSC medium reduced H/R-induced cardiomyocyte apoptosis, increased the Bcl-2/Bax ratio, and reduced the release of cytochrome C and AIF from mitochondria into the cytosol. Conclusion  MSCs protected the cardiomyocytes from H/R-induced apoptosis through a mitochondrial pathway in a paracrine manner. Project supported by the National Natural Science Foundation of China (No. 30670868) and the Natural Science Foundation of Zhejiang Province, China (No. R206007)     

冷凝或联合小梁切除术治疗新生血管性青光眼

[目的]探讨冷凝联合小梁切除术治疗新生血管性青光眼的疗效.[方法]A组12例14眼新生血管性青光眼,施行180°视网膜周边部巩膜外和睫状体冷凝;B组20例22眼,除上述冷凝外,又联合小梁切除术治疗.[结果]随访3～6个月,两组术后视力均无明显改善,A组平均眼压自57.48mmHg降至32mmHg,需要再做冷凝或手术减压;B组随访的15例(16眼)有14眼的眼压<21mmHg,2眼需加用0.1%噻吗心安滴眼治疗,两组术后短期内眼压控制有明显的差异(P<0.05).[结论]冷凝或联合小梁切除术对新生血管性青光眼治疗有较好的短期疗效.     

Protective effect of recombinant Lactobacillus plantarum against H2O2-induced oxidative stress in HUVEC cells

This study probed the protective effect of recombinant Lactobacillus plantarum against hydrogen peroxide(H_2O_2)-induced oxidative stress in human umbilical vein endothelial cells(HUVECs). We constructed a new functional L. plantarum(NC8-p SIP409-alr-angiotensin-converting enzyme inhibitory peptide(ACEIP)) with a double-gene-labeled non-resistant screen as an expression vector. A 3-(4, 5-dimethyl-2-thiazolyl)-2, 5-diphenyl-2 H-tetrazolium bromide(MTT) colorimetric assay was carried out to determine the cell viability of HUVEC cells following pretreatment with NC8-p SIP409-alr-ACEIP. Flow cytometry(FCM) was used to determine the apoptosis rate of HUVEC cells. Cysteinyl aspartate specific proteinase(caspase)-3/8/9 activity was also assayed and western blotting was used to determine protein expression of B-cell lymphoma 2(Bcl-2), Bcl-2-associated X protein(Bax), inducible nitric oxide synthase(i NOS), nicotinamide adenine dinucleotide phosphate oxidase 2(gp91 phox), angiotensin II(Ang II), and angiotensin-converting enzyme 2(ACE2), as well as corresponding indicators of oxidative stress, such as reactive oxygen species(ROS), mitochondrial membrane potential(MMP), malondialdehyde(MDA),and superoxide dismutase(SOD). NC8-p SIP409-alr-ACEIP attenuated H_2O_2-induced cell death, as determined by the MTT assay. NC8-p SIP409-alr-ACEIP reduced apoptosis of HUVEC cells by FCM. In addition, compared to the positive control, the oxidative stress index of the H_2O_2-induced HUVEC(Hy-HUVEC), which was pretreated by NC8-p SIP409-alr-ACEIP, i NOS,gp91 phox, MDA, and ROS, was decreased obviously; SOD expression level was increased; caspase-3 or-9 was decreased, but caspase-8 did not change; Bcl-2/Bax ratio was increased; permeability changes of mitochondria were inhibited; and loss of transmembrane potential was prevented. Expression of the hypertension-related protein(Ang II protein) in HUVEC cells protected by NC8-p SIP409-alr-ACEIP decreased and expression of ACE2 protein increased. These plantarum results suggested that NC8-p SIP409-alr-ACEIP protects against H_2O_2-induced injury in HUVEC cells. The mechanism for this effect is related to enhancement of antioxidant capacity and apoptosis.     

钙拮抗剂治疗腹泻型及便秘型肠易激综合征临床观察

[目的]探讨钙拮抗剂治疗不同类型肠易激综合征疗效.[方法]选择腹泻型肠易激综合征与便秘型肠易激综合征患者分别分成治疗组和对照组,治疗后进行疗效观察,各型组间进行统计学分析.[结果]显示钙拮抗剂对腹泻型肠易激综合征腹痛及腹部不适有明显效果(P<0.05),便秘型肠易激综合征腹痛疗效不显著(P>0.05).[结论]钙拮抗剂治疗不同类型肠易激综合征疗效有差异.     

Pure total flavonoids from Citrus paradisi Macfadyen actmsynergistically with arsenic trioxide in inducingmapoptosis of Kasumi-1 leukemia cells in vitro

To investigate the potential effects of pure total flavonoid compounds (PTFCs) from Citrus paradisi Macfadyen separately or combined with arsenic trioxide on the proliferation of human myeloid leukemia cells and the mechanisms underlying the action of PTFCs. The effects of PTFCs separately or combined with arsenic trioxide on the proliferation and apoptosis of leukemia cells were determined by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT), fluorescence microscopy, and flow cytometry. Their effects on the expression levels of apoptosis-related regulators were determined by Western blot assay. PTFCs combined with arsenic trioxide significantly inhibited the growth of Kasumi-1 cells, and apoptosis was confirmed by flow cytometry analysis. Hoechst 33258 staining showed more significant morphological changes and more apoptosis following the combined treatment. Western blots showed changes in the expression of genes for poly ADP-ribose polymerase (PARP), caspase 3/9, and P65. The results indicated that PTFCs separately or combined with arsenic trioxide inhibited proliferation of leukemia cells in vitro and induced their apoptosis by modulating the expression of apoptosis-related regulator genes.     

Protective mechanisms of hypaconitine and glycyrrhetinic acid compatibility in oxygen and glucose deprivation injury

This study investigated the protective effect of the compatibility of hypaconitine (HA) and glycyrrhetinic acid (GA) on H9c2 cells under oxygen and glucose deprivation (OGD)-induced injury, and the possible mechanisms. We found that HA+GA significantly improved pathology and morphology of the nucleus and ultrastructure of H9c2 cells under OGD as determined by Hoechst 33342 staining and transmission electron microscopy (TEM) tests. It also reduced the releases of lactate dehydrogenase (LDH), creatine kinase-myocardial band isoenzyme (CK-MB), and aspartate transaminase (AST) from the cultured supernatant of H9c2 cells, which were tested by enzyme-linked immune sorbent assay (ELISA) kits. In addition, it lessened the apoptotic rate as determined by a fluorescein isothiocyanate-annexin V/propidium iodide (FITC-AV/PI) double staining assay. It was also found that HA+GA might regulate the protein expression associated with the phosphatidylinositol 3-kinase (PI3K)/Akt signaling pathway. Overall, the study demonstrated that HA+GA protected H9c2 cells against OGD-induced injury, and the signaling mechanism might be related to the PI3K/Akt signaling pathway.     

炎性乳腺癌的诊断和治疗

[目的]:探索炎性乳腺癌的诊断和治疗.[方法]:10例经病理或细胞学检查确诊的炎性乳腺癌先给2~3个周期的CAF(阿霉素+环磷先胺+氟尿嘧啶)方案化疗,然后行总量的2/3量的乳腺及同侧腋窝的放射治疗,放疗结束后休息2周即行乳癌根治切除术,术后切口愈合后再给剩余1/3量的放射治疗,照射胸壁及同侧腋窝和锁骨上使放射总量达到60~70 Gy/野,在放疗前后穿插化疗(CAF)4个周期.[结果]:1例患者未完成治疗出院后1个月死亡.9例患者完成上述序惯治疗,全部病例乳腺红肿、高张消退,乳腺内肿块及同侧腋窝淋巴结明显缩小.手术切除标本检查有5例乳腺内找到癌灶,其中典型炎性癌1例、单纯癌2例、硬癌2例,4例乳腺组织内未找到癌灶.同侧腋窝淋巴结见癌转移7例,同侧锁骨上淋巴结见癌转移2例.生存时间1~28个月,中位生存时间12个月.[结论]:炎性乳腺癌通过序惯治疗可以提高治疗的有效性、延长生命,提高生存质量.