鱼肝油酸钠经皮瘤内注射治疗肝癌的实验研究及机制探讨

目的以肝细胞癌裸鼠动物模型为对象,研究鱼肝油酸钠经皮瘤内注射治疗肝癌的疗效及其机制.方法肝癌模型裸鼠28只,随机分成两组,分别给予鱼肝油酸钠及生理盐水局部浸润注射治疗.每隔2天局部注射1次,共3次.治疗前后分别测肝、肾功能及白细胞计数.第一次注射后两组各随机选取4只裸鼠,分别依次切取肿瘤组织进行病理组织学检查.完成实验,对所有动物行解剖病理检查,明确有无内脏及淋巴结转移情况.结果第一次注射后,治疗组病理切片报告较对照组有不同程度的血管内血栓形成.实验结束时,治疗组肿瘤全部坏死脱落,与对照组肿瘤体积比较有显著性差异(P＜0.001).治疗前后两组肝、肾功能检查及白细胞计数未见统计学差异(P＞0.05).结论鱼肝油酸钠瘤内注射治疗具有显著效果,肿瘤血供障碍是一重要作用机制.并且该药无明显肝、肾毒性.     

瘤内注射复方中药"99-克星"的活性组分抗肝癌作用及机制

目的 探讨超声引导瘤内注射复方中药"99-克星"的活性组分AG-05体内外抗肝癌作用及机制.方法 观察AG-05瘤内注射对小鼠肝癌H22肿瘤生长抑制率的影响;应用磺酰罗丹明(SRB)法检测AG-05对体外培养肝癌HepG2细胞生长的抑制作用,透射电镜观察细胞形态学变化,流式细胞术分析细胞凋亡率.结果 AG-05具有较强的抑制小鼠肝癌H22生长的作用,2 mg/kg、1 mg/kg肿瘤生长抑制率分别达62.8%、37.4%;AG-05对HepG2细胞的生长具有明显的抑制作用,且呈一定的量效、时效关系;AG-05作用48 h后癌细胞出现明显的形态学变化,大部分细胞肿胀坏死,部分细胞出现染色质边集、凋亡小体形成等典型的凋亡形态学改变;流式细胞术检测癌细胞凋亡率实验组20 μg/ml、10 μg/ml两剂量组细胞凋亡率明显升高,分别为 (20.1±4.3)%和(13.5±2.1)%.结论 复方中药 "99-克星"活性组分AG-05具有明显的抗肝癌作用,其作用机理与直接破坏肿瘤细胞及诱导肿瘤细胞凋亡有关.     

重组腺病毒ADV-TK对肝癌抑制作用的实验研究

目的观察已构建的含胸苷激酶（TK）自杀基因的重组腺病毒（ADV-TK）对肝癌细胞的体外杀伤作用和对肝癌裸鼠移植瘤的治疗效果。方法将ADV-TK体外感染人肝癌细胞株SMMC-7721,噻唑蓝（MTT）法检测受感染的SMMC-7721细胞被不同浓度更昔洛韦（GCV）作用后的细胞存活率情况。构建肝癌SMMC-7721裸鼠移植瘤模型,观察肿瘤注射重组腺病毒ADV-TK结合GCV治疗移植瘤的变化。结果相同滴度的重组腺病毒与不同浓度的GCV作用于肝癌细胞株SMMC-7721后,MTT法检测到细胞的存活率随着GCV浓度的增加而不断降低。动物实验中ADV-TK治疗组肿瘤体积明显小于对照组（ADV-null及NS）（P〈0.01）。结论重组腺病毒ADV-TK对肝癌SMMC-7721细胞的体外增殖和裸鼠体内的移植瘤生长均有明显的抑制作用。     

肝细胞刺激因子与阿霉素协同诱导肝癌细胞凋亡的实验研究

目的:研究肝细胞刺激因子(HSS)与阿霉素(ADM)联合用药对裸鼠肝癌移植瘤细胞的凋亡作用.方法:建立裸鼠肝癌移植瘤模型;将荷瘤裸鼠分组,分别腹腔内注射不同剂量HSS及ADM;4周后解剖取瘤计算抑瘤率;病理组织学检查;采用Annexin V/PI流式细胞分析法检测肝癌细胞凋亡率.结果:与对照组相比,三种剂量联合用药组抑瘤作用明显(P＜0.01).HSS组诱导细胞凋亡作用不明显(P＞0.05),ADM组细胞凋亡作用明显(P＜0.01),但裸鼠有明显的不良反应.中小剂量联合用药组细胞凋亡率明显增加(P＜0.01),与ADM组相比也表现出明显差异(P＜0.01).大剂量联合用药组诱导细胞凋亡作用明显(P＜0.01),与ADM组相比差异同样有显著性(P＜0.01),但裸鼠表现出强烈的毒副反应.结论:中小剂量HSS和ADM联合用药抑瘤作用显著,对诱导裸鼠肝癌细胞的凋亡具有协同作用.     

超声引导瘤内注射亲细胞非均质分子脂质治疗肝癌新方法的临床应用

目的探讨超声引导下不饱和脂肪酸类药物--亲细胞非均质分子脂质(CHML)肝癌瘤内注射临床应用效果.方法总结经自动活检病理证实肝细胞肝癌15例注射CHML治疗后的临床疗效,观察治疗前后AFP、CDFI和CT局部肿瘤的改变,并计算半年生存率.结果15例肝细胞肝癌(22结节)用CHML瘤内注射后,瘤体体积均无增大或者明显减小,8例(8/9)AFP下降,7例行CT复查瘤体缩小,增强扫描无强化(7/7),9例注射CHML 2～3次后穿刺病理复查发现肿瘤细胞变性、坏死伴瘤周纤维组织增生,血管内血栓形成,但无血管扩张及增生.CHML注射治疗前后肝、肾、肺功能无明显改变(P＞0.05).结论CHML作为一种局部硬化剂具有明显的抗肿瘤作用,近期临床效果较好.其作用机制可能为造成肿瘤血管内血栓形成,导致肿瘤血供障碍,初步观察认为,CHML与无水乙醇混合应用效果可更佳.     

《临床技术操作规范——超声医学分册》出版

目的探讨超声引导下不饱和脂肪酸类药物--亲细胞非均质分子脂质(CHML)肝癌瘤内注射临床应用效果.方法总结经自动活检病理证实肝细胞肝癌15例注射CHML治疗后的临床疗效,观察治疗前后AFP、CDFI和CT局部肿瘤的改变,并计算半年生存率.结果15例肝细胞肝癌(22结节)用CHML瘤内注射后,瘤体体积均无增大或者明显减小,8例(8/9)AFP下降,7例行CT复查瘤体缩小,增强扫描无强化(7/7),9例注射CHML 2～3次后穿刺病理复查发现肿瘤细胞变性、坏死伴瘤周纤维组织增生,血管内血栓形成,但无血管扩张及增生.CHML注射治疗前后肝、肾、肺功能无明显改变(P＞0.05).结论CHML作为一种局部硬化剂具有明显的抗肿瘤作用,近期临床效果较好.其作用机制可能为造成肿瘤血管内血栓形成,导致肿瘤血供障碍,初步观察认为,CHML与无水乙醇混合应用效果可更佳.     

复方中药99-克星瘤内注射治疗裸鼠肝癌的实验研究

目的探讨超声引导瘤内注射复方中药99-克星(99-克星)治疗裸鼠肝癌的作用机制、抗癌效果及其对机体的影响.方法选取人肝癌SMMC-7721皮下移植的实验裸鼠及肝内原位移植的实验裸鼠模型各14只,随机分为3组,其中99-克星治疗组14只,无水乙醇与生理盐水治疗组各7只.全部裸鼠在接种10 d后每隔5 d向肿瘤内注入治疗药物共4次后处死,治疗前后分别采用高频超声测量肿瘤三径并计算肿瘤的生长指数.采用放射免疫法(RIA)测定治疗后荷瘤鼠血清白介素2(IL-2)和肿瘤坏死因子(TNF-α)含量,采用末端脱氧核苷酸转移酶介导的dUTP切口末端标记法(TUNEL)对细胞凋亡现象进行原位观察并观察血清TNF-α水平与肝癌细胞凋亡的关系.治疗前后分别测血天门冬氨酸氨基转移酶(AST)、丙氨酸氨基转移酶(ALT)、碱性磷酸酶(ALP)、尿素氮(BUN)及白细胞计数.实验完成后解剖取检28只实验裸鼠心、肝、肾、脾、肺、脑等器官行病理形态学观察.实验均在双盲条件下进行.结果治疗后克星组肿瘤生长指数(0.080±0.024)明显低于盐水组(4.842±1.293)(P＜0.01),并且低于乙醇组(0.083±0.026),但差异无统计学意义(P＞0.05);治疗后克星组血清IL-2和TNF-α水平明显高于盐水组及乙醇组(P＜0.05);克星组凋亡指数(AI)高达(45.08±5.39)%,显著高于乙醇组(13.45±2.54)%和盐水组(12.38±3.91)%(P＜0.01);治疗前后ALT、AST、BUN及白细胞计数均无明显变化,3组间比较无统计学差异;血清TNF-α含量及肝癌细胞凋亡指数间呈正相关(r=0.494,P＜0.05);3组实验裸鼠心、肝、肾、脾、肺、脑等器官均无病理形态学异常改变.结论99-克星具有明显破坏和抑制肿瘤细胞生长的作用,而无抑制骨髓及损害肝肾功能等毒副作用,其作用机制不同于无水乙醇,主要通过提高血清细胞因子IL-2和TNF-α水平激发机体细胞免疫功能以及诱导癌细胞凋亡,从而达到抑制肿瘤生长的目的.     

亲细胞非均质分子脂质局部注射治疗乳癌的实验研究

目的阐明CHML经皮瘤内注射治疗乳癌的效果并探讨其机理,明确CHML有无骨髓抑制及肝肾功能损伤等副作用.方法5周龄裸鼠共30只,分别于右腋后皮下直接种植人MCF-7乳癌组织块(0.2 cm×0.2 cm×0.2 cm),制成乳癌动物模型.2周后,肿瘤直径长至0.2～0.8 cm,随机分成2组,分别给予CHML及生理盐水局部浸润注射.治疗组CHML剂量按瘤体大小为50 mg/cm3,隔2 d局部注射1次,共3次.对照组给予相应容量的生理盐水.治疗前及治疗后分别测ALT、AST、ALP、BUN及白细胞数,以明确有无肝肾功能损伤和骨髓抑制.第1次注射后48h,两组分别处死5只裸鼠,对肿瘤进行原位末端标记(TUNEL),以明确有无凋亡.完成局部注射后2周,处死所有裸鼠,解剖并行病理检查,明确两组肿瘤愈合及淋巴结转移情况.结果(1)局部注射后48h治疗组5只裸鼠的肿瘤均出现不同程度的凋亡( ～ ),而对照组仅有个别细胞凋亡(-).(2)实验结束时,治疗组肿瘤全部脱落,且无残存癌细胞;而对照组肿瘤明显增大,瘤体直径平均(0.94±0.34)cm,两组有显著统计学差异(P＜0.001).(3)治疗前后ALT、AST、ALP、BUN及白细胞均无明显变化,两组比较无统计学差异.结论CHML瘤内局部注射可有效杀灭乳腺癌细胞,并可诱导凋亡,且无抑制骨髓和损伤肝肾功能的副作用.     

三氧化二砷局部注射治疗原发性肝癌的护理38例

临床确诊时的原发性肝癌大多为晚期 ,恶性程度高 ,手术切除机会少。因此 ,失去手术机会的原发性肝癌主要以肝动脉栓塞介入治疗为主。大量的基础研究表明 ,三氧化二砷(As2 O3)对肝癌细胞具有诱导凋亡等作用 ,经皮肝穿病灶局部中心注射As2 O3充分利用As2 O3诱导肝细胞分化、促进凋亡的作用 ,达到抑制及消灭肝癌细胞的目的。 2 0 0 1年开始我院应用在B超引导下病灶局部注射As2 O3治疗原发性肝癌38例 ,收到了较好的疗效 ,现将术前、术中及术后护理介绍如下。1　临床资料1 1　一般资料选择均经病理证实的原发性肝癌 38例 ,其中原发性肝细胞癌…     

超声评价三氧化二砷量与肝癌细胞凋亡关系的实验性研究

目的探讨三氧化二砷(As_2O_3)体外诱导肝癌细胞凋亡作用,利用超声观察AszO_3对小鼠肝癌的抑瘤效果,探讨其应用价值。方法 (1)以肝癌细胞株HepG2为研究对象,在体外用浓度分别为0、1、2、4、5、6、7 μmol/L的As_2P_3。作用48 h,显微镜观察细胞形态学变化,用MTT法和流式细胞术检测As_2O_3对HepG2的抑制作用;(2)裸鼠30只,随机分成2组。皮下注射HepG2细胞悬液,制作肿瘤模型,用0.1ml的PBS和不同浓度的As_2O_3进行多点注射。观察肿瘤的超声影像改变,病理检查肿瘤坏死情况。结果 (1)As_2O_3能抑制肝癌细胞生长,呈现剂量依赖性。(2)As_2O_3根据干预浓度不同,抑瘤效果不一致,超声观察其肿瘤大小及体积之间差异有统计学意义(P0.01)。结论 As_2O_3能抑制HepG2生长,促进其凋亡,超声作为一种检查手段,可观察肿瘤生长的情况,为抑瘤效果的评价提供有价值的信息。     

Shenfu injection for improving cellular immunity and clinical outcome in patients with sepsis or septic shock

Purpose

To assess the efficacy of Shenfu injection (SFI) for enhancing cellular immunity and improving the clinical outcomes of patients with septic shock.

131I美妥昔单抗注射液介入治疗肝细胞癌的护理

肝细胞癌是我国高发的恶性肿瘤,就死亡率而言,已位居第3位.其病程发展很快,病人就诊时往往已属中晚期.经导管肝动脉灌注(TAI)及栓塞术(TAE)是肝细胞癌非手术治疗的最有效方法之一[1],但远期疗效不佳.     

Charge injection based electrical stimulation on polypyrrole planar electrodes to regulate cellular osteogenic differentiation

In this study, polypyrrole (Ppy) electrodes were prepared to support an electrical stimulation to MC3T3-E1 cells for regulating their osteogenic differentiation. The charge injection capacity (C_Q) of the Ppy electrodes could be adjusted by the Ppy thickness, and a higher C_Q could make the electrode able to produce a higher charge injection quantity (Q_inj) at applied voltage. The Q_inj onto electrode could be considered as the intensity of the stimulation pulse to cells, and the pulse frequency means the number of electric stimulation with Q_inj at one second. Hence, we conducted the present work in the view of Q_inj. When the cells were electrically stimulated for 1 hour per day, the electrodes with Q_inj ranged in 0.08–0.15 μQ had an obvious role in enhancing cellular osteogenic differentiation whereas Q_inj of lower than 0.03 μQ or more than 0.30 μQ gave the stimulations with no or negative effects. And the stimulation with 1 or 25 Hz showed to enhance the differentiation, whereas the stimulation with 50 Hz gave an inhibiting effect. We further found the osteogenic differentiation potential triggered by electrical simulation was related to cell growth stage, and the stimulation carried out at early stage (day 2–5) during 8 days cell culture showed more contribution to enhancing osteogenic differentiation than that at later stage (day 6–8). It is proposed that the desired stimulation effects require that an appropriate voltage-gated calcium ion channel and efficient intracellular calcium ion oscillation are well activated. This work therefore reveals Q_inj as an important electrode parameter to decide effective simulations and provides an insight into understanding of the role of electrode material characters in regulating cellular osteogenic differentiation during stimulation.

This study reveals that the Q_inj on electrodes is a more significant factor than applied voltage for electrical stimulation to regulate cellular osteogenic differentiation, and the charge injection capacity can be tuned by thickness of Ppy.     

Plasmid DNA vaccines: investigation of integration into host cellular DNA following intramuscular injection in mice

The primary safety concern for DNA vaccines is their potential to integrate into the host cell genome. We describe an integration assay based on purification of high-molecular-weight genomic DNA away from free plasmid using gel electrophoresis, such that the genomic DNA can then be assayed for integrated plasmid using a sensitive PCR method. The assay sensitivity was approximately 1 plasmid copy/microg DNA (representing approximately 150,000 diploid cells). Using this assay, we carried out integration studies of three different plasmid DNA vaccines, containing either the influenza hemagglutinin, influenza matrix or HIV gag gene. Six weeks after intramuscular injection, free plasmid was detected in treated muscle at levels ranging from approximately 1,000 to 4,000 copies/microg DNA. At 6 months, the plasmid levels ranged between 200 and 800 copies/microg DNA. Gel purification of genomic DNA revealed that essentially all of the detectable plasmid in treated quadriceps was extrachromosomal. If integration had occurred, the frequency was 相似文献   

黄芪注射液雾化吸入对反复呼吸道感染患儿细胞免疫功能的影响

目的观察黄芪注射液雾化吸入治疗对反复呼吸道感染(RRTI)患儿细胞免疫功能的影响。方法将55例RRTI患儿分为A组30例和B组25例,另设C组正常对照组20例。A组在常规治疗基础上加用黄芪注射液雾化吸入治疗,2次/d,连用10 d为1个疗程。采用流式细胞仪观察A、B组于治疗前后、C组于健康体检时外周血T细胞亚群(CD3、CD4、CD4/CD8)量的变化。结果 A组黄芪治疗前与C组正常对照组比较,CD3、CD4、CD4/CD8均明显低于对照组(P<0.01),A组黄芪治疗前及治疗后比较,CD3、CD4、CD4/CD8均明显回升(P<0.01),而A、B组治疗后比较差异有统计学意义。结论黄芪注射液雾化吸入给药可提高RRTI患儿细胞免疫功能,且操作简单方便,疗效显著,不良反应小,迅速发挥治疗作用。     

Phagocytosis and cellular metabolism

Summary The uptake of foreign particles by mouse and human macrophages influenced by various metabolic inhibitors was examined in order to obtain further informations about the energy-dependent mechanisms which are involved in the phagocytic process. The inhibitors employed were iodoacetate, fluoroacetate, fluoride, malonate, sodium azide, 2-4-dinitrophenol, cycloheximide and ouabain. These substances were rested on monolayer cultures and the phagocytosis assay was performed by using zymosan suspension in the nutrient media. The quantitation of phagocytosis was obtained by the direct count of intracellular zymosan particles (immersion microscopy, 100x) and the results were evaluated and compared by biometrical analysis. The effects of these inhibitors on phagocytosis and their relation with the metabolic intracellular pathways are discussed.