Intraperitoneal perfusion of cytokine-induced killer cells with local hyperthermia for advanced hepatocellular carcinoma

AIM:To study the effect and tolerance of intraperitoneal perfusion of cytokine-induced killer(CIK) cells in combination with local radio frequency(RF) hyperthermia in patients with advanced primary hepatocellular carcinoma(HCC).METHODS:Patients with advanced primary HCC were included in this study.CIK cells were perfused intraperitoneal twice a week,using 3.2 × 10 9 to 3.6 × 10 9 cells each session.Local RF hyperthermia was performed 2 h after intraperitoneal perfusion.Following an interval of one month,the next course of treatment was administered.Patients received treatment until disease progression.Tumor size,immune indices(CD3 +,CD4 +,CD3 + CD8 +,CD3 + CD56 +),alpha-fetoprotein(AFP) level,abdominal circumference and adverse events were recorded.Time to progression and overall survival(OS) were calculated.RESULTS:From June 2010 to July 2011,31 patients diagnosed with advanced primary HCC received intraperitoneal perfusion of CIK cells in combination with local RF hyperthermia in our study.Patients received an average of 4.2 ± 0.6 treatment courses(range,1-8 courses).Patients were followed up for 8.3 ± 0.7 mo(range,2-12 mo).Following combination treatment,CD4 +,CD3 + CD8 + and CD3 + CD56 + cells increased from 35.78% ± 3.51%,24.61% ± 4.19% and 5.94% ± 0.87% to 45.83% ± 2.48%(P = 0.016),39.67% ± 3.38%(P = 0.008) and 10.72% ± 0.67%(P = 0.001),respectively.AFP decreased from 167.67 ± 22.44 to 99.89 ± 22.05 ng/mL(P = 0.001) and abdominal circumference decreased from 97.50 ± 3.45 cm to 87.17 ± 4.40 cm(P = 0.002).The disease control rate was 67.7%.The most common adverse events were low fever and slight abdominal erubescence,which resolved without treatment.The median time to progression was 6.1 mo.The 3-,6-and 9-mo and 1-year survival rates were 93.5%,77.4%,41.9% and 17.4%,respectively.The median OS was 8.5 mo.CONCLUSION:Intraperitoneal perfusion of CIK cells in combination with local RF hyperthermia is safe,can efficiently improve immunological status,and may prolong survival in HCC patients.

Human induced pluripotent stem cells labeled with fluorescent magnetic nanoparticles for targeted imaging and hyperthermia therapy for gastric cancer

Objective: Human induced pluripotent stem(i PS) cells exhibit great potential for generating functional human cells for medical therapies. In this paper, we report for use of human i PS cells labeled with fluorescent magnetic nanoparticles(FMNPs) for targeted imaging and synergistic therapy of gastric cancer cells in vivo. Methods: Human i PS cells were prepared and cultured for 72 h. The culture medium was collected, and then was coincubated with MGC803 cells. Cell viability was analyzed by the MTT method. FMNP-labeled human i PS cells were prepared and injected into gastric cancer-bearing nude mice. The mouse model was observed using a small-animal imaging system. The nude mice were irradiated under an external alternating magnetic field and evaluated using an infrared thermal mapping instrument. Tumor sizes were measured weekly. Results: iP S cells and the collected culture medium inhibited the growth of MGC803 cells. FMNP-labeled human iP S cells targeted and imaged gastric cancer cells in vivo, as well as inhibited cancer growth in vivo through the external magnetic field. Conclusion: FMNP-labeled human i PS cells exhibit considerable potential in applications such as targeted dual-mode imaging and synergistic therapy for early gastric cancer.

Effect of Tirapazamine and Mild Temperature Hyperthermia on the Recovery from Radiation-Induced Damage in Pimonidazole-Unlabeled Quiescent Tumor Cell Population

The aim in this study is to examine the effect of tirapazamine (TPZ) and mild temperature hyperthermia (MTH) on the repair of radiation-induced damage in pimonidazole-unlabeled quiescent (Q) tumor cells. Labeling of proliferating (P) cells in C57BL/6J mice bearing EL4 tumors was achieved by continuous administration of 5-bromo-2-deoxyuridine (BrdU). Tumors were irradiated with γ-rays at 1 h after the administration of pimonidazole followed by TPZ treatment or MTH. Twenty-four hours later, assessment of the responses of Q and total (= P + Q) cells were based on the frequencies of micronucleation and apoptosis using immunofluorescence staining for BrdU. The response of the pimonidazole-unlabeled tumor cell fractions was assessed by means of apoptosis frequency using immunofluorescence staining for pimonidazole. With γ-rays only, the pimonidazole-unlabeled cell fraction showed significantly enhanced radio-sensitivity compared with the whole cell fraction more remarkably in Q cells than total cells. However, a significantly greater decrease in radio-sensitivity in the pimonidazole-unlabeled than the whole cell fraction, evaluated using a delayed assay, was more clearly observed in Q cells than total cells. Post-irradiation MTH more remarkably repressed the decrease in radio-sensitivity in the Q cell than the total cells. Post-irradiation TPZ administration produced a large radio-sensitizing effect on both total and Q cells, especially on Q cells. On the other hand, in pimonidazole-unlabeled cell fractions in both total and Q cells, TPZ suppressed the reduction in sensitivity due to delayed assay much more efficiently than MTH, whereas no radio-sensitizing effect was produced. Not only through suppressing the recovery from radiation-induced damage but also through radio-sensitizing effect, post-irradiation TPZ administration is very useful for repressing the increase in the difference in radio-sensitivity due to the delayed assay not only between total and Q tumor cells but also between the pimonidazole-unlabeled and the whole cell fractions within the total and Q tumor cells.

A DYNAMIC STUDY OF THE CYTOTOXIC EFFECTS OF HYPERTHERMIA COMBINED WITH CIS-DIAMINE DICHLOROLPLATINUM(DDP) ON HUMAN GASTRIC CANCER CELL LINES MKN_(28) AND MKN_(45) IN VITRO

Cytotoxic effects of hyperthermia combined with DDP on MKN28 and MKN45 cells were studied by MTT assay according to a nested design. The results showed:hyperthermia alone above 43℃ for 30 mins was cytotoxic;hyperthermia at temperature lower than 43℃ for 30 mins could increase sensitivity of cancer cells to DDP. The cytotoxic effect of simultaneous use of hyperthermia and DDP was more marked than that of sequential use of the 2 treatments. Hyperthermia combined with DDP could inhibit growth of human gastric adenocarcinoma cells regardless of their degree of differentiation.

Effect of hyperthermia in combination with cryotherapy on sarcoma 180 in mice

The significant decrease in tumor recurrence caused by combined hyperthermia (local water bath) with cryotherapy has previously been shown. This study demonstrated an in vivo additive interaction of local microwave diathermy (hyperthermia, 42±0.5℃, 20 min. ) and liquid nitrogen treatment (cryotherapy,-180℃, 3 min. ) against intraperitoneally implanted sarcoma 180 in ICR mice. Local hyperthermia was produced by applicating 2450 MHz microwave to the region of the tumor without induction of significant whole body hyperthermia.Both hyperthermia and cryotherapy were delivered as a single dose individually on day 1 and day 2. The analysis of the tumor size curves showed that all of the treatments had more significant effect on sarcoma 180 than on the control group (P < 0. 0001 ). The optimal sequence of the combined therapies found in the experiment was cryotherapy→hyperthermia (CH) (P<0. 0001). Weights of the tumors excised from the mice on day 11 showed that the combined therapies (cryotherapy→hyperthermia or hyperthermia→cryotherapy) were more effective on sarcoma 180 than hyperthermia or cryotherapy used alone(P<0.01).

基于SVMD-PSR-MNF的海洋MT数据噪声压制方法

海洋大地电磁法是一种天然源方法,其采集的大地电磁信号具有场强微弱、易受干扰等特点.在浅水环境下,海浪运动引起的复杂强烈的感应电磁场是大地电磁信号主要干扰源,极大降低了大地电磁数据的信噪比,致使视电阻率和相位畸变.针对这种复杂的强能量噪声,本文提出SVMD-PSR-MNF方法对其进行压制.通过连续变分模态分解(Successive Variational Mode Decomposition, SVMD)实现噪声预估计,然后利用相空间重构(Phase Space Reconstruction,PSR)将时间序列转化为矩阵,在此基础上,结合最小噪声分离(Minimum Noise Fraction,MNF)将含噪MT数据分解为一系列MNF成分,最后通过MNF成分的选取重构实现信噪分离.针对实测数据去噪效果评价难的问题,本文基于大地电磁信号的非平稳特性和海浪感应磁噪声的周期性,提出基于自相关函数(Autocorrelation Function,ACF)的去噪效果评价方法.为了验证提出方法的有效性,将其应用于合成数据和南黄海实测数据的处理中.本文基于一维各向异性模型模拟连续频谱的MT信号,并采用三维随机海浪感应磁场公式合成符合实际情况的含噪MT数据.合成数据和南黄海实测数据的处理结果表明,SVMD-PSR-MNF方法可在压制噪声的同时较好地恢复有效信号,校正噪声导致的视电阻率和相位的畸变,提高MT数据质量;ACF可评价海浪感应磁噪声的压制效果.

Distinct Cytoskeletal Injuries Induced by As,Cd,Co,Cr,and Ni Compounds

The risks of metal compounds to human health are highlighted by the ubiquity of exposure and their persistence in the environment.Although compounds of As,Cd,Co,Cr,and Ni are known or“reasonably anticipated”to be carcinogenic to humans and/or experimental animals, the cellular targets of these health hazards and the underlying mechanisms of their carcinogenic- ity are still unclear.We show in this report that dramatic,time-and dose-dependent cytoskeletal perturbations,especially in the distribution and organization patterns of microtubules and mi- crofilaments,two of the principal components of the cytoskeleton,occurred in 3T3 cells upon exposure to these metal salts.Each metal salt appeared to induce a different,typical pattern of cytoskeletal injury,probably reflecting the specificity of action of each metal ion.These results suggest that the cytoskeleton can indeed act as a target for injury by epigenetic carcinogenic metal compounds in the environment.These findings should help our efforts to understand the mechanisms of action of metal compounds at the subcellular and molecular levels.1989 Academic Press,Inc.

由MT资料反演真谱参数的基本原理

常规的大地电磁测深反演方法没有考虑岩矿石激电效应。基于岩矿石激电响应的Dias新模型,分别计算了3层模型的中间层在有极化与无极化2种情况下的视电阻率和相位曲线。结果表明,2种情况下的视电阻率和相位曲线在高频段和低频段基本上重叠,而在中频段有明显的差异。进而研究了从大地电磁测深复视电阻率资料反演地层的真谱参数的方法,推导了层状模型反演的基本公式并给出了相应的具体反演算法。该算法能从MT资料中提取激电谱参数,为解释提供更多的依据。

军事噪声性听力损失群体中mtDNA和GJB2基因变异研究

目的探讨线粒体基因及GJB2基因突变与军事噪声性听力损失(noise induced hearing loss,NI HL)易感性的关系,为易感个体的基因筛查及相关分子流行病学研究提供科学依据。方法调查了北京某部349名接触军事噪声的官兵,收集军事噪声性听力损失易感者和耐受者外周血标本182份,提取DNA,PCR扩增线粒体DNA(mitochondrial DNA,mtDNA)目的片段及GJB2编码区,产物直接基因测序。结果基因序列分析共发现98种mtDNA和12种GJB2变异基因型,其中41种存在于12SrRNA;在易感者中发现4例mtDNA突变均为T1095C合并G7642A,在耐受者中未发现该突变;另有3例耐受者均为961delT+insC(其中一人合并235delC杂合),而易感者中未发现该突变。结论证实12SrRNA确为线粒体高突变区。T1095C合并G7642A突变在这些无相同遗传背景但受相同环境因素影响的人群中集中出现,强烈提示该突变可能为导致NI HL的致病性突变。3例有961delT+in-sC的耐受者均长期暴露于噪声环境,这与该突变应为条件致病性突变的推测相符,但与NI HL无明显相关性。

MT启动子控制的GFP基因在CHO细胞中的诱导表达

用羊金属硫蛋白(MT)启动子构建含绿色荧光蛋白(GFP)报告基因的真核表达载体,经脂质体介导在培养的中国仓鼠卵巢(CHO)细胞中进行诱导性表达。结果:构建的GFP基因表达载体能在CHO细胞中进行表达,用硫酸锌诱导可提高GFP的表达量。

EICP联合Na-Mt固化粉砂抗剪特性

针对传统酶诱导碳酸盐沉淀(enzyme-induced carbonate precipitation,EICP)固化土体时,生成的碳酸钙分布零散、不聚集,进而导致土体固化效果不佳的问题,提出将钠基蒙脱石(Na-montmorillonite,Na-Mt)与EICP技术相结合,对黄泛区粉砂进行固化。通过直剪试验研究固化粉砂抗剪特性的变化,借助扫描电子显微镜(scanning electron microscope,SEM)并结合Photoshop软件分析固化粉砂孔隙率的变化,以揭示EICP联合Na-Mt固化粉砂的机理。结果表明:与传统EICP技术处理粉砂相比,EICP联合Na-Mt固化粉砂的抗剪强度、黏聚力最大可分别提高41.4%、147.7%,孔隙率降低27.97%;决定固化粉砂抗剪强度的关键因素并非孔隙率,而是“Na-Mt-CaCO_(3)-土颗粒”单元结构的建立。与传统EICP技术相比,EICP联合Na-Mt固化粉砂可形成大体积的碳酸钙,增强粉砂的骨架效应。EICP联合Na-Mt是一种高效的细粒土体固化技术。

Metallothionein Genes in the Nematode Caenorhabditis elegans and Metal Inducibility in Mammalian Culture Cells

Genomic DNAs of metallothionein Ⅰ and Ⅱ in Caenorhabditis elegans (CeMT-Ⅰand CeMT-Ⅱ) were isolated by YAC library/polytene filter hybridization followed by subcloning of corresponding cosmid clones. Both genes are mapped at chromosome V. Although the similarities of 5'-flanking regions and coding regions have shown only 55-58%, the introns are split at the same position in both genes, indicating that these two genes are originally from the same gene. While several metal responsive elements are conserved among eukaryotes, only one metal responsive element was found in the promoter region in CeMT-Ⅱ and not in CeMT-Ⅰ. Indced, neither of 5'-flanking regions of CeMT-Ⅰ nor CeMT-Ⅱ connected to chloramphenicol acetyltransferase reporter gene is responsive to heavy metals in mammalian culture cells by transient transfection analysis. These results would suggest that the metal regulatory factors in C.elegans might be different from those conserved in invertebrates and vertebrates, although the MTs in C elegans revealed the similarities to mammalian MTs in several points

高温和脉冲噪声联合作用对职业人群心血管系统的影响

目的探讨高温、脉冲噪声联合暴露对职业人群心血管系统的影响。方法采用现场环境检测方法,对某汽车制造企业从事高温脉冲噪声作用工人326人、高温稳态噪声作业工人311人进行调查,同时选择接触低噪声、常温作业的319人作为对照。结果高温脉冲噪声组和高温稳态噪声组的高血压患病率、心电图异常率与对照组间差异有统计学意义(P<0.01),并与工作年限有一定的关系。吸烟可使高温脉冲噪声暴露人员高血压和心电图异常的危险性增加。结论高温与脉冲噪声联合作用比高温与稳态噪声联合作用对心血管系统的影响更大,且随工龄延长,其效应更明显。

热疗联合同步放化疗治疗中晚期食管癌的临床研究

目的观察热疗联合同步放化疗治疗中晚期食管癌的近期疗效、不良反应。方法 66例不能手术或不同意手术的中晚期食管癌患者,随机分成两组,治疗组应用深部热疗联合同步放化疗,对照组应用同步放化疗,比较两组近期疗效和不良反应。结果治疗组近期疗效高于对照组,差异有统计学意义(P<0.05);治疗组在骨髓抑制、胃肠道反应及放射性食管炎发生率方面均低于对照组,差异有统计学意义(P<0.05)。结论热疗联合同步放化疗治疗中晚期食管癌疗效优于同步放化疗,并且降低了放化疗的不良反应,值得进一步探讨。

热疗促进HSP70表达对免疫效应细胞杀伤肿瘤的影响

目的: 探讨热疗通过促进乳腺癌药物敏感细胞株与多药耐药细胞株表达热休克蛋白 (HSP)70,提高免疫效应细胞对靶细胞的杀伤活性。为开展基于热疗的细胞免疫治疗提供实验依据。方法: 采用IFNγ、CD3单抗、IL 1和IL 2等细胞因子诱导并扩增免疫效应细胞。分别给予乳腺癌药物敏感株MCF7和耐受株MCF7 /ADR非致死温度热冲击 42℃ 1h,继续培养 4h, 24h, 48h后收获细胞,用流式细胞术检测靶细胞上HSP70的表达。用四甲基偶氮唑蓝(MTT)法测定靶细胞增殖活性以及免疫效应细胞对靶细胞的杀伤活性。结果: HSP70在两种靶细胞上的表达有差异,加热前MCF7 /ADR细胞HSP70表达明显高于MCF7,加热后 4h,HSP70在MCF7 /ADR表达提高了 6倍,在MCF7表达提高了 22倍,超过了耐药株。药物敏感株MCF7增殖的热抑制率低于耐药株MCF7 /ADR。热冲击前免疫效应细胞对MCF7细胞的杀伤活性低于MCF7 /ADR细胞。热冲击后免疫效应细胞对MCF7的杀伤活性提高了 86. 23%,超过了对MCF7 /ADR的杀伤活性 (提高了 30. 32% )。结论: 热疗明显提高HSP70在乳腺癌药物敏感与耐受细胞系上的表达,增加了免疫效应细胞对这两种靶细胞的杀伤活性。HSP70在乳腺癌药物敏感与耐受细胞系上的差异性表达对免疫效应细胞的杀伤活性有影响。

肿瘤热疗用F_2O_3纳米磁性粒子的生物相容性研究

目的 :研究将要用于肿瘤细胞内热疗的自制纳米级F2 O3磁性粒子的生物相容性。方法 :MTT法评价其体外细胞毒性 ,溶血试验评价其有无溶血作用 ,小鼠腹腔注射其无菌生理盐水混悬液以测定其LD50 ,微核试验评价其有无致畸、致突变作用等。结果 :该材料对L 92 9细胞毒性为 0～ 1级 ;无溶血作用 ;昆明小鼠腹腔注射该材料混悬液 1ml ,其LD50 为 5 .45g·kg- 1 ;微核实验结果表明无致畸、致突变作用。结论 :就目前所取得的实验结果来看 ,可认为该材料是一种具有良好生物相容性的材料 ,对于肿瘤热疗可能具有广阔的应用前景。

插入式微波天线阵列诱导高温原位灭活治疗肢体恶性或侵袭性骨肿瘤

评价插入式微波天线阵列诱导高温原位灭活治疗肢体恶性或侵袭性骨肿瘤保肢手术的临床应用．方法：应用插入式微波天线阵列诱导高温原位灭活治疗肢体恶性或侵袭性骨肿瘤19o例．从手术技术、肿瘤学、关节功能、并发症等方面全面综合评价此方法的临床应用效果．结果：恶性骨肿瘤134例，术后平均随访47mo，因远隔脏器转移死亡者32例，另有3例虽有肺部转移，但经转移病灶切除后现仍无瘤存活；存活率为74．2％；IO2例存活者中肢体关节功能优者72例，良者14例．骨巨细胞瘤56例，术后平均随访49mo，l例死于肺转移；3例局部复发，经行瘤段截除大段异体骨移植后治愈；35例关节功能属优，21例为良．结论：插入式微波天线阵列诱导高温原位灭活治疗骨肿瘤技术是一种创新的、有效安全的、临床效果显著的、治疗肢体恶性或侵袭性骨肿瘤的新手段，值得继续应用、改进和评价这一治疗体系．

微波高温作用骨组织

目的：确定微波（2450MHz）灭活犬骨的需要温度和时间．方法：采用本地健康犬．分为即时，2wk组．即时组采用不同功率时间辐射犬骨自辐射点向外等距测温．分别于即时和2wk后收获标本分别进行光镜，AgNO3染色，电镜，骨组织碱性磷酸酶组化检查．结果：①功率越大，时间越长，辐射距离越长．②50℃10min即可使骨坏死，而42℃～50℃使骨不同程度损伤．结论：①临床对正常骨的加热应低于50℃．②选用合适的功率和时间作用骨组织．

高温诱导Hep-2细胞凋亡及其对细胞周期进程的影响

目的 :通过高温诱导 Hep- 2细胞凋亡探讨细胞凋亡和细胞周期的关系。方法 :应用 MTT比色法和流式细胞仪检测高温诱导 Hep- 2细胞凋亡和细胞周期时相的改变。结果 :高温组与对照组之间 Hep- 2细胞凋亡率差异有显著性 (P<0 .0 1 )。 MTT比色法高温组与对照组之间 ,Hep- 2细胞增殖抑制率差异亦有显著性。结论 :高温可提高 Hep- 2细胞的凋亡率 ,抑制 Hep- 2细胞的增殖 ,抑制 Hep- 2细胞 G1 期向

具有磁感应定向加热治疗肿瘤作用的锰锌铁氧体纳米粒子的制备及其特性检测

采用改良的化学共沉淀法制备Mn0.5Zn0.5Fe2O4纳米磁性材料，用透射电镜、分析仪及热分析系统等进行表征及特性检测。将Mn0.5Zn0.5Fe2O4纳米粒子以及其不同浓度的磁流体置于200KHz的交变磁场中照射，检测其磁感应自控加热作用；用Mn0.5Zn0.5；Fe2O4浸渍液体外干预小鼠L929成纤维细胞，通过检测细胞增值率来评价其细胞毒性。结果表明，制备的锰锌铁氧体纳米粒子为圆形，约40nm，并经检索X-线粉末衍射图库证实；纳米锰锌铁氧体磁流体能磁感应加热而升温到40℃～51℃，且最终温度能稳定控制不变；纳米锰锌铁氧体浸渍液对细胞未见明显毒性作用。