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.

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.

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.

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).

Streptozotocin induced Alzheimer's disease like changes and the underlying neural degeneration and regeneration mechanism

Alzheimer’s disease（AD）is a neurodegenerative disorder which is remarkably characterized by pathological hallmarks that include neurofibrillary tangles,neuronal loss extracellular senile plaques containing aggregated amyloid beta（Aβ）,and neurofibrillary tangles composed of the hyperphosphorylated form of the microtubule protein tau.It is the most common form of dementia which is characterized by severe neurodegenerative changes such as loss of neurons and synapses in brain（Kamat et al.,2014）.

Hydrodynamics of A Flexible Riser Undergoing the Vortex-Induced Vibration at High Reynolds Number

This study proposed a method to obtain hydrodynamic forces and coefficients for a flexible riser undergoing the vortex-induced vibration(VIV), based on the measured strains collected from the scale-model testing with the Reynolds numbers ranging from 1.34 E5 to 2.35 E5. The riser is approximated as a tensioned spatial beam, and an inverse method based on the FEM of spatial beam is adopted for the calculation of hydrodynamic forces in the cross flow(CF) and inline(IL) directions. The drag coefficients and vortex-induced force coefficients are obtained through the Fourier Series Theory. Finally, the hydrodynamic characteristics of a flexible riser model undergoing the VIV in a uniform flow are carefully investigated. The results indicate that the VIV amplifies the drag coefficient, and the drag coefficient does not change with time when the CF VIV is stable. Only when the VIVs in the CF and IL directions are all steady vibrations, the vortex-induced force coefficients keep as a constant with time, and under"lock-in" condition, whether the added-mass coefficient changes with time or not, the oscillation frequency of the VIV keeps unchanged. It further shows that the CF excitation coefficients at high frequency are much smaller than those at the dominant frequency, while, the IL excitation coefficients are in the same range. The axial distributions of the excitation and damping region at the dominant frequency and high frequency are approximately consistent in the CF direction, while, in the IL direction, there exists a great difference.

Maternal Lead Exposure Induces Down-regulation of Hippocampal Insulin-degrading Enzyme and Nerve Growth Factor Expression in Mouse Pups

Lead exposure is a known potential risk factor for neurodegenerative diseases such as Alzheimer’s disease （AD）. Exposure to lead during the critical phase of brain development has been linked with mental retardation and hypophrenia in later life.

加热对人肝癌耐药细胞模型-7721/Adm细胞内药物浓度的影响

目的 探讨比较 43℃加热前后人肝癌细胞 - 772 1(以下简称 772 1细胞 )和耐药人肝癌细胞模型 - 772 1/ Adm(以下简称 772 1/ Adm细胞 )细胞内阿霉素 (ADM)药物浓度的变化 .方法 以体外培养的人肝癌细胞 - 772 1和作者自行培养建立的人肝癌细胞模型 - 772 1/ Adm为研究对象 ,采用水浴加温法、流式细胞荧光技术观察阿霉素化疗加热前后 772 1和772 1/ Adm细胞胞内阿霉素 (ADM)浓度的变化 .结果 加热后 772 1细胞组提高 30 .8% ,HCC- 772 1/ Adm组提高 5 1% .结论 加热可明显提高这两种细胞内的阿霉素浓度 ,从而提高这两种细胞的化疗敏感性 ,为临床克服多药耐药问题提供了重要依据 .

重组大肠杆菌利用乳糖自诱导系统融合表达天蚕素AD和蛙BuforinⅡ

以包含质粒pET-Trx-CAD-BuforinⅡ的大肠杆菌BL21(DE3)为研究对象,开发了一种利用乳糖自诱导方式表达天蚕素AD和蛙BuforinⅡ的表达系统。首先,在LB培养基中研究了乳糖浓度、诱导时机和诱导时间对目的蛋白表达的影响。然后,利用"分解代谢物阻遏"原理构建乳糖自诱导表达系统,以"葡萄糖+主碳源"两种碳源的依次代谢将蛋白表达过程分为两个阶段:第一阶段为菌体生长阶段,菌体利用葡萄糖生长,乳糖无法进入细胞诱导蛋白表达;第二阶段为蛋白表达阶段,葡萄糖耗尽,菌体开始利用主碳源生长;同时乳糖进入细胞诱导蛋白表达。最后,利用优选的"葡萄糖+木糖"组合自诱导表达天蚕素AD和BuforinⅡ的融合蛋白,得到的目的蛋白占全菌总蛋白含量的45.2%。

Evidence based tools to improve efficiency of currently administered oncotherapies for tumors of the hepatopancreatobiliary system

Hepatopancreatobiliary tumors are challenging to treat,and the advanced or metastatic forms have a very low 5-year survival rate.Several drug combinations have been tested,and new therapeutic approaches have been introduced in the last decades,including radiofrequency and heat based methods.Hyperthermia is the artificial heating of tumors by various biophysical methods that may possess immunostimulant,tumoricidal,and chemoradiotherapy sensitizer effects.Both whole-body and regional hyperthermia studies have been conducted since the 1980s after the introduction of deep-seated tumor hyperthermia techniques.Results of the effects of hyperthermia in hepatocellular and pancreatic cancer are known from several studies.Hyperthermia in biliary cancers is a less investigated area.High local and overall responses to treatment,increased progression-free and overall survival,and improved laboratory and quality-of-life results are associated with hyperthermia in all three tumor types.With the evolution of chemotherapeutic agents and the introduction of newer techniques,the combination of adjuvant hyperthermia with those therapies is advantageous and has not been associated with an increase in alarming adverse effects.However,despite the many positive effects of hyperthermia,its use is still only known at the experimental level,and its concomitant utilization in routine cancer treatment is not certain because of the lack of thorough clinical studies.

ADIG诱导牛成肌细胞转分化及相关基因的表达分析

旨在通过在牛成肌细胞中过表达ADIG基因,诱导成肌细胞向成脂转分化,探讨该牛脂肪分化转录因子ADIG基因在脂类代谢过程中的功能,为阐明牛脂肪细胞分化转录因子ADIG基因在牛脂肪组织生长发育过程中发挥的作用提供证据。本研究用AD-ADIG腺病毒原液诱导转分化初生荷斯坦牛的成肌细胞,并在诱导分化后第2、4、6、8、10、12、14天进行油红O染色观察诱导成脂情况,同时收集细胞提取总RNA,利用实时荧光定量PCR分别检测ADIG、PPARγ、C/EBPα、SREBPF1、FABP4、FAS、MyoD基因的表达量变化。实时荧光定量PCR检测结果表明,ADIG基因在第2、4、6、8、10、12、14天均有高水平表达,且随着诱导天数增加脂滴明显增多。成脂相关的基因PPARγ、SREBPF1和FAS在后期AD-ADIG组中表达量均高于空白组,成肌相关的基因MyoD相对表达量在初期就显著下降,且AD-ADIG组明显低于对照组。综合脂滴染色和相关基因表达量方面的研究结果,说明ADIG基因在诱导牛成肌细胞分化向成脂转分化过程中发挥着重要作用,而且牛成肌细胞有向脂肪细胞分化的倾向。

Variation of Added Mass and Its Application to the Calculation of Amplitude Response for A Circular Cylinder

In the present study, analyzed are the variation of added mass for a circular cylinder in the lock-in （synchronization） range of vortex-induced vibration （VIV） and the relationship between added mass and natural frequency. A theoretical minimum value of the added mass coefficient for a circular cylinder at lock-in is given. Developed are semi-empirical formulas for the added mass of a circular cylinder at lock-in as a function of flow speed and mass ratio. A comparison between experiments and numerical simulations shows that the semi-empirical formulas describing the variation of the added mass for a cireular cylinder at lock-in are better than the ideal added mass. In addition, computation models such as the wake oscillator model using the present formulas can predict the amplitude response of a circular cylinder at lock-in more accurately than those using the ideal added mass.

深部热疗对AD离体细胞治疗模式的探索及疗效

目的:探索深部热疗对AD离体细胞的最佳治疗模式并评估治疗效果。方法:制备离体AD细胞模型,随机分成对照组、对照组热疗组、AD模型组、AD模型热疗组,分别置于41.5℃加热30、45、60、75、90min,于600nm处测吸光度(A)值,计算最佳热疗时间。造模成AD细胞后置于41.5℃加热60min,加热后继续置于37℃恒温孵育24h,分别于加热后2、4、8、12、24、72h取出,使用流式细胞仪行活细胞膜HSP70表达量,计算最佳热疗间隔时间。4组细胞热疗后分别加入20μmol/L的Aβ25-35溶液培养24h,应用流式细胞仪检测细胞活力,评估热疗治疗AD效果。结果:对照组和AD模型组分别给予不同时间41.5℃热疗,两组细胞从加热60min开始细胞增殖趋势与非加热细胞相比开始下降(P<0.05),这种下降趋势在75min达到最大,确定细胞加热的安全有效时间为60min,同时对照热疗组与AD模型热疗组对比,两组细胞随温度变化的细胞增殖趋势基本相同,两者比较无统计学差异(P>0.05),表明两组细胞的最佳热疗时间均为60min。AD模型组细胞在热疗后2、4、8、12、24及72h各时间段监测的HSP70表达量均不同,AD细胞在24h HSP70表达量均达到最大值(P<0.05),其后HSP70表达量开始下降,表明24h是最佳的热疗间隔时间。4组细胞与Aβ作用后加热,MTT检测表明4组细胞活力呈温度依赖性上升趋势(P<0.05),表明热疗可有效的减少Aβ的聚集,促进Aβ的清除,保护AD细胞。结论:深部热疗对AD离体细胞的最佳热疗时间为60min,最佳热疗间隔时间为24h,深部热疗可以有效的清除Aβ,促进神经元细胞修复。

Theories behind Bacillus Calmette-Guerin failure in high-risk non-muscle-invasive bladder cancer and update on current management

Bladder cancer encapsulates a wide spectrum of disease severities,with non-muscle invasive bladder cancer(NMIBC)representing an entirely different entity from muscle-invasive disease.Bacillus Calmette-Guerin(BCG)is one of the most successful intravesical treatment methods for patients diagnosed.However,a considerable pro-portion of patients fail to respond to BCG treatment.Given the propensity for recurrence in patients with high-risk bladder cancer,these patients present with surgical dilemmas.There is currently no gold standard for salvage treatment post-BCG failure or unified definition as to what that means.In this review,we discuss the mechanisms of action and pathophysiology of BCG,potential theories behind BCG failure,and the scope of novel treatments forthis surgical conundrum.

基于生物信息学方法双芯片联合分析阿尔茨海默病海马相关差异基因的研究

目的利用生物信息学筛选阿尔茨海默病(Alzheimer’s disease,AD)海马组织的差异表达基因(DEGs),分析DEGs的生物学功能和信号通路,为AD的诊断、治疗靶点的筛选提供参考。方法利用GEO(gene expression omnibus)数据库下载基因芯片GSE48350、GSE5281,使用数据库GEO2R工具在线分析,以P<0.05,|logFC>1|为条件筛选差异基因,筛选出与AD海马组织相关的DEGs,通过制作维恩图取交集获得两个芯片共同DEGs;使用R语言enrichGO函数对筛选出来的DEGs进行基因本体论(GO)功能富集分析;在Metascape网站做京都基因与基因组百科全书(KEGG)通路分析;使用STRING数据库和Cytoscape3.9.1软件进行蛋白-蛋白相互作用(PPI)网络分析。把筛选出来的共同DEGs作为关键基因,通过Cytoscape3.9.1软件中插件Mcode和CytoHubba分析后,按照节点度值大小排序,将前四个基因视为核心基因。最后,通过TargetScan与miRanda数据库评估核心基因相关的miRNA,并选用基因芯片GSE129053进行验证,构建核心基因的miRNA-mRNA调控网络。结果两个基因芯片共有67个DEGs,其中上调基因36个,下调基因31个。GO分析结果显示,主要富集在生物过程中为突触囊泡介导的运输、神经元突触囊泡循环、网格蛋白介导的突触囊泡内吞机制等;在细胞定位中主要的过程有胞外囊泡、突触小泡等;分子功能主要过程有网格蛋白结合、葡萄糖的跨膜转运活性等。KEGG通路富集分析结果显示,DEGs富集在甲状腺激素信号通路(hsa04919);DEGs编码蛋白的PPI网络含节点10个;运用Cytoscape中CytoHubba插件和Mcode插件,把筛选出来的共同基因作为关键基因,前四个基因视为核心基因,分别为SNAP-25、SNAP-91、STMN2、GAP43,可以发现hsa-miR-153-3p关联的靶基因最多。结论AD患者海马组织中关键的核心基因为SNAP-25、SNAP-91、STMN2、GAP43,hsa-miR-153-3p可能通过调控其靶基因SNAP-25、STMN2在AD患者海马组织中发挥重要作用,为AD的诊断和治疗提供了候选靶点。

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。