Extensive pectoral muscle necrosis after defibrillation: nonthermal skeletal muscle damage caused by electroporation

A 37-year-old Italian male developed a myocardial infarct with subsequent ventricular fibrillation. He was defibrillated seven times with up to 360 Joules. Thirteen days later the patient died of recurrent myocardial infarct due to thrombotic occlusion of the left circumflex coronary artery. At autopsy, necrosis of the right pectoralis muscle was observed. Electroporation is the pathogenetic mechanism of skeletal muscle damage due to multiple defibrillations with high energy levels. Received: 8 January 1998 Accepted: 15 April 1998     

电脉冲致离体S-180细胞电穿孔的观察与研究

扫描电镜下我们观察到了一定强度电脉冲致 S- 180细胞电穿孔的现象。同时分别改变电压、电容、脉冲个数 ,对 S- 180离体细胞进行电脉冲作用 ,通过苔盼蓝追踪 ,发现在一定的条件下 ,随着对 S- 180离体细胞电穿孔的电压越大、电容越小、脉冲个数越多 ,其穿孔百分率越大。     

强电脉冲治疗肿瘤提高抗癌药物疗效的研究

报道了用强电脉冲结合抗癌药物环磷酰胺作用于昆明小鼠S-180肉瘤。实验结果表明，电场 药物组的治疗效果最佳，与对照组相比，呈显著差异。同时观察到电场的作用可以抑制肿瘤的微血管形成，这样就减少了对癌细胞的营养供应、降低其新陈代谢，从而抑制了肿瘤的生长。     

Prime-boost vaccination regimens with INO-4800 and INO-4802 augment and broaden immune responses against SARS-CoV-2 in nonhuman primates

The enhanced transmissibility and immune evasion associated with emerging SARS-CoV-2 variants demands the development of next-generation vaccines capable of inducing superior protection amid a shifting pandemic landscape. Since a portion of the global population harbors some level of immunity from vaccines based on the original Wuhan-Hu-1 SARS-CoV-2 sequence or natural infection, an important question going forward is whether this immunity can be boosted by next-generation vaccines that target emerging variants while simultaneously maintaining long-term protection against existing strains. Here, we evaluated the immunogenicity of INO-4800, our synthetic DNA vaccine candidate for COVID-19 currently in clinical evaluation, and INO-4802, a next-generation DNA vaccine designed to broadly target emerging SARS-CoV-2 variants, as booster vaccines in nonhuman primates. Rhesus macaques primed over one year prior with the first-generation INO-4800 vaccine were boosted with either INO-4800 or INO-4802 in homologous or heterologous prime-boost regimens. Both boosting schedules led to an expansion of T cells and antibody responses which were characterized by improved neutralizing and ACE2 blocking activity across wild-type SARS-CoV-2 as well as multiple variants of concern. These data illustrate the durability of immunity following vaccination with INO-4800 and additionally support the use of either INO-4800 or INO-4802 in prime-boost regimens.     

重组质粒导入真核细胞的两种方法比较

目的：比较糖化多聚赖氨酸介导和电穿孔两种方法将重组质粒导入真核细胞的优缺点。方法：用最佳电压、电容、温度、DNA浓度进行电穿孔转染,或者先使重组质粒与糖化多聚赖氨酸电性结合后,再与2.2.15细胞共同温育的方法,分别将pcEP4-aC重组质粒导入2.2.15细胞,经潮霉素筛选,比较两种方法的优劣。结果：两种方法都能成功地将重组质粒导入2.2.15细胞,但糖化多聚赖氨酸介导者,细胞生存时间较长。结论：糖化多聚赖氨酸导向配体具有良好的导入重组质粒进入肝细胞的功能,且更适合于应用。     

甲状腺癌细胞系方波电脉冲基因转染条件的探讨

目的：探讨方波脉冲基因电转染对人甲状腺癌细胞系的转染条件和效果。方法：选用pEGFP-Cl作为外源基因与方波电脉冲相结合，以人甲状腺癌细胞系TA-K为导入对象，探讨方波脉冲基因电转染对人甲状腺癌细胞系转染的脉冲幅度，脉冲时值，脉冲次数和反应体系大小。结果：当脉冲幅度在60V／mm时，开始出现阳性细胞，脉冲时值20ms时，基因转染效率可高达35％，细胞生存分数55％，脉冲时值20ms、次数1次转染效率高、细胞死亡率低。200μl混合液电转染后观察，对细胞生存影响不大。结论：脉冲幅度在60V／mm，脉冲时值20ms，脉冲次数1次，反应体系200μl，可以为人甲状腺癌细胞系的转染提供良好效果。     

Workshop report: Nucleic acid delivery devices for HIV vaccines: Workshop proceedings,National Institute of Allergy and Infectious Diseases,Bethesda, Maryland,USA, May 21, 2015

On May 21st, 2015, the U.S. National Institute of Allergy and Infectious Diseases (NIAID) convened a workshop on delivery devices for nucleic acid (NA) as vaccines in order to review the landscape of past and future technologies for administering NA (e.g., DNA, RNA, etc.) as antigen into target tissues of animal models and humans. Its focus was on current and future applications for preventing and treating human immunodeficiency virus (HIV) infection and acquired immune deficiency syndrome (AIDS) disease, among other infectious-disease priorities. Meeting participants presented the results and experience of representative clinical trials of NA vaccines using a variety of alternative delivery devices, as well as a broader group of methods studied in animal models and at bench top, to improve upon the performance and/or avoid the drawbacks of conventional needle-syringe (N–S) delivery. The subjects described and discussed included (1) delivery targeted into oral, cutaneous/intradermal, nasal, upper and lower respiratory, and intramuscular tissues; (2) devices and techniques for jet injection, solid, hollow, and dissolving microneedles, patches for topical passive diffusion or iontophoresis, electroporation, thermal microporation, nasal sprayers, aerosol upper-respiratory and pulmonary inhalation, stratum-corneum ablation by ultrasound, chemicals, and mechanical abrasion, and kinetic/ballistic delivery; (3) antigens, adjuvants, and carriers such as DNA, messenger RNA, synthesized plasmids, chemokines, wet and dry aerosols, and pollen-grain and microparticle vectors; and (4) the clinical experience and humoral, cellular, and cytokine immune responses observed for many of these target tissues, technologies, constructs, and carriers. This report summarizes the presentations and discussions from the workshop (https://web.archive.org/web/20160228112310/https://www.blsmeetings.net/NucleicAcidDeliveryDevices/), which was webcast live in its entirety and archived online (http://videocast.nih.gov/summary.asp?live=16059).     

产黄青霉简单高效遗传转化法——电激转化法

目的 研究产黄青霉(P.chrysogenum)电激转化方法,建立产黄青霉快速、高效的遗传转化方法.方法 应用美国Bio-Rad电转仪,将外源DNA转化入产黄青霉菌丝中,研究了菌丝培养时间、电场强度、DNA类型等对遗传转化的影响.结果菌丝培养24h,处于旺盛生长阶段,电场强度为6000V/cm,电阻200Ω,电容20μF,环状质粒DNA较线状能获得转化效率稍高,1μg DNA获得的克隆数可达500个.结论 应用电激转化方法对产黄青霉进行遗传转化属首次报道,特别是可直接将目的片段与T-DNA相融合,转化入产黄青霉中,该方法操作简单,快速高效.     

Irreversible electroporation of pancreatic cancer – Effect on quality of life and pain perception

BackgroundMost pancreatic cancer (PC) patients are incurable and may need palliative treatment at some point in time. Irreversible electroporation (IRE) is a novel ablative treatment, which aims to provide local tumor control. The aim of this study was to examine how consolidative treatment with IRE affects quality of life (QOL) and pain perception (PP) in patients with non-metastatic pancreatic cancer.MethodsSecondary outcomes were extracted from a prospective cohort of non-metastatic PC patients treated with IRE from 2013 to 2019. Patients filled in two questionnaires examining QOL and PP at different timepoints during treatment and follow-up. Data from a selected panel of subscales were extracted and analyzed using a mixed random intercept regression model.ResultsSubscales from 41 patients at four different timepoints were included in the model. Global health status, physical functioning, fatigue, nausea and vomiting, appetite loss and mean pain interference were negatively impacted (p < 0.05) in the short- and mid-term, corresponding to a low or moderate clinical effect size. However, all negative effects showed a tendency to dissipate over time.ConclusionsIRE treatment negatively impacted QOL and PP in the short- and mid-term. No positive long-term effects of IRE were found.     

电穿孔法介导醛脱氢酶基因与多药耐药基因的转移和表达

背景与目的：将不同类型的耐药基因同时导入造血细胞,以扩大耐药范围和进行体内选择是基因治疗的重要策略,这类载体基因片段较长,进行基因转移有一定的难度。本研究旨在寻找安全有效的长片段基因转移G1Na-AIM以NdeⅠ酶切线性化,电穿孔法转导PA317细胞,经长春新碱（VCR）和4－氢过氧化环磷酰胺（4－HC）选择后,应用Southern blot法确定原病毒的整合,逆转录聚合酶链反应（RT－PCR）和流式细胞术（FCM）分析转移基因的表达,筑巢式聚合酶链反应（nested PCR)检验转移系统的安全性。结果：电穿孔法有效介导了ALDH1与MDR1基因的同时转移,Southern blot法证实ALDH1与MDR1基因稳定整合至宿主细胞基因组,RT－PCR检测到转移基因的转录,FCM测定下游基因MDR1蛋白表达增高4倍,转导率达98％。nested PCR未检测到辅助病毒（env)。结论：电穿孔法安全有效地介导了ALDH1与MDR1基因的共移和高表达。