Green fluorescent protein (GFP) gene was successfully transferred into the isolated zygotes and early proembryos of wheat (Triticum aestivum L.) by electroporation. A frequency, as high as 46.7% of GFP gene transient ...Green fluorescent protein (GFP) gene was successfully transferred into the isolated zygotes and early proembryos of wheat (Triticum aestivum L.) by electroporation. A frequency, as high as 46.7% of GFP gene transient expression in early proembryos, was achieved under 150 V/cm electric field strength, 25 muF capacitor, 200 mug/mL of linear plasmid DNA and an electroporation buffer at pH 7.2. Compared with five-day-old proembryos, the zygotes and early proembryos needed lower optimum strength of electric field. After culturing in KM8p medium, the electroporated early proembryos divided and GFP gene expression was observed in daughter cells and subsequent divisions. There was no mosaicism of gene expression in the zygotes and 2-, 4- and 8-celled proembryos.展开更多
Bitter receptors were initially identified within the gustatory system.In recent years,bitter receptors have been found in various non-gustatory tissues,including the cardiovascular system,where they participate in di...Bitter receptors were initially identified within the gustatory system.In recent years,bitter receptors have been found in various non-gustatory tissues,including the cardiovascular system,where they participate in diverse physiological processes.To investigate the electrophysiological and potential therapeutic implications of bitter receptors,we have developed a highly sensitive,multifunctional planar-electroporated cell biosensor(PECB)for high-throughput evaluation of the effects of bitter substances on cardiomyocytes.The PECB demonstrated the capability for highthroughput,stable,and reproducible detection of intracellular action potentials(IAPs).In comparison to conventional biosensors that utilize extracellular action potentials(EAPs)for data analysis,the IAPs recorded by the PECB provided high-resolution insights into action potentials,characterized by increased amplitudes and an enhanced signal-to-noise ratio(SNR).The PECB successfully monitored IAPs induced by the activation of bitter receptors by using three bitter substances:diphenidol,denatonium benzoate,and arbutin in cardiomyocytes.To further assess the drug development ability of our PECB,we established an in vitro long QT syndrome(LQTS)model to investigate the potential therapeutic effects of arbutin.The results indicated that arbutin altered the electrophysiological properties of cardiomyocytes and significantly shortened the repolarization time in the LQTS model.Moreover,it demonstrated its potential mechanistic pathway by activating bitter receptors to modulate cardiac ion channel activities.The developed PECB provides an effective platform for high-throughput screening of substrates of bitter receptors for the treatment of heart disease,presenting new opportunities for the development of antiarrhythmic therapies.展开更多
BACKGROUND Duodenal mucosal ablation(DMA)using irreversible electroporation(IRE)with a glucagon-like peptide-1 receptor agonist has been clinically shown to reduce liver lipid deposition in non-alcoholic fatty liver d...BACKGROUND Duodenal mucosal ablation(DMA)using irreversible electroporation(IRE)with a glucagon-like peptide-1 receptor agonist has been clinically shown to reduce liver lipid deposition in non-alcoholic fatty liver disease(NAFLD).However,the specific metabolic contributions of DMA using IRE in NAFLD remain unclear.AIM To assess the feasibility and effectiveness of DMA using IRE in NAFLD rat models.METHODS Seven-week-old male Sprague-Dawley rats underwent DMA using IRE after 8 weeks on a high-fat diet.Two weeks post-treatment,duodenal and liver tissues and blood samples were collected.We evaluated differences in the duodenal wall structure,liver lipid deposition,enteroendocrine,claudin,and zonula ocludens-1 in the duodenal mucosa.RESULTS DMA using IRE could be safely performed in rats with NAFLD without duodenal bleeding,perforation,or stenosis.The duodenum healed well 2 weeks after DMA and was characterized by slimmer villi,narrower and shallower crypts,and thicker myenterons compared with the sham-control setting.Liver lipid deposition was reduced and serum lipid index parameters were considerably improved in the DMA setting.However,these improvements were independent of food intake and weight loss.In addition,enteroendocrine parameters,such as claudin,and zonula ocludens-1 levels in the duodenal mucosa,differed between the different settings in the DMA group.CONCLUSION By altering enteroendocrine and duodenal permeability,simple DMA using IRE ameliorated liver lipid deposition and improved serum lipid parameters in NAFLD rats.展开更多
BACKGROUND Irreversible electroporation(IRE)is a novel local tumor ablation approach with the potential to activate the host’s immune system.However,this approach is insufficient to prevent cancer progression,and com...BACKGROUND Irreversible electroporation(IRE)is a novel local tumor ablation approach with the potential to activate the host’s immune system.However,this approach is insufficient to prevent cancer progression,and complementary approaches are required for effective immunotherapy.AIM To assess the immunomodulatory effects and mechanism of IRE combined antiprogrammed cell death protein 1(PD-1)treatment in subcutaneous pancreatic cancer models.METHODS C57BL-6 tumor-bearing mice were randomly divided into four groups:Control group;IRE group;anti-PD-1 group;and IRE+anti-PD-1 group.Tumor-infiltrating T,B,and natural killer cell levels and plasma concentrations of T helper type 1 cytokines(interleukin-2,interferon-γ,and tumor necrosis factor-α)were evaluated.Real-time PCR was used to determine the expression of CD8(marker of CD8+T cells)in tumor tissues of the mice of all groups at different points of time.The growth curves of tumors were drawn.RESULTS The results demonstrated that the IRE+anti-PD-1 group exhibited significantly higher percentages of T lymphocyte infiltration,including CD4+and CD8+T cells compared with the control group.Additionally,the IRE+anti-PD-1 group showed increased infiltration of natural killer and B cells,elevated cytokine levels,and higher CD8 mRNA expression.Tumor volume was significantly reduced in the IRE+anti-PD-1 group,indicating a more pronounced therapeutic effect.CONCLUSION The combination of IRE and anti-PD-1 therapy promotes CD8+T cell immunity responses,leading to a more effective reduction in tumor volume and improved therapeutic outcomes,which provides a new direction for ablation and immunotherapy of pancreatic cancer.展开更多
Irreversible electroporation is a promising non-thermal ablation method that has been shown to increase overall survival in locally advanced pancreatic cancer in some studies.However,higher quality studies with proper...Irreversible electroporation is a promising non-thermal ablation method that has been shown to increase overall survival in locally advanced pancreatic cancer in some studies.However,higher quality studies with proper controls and randomization are required to establish its superiority when added with neoadjuvant chemotherapy over the current management of choice,which is chemotherapy alone.Further studies are required before establishment of any survival benefit in metastatic pancreatic carcinoma,and such evidence is lacking at present.展开更多
1.Introduction Irreversible electroporation(IRE)has emerged as a promising non-thermal ablation technique for the treatment of malignant tumors.^([1-3])By applying high-voltage electrical pulses to tumor tissue,IRE cr...1.Introduction Irreversible electroporation(IRE)has emerged as a promising non-thermal ablation technique for the treatment of malignant tumors.^([1-3])By applying high-voltage electrical pulses to tumor tissue,IRE creates irrecoverable pores in the cell membrane,thus disrupting the balance between the intracellular and extracellular environments,and ultimately causing cell death.^([1])展开更多
Objective:To observed the effect of a curcumin-based vaginal gel combined with electroporation for the treatment of vulvovaginal candidiasis(VVC)caused by Candida albicans.Methods:Temperature-sensitive in situ gels(IS...Objective:To observed the effect of a curcumin-based vaginal gel combined with electroporation for the treatment of vulvovaginal candidiasis(VVC)caused by Candida albicans.Methods:Temperature-sensitive in situ gels(ISG)were prepared using poloxamers 407 and 188 as matrices.The mass ratio of poloxamer 407 and poloxamer 188 was 7:1 with a gelation temperature of approximately 29℃ and gelation time of 2.5 min.Results:Electroporation increased the transmucosal permeability of the model drug,doxorubicin and improved the antifungal effects of curcumin.In vitro antifungal experiments showed that the number of fungal colonies in curcumin ISG combined with electroporation was lower than that in pure curcumin ISG.In vivo pharmacodynamic experiments showed that,compared to the model group,curcumin ISG with electroporation inhibited the growth of C.albicans,alleviated vaginal mucosal edema,and reduced the inflammatory response.Conclusion:Curcumin ISG combined with electroporation has substantial potential for the efficient clinical treatment of VVC.展开更多
BACKGROUND Irreversible electroporation(IRE)is a novel non-thermal ablation technology for unresectable tumors.Hypertension and tachycardia usually occur during the IRE.To date,there has been little explanation about ...BACKGROUND Irreversible electroporation(IRE)is a novel non-thermal ablation technology for unresectable tumors.Hypertension and tachycardia usually occur during the IRE.To date,there has been little explanation about this phenomenon.AIM To investigate the reasons of hypertension and tachycardia and appropriate preventive measures.METHODS IRE was performed under general anesthesia and neuromuscular blockade.Systolic blood pressures,diastolic blood pressures,heart rate,and the distance of the electrode from abdominal aorta and adrenal gland during IRE were recorded.RESULTS All of 78 patients underwent 96 IRE sessions,44(56.4%)patients occurred hypertension when the electrode was close to the abdominal aorta(<2.0 cm).The median systolic blood pressures and diastolic blood pressures was 194 and 108 mmHg.Furthermore,11(14.1%)patients occurred tachycardia when the electrode was close to the adrenal gland(<1.3 cm).The median heart rate of patients with tachycardia was 114 beats per minute.Furthermore,hypertension and tachycardia can be prevented with nicardipine and esmolol before treatment.CONCLUSION Intraoperative hypertension and tachycardia occur because electrodes close to the abdominal aorta(<2.0 cm)and adrenal glands(<1.3 cm),which can be prevented by preoperative treatment of vasoactive drugs.展开更多
Ultrasound-guided percutaneous thermal ablation has gained popularity as treatment for malignant hepatic tumors.It was first introduced as ablation therapy for hepatocellular carcinoma and cirrhosis comorbidity.Recent...Ultrasound-guided percutaneous thermal ablation has gained popularity as treatment for malignant hepatic tumors.It was first introduced as ablation therapy for hepatocellular carcinoma and cirrhosis comorbidity.Recently,this technique has also been used in the treatment of intrahepatic cholangiocarcinoma for patients who are not eligible for surgical resection.There are several types of thermal ablation techniques.Radiofrequency ablation and microwave ablation are two common methods that induce necrosis of the lesions.Irreversible electroporation is a relatively new non-thermal technique and is suitable in cases where thermal ablation would be ineffective or dangerous(e.g.,malignant tumors close to vascular or biliary structures).Irreversible electroporation can induce tumoral necrosis without damage to vascular and biliary structures.The aim of this minireview was to describe the safety,efficacy,and clinical indications of these techniques in the treatment of patients with intrahepatic cholangiocarcinoma who are ineligible for surgery.展开更多
Acute mitochondrial damage and the energy crisis following axonal injury highlight mitochondrial transport as an important target for axonal regeneration.Syntaphilin(Snph),known for its potent mitochondrial anchoring ...Acute mitochondrial damage and the energy crisis following axonal injury highlight mitochondrial transport as an important target for axonal regeneration.Syntaphilin(Snph),known for its potent mitochondrial anchoring action,has emerged as a significant inhibitor of both mitochondrial transport and axonal regeneration.Therefore,investigating the molecular mechanisms that influence the expression levels of the snph gene can provide a viable strategy to regulate mitochondrial trafficking and enhance axonal regeneration.Here,we reveal the inhibitory effect of microRNA-146b(miR-146b)on the expression of the homologous zebrafish gene syntaphilin b(snphb).Through CRISPR/Cas9 and single-cell electroporation,we elucidated the positive regulatory effect of the miR-146b-snphb axis on Mauthner cell(M-cell)axon regeneration at the global and single-cell levels.Through escape response tests,we show that miR-146b-snphb signaling positively regulates functional recovery after M-cell axon injury.In addition,continuous dynamic imaging in vivo showed that reprogramming miR-146b significantly promotes axonal mitochondrial trafficking in the pre-injury and early stages of regeneration.Our study reveals an intrinsic axonal regeneration regulatory axis that promotes axonal regeneration by reprogramming mitochondrial transport and anchoring.This regulation involves noncoding RNA,and mitochondria-associated genes may provide a potential opportunity for the repair of central nervous system injury.展开更多
One of the main causes of liver fibrosis and cancer,non-alcoholic fatty liver disease(NAFLD)is becoming more common every year.The novel work by Yu et al,which evaluates the viability and efficacy of duodenal mucosal ...One of the main causes of liver fibrosis and cancer,non-alcoholic fatty liver disease(NAFLD)is becoming more common every year.The novel work by Yu et al,which evaluates the viability and efficacy of duodenal mucosal ablation(DMA)with irreversible electroporation(IRE)in NAFLD rat models,is examined in this article.When DMA was used with IRE to small rodents,the study found that the duodenum healed successfully two weeks later and had thicker myenterons,narrower and shallower crypts,and slimmer villi than in the sham-control group.When DMA with IRE were used,liver lipid deposition and serum lipid index values decreased;these improvements occurred regardless of food consumption or weight loss.Furthermore,the DMA group's enteroendocrine parameters varied among the various duodenal areas,including claudin and zonula ocludens-1 Levels in the duodenal mucosa.As a result,DMA with IRE in rodents demonstrated no duodenal bleeding or perforation following ablation,providing a promising path for more advanced NAFLD treatment approaches.In order to improve approach outcomes,this paper addresses the implications of extending the study length and animal size,analyzing inflammatory marker studies,and measuring intestinal lipid indexes and endocrine parameters on a weekly basis.展开更多
We read with great interest the recent article by Xing et al,which describes the synergy between irreversible electroporation and anti-programmed death-1 therapy in a murine hepatocellular carcinoma model.The study of...We read with great interest the recent article by Xing et al,which describes the synergy between irreversible electroporation and anti-programmed death-1 therapy in a murine hepatocellular carcinoma model.The study offers valuable mechanistic insights into local ablation,enhancing the efficacy of immune checkpoint blockade.However,critical methodological limitations and an overstatement of mechanistic conclusions warrant cautious interpretation.We recommend clarifying experimental details,optimizing murine models,applying more robust statistical analyses,and tempering conclusions to reflect the correlative nature of the findings.Further work should investigate immune mechanisms,durability of response,and safety in clinically relevant models to maximize translational potential.These refinements will strengthen the study’s impact in advancing ablation-immunotherapy strategies in hepatocellular carcinoma.展开更多
BACKGROUND Irreversible electroporation(IRE)represents an innovative localized technique for tumor ablation,possessing the capacity to activate the immune response of the host.However,this method alone is inadequate t...BACKGROUND Irreversible electroporation(IRE)represents an innovative localized technique for tumor ablation,possessing the capacity to activate the immune response of the host.However,this method alone is inadequate to halt cancer progression,necessitating the integration of additional strategies to achieve effective immuno-therapy.AIM To investigate the effects and underlying mechanisms of antitumor immunity derived from the synergistic application of IRE and anti-programmed cell death protein 1(PD-1)therapy within a murine model of hepatocellular carcinoma.METHODS C57BL-6 mice with tumor growth were divided into four separate cohorts:Control group;IRE group;Anti-PD-1 group;And IRE+anti-PD-1 group.The infiltration levels of T,B,and natural killer cells within the tumors,as well as the plasma concentrations of T helper type 1 cytokines(interleukin-2,interferon-γ,and tumor necrosis factor-β),were evaluated.Real-time polymerase chain reaction was utilized to quantify the expression of cluster of differentiation(CD)8(a marker indicative of CD8+T cells)in the tumor specimens of the mice at various temporal intervals.Tumor growth trajectories were charted.RESULTS The results indicated that the IRE+anti-PD-1 group exhibited significantly heightened percentages of T lymphocyte infiltration,particularly CD4+and CD8+T cells,when compared to the control cohort.Additionally,this group displayed increased infiltration of natural killer and B cells,augmented cytokine levels,and elevated CD8 messenger RNA expression.A marked decrease in tumor volume was noted in the IRE+anti-PD-1 group,indicating enhanced therapeutic efficacy.CONCLUSION The combined application of IRE and checkpoint blockade elicits an antitumor immune response,leading to a more substantial reduction in tumor volume and improved therapeutic outcomes,thereby establishing a novel avenue for the ablation and immunotherapy of hepatocellular carcinoma.展开更多
Metallothionein gene (MT) has been transferred into mushroom protoplasts by electroporation. It is a low molecular weight, cysteine-rich and metal-binding protein. MT can bind metals. Its synthesis is induced by Zn io...Metallothionein gene (MT) has been transferred into mushroom protoplasts by electroporation. It is a low molecular weight, cysteine-rich and metal-binding protein. MT can bind metals. Its synthesis is induced by Zn ion. Thus the expression of MT gene in mushroom can improve the accumulation of Zn in this fungus. This transgenic mushroom, consumed as a kind of vegetable, can supply the necessary Zn to people who are short of the element. When protoplasts were prepared, the concentration (C) of protoplasts is 6.745 x 10(6) /mL. After protoplast electroporation, the transformation rate of protoplasts is 0.01 %. Polymerase chain reaction (PCR) analysis showed that the gene had been integrated into the mushroom chromosome, SDS-PAGE, Western blot analysis indicated that the MT gene had been expressed in the transgenic mushroom. The expressing level, detected by ELISA, is 0.6 % - 0.8 %. Tested for metal resistance, the wild-type mushroom growth was inhibited on die medium containing 1.0 - 1.2 mmoL/L ZnSO4. While the transgenic mushroom was inhibited on the medium containing 1. 5 - 2.0 mmol/L ZnSO4. The mycelium can develop into hymenophore in the medium of rice bran: sawdust = 1: 3, and not in the medium of rice bran: sawdust = 1: 4.展开更多
Aim To induce and express the T4 lysozyme in Pichia pastoris and test the antibacterial activity of the protein. Methods T4 lysozyme gene was inserted into expression vector pPIC9K of Pichia pastoris with the fusion a...Aim To induce and express the T4 lysozyme in Pichia pastoris and test the antibacterial activity of the protein. Methods T4 lysozyme gene was inserted into expression vector pPIC9K of Pichia pastoris with the fusion at N terminal. The recombinant plasmid was digested by Sal I and then introduced into prepared GS115 competent cells by electroporation. Positive clone and multiple inserts were screened. The secreted proteins in the supernatants were tested. In the agar holes diffusion assay, our expressed protein showed significant antibacterial circles. Results T4 lysozyme protein inhibited the growth of staphylococcus aureus and streptococcus Pneumoniae. There was no difference in the bactericidal activity and the amount of protein expression between the single and multiple copies. The antibacterial activity of expressed protein remained the same during the heat stability test. Conclusion T4 lysozyme was successfully induced and expressed in Pichia pastoris. There is no relationship between copy number and expression. T4 lysozyme protein is heat stable.展开更多
A retroviral vector pINC-lacZ containing an E coli β-galactosidase(β-gal)gene was constructed and introduced into the MESPU-13 cells by electroporation.Four G418-resistant colonies were obtained from 1×107,elec...A retroviral vector pINC-lacZ containing an E coli β-galactosidase(β-gal)gene was constructed and introduced into the MESPU-13 cells by electroporation.Four G418-resistant colonies were obtained from 1×107,electroporated MESPU-13 cells. Histochemical staining for β-gal activity showed that lacZ gene was expressed in at least three of the four colonies.Southern analysis proved that one copy of foreign gene was integrated in the chromosome of one of the transformed lines(MC15).These results showed that the expression of lacZ gene driven by SiCMVIE promoter can be detected in the transformed MESPU-13 cells.展开更多
Green fluorescent protein ( GFP ) gene was expressed transiently in 2-3 d old rice embryos by electroporation with the aid of a specially designed loading net. Under suitable conditions (500 μF capacitance, 300 V/c...Green fluorescent protein ( GFP ) gene was expressed transiently in 2-3 d old rice embryos by electroporation with the aid of a specially designed loading net. Under suitable conditions (500 μF capacitance, 300 V/cm Voltage, 100 μg/mL plasmid DNA), the percentage of embryos expressing GFP was up to 35%. The highest electroporation efficiency (40%) was obtained at pH 5.8 of the electroporation buffer. The GFP gene driven by the Ubi promoter produced the highest efficiency. Thus, on the basis of optimizing electroporation conditions, a transformation system has been developed for young embryos in rice. The electroporated 4-6 d old embryos regenerated plantlets under the controlled cultural conditions. Fluorescence microscopic observations indicated that GFP gene expressed in their calli and R0 plantlets.展开更多
Various therapeutic modalities including radiofrequency ablation, cryoablation, microwave ablation, and irreversible electroporation have attracted attention as energy sources for effective locoregional treatment of h...Various therapeutic modalities including radiofrequency ablation, cryoablation, microwave ablation, and irreversible electroporation have attracted attention as energy sources for effective locoregional treatment of hepatocellular carcinoma(HCC); these are accepted non-surgical treatments that provide excellent local tumor control and favorable survival. However, in contrast to surgery, tumor location is a crucial factor in the outcomes of locoregional treatment because such treatment is mainly performed using a percutaneous approach for minimal invasiveness; accordingly, it has a limited range of ablation volume. When the index tumor is near large blood vessels, the blood flow drags thermal energy away from the targeted tissue, resulting in reduced ablation volume through a socalled "heat-sink effect". This modifies the size and shape of the ablation zone considerably. In addition, serious complications including infarction or aggressive tumor recurrence can be observed during follow-up after ablation for perivascular tumors by mechanical or thermal damage. Therefore, perivascular locations of HCC adjacent to large intrahepatic vessels can affect post-treatment outcomes. In this review, we primarily focus on physical properties of perivascular tumor location, characteristics of perivascular HCC,potential complications, and clinical outcomes after various locoregional treatments; moreover, we discuss the current status and future perspectives regarding percutaneous ablation for perivascular HCC.展开更多
Unresectable locally advanced pancreatic cancer with or without metastatic disease is associated with a very poor prognosis.Current standard therapy is limited to chemotherapy or chemoradiotherapy.Few regimens have be...Unresectable locally advanced pancreatic cancer with or without metastatic disease is associated with a very poor prognosis.Current standard therapy is limited to chemotherapy or chemoradiotherapy.Few regimens have been shown to have a substantial survival advantage and novel treatment strategies are urgently needed.Thermal and laser based ablative techniques are widely used in many solid organ malignancies.Initial studies in the pancreas were associated with significant morbidity and mortality,which limited widespread adoption.Modifications to the various applications,in particular combining the techniques with high quality imaging such as computed tomography and intraoperative or endoscopic ultrasound has enabled real time treatment monitoring and significant improvements in safety.We conducted a systematic review of the litera-ture up to October 2013.Initial studies suggest that ablative therapies may confer an additional survival benefit over best supportive care but randomised studies are required to validate these findings.展开更多
AIM:To investigate the effect of transgenic expression of kallistatin(Kal) on carbon tetrachloride(CCl 4)induced liver injury by intramuscular(im) electrotransfer of a Kal-encoding plasmid formulated with poly-Lglutam...AIM:To investigate the effect of transgenic expression of kallistatin(Kal) on carbon tetrachloride(CCl 4)induced liver injury by intramuscular(im) electrotransfer of a Kal-encoding plasmid formulated with poly-Lglutamate(PLG).METHODS:The pKal plasmid encoding Kal gene was formulated with PLG and electrotransferred into mice skeletal muscle before the administration of CCl 4.The expression level of Kal was measured.The serum biomarker levels of alanine aminotransferase(ALT),aspartate aminotransferase(AST),malonyldialdehyde(MDA),and tumor necrosis factor(TNF)-α were monitored.The extent of CCl 4-induced liver injury was analyzed histopathologically.RESULTS:The transgene of Kal was sufficiently expressed after an im injection of plasmid formulated with PLG followed by electroporation.In the Kal gene-transferred mice,protection against CCl 4-induced liver injury was reflected by significantly decreased serum ALT,AST,MDA and TNF-α levels compared to those in control mice(P < 0.01 to 0.05 in a dose-dependent manner).Histological observations also revealed that hepatocyte necrosis,hemorrhage,vacuolar change and hydropic degeneration were apparent in mice after CCl 4 administration.In contrast,the damage was markedly attenuated in the Kal gene-transferred mice.The expression of hepatic fibrogenesis marker transforming growth factor-β1 was also reduced in the pKal transferred mice.CONCLUSION:Intramuscular electrotransfer of plasmid pKal which was formulated with PLG significantly alleviated the CCl 4-induced oxidative stress and inflammatory response,and reduced the liver damage in a mouse model.展开更多
文摘Green fluorescent protein (GFP) gene was successfully transferred into the isolated zygotes and early proembryos of wheat (Triticum aestivum L.) by electroporation. A frequency, as high as 46.7% of GFP gene transient expression in early proembryos, was achieved under 150 V/cm electric field strength, 25 muF capacitor, 200 mug/mL of linear plasmid DNA and an electroporation buffer at pH 7.2. Compared with five-day-old proembryos, the zygotes and early proembryos needed lower optimum strength of electric field. After culturing in KM8p medium, the electroporated early proembryos divided and GFP gene expression was observed in daughter cells and subsequent divisions. There was no mosaicism of gene expression in the zygotes and 2-, 4- and 8-celled proembryos.
基金supported by the National Key Research and Development Program of China(2024YFB3212300)Key Project of Zhejiang Province(2023C03104,2024C03146)+4 种基金National Natural Science Foundation of China(32201082,62301481,62401505)Scientific Research Fund of Zhejiang Provincial Education Department(Y202353232)the Postdoctoral Fellowship Program of CPSF(GZC20232333)Postdoctoral Science Foundation Funded Project(BX2021265,2021M702859)the Fundamental Research Funds for the Central Universities(226-2024-00059).
文摘Bitter receptors were initially identified within the gustatory system.In recent years,bitter receptors have been found in various non-gustatory tissues,including the cardiovascular system,where they participate in diverse physiological processes.To investigate the electrophysiological and potential therapeutic implications of bitter receptors,we have developed a highly sensitive,multifunctional planar-electroporated cell biosensor(PECB)for high-throughput evaluation of the effects of bitter substances on cardiomyocytes.The PECB demonstrated the capability for highthroughput,stable,and reproducible detection of intracellular action potentials(IAPs).In comparison to conventional biosensors that utilize extracellular action potentials(EAPs)for data analysis,the IAPs recorded by the PECB provided high-resolution insights into action potentials,characterized by increased amplitudes and an enhanced signal-to-noise ratio(SNR).The PECB successfully monitored IAPs induced by the activation of bitter receptors by using three bitter substances:diphenidol,denatonium benzoate,and arbutin in cardiomyocytes.To further assess the drug development ability of our PECB,we established an in vitro long QT syndrome(LQTS)model to investigate the potential therapeutic effects of arbutin.The results indicated that arbutin altered the electrophysiological properties of cardiomyocytes and significantly shortened the repolarization time in the LQTS model.Moreover,it demonstrated its potential mechanistic pathway by activating bitter receptors to modulate cardiac ion channel activities.The developed PECB provides an effective platform for high-throughput screening of substrates of bitter receptors for the treatment of heart disease,presenting new opportunities for the development of antiarrhythmic therapies.
基金Supported by the National Key Research and Development Program,No.2023YFF0713700 and No.2023YFF0713705Common Technology R&D Platform of Shaanxi Province,No.2023GXJS-01-1-2the Cyrus Tang Foundation Chung Ying Young Scholars Program.
文摘BACKGROUND Duodenal mucosal ablation(DMA)using irreversible electroporation(IRE)with a glucagon-like peptide-1 receptor agonist has been clinically shown to reduce liver lipid deposition in non-alcoholic fatty liver disease(NAFLD).However,the specific metabolic contributions of DMA using IRE in NAFLD remain unclear.AIM To assess the feasibility and effectiveness of DMA using IRE in NAFLD rat models.METHODS Seven-week-old male Sprague-Dawley rats underwent DMA using IRE after 8 weeks on a high-fat diet.Two weeks post-treatment,duodenal and liver tissues and blood samples were collected.We evaluated differences in the duodenal wall structure,liver lipid deposition,enteroendocrine,claudin,and zonula ocludens-1 in the duodenal mucosa.RESULTS DMA using IRE could be safely performed in rats with NAFLD without duodenal bleeding,perforation,or stenosis.The duodenum healed well 2 weeks after DMA and was characterized by slimmer villi,narrower and shallower crypts,and thicker myenterons compared with the sham-control setting.Liver lipid deposition was reduced and serum lipid index parameters were considerably improved in the DMA setting.However,these improvements were independent of food intake and weight loss.In addition,enteroendocrine parameters,such as claudin,and zonula ocludens-1 levels in the duodenal mucosa,differed between the different settings in the DMA group.CONCLUSION By altering enteroendocrine and duodenal permeability,simple DMA using IRE ameliorated liver lipid deposition and improved serum lipid parameters in NAFLD rats.
基金Science and Technology Program of Guangzhou,No.202102010077International Science Foundation of Guangzhou Fuda Cancer Hospital,No.Y2020-ZD-03.
文摘BACKGROUND Irreversible electroporation(IRE)is a novel local tumor ablation approach with the potential to activate the host’s immune system.However,this approach is insufficient to prevent cancer progression,and complementary approaches are required for effective immunotherapy.AIM To assess the immunomodulatory effects and mechanism of IRE combined antiprogrammed cell death protein 1(PD-1)treatment in subcutaneous pancreatic cancer models.METHODS C57BL-6 tumor-bearing mice were randomly divided into four groups:Control group;IRE group;anti-PD-1 group;and IRE+anti-PD-1 group.Tumor-infiltrating T,B,and natural killer cell levels and plasma concentrations of T helper type 1 cytokines(interleukin-2,interferon-γ,and tumor necrosis factor-α)were evaluated.Real-time PCR was used to determine the expression of CD8(marker of CD8+T cells)in tumor tissues of the mice of all groups at different points of time.The growth curves of tumors were drawn.RESULTS The results demonstrated that the IRE+anti-PD-1 group exhibited significantly higher percentages of T lymphocyte infiltration,including CD4+and CD8+T cells compared with the control group.Additionally,the IRE+anti-PD-1 group showed increased infiltration of natural killer and B cells,elevated cytokine levels,and higher CD8 mRNA expression.Tumor volume was significantly reduced in the IRE+anti-PD-1 group,indicating a more pronounced therapeutic effect.CONCLUSION The combination of IRE and anti-PD-1 therapy promotes CD8+T cell immunity responses,leading to a more effective reduction in tumor volume and improved therapeutic outcomes,which provides a new direction for ablation and immunotherapy of pancreatic cancer.
基金Supported by Department of Biotechnology,Government of India,No.RLS/BT/Re-entry/05/2012.
文摘Irreversible electroporation is a promising non-thermal ablation method that has been shown to increase overall survival in locally advanced pancreatic cancer in some studies.However,higher quality studies with proper controls and randomization are required to establish its superiority when added with neoadjuvant chemotherapy over the current management of choice,which is chemotherapy alone.Further studies are required before establishment of any survival benefit in metastatic pancreatic carcinoma,and such evidence is lacking at present.
文摘1.Introduction Irreversible electroporation(IRE)has emerged as a promising non-thermal ablation technique for the treatment of malignant tumors.^([1-3])By applying high-voltage electrical pulses to tumor tissue,IRE creates irrecoverable pores in the cell membrane,thus disrupting the balance between the intracellular and extracellular environments,and ultimately causing cell death.^([1])
基金supported by the Beijing Natural Science Foundation(L222126).
文摘Objective:To observed the effect of a curcumin-based vaginal gel combined with electroporation for the treatment of vulvovaginal candidiasis(VVC)caused by Candida albicans.Methods:Temperature-sensitive in situ gels(ISG)were prepared using poloxamers 407 and 188 as matrices.The mass ratio of poloxamer 407 and poloxamer 188 was 7:1 with a gelation temperature of approximately 29℃ and gelation time of 2.5 min.Results:Electroporation increased the transmucosal permeability of the model drug,doxorubicin and improved the antifungal effects of curcumin.In vitro antifungal experiments showed that the number of fungal colonies in curcumin ISG combined with electroporation was lower than that in pure curcumin ISG.In vivo pharmacodynamic experiments showed that,compared to the model group,curcumin ISG with electroporation inhibited the growth of C.albicans,alleviated vaginal mucosal edema,and reduced the inflammatory response.Conclusion:Curcumin ISG combined with electroporation has substantial potential for the efficient clinical treatment of VVC.
基金Supported by International Science Foundation of Guangzhou Fuda Cancer Hospital,No.Y2022-MS-02.
文摘BACKGROUND Irreversible electroporation(IRE)is a novel non-thermal ablation technology for unresectable tumors.Hypertension and tachycardia usually occur during the IRE.To date,there has been little explanation about this phenomenon.AIM To investigate the reasons of hypertension and tachycardia and appropriate preventive measures.METHODS IRE was performed under general anesthesia and neuromuscular blockade.Systolic blood pressures,diastolic blood pressures,heart rate,and the distance of the electrode from abdominal aorta and adrenal gland during IRE were recorded.RESULTS All of 78 patients underwent 96 IRE sessions,44(56.4%)patients occurred hypertension when the electrode was close to the abdominal aorta(<2.0 cm).The median systolic blood pressures and diastolic blood pressures was 194 and 108 mmHg.Furthermore,11(14.1%)patients occurred tachycardia when the electrode was close to the adrenal gland(<1.3 cm).The median heart rate of patients with tachycardia was 114 beats per minute.Furthermore,hypertension and tachycardia can be prevented with nicardipine and esmolol before treatment.CONCLUSION Intraoperative hypertension and tachycardia occur because electrodes close to the abdominal aorta(<2.0 cm)and adrenal glands(<1.3 cm),which can be prevented by preoperative treatment of vasoactive drugs.
文摘Ultrasound-guided percutaneous thermal ablation has gained popularity as treatment for malignant hepatic tumors.It was first introduced as ablation therapy for hepatocellular carcinoma and cirrhosis comorbidity.Recently,this technique has also been used in the treatment of intrahepatic cholangiocarcinoma for patients who are not eligible for surgical resection.There are several types of thermal ablation techniques.Radiofrequency ablation and microwave ablation are two common methods that induce necrosis of the lesions.Irreversible electroporation is a relatively new non-thermal technique and is suitable in cases where thermal ablation would be ineffective or dangerous(e.g.,malignant tumors close to vascular or biliary structures).Irreversible electroporation can induce tumoral necrosis without damage to vascular and biliary structures.The aim of this minireview was to describe the safety,efficacy,and clinical indications of these techniques in the treatment of patients with intrahepatic cholangiocarcinoma who are ineligible for surgery.
基金supported by the core facility Center for Life Sciences,University of Science and Technology of China,Research Funds of the Center for Advanced Interdisciplinary Science and Biomedicine of IHM(QYZD20220002)the National Natural Science Foundation of China(82071357)the Ministry of Science and Technology of China(2019YFA0405600).
文摘Acute mitochondrial damage and the energy crisis following axonal injury highlight mitochondrial transport as an important target for axonal regeneration.Syntaphilin(Snph),known for its potent mitochondrial anchoring action,has emerged as a significant inhibitor of both mitochondrial transport and axonal regeneration.Therefore,investigating the molecular mechanisms that influence the expression levels of the snph gene can provide a viable strategy to regulate mitochondrial trafficking and enhance axonal regeneration.Here,we reveal the inhibitory effect of microRNA-146b(miR-146b)on the expression of the homologous zebrafish gene syntaphilin b(snphb).Through CRISPR/Cas9 and single-cell electroporation,we elucidated the positive regulatory effect of the miR-146b-snphb axis on Mauthner cell(M-cell)axon regeneration at the global and single-cell levels.Through escape response tests,we show that miR-146b-snphb signaling positively regulates functional recovery after M-cell axon injury.In addition,continuous dynamic imaging in vivo showed that reprogramming miR-146b significantly promotes axonal mitochondrial trafficking in the pre-injury and early stages of regeneration.Our study reveals an intrinsic axonal regeneration regulatory axis that promotes axonal regeneration by reprogramming mitochondrial transport and anchoring.This regulation involves noncoding RNA,and mitochondria-associated genes may provide a potential opportunity for the repair of central nervous system injury.
文摘One of the main causes of liver fibrosis and cancer,non-alcoholic fatty liver disease(NAFLD)is becoming more common every year.The novel work by Yu et al,which evaluates the viability and efficacy of duodenal mucosal ablation(DMA)with irreversible electroporation(IRE)in NAFLD rat models,is examined in this article.When DMA was used with IRE to small rodents,the study found that the duodenum healed successfully two weeks later and had thicker myenterons,narrower and shallower crypts,and slimmer villi than in the sham-control group.When DMA with IRE were used,liver lipid deposition and serum lipid index values decreased;these improvements occurred regardless of food consumption or weight loss.Furthermore,the DMA group's enteroendocrine parameters varied among the various duodenal areas,including claudin and zonula ocludens-1 Levels in the duodenal mucosa.As a result,DMA with IRE in rodents demonstrated no duodenal bleeding or perforation following ablation,providing a promising path for more advanced NAFLD treatment approaches.In order to improve approach outcomes,this paper addresses the implications of extending the study length and animal size,analyzing inflammatory marker studies,and measuring intestinal lipid indexes and endocrine parameters on a weekly basis.
文摘We read with great interest the recent article by Xing et al,which describes the synergy between irreversible electroporation and anti-programmed death-1 therapy in a murine hepatocellular carcinoma model.The study offers valuable mechanistic insights into local ablation,enhancing the efficacy of immune checkpoint blockade.However,critical methodological limitations and an overstatement of mechanistic conclusions warrant cautious interpretation.We recommend clarifying experimental details,optimizing murine models,applying more robust statistical analyses,and tempering conclusions to reflect the correlative nature of the findings.Further work should investigate immune mechanisms,durability of response,and safety in clinically relevant models to maximize translational potential.These refinements will strengthen the study’s impact in advancing ablation-immunotherapy strategies in hepatocellular carcinoma.
基金Supported by the Science and Technology Program of Guangzhou,No.202201020024.
文摘BACKGROUND Irreversible electroporation(IRE)represents an innovative localized technique for tumor ablation,possessing the capacity to activate the immune response of the host.However,this method alone is inadequate to halt cancer progression,necessitating the integration of additional strategies to achieve effective immuno-therapy.AIM To investigate the effects and underlying mechanisms of antitumor immunity derived from the synergistic application of IRE and anti-programmed cell death protein 1(PD-1)therapy within a murine model of hepatocellular carcinoma.METHODS C57BL-6 mice with tumor growth were divided into four separate cohorts:Control group;IRE group;Anti-PD-1 group;And IRE+anti-PD-1 group.The infiltration levels of T,B,and natural killer cells within the tumors,as well as the plasma concentrations of T helper type 1 cytokines(interleukin-2,interferon-γ,and tumor necrosis factor-β),were evaluated.Real-time polymerase chain reaction was utilized to quantify the expression of cluster of differentiation(CD)8(a marker indicative of CD8+T cells)in the tumor specimens of the mice at various temporal intervals.Tumor growth trajectories were charted.RESULTS The results indicated that the IRE+anti-PD-1 group exhibited significantly heightened percentages of T lymphocyte infiltration,particularly CD4+and CD8+T cells,when compared to the control cohort.Additionally,this group displayed increased infiltration of natural killer and B cells,augmented cytokine levels,and elevated CD8 messenger RNA expression.A marked decrease in tumor volume was noted in the IRE+anti-PD-1 group,indicating enhanced therapeutic efficacy.CONCLUSION The combined application of IRE and checkpoint blockade elicits an antitumor immune response,leading to a more substantial reduction in tumor volume and improved therapeutic outcomes,thereby establishing a novel avenue for the ablation and immunotherapy of hepatocellular carcinoma.
文摘Metallothionein gene (MT) has been transferred into mushroom protoplasts by electroporation. It is a low molecular weight, cysteine-rich and metal-binding protein. MT can bind metals. Its synthesis is induced by Zn ion. Thus the expression of MT gene in mushroom can improve the accumulation of Zn in this fungus. This transgenic mushroom, consumed as a kind of vegetable, can supply the necessary Zn to people who are short of the element. When protoplasts were prepared, the concentration (C) of protoplasts is 6.745 x 10(6) /mL. After protoplast electroporation, the transformation rate of protoplasts is 0.01 %. Polymerase chain reaction (PCR) analysis showed that the gene had been integrated into the mushroom chromosome, SDS-PAGE, Western blot analysis indicated that the MT gene had been expressed in the transgenic mushroom. The expressing level, detected by ELISA, is 0.6 % - 0.8 %. Tested for metal resistance, the wild-type mushroom growth was inhibited on die medium containing 1.0 - 1.2 mmoL/L ZnSO4. While the transgenic mushroom was inhibited on the medium containing 1. 5 - 2.0 mmol/L ZnSO4. The mycelium can develop into hymenophore in the medium of rice bran: sawdust = 1: 3, and not in the medium of rice bran: sawdust = 1: 4.
文摘Aim To induce and express the T4 lysozyme in Pichia pastoris and test the antibacterial activity of the protein. Methods T4 lysozyme gene was inserted into expression vector pPIC9K of Pichia pastoris with the fusion at N terminal. The recombinant plasmid was digested by Sal I and then introduced into prepared GS115 competent cells by electroporation. Positive clone and multiple inserts were screened. The secreted proteins in the supernatants were tested. In the agar holes diffusion assay, our expressed protein showed significant antibacterial circles. Results T4 lysozyme protein inhibited the growth of staphylococcus aureus and streptococcus Pneumoniae. There was no difference in the bactericidal activity and the amount of protein expression between the single and multiple copies. The antibacterial activity of expressed protein remained the same during the heat stability test. Conclusion T4 lysozyme was successfully induced and expressed in Pichia pastoris. There is no relationship between copy number and expression. T4 lysozyme protein is heat stable.
文摘A retroviral vector pINC-lacZ containing an E coli β-galactosidase(β-gal)gene was constructed and introduced into the MESPU-13 cells by electroporation.Four G418-resistant colonies were obtained from 1×107,electroporated MESPU-13 cells. Histochemical staining for β-gal activity showed that lacZ gene was expressed in at least three of the four colonies.Southern analysis proved that one copy of foreign gene was integrated in the chromosome of one of the transformed lines(MC15).These results showed that the expression of lacZ gene driven by SiCMVIE promoter can be detected in the transformed MESPU-13 cells.
文摘Green fluorescent protein ( GFP ) gene was expressed transiently in 2-3 d old rice embryos by electroporation with the aid of a specially designed loading net. Under suitable conditions (500 μF capacitance, 300 V/cm Voltage, 100 μg/mL plasmid DNA), the percentage of embryos expressing GFP was up to 35%. The highest electroporation efficiency (40%) was obtained at pH 5.8 of the electroporation buffer. The GFP gene driven by the Ubi promoter produced the highest efficiency. Thus, on the basis of optimizing electroporation conditions, a transformation system has been developed for young embryos in rice. The electroporated 4-6 d old embryos regenerated plantlets under the controlled cultural conditions. Fluorescence microscopic observations indicated that GFP gene expressed in their calli and R0 plantlets.
基金Supported by Nano-Convergence Foundation,the Ministry of Science,ICT and Future Planning(MSIP,South Korea)the Ministry of Trade,Industry and Energy(MOTIE,South Korea),No.R201603110
文摘Various therapeutic modalities including radiofrequency ablation, cryoablation, microwave ablation, and irreversible electroporation have attracted attention as energy sources for effective locoregional treatment of hepatocellular carcinoma(HCC); these are accepted non-surgical treatments that provide excellent local tumor control and favorable survival. However, in contrast to surgery, tumor location is a crucial factor in the outcomes of locoregional treatment because such treatment is mainly performed using a percutaneous approach for minimal invasiveness; accordingly, it has a limited range of ablation volume. When the index tumor is near large blood vessels, the blood flow drags thermal energy away from the targeted tissue, resulting in reduced ablation volume through a socalled "heat-sink effect". This modifies the size and shape of the ablation zone considerably. In addition, serious complications including infarction or aggressive tumor recurrence can be observed during follow-up after ablation for perivascular tumors by mechanical or thermal damage. Therefore, perivascular locations of HCC adjacent to large intrahepatic vessels can affect post-treatment outcomes. In this review, we primarily focus on physical properties of perivascular tumor location, characteristics of perivascular HCC,potential complications, and clinical outcomes after various locoregional treatments; moreover, we discuss the current status and future perspectives regarding percutaneous ablation for perivascular HCC.
基金Supported by National Institutes of Health Grant PO1CA84203The work was undertaken at UCLH/UCL,which receives a proportion of funding from the Department of Health’s National Institute for Health Research Biomedical Research Centres fundingschemeA CRUK research bursary to Keane MG
文摘Unresectable locally advanced pancreatic cancer with or without metastatic disease is associated with a very poor prognosis.Current standard therapy is limited to chemotherapy or chemoradiotherapy.Few regimens have been shown to have a substantial survival advantage and novel treatment strategies are urgently needed.Thermal and laser based ablative techniques are widely used in many solid organ malignancies.Initial studies in the pancreas were associated with significant morbidity and mortality,which limited widespread adoption.Modifications to the various applications,in particular combining the techniques with high quality imaging such as computed tomography and intraoperative or endoscopic ultrasound has enabled real time treatment monitoring and significant improvements in safety.We conducted a systematic review of the litera-ture up to October 2013.Initial studies suggest that ablative therapies may confer an additional survival benefit over best supportive care but randomised studies are required to validate these findings.
基金Supported by The State High Technology Research and Development Program of China (863 Program),No.2008AA02Z135the Important National Science and Technology Specific Projects,No.2009ZX09103-643the Natural Science Foundation of China,No.30973591
文摘AIM:To investigate the effect of transgenic expression of kallistatin(Kal) on carbon tetrachloride(CCl 4)induced liver injury by intramuscular(im) electrotransfer of a Kal-encoding plasmid formulated with poly-Lglutamate(PLG).METHODS:The pKal plasmid encoding Kal gene was formulated with PLG and electrotransferred into mice skeletal muscle before the administration of CCl 4.The expression level of Kal was measured.The serum biomarker levels of alanine aminotransferase(ALT),aspartate aminotransferase(AST),malonyldialdehyde(MDA),and tumor necrosis factor(TNF)-α were monitored.The extent of CCl 4-induced liver injury was analyzed histopathologically.RESULTS:The transgene of Kal was sufficiently expressed after an im injection of plasmid formulated with PLG followed by electroporation.In the Kal gene-transferred mice,protection against CCl 4-induced liver injury was reflected by significantly decreased serum ALT,AST,MDA and TNF-α levels compared to those in control mice(P < 0.01 to 0.05 in a dose-dependent manner).Histological observations also revealed that hepatocyte necrosis,hemorrhage,vacuolar change and hydropic degeneration were apparent in mice after CCl 4 administration.In contrast,the damage was markedly attenuated in the Kal gene-transferred mice.The expression of hepatic fibrogenesis marker transforming growth factor-β1 was also reduced in the pKal transferred mice.CONCLUSION:Intramuscular electrotransfer of plasmid pKal which was formulated with PLG significantly alleviated the CCl 4-induced oxidative stress and inflammatory response,and reduced the liver damage in a mouse model.
