Tumor-directed gene therapy in mice using a composite nonviral gene delivery system consisting of the piggyBac transposon and polyethylenimine

<p>Abstract</p> <p>Background</p> <p>Compared with viral vectors, nonviral vectors are less immunogenic, more stable, safer and easier to replication for application in cancer gene therapy. However, nonviral gene delivery system has not been extensively used because of the low transfection efficiency and the short transgene expression, especially <it>in vivo</it>. It is desirable to develop a nonviral gene delivery system that can support stable genomic integration and persistent gene expression <it>in vivo</it>. Here, we used a composite nonviral gene delivery system consisting of the <it>piggyBac </it>(PB) transposon and polyethylenimine (PEI) for long-term transgene expression in mouse ovarian tumors.</p> <p>Methods</p> <p>A recombinant plasmid PB [Act-RFP, HSV-tk] encoding both the herpes simplex thymidine kinase (HSV-tk) and the monomeric red fluorescent protein (mRFP1) under PB transposon elements was constructed. This plasmid and the PBase plasmid were injected into ovarian cancer tumor xenografts in mice by <it>in vivo </it>PEI system. The antitumor effects of HSV-tk/ganciclovir (GCV) system were observed after intraperitoneal injection of GCV. Histological analysis and TUNEL assay were performed on the cryostat sections of the tumor tissue.</p> <p>Results</p> <p>Plasmid construction was confirmed by PCR analysis combined with restrictive enzyme digestion. mRFP1 expression could be visualized three weeks after the last transfection of pPB/TK under fluorescence microscopy. After GCV admission, the tumor volume of PB/TK group was significantly reduced and the tumor inhibitory rate was 81.96% contrasted against the 43.07% in the TK group. Histological analysis showed that there were extensive necrosis and lymphocytes infiltration in the tumor tissue of the PB/TK group but limited in the tissue of control group. TUNEL assays suggested that the transfected cells were undergoing apoptosis after GCV admission <it>in vivo</it>.</p> <p>Conclusion</p> <p>Our results show that the nonviral gene delivery system coupling PB transposon with PEI can be used as an efficient tool for gene therapy in ovarian cancer.</p

XRCC1 codon 399Gln polymorphism is associated with radiotherapy-induced acute dermatitis and mucositis in nasopharyngeal carcinoma patients

BACKGROUND: To evaluate the association between single nucleotide polymorphisms (SNPs) at the 194 and 399 codons of XRCC1, and the risk of severe acute skin and oral mucosa reactions in nasopharyngeal carcinoma patients in China. METHODS: 114 patients with nasopharyngeal carcinoma were sequentially recruited in this study. Heparinized peripheral blood samples were taken for SNPs analysis before the start of radiation treatment. SNPs in XRCC1 (194Arg/Trp and 399Arg/Gln) gene were analyzed by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP). Dermatitis at upper neck and oral mucositis were clinically recorded according to the Common Terminology Criteria for Adverse Events v.3.0. RESULTS: The variant allele frequencies were 0.289 for XRCC1 194Trp and 0.263 for XRCC1 399Gln. Of the 114 patients, 24 experienced grade 3 acute dermatitis and 48 had grade 3 acute mucositis. The XRCC1 399Arg/Gln was significantly associated with the development of grade 3 dermatitis (Odds Ratio, 2.65; 95% CI, 1.04–6.73; p = 0.037, χ2 = 4.357). In addition, it was also associated with higher incidence of grade 3 mucositis with a borderline statistical significance (Odds Ratio, 2.11; 95% CI, 0.951–4.66; p = 0.065, χ2 = 3.411). The relationship between XRCC1 194Arg/Trp and acute dermatitis, and mucositis was not found. CONCLUSIONS: Our investigation shows, for the first time, that patients with the XRCC1 399Arg/Gln genotype were more likely to experience severe acute dermatitis and oral mucositis. With further validation, the information can be used to determine personalized radiotherapy strategy

Isolation of Robinsoniella peoriensis from the fecal material of the endangered Yangtze finless porpoise, Neophocaena asiaeorientalis asiaeorientalis

The aim of this study was to determine the causative agent of diarrhea in an endangered Yangtze finless porpoise (Neophocaena asiaeorientalis asiaeorientalis). From the fecal material collected from this porpoise Robinsoniella peoriensis was isolated. (C) 2013 Elsevier Ltd. All rights reserved.The aim of this study was to determine the causative agent of diarrhea in an endangered Yangtze finless porpoise (Neophocaena asiaeorientalis asiaeorientalis). From the fecal material collected from this porpoise Robinsoniella peoriensis was isolated. (C) 2013 Elsevier Ltd. All rights reserved

The proportion of state-owned shares and capacity sharing with constraints and prices in a mixed oligopoly

This study constructs an oligopoly model composed of mixed-ownership and private enterprises, examining the equilibrium results of three cases: when two enterprises compete with sufficient capacity (Model AA), insufficient capacity and overcapacity coexist without sharing (Model IA), and sharing (Model IS). This study also explores the effects of the proportion of state-owned shares, capacity constraints, and capacity prices. The realisation conditions and impacts of capacity sharing are further analysed. The results show that the efficiency of state-owned capital affects the effects of state-owned shares on the equilibrium results. An optimal capacity price exists for the capacity provider (private enterprise). Capacity sharing can effectively allocate resources and increase profits; however, consumers and society do not necessarily benefit from it. Full privatisation and the highest proportion of state-owned shares may be the best choice for the government under certain conditions. The government can intervene in enterprises’ capacity decision-making through subsidies to promote social welfare and realise capacity sharing simultaneously. Moreover, the government subsidises different enterprises when the proportions of state-owned shares and capacity prices are within different ranges

Regulating Cytoplasmic Calcium Homeostasis Can Reduce Aluminum Toxicity in Yeast

Our previous study suggested that increased cytoplasmic calcium (Ca) signals may mediate aluminum (Al) toxicity in yeast (Saccharomyces cerevisiae). In this report, we found that a yeast mutant, pmc1, lacking the vacuolar calcium ion (Ca2+) pump Ca2+-ATPase (Pmc1p), was more sensitive to Al treatment than the wild-type strain. Overexpression of either PMC1 or an anti-apoptotic factor, such as Bcl-2, Ced-9 or PpBI-1, decreased cytoplasmic Ca2+ levels and rescued yeast from Al sensitivity in both the wild-type and pmc1 mutant. Moreover, pretreatment with the Ca2+ chelator BAPTA-AM sustained cytoplasmic Ca2+ at low levels in the presence of Al, effectively making the cells more tolerant to Al exposure. Quantitative RT-PCR revealed that the expression of calmodulin (CaM) and phospholipase C (PLC), which are in the Ca2+ signaling pathway, was down-regulated under Al stress. This effect was largely counteracted when cells overexpressed anti-apoptotic Ced-9 or were pretreated with BAPTA-AM. Taken together, our results suggest that the negative regulation of Al-induced cytoplasmic Ca signaling is a novel mechanism underlying internal resistance to Al toxicity

The efficacy and safety of Niaoduqing granules in the treatment of diabetic kidney disease: a systematic review and meta-analysis

Background: Diabetic nephropathy (DN) is the main cause of chronic kidney disease (CKD) and end-stage renal failure (ESRF), and the control of disease progression and adverse events during treatment needs to be improved.Objective: This study aimed to systematically evaluate the clinical efficacy and safety of Niaoduqing granules (NDQG) in the treatment of diabetic kidney disease (DKD).Method: Randomized controlled trials (RCTs) of NDQG for DKD from Chinese and English databases up to 31 August 2022 were included. The quality of the literature was assessed using the risk of bias tool of the Cochrane Handbook. At a 95% confidence interval (CI), relative risk (RR) and Cohen’s d were used for the categorical and continuous variables, respectively, and Stata 16.0 software was used for statistical analysis. A funnel plot and Egger’s tests were used to assess publication bias.Result: A total of 4,006 patients were included in 52 RCTs, including 1,987 cases in the control group and 2,019 cases in the treatment group. Compared with conventional treatment (CT), combined NDQG therapy is more effective in improving clinical efficiency [RR = 1.23, 95% confidence interval (1.17, 1.29), p &lt; 0.001, I2 = 53.17%], kidney function (urinary albumin excretion rate [SMD = −0.90, 95% CI (−1.14, −0.66), p &lt; 0.001, I2 = 78.19%], 24hUTP levels [SMD = −0.81, 95% CI (−1.08, −0.55), p &lt; 0.001, I2 = 87.08%], blood urea nitrogen [SMD = −0.54, 95% CI (−0.69, −0.39), p &lt; 0.01, I2 = 77.01%], SCr [SMD = −0.68, 95% CI (−0.90, −0.45), p &lt; 0.001, I2 = 89.97%], CCr [SMD = 0.76, 95% CI (0.10,1.42), p = 0.02, I2 = 95.97%], and Cys-C [SMD = −1.32, 95% CI (−2.25, −0.40), p = 0.01, I2 = 93.44%]), the level of glucose metabolism (fasting blood glucose [SMD = −0.18, 95% CI (−0.38, 0.03), p = 0.10, I2 = 71.18%] and HbA1c [SMD = −0.42, 95% CI (−0.86, −0.02), p = 0.06, I2 = 81.64%]), the level of lipid metabolism (total cholesterol [SMD = −0.70, 95% CI (−1.01, −0.39), p &lt; 0.001, I2 = 86.74%] and triglyceride [SMD = −0.61, 95% CI (−0.87,−0.36), p &lt; 0.001, I2 = 80.64%]), inflammatory factors (Hs-CRP [SMD = −1.00, 95% CI (−1.54, −0.46), p &lt; 0.001, I2 = 86.81%], IL-18 [SMD = −1.25, 95% CI (−1.58, −0.92), p &lt; 0.001, I2 = 0], and TNF-α [SMD = −1.28, 95% CI (−1.64, −0.91), p &lt; 0.001, I2 = 75.73%]), and indicators of oxidative stress (malondialdehyde [SMD = −0.88, 95% CI (−1.22, −0.54), p &lt; 0.001, I2 = 66.01%] and advanced oxidation protein products [SMD = −0.92, 95% CI (−1.85, 0.00), p &lt; 0.001, I2 = 90.68%]). In terms of improving uric acid [SMD = −1.59, 95% CI (−3.45, 0.27), p = 0.09, I2 = 94.67%], 2hPG [SMD = −0.04, 95% CI (−0.61, 0.53), p = 0.89, I2 = 84.33%], HDL-C [SMD = 0.71, 95% CI (0.02, 1.40), p = 0.04, I2 = 87.43%], Hb [SMD = 0.11, 95% CI (−0.10, 0.32), p = 0.32, I2 = 0.00]), and superoxide dismutase [SMD = 1.32, 95% CI (0.44, 2.20), p &lt; 0.001, I2 = 93.48%], the effect is not obvious. Adjuvant treatment with NDQG did not increase the incidence of adverse reactions in the control group [SMD = 0.98, 95% CI (0.71, 1.34), p = 0.89, I2 = 1.59%]. Obvious publication bias was detected by funnel plot and Egger’s test.Conclusion: Our meta-analysis showed that adjuvant treatment with NDQG has more advantages than conventional treatment alone in the DKD treatment, which could improve clinical efficiency, kidney function, the level of glucose metabolism, the level of lipid metabolism, inflammatory factors, and oxidative stress indicators. At the same time, it also showed that NDQG are relatively safe. However, more high-quality studies are needed to provide more reliable evidence for clinical use.Systematic Review Registration: https://www.crd.york.ac.uk/prospero/display_record.php?ID=CRD42022373726, identifier CRD42022373726

Research progress on extracellular vesicles in the renal tubular injury of diabetic kidney disease

Diabetic kidney disease (DKD) is a severe microvascular complication of diabetes and is a chronic progressive condition. It is also a common cause of end-stage renal disease (ESRD), which is characterized by proteinuria or a progressive decline in the glomerular filtration rate. Due to their dependence on high-energy and aerobic metabolism, renal tubules are more susceptible to the metabolic disturbances associated with DKD, leading to inflammation and fibrosis. Consequently, tubular injury has become a recent research focus, and significant advancements have been made in studying the role of extracellular vesicles in DKD-associated tubular injury. This review aimed to elucidate the mechanisms and potential applications of different types of extracellular vesicles in tubular injury in DKD to provide new insights for the prevention and treatment of DKD

Net exchanges of CO2, CH4, and N2O between China's terrestrial ecosystems and the atmosphere and their contributions to global climate warming

Author Posting. © American Geophysical Union, 2011. This article is posted here by permission of American Geophysical Union for personal use, not for redistribution. The definitive version was published in Journal of Geophysical Research 116 (2011): G02011, doi:10.1029/2010JG001393.China's terrestrial ecosystems have been recognized as an atmospheric CO2 sink; however, it is uncertain whether this sink can alleviate global warming given the fluxes of CH4 and N2O. In this study, we used a process-based ecosystem model driven by multiple environmental factors to examine the net warming potential resulting from net exchanges of CO2, CH4, and N2O between China's terrestrial ecosystems and the atmosphere during 1961–2005. In the past 45 years, China's terrestrial ecosystems were found to sequestrate CO2 at a rate of 179.3 Tg C yr−1 with a 95% confidence range of (62.0 Tg C yr−1, 264.9 Tg C yr−1) while emitting CH4 and N2O at rates of 8.3 Tg C yr−1 with a 95% confidence range of (3.3 Tg C yr−1, 12.4 Tg C yr−1) and 0.6 Tg N yr−1 with a 95% confidence range of (0.2 Tg N yr−1, 1.1 Tg N yr−1), respectively. When translated into global warming potential, it is highly possible that China's terrestrial ecosystems mitigated global climate warming at a rate of 96.9 Tg CO2eq yr−1 (1 Tg = 1012 g), substantially varying from a source of 766.8 Tg CO2eq yr−1 in 1997 to a sink of 705.2 Tg CO2eq yr−1 in 2002. The southeast and northeast of China slightly contributed to global climate warming; while the northwest, north, and southwest of China imposed cooling effects on the climate system. Paddy land, followed by natural wetland and dry cropland, was the largest contributor to national warming potential; forest, followed by woodland and grassland, played the most significant role in alleviating climate warming. Our simulated results indicate that CH4 and N2O emissions offset approximately 84.8% of terrestrial CO2 sink in China during 1961–2005. This study suggests that the relieving effects of China's terrestrial ecosystems on climate warming through sequestering CO2 might be gradually offset by increasing N2O emission, in combination with CH4 emission.This study has been supported by NASA LCLUC Program (NNX08AL73G_S01) , NASA IDS Program (NNG04GM39C), and China’s Ministry of Science and Technology (MOST) 973 Program (2002CB412500)

Research progress on exosomes in podocyte injury associated with diabetic kidney disease

Diabetic kidney disease (DKD), a common cause of end-stage renal disease, is a serious complication that develops with the progression of chronic diabetes. Its main clinical manifestations are persistent proteinuria and/or a progressive decline in the estimated glomerular filtration rate. Podocytes, terminally differentiated glomerular visceral epithelial cells, constitute the glomerular filtration barrier together with the basement membrane and endothelial cells, and the structural and functional barrier integrity is closely related to proteinuria. In recent years, an increasing number of studies have confirmed that podocyte injury is the central target of the occurrence and development of DKD, and research on exosomes in podocyte injury associated with DKD has also made great progress. The aim of this review is to comprehensively describe the potential diagnostic value of exosomes in podocyte injury associated with DKD, analyze the mechanism by which exosomes realize the communication between podocytes and other types of cells and discuss the possibility of exosomes as targeted therapy drug carriers to provide new targets for and insights into delaying the progression of and treating DKD