Recombinant Newcastle disease virus (NDV/Anh-IL-2) expressing human IL-2 as a potential candidate for suppresses growth of hepatoma therapy

AbstractNewcastle disease virus (NDV) have shown oncolytic therapeutic efficacy in preclinical study and are currently approved for clinical trials. NDV Anhinga strain which is a mesogenic strain should be classified as lytic strain and has a therapeutic efficacy in hepatocellular cancer. In this study, we evaluated the capacity of NDV Anhinga strain to elicit immune reaction in vivo and the possibility for using as a vaccine vector for expressing tumor therapeutic factors. Interleukin-2 (IL-2) could boost the immune response against the tumor cells. Therefore, we use NDV Anhinga strain as backbone to construct a recombinant virus (NDV/Anh-IL-2) expressing IL-2. The virus growth curve showed that the production of recombinant NDV/Anh-IL-2 was slightly delayed compared to the wild type. The NDV/Anh-IL-2 strain could express soluble IL-2 and effectively inhibit the growth of hepatocellular carcinoma in vivo. 60 days post-treatment, mice which were completely cured by previous treatment were well protected when rechallenged with the same tumor cell. From the H&E-stained sections, intense infiltration of lymphocyte was observed in the NDV Anhinga strain treated group, especially in NDV/Anh-IL-2 group. The NDV Anhinga strain could not only kill the tumor directly, but could also elicit immune reaction and a potent immunological memory when killing tumor in vivo. In conclusion, the Anhinga strain could be an effective vector for tumor therapy; the recombinant NDV/Anh-IL-2 strain expressing soluble IL-2 is a promising candidate for hepatoma therapy

Evaluating the sustainable cultivation of 'Fuji' apples: suitable crop load and the impact of chemical thinning agents on fruit quality and transcription

The optimal load capacity provides sustainable production and high quality in 'Fuji' apple production. Determining optimal crop loads with simple and affordable thinning methods is critical for apple. We set different load capacities on different apple tree rootstocks and evaluated their effects on tree development, fruit yield and quality. The optimal load capacity for vigorous stock, dwarfing rootstock, and dwarfing interstock were 240, 90, and 100 for apples above 80 mm in diameter, respectively. To explore the optimal strategies of diverse fruit thinning agents (carbaryl, 6-BA, NAA, and Metamitron) used in the 'Fuji' apple tree, we assess from thinning agents' types, varying spraying concentrations, and application time in the most widely used dwarfing interstock. The results showed that the best use of fruit thinning agents is 2,000 mg/L carbaryl at 10 d after flowering could significantly reduce fruit set rate and improve the fruit quality in the dwarfing interstock. During the research, only the fruit thinner NAA treatment suppresses fruit development. Thus, we performed the transcriptome analysis on the NAA-treated and control fruits at 60 (FS1), 90 (FS2), 120 (FS3), and 150 (FS4) days after flowering stages to investigate the potential transcriptional regulations of NAA on fruit development and ripening. Transcriptome results showed that genes related to fruit expansion (expansin A15, expansin B3), phytohormone-related genes (HVA22C, PRE1, AHP1, etc.), fruit coloring-related genes (PAL), and many ripening-related transcription factors (EIN3, ERF, ARF, etc.) might be regulated by exogenous NAA. Our results provide an important reference for the sustainable production and optimal use of fruit thinning agents

Reduced macular thickness and vascular density in abnormal glucose metabolism patients: A meta-analysis of optical coherence tomography (OCT) and OCT angiography studies

Abstract. Background:. Alterations in macular thickness and vascular density before clinically visible diabetic retinopathy (DR) remain inconclusive. This study aimed to determine whether retinal manifestations in abnormal glucose metabolism (AGM) patients differ from those in the healthy individuals. Methods:. PubMed, Embase, and Web of Science were searched between 2000 and 2021. The eligibility criteria were AGM patients without DR. Primary and secondary outcomes measured by optical coherence tomography (OCT) and OCT angiography (OCTA) were analyzed and expressed as standardized mean differences (SMDs) with 95% confidence intervals (CIs). A random-effects model was used in the data synthesis. The potential publication bias for the variables was evaluated using Egger’s test. Results:. A total of 86 observational studies involving 13,773 participants and 15,416 eyes were included. OCT revealed that compared to healthy controls, the total macular thickness of AGM patients was thinner, including the thickness of fovea (–0.24, 95% CI [–0.39, –0.08]; P = 0.002, I2 = 87.7%), all regions of parafovea (–0.32, 95% CI [–0.54, –0.11]; P = 0.003; I2 = 71.7%) and the four quadrants of perifovea; the thickness of peripapillary retinal nerve fiber layer (pRNFL), macular retinal nerve fiber layer (mRNFL), and ganglion cell layer (GCL) also decreased. OCTA indicated that the superficial and deep vascular density decreased, the foveal avascular zone (FAZ) area enlarged, and the acircularity index (AI) reduced in AGM individuals. Conclusions:. Retinal thinning and microvascular lesions have occurred before the advent of clinically detectable DR; OCT and OCTA may have the potential to detect these preclinical changes. Registration:. PROSPERO; http://www.crd.york.ac.uk/prospero/; No. CRD42021269885

Dynamic Characteristics and Successive Start-Up Control Strategy Optimization of Pumped Storage Units under Low-Head Extreme Conditions

With inherent ‘S’ characteristics and the one-tunnel-with-two-units arrangement of the pump-turbine, hydraulic transient changes in the successive start-up process are complex, and the optimal control is difficult. This paper aims to study the dynamic characteristics and successive start-up control strategy optimization of two hydraulic couplings pumped storage units (PSUs) under low-head extreme conditions. Firstly, an accurate model of two hydraulic coupling PSUs’ successive start-up is established. Based on this model, the influence of the interval time of successive start-up on the dynamic characteristics of PSUs is carried out. It is shown that the change of the interval time of the successive start-up (ΔT) of the two PSUs has a significant impact on the dynamic response stability of the low-head start-up. If ΔT is more than 40 s, the hydraulic oscillation and speed fluctuation of the PSUs deteriorate. Secondly, with the different controller parameters for the two PSUs, a novel multi-objective optimization scheme with fractional order PID controller (FOPID) is proposed to figure out the best control scheme for the successive start-up. Furthermore, selecting the sum of the rise time (Tr) of the rotating speed of two PSUs and the sum of the integral time absolute error (ITAE) of two PSUs is the objective. Meanwhile, the optimization scheme of PID with different parameters (PIDDP) is used to compare and verify the optimization method proposed in this paper. The results for this extreme condition indicate that FOPID has more significant advantages in optimizing the instability of the successive start-up process, with the better Pareto front, and the optimized scheme has a more stable dynamic transition process of flow, water hammer pressure, and rotational speed

Environmental exposure to perchlorate, nitrate, and thiocyanate in relation to chronic kidney disease in the general US population, NHANES 2005–2016

Abstract. Background:. Few studies have explored the impact of perchlorate, nitrate, and thiocyanate (PNT) on kidney function. This study aimed to evaluate the association of urinary levels of PNT with renal function as well as the prevalence of chronic kidney disease (CKD) among the general population in the United States. Methods:. This analysis included data from 13,373 adults (≥20 years) from the National Health and Nutrition Examination Survey 2005 to 2016. We used multivariable linear and logistic regression, to explore the associations of urinary PNT with kidney function. Restricted cubic splines were used to assess the potentially non-linear relationships between PNT exposure and outcomes. Results:. After traditional creatinine adjustment, perchlorate (P-traditional) was positively associated with estimated glomerular filtration rate (eGFR) (adjusted β: 2.75; 95% confidence interval [CI]: 2.25 to 3.26; P < 0.001), and negatively associated with urinary albumin-to-creatinine ratio (ACR) (adjusted β: −0.05; 95% CI: −0.07 to −0.02; P = 0.001) in adjusted models. After both traditional and covariate-adjusted creatinine adjustment, urinary nitrate and thiocyanate were positively associated with eGFR (all P values <0.05), and negatively associated with ACR (all P values <0.05); higher nitrate or thiocyanate was associated with a lower risk of CKD (all P values <0.001). Moreover, there were L-shaped non-linear associations between nitrate, thiocyanate, and outcomes. In the adjusted models, for quartiles of PNT, statistically significant dose–response associations were observed in most relationships. Most results were consistent in the stratified and sensitivity analyses. Conclusions:. Exposures to PNT might be associated with kidney function, indicating a potential beneficial effect of environmental PNT exposure (especially nitrate and thiocyanate) on the human kidney

Use of viral vector to deliver IL-15 for cancer therapy: An Overview

97-108One of the most important goals of gene therapy is to create delivery systems that are capable of effectively targeting the tumor cells with limited harmful side effects. The immune stimulatory cytokine interleukin-15 (IL-15) was recognized as one of the most promising drugs for cancer treatment and is currently applied in clinical trials alone or as an adjuvant for certain types of metastatic solid tumors.But the administration of IL-15 alone has faced some hurdles that made its anti-tumor effects not very satisfying, here comes the need to establish new approaches for demonstrating IL-15 in a more effective way that can overcome these obstacles and get maximum benefit from its effects in vivo. One of the alternative approaches is the use of viruses as carriers to deliver foreign genes (virotherapy) which has been used almost two decades ago. Since the 1990s when the first genetically engineered virus was described to selectively replicate in tumor cells, several preclinical and clinical studies with different viruses were carried out. In this review we will shed the light on the most widely used vectors to deliver IL-15 and discuss their results to grasp the benefits of using different vectors delivering IL-15 in cancer therapy, as we will mention in brief the disadvantages of non-viral delivery systems to overcome IL-15 obstacles, then discuss some of the successful studies with promising results to deliver IL-15 with overcome the obstacles and get better achievements in fighting cancer battle

Direct reprogramming of fibroblasts into functional hepatocytes via CRISPRa activation of endogenous Gata4 and Foxa3

Abstract. Background:. The ability to generate functional hepatocytes without relying on donor liver organs holds significant therapeutic promise in the fields of regenerative medicine and potential liver disease treatments. Clustered regularly interspaced short palindromic repeats (CRISPR) activator (CRISPRa) is a powerful tool that can conveniently and efficiently activate the expression of multiple endogenous genes simultaneously, providing a new strategy for cell fate determination. The main purpose of this study is to explore the feasibility of applying CRISPRa for hepatocyte reprogramming and its application in the treatment of mouse liver fibrosis. Method:. The differentiation of mouse embryonic fibroblasts (MEFs) into functional induced hepatocyte-like cells (iHeps) was achieved by utilizing the CRISPRa synergistic activation mediator (SAM) system, which drove the combined expression of three endogenous transcription factors—Gata4, Foxa3, and Hnf1a—or alternatively, the expression of two transcription factors, Gata4 and Foxa3. In vivo, we injected adeno-associated virus serotype 6 (AAV6) carrying the CRISPRa SAM system into liver fibrotic Col1a1-CreER; Cas9fl/fl mice, effectively activating the expression of endogenous Gata4 and Foxa3 in fibroblasts. The endogenous transcriptional activation of genes was confirmed using real-time quantitative polymerase chain reaction (RT-qPCR) and RNA-seq, and the morphology and characteristics of the induced hepatocytes were observed through microscopy. The level of hepatocyte reprogramming in vivo is detected by immunofluorescence staining, while the improvement of liver fibrosis is evaluated through Sirius red staining, alpha-smooth muscle actin (α-SMA) immunofluorescence staining, and blood alanine aminotransferase (ALT) examination. Results:. Activation of only two factors, Gata4 and Foxa3, via CRISPRa was sufficient to successfully induce the transformation of MEFs into iHeps. These iHeps could be expanded in vitro and displayed functional characteristics similar to those of mature hepatocytes, such as drug metabolism and glycogen storage. Additionally, AAV6-based delivery of the CRISPRa SAM system effectively induced the hepatic reprogramming from fibroblasts in mice with live fibrosis. After 8 weeks of induction, the reprogrammed hepatocytes comprised 0.87% of the total hepatocyte population in the mice, significantly reducing liver fibrosis. Conclusion:. CRISPRa-induced hepatocyte reprogramming may be a promising strategy for generating functional hepatocytes and treating liver fibrosis caused by hepatic diseases