Efficacy of Endoscopic Tissue Adhesive in Patients with Gastrointestinal Tumor Bleeding.

BACKGROUND: Gastrointestinal tumors bleeding remains a significantly clinical challenge due to its resistance to conventional endoscopic hemostasis methods. While the efficacy of endoscopic tissue adhesives (ETA) in variceal bleeding has been established, its role in gastrointestinal tumor bleeding (GITB) remains ambiguous. AIMS: This study aims to assess the feasibility and effectiveness of ETA in the treatment of GITB. METHODS: The study enrolled 30 patients with GITB who underwent hemostasis through Histoacryl® tissue glue injection. Hemostasis success rates, ETA-related adverse events, and re-bleeding rates were evaluated. RESULTS: ETA application achieved successful hemostasis at all tumor bleeding sites, with immediate hemostasis observed in all 30 (100.0%) patients. Among the initially hemostasis cases, 5 patients (17.0%) experienced re-bleeding within 30 days, and the 60 day re-bleeding rate was 20.0% (6/30). Expect for one case of vascular embolism, no adverse events related with ETA application were reported. The 6 month survival was 93%. CONCLUSION: ETA demonstrated excellent immediate hemostasis success rate in GITB cases and showed promising outcomes in prevention re-bleeding.

The predictive value of monocyte-related inflammatory factors for recurrence of atrial fibrillation after cryoablation.

Our objective was to investigate the predictive value of monocyte-related inflammatory factors, including monocyte to high-density lipoprotein cholesterol ratio (MHR) and monocyte to lymphocyte ratio (MLR), for the recurrence of atrial fibrillation (AF) after cryoablation in AF patients. The 570 patients who underwent cryoablation were divided into AF recurrence group and non-recurrence group based on follow-up results. The multivariable logistic regression analysis was used to evaluate the effect of MHR and MLR on AF patients. The AF-free survival status of patients was tested by Kaplan-Meier method. ROC analysis was performed to assess the predictive value of MHR and MLR for post-ablation recurrence of AF. A total of 113 (19.8%) patients relapsed, while 457 patients (80.2%) had no AF recurrence during follow up. Patients with AF recurrence had higher MHR values (0.37±0.14 vs. 0.33±0.14; P=0.004) and higher MLR values (0.49±0.32 vs. 0.18±0.07; P<0.001) compared to those without AF recurrence. MHR≥0.34 combined with MLR≥0.24 (HR=9.979, 95% CI: 6.070-16.407, P<0.001) was an independent factor for predicting AF recurrence after cryoablation in patients by logistic regression analysis. The ROC analysis showed that the AUC for the combination of the MHR and MLR variables was 0.974 (95% CI: 0.962-0.985) and had the highest diagnostic sensitivity (97.4%). Elevated baseline values of the monocyte-related inflammatory factors, MHR and MLR, have a certain predictive value for increased AF recurrence after cryoablation.

A Study of the Detection of SARS-CoV-2 by the Use of Electrochemiluminescent Biosensor Based on Asymmetric Polymerase Chain Reaction Amplification Strategy.

A new and reliable method has been constructed for detecting severe acute respiratory syndrome coronavirus type 2 (SARS-CoV-2) open reading frames 1ab (ORF1ab) gene via highly sensitive electrochemiluminescence (ECL) biosensor technology based on highly efficient asymmetric polymerase chain reaction (asymmetric PCR) amplification strategy. This method uses magnetic particles coupled with biotin-labeled one complementary nucleic acid sequence of the SARS-CoV-2 ORF1ab gene as the magnetic capture probes, and [Formula: see text]-labeled amino-modified another complementary nucleic acid sequence as the luminescent probes, and then a detection model of magnetic capture probes-asymmetric PCR amplification nucleic acid products-[Formula: see text]-labeled luminescent probes is formed, which combines the advantages of highly efficient asymmetric PCR amplification strategy and highly sensitive ECL biosensor technology, enhancing the method sensitivity of detecting the SARS-CoV-2 ORF1ab gene. The method enables the rapid and sensitive detection of the ORF1ab gene and has a linear range of 1-[Formula: see text] copies/[Formula: see text], a regression equation of [Formula: see text] = [Formula: see text] + 2919.301 ([Formula: see text] = 0.9983, [Formula: see text] = 7), and a limit of detection (LOD) of 1 copy/[Formula: see text]. In summary, it can meet the analytical requirements for simulated saliva and urine samples and has the benefits of easy operation, reasonable reproducibility, high sensitivity, and anti-interference abilities, which can provide a reference for developing efficient field detection methods for SARS-CoV-2.

A Study of the Detection of SARS-CoV-2 ORF1ab Gene by the Use of Electrochemiluminescent Biosensor Based on Dual-Probe Hybridization.

To satisfy the need to develop highly sensitive methods for detecting the severe acute respiratory syndrome coronavirus type 2 (SARS-CoV-2) and further enhance detection efficiency and capability, a new method was created for detecting SARS-CoV-2 of the open reading frames 1ab (ORF1ab) target gene by a electrochemiluminescence (ECL) biosensor based on dual-probe hybridization through the use of a detection model of "magnetic capture probes-targeted nucleic acids-Ru(bpy)32+ labeled signal probes". The detection model used magnetic particles coupled with a biotin-labeled complementary nucleic acid sequence of the SARS-CoV-2 ORF1ab target gene as the magnetic capture probes and Ru(bpy)32+ labeled amino modified another complementary nucleic acid sequence as the signal probes, which combined the advantages of the highly specific dual-probe hybridization and highly sensitive ECL biosensor technology. In the range of 0.1 fM~10 µM, the method made possible rapid and sensitive detection of the ORF1ab gene of the SARS-CoV-2 within 30 min, and the limit of detection (LOD) was 0.1 fM. The method can also meet the analytical requirements for simulated samples such as saliva and urine with the definite advantages of a simple operation without nucleic acid amplification, high sensitivity, reasonable reproducibility, and anti-interference solid abilities, expounding a new way for efficient and sensitive detection of SARS-CoV-2.

CD205+ polymorphonuclear myeloid-derived suppressor cells suppress antitumor immunity by overexpressing GLUT3.

Myeloid-derived suppressor cells (MDSCs) are responsible for antitumor immunodeficiency in tumor-bearing hosts. Primarily, MDSCs are classified into 2 groups: monocytic (M)-MDSCs and polymorphonuclear (PMN)-MDSCs. In most cancers, PMN-MDSCs (CD11b+ Ly6Clow Ly6G+ cells) represent the most abundant MDSC subpopulation. However, the functional and phenotypic heterogeneities of PMN-MDSC remain elusive, which delays clinical therapeutic targeting decisions. In the 4T1 murine tumor model, CD11b+ Ly6Glow PMN-MDSCs were sensitive to surgical and pharmacological interventions. By comprehensively analyzing 64 myeloid cell-related surface molecule expression profiles, cell density, nuclear morphology, and immunosuppressive activity, the PMN-MDSC population was further classified as CD11b+ Ly6Glow CD205+ and CD11b+ Ly6Ghigh TLR2+ subpopulations. The dichotomy of PMN-MDSCs based on CD205 and TLR2 is observed in 4T07 murine tumor models (but not in EMT6). Furthermore, CD11b+ Ly6Glow CD205+ cells massively accumulated at the spleen and liver of tumor-bearing mice, and their abundance correlated with in situ tumor burdens (with or without intervention). Moreover, we demonstrated that CD11b+ Ly6Glow CD205+ cells were sensitive to glucose deficiency and 2-deoxy-d-glucose (2DG) treatment. Glucose transporter 3 (GLUT3) knockdown by siRNA significantly triggered apoptosis and reduced glucose uptake in CD11b+ Ly6Glow CD205+ cells, demonstrating the dependence of CD205+ PMN-MDSCs survival on both glucose uptake and GLUT3 overexpression. As GLUT3 has been recognized as a target for the rescue of host antitumor immunity, our results further directed the PMN-MDSC subsets into the CD205+ GLUT3+ subpopulation as future targeting therapy.

Recurrent atrial fibrillation after initial cryoballoon ablation: New perspectives for intensive ablation in right superior pulmonary vein ostium for atrial fibrillation.

A significant proportion of patients with recurrent atrial fibrillation (AF) require repeat radiofrequency (RF) ablation after cryoballoon (CB) ablation. However, little is known about the pulmonary vein (PV) potential reconnection to left atrium and localization of gaps in the initial lesion sets following cryoablation in patients with recurrent AF. The data of 29 consecutive patients with repeat RF ablation for recurrent AF were analyzed. During the second ablation procedures, PV foci of AF were explored in 116 PVs by the CARTO system. All patients had complete PV isolation from initial cryoablation procedure. The fluoroscopy duration, mean cryoablation time and mean cryoablation frequency were lowest for the right superior pulmonary vein (RSPV) (58.69 ± 9.18 s, 185.10 ± 49.25 s and 1.07 ± 0.26; p = 0.024, p = 0.042 and p = 0.032). A significantly higher incidence of conduction gaps per patient was found for the RSPVs compared to the other PVs (p < 0.05 or p < 0.01). For RSPVs, it seemed that gaps were predominantly located at the anterior segment (22 gaps) and inferior segment (22 gaps). RSPV reconnection was independently related to a lower risk of major adverse events after the second ablation during follow up in the study patients (HR 0.275, 95%CI 0.078-0.967, p = 0.044). AF recurrence in patients after cryoablation is significantly associated with conduction gaps in the anterior and inferior segments of RSPVs. Various ablation strategies of close touch of CB on anterior and inferior segments of RSPV ostium, more freezing time and frequency for RSPV may help achieving durable PV isolation during follow up.

Subgroup Identification with Gene Expression Profiles of Adipose Tissue in Patients with Coronary Artery Disease.

Among many diseases, coronary artery disease (CAD) is the primary cause of mortality and morbidity worldwide. With the aim of revealing the underlying genetic characteristics of the CAD subtypes, we recruited patients with CAD and categorized them into subgroups according to the transcriptome expression profiles of the adipose tissue.With the removal of the batch effect, consensus clustering was employed to determine the subgroup numbers. Subgroup-specific genes were determined to conduct analysis of Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG). Weighted gene co-expression network analysis (WGCNA) revealed the subgroup-specific WGCNA modules. Moreover, gene set enrichment analysis (GSEA) was conducted. Overrepresentation enrichment analysis (OEA) of subgroup-specific signatures was also conducted to reveal the significant gene module associated with the corresponding clinical characteristics.After the removal of the batch effect, 77 CAD objects were divided into three subgroups. It was observed that the patients in subgroup III tended to be fat. After analyzing the dominant pathways of each subgroup, we discovered that the protein digestion and absorption pathway was specifically upregulated in subgroup I, which might result from the lowest proportion of the epicardial adipose tissue (EAT) sample. Moreover, subgroup II patients had genetic characteristics of high expression of complement and coagulation cascades and TNF signaling pathway. Furthermore, Th17 cell differentiation was significantly upregulated in subgroup III, indicating that Th17 cell differentiation is related to the clinical characteristics of body mass index (BMI).In conclusion, the genetic classification of CAD subjects indicated that subjects from different subgroups may exhibit specific gene expression patterns, suggesting that more personalized treatment should be applied to patients in each subgroup.

Quantitative Metabolomic Profiling of Plasma, Urine, and Liver Extracts by 1H NMR Spectroscopy Characterizes Different Stages of Atherosclerosis in Hamsters.

Atherosclerosis (AS) is a progressive disease that contributes to cardiovascular disease and shows a complex etiology, including genetic and environmental factors. To understand systemic metabolic changes and to identify potential biomarkers correlated with the occurrence and perpetuation of diet-induced AS, we applied 1H NMR-based metabolomics to detect the time-related metabolic profiles of plasma, urine, and liver extracts from male hamsters fed a high fat and high cholesterol (HFHC) diet. Conventional biochemical assays and histopathological examinations as well as protein expression analyses were performed to provide complementary information. We found that diet treatment caused obvious aortic lesions, lipid accumulation, and inflammatory infiltration in hamsters. Downregulation of proteins related to cholesterol metabolism, including hepatic SREBP2, LDL-R, CYP7A1, SR-BI, HMGCR, LCAT, and SOAT1 was detected, which elucidated the perturbation of cholesterol homeostasis during the HFHC diet challenge. Using "targeted analysis", we quantified 40 plasma, 80 urine, and 60 liver hydrophilic extract metabolites. Multivariate analyses of the identified metabolites elucidated sophisticated metabolic disturbances in multiple matrices, including energy homeostasis, intestinal microbiota functions, inflammation, and oxidative stress coupled with the metabolisms of cholesterol, fatty acids, saccharides, choline, amino acids, and nucleotides. For the first time, our results demonstrate a time-dependent metabolic progression of multiple biological matrices in hamsters from physiological status to early AS and further to late-stage AS, demonstrating that 1H NMR-based metabolomics is a reliable tool for early diagnosis and monitoring of the process of AS.

1H nuclear magnetic resonance-based metabolomic study on efficacy of Qingrehuatan decoction against abundant phlegm-heat syndrome in young adults with essential hypertension.

OBJECTIVE: To observe the influence of Qingrehuatan decoction (QRHT) on serum metabolic profile in young essential hypertension (YEH) patients with abundant phlegm-heat syndrome and provide a basis for treatment with the decoction. METHODS: Twelve male YEH patients were randomly selected and serum samples were collected for examination before and after 4 weeks of the treatment with QRHT. Twelve healthy males were randomly selected and their serum samples were collected as a control. All serum samples were detected using metabolomic technology with 1H nuclear magnetic resonance. Differences in metabolites were studied by principal component analysis and partial least squares-discriminate analysis, which produced scores and loadings plots. RESULTS: After 4 weeks of treatment, serum substances could be distinguished between the YEH patients with abundant phlegm-heat syndrome and the control patients. The specific serum endog- enous metabolites tended to improve after the treatment. QRHT can appropriately increase the levels of glucose, lactic acid, citric acid, high-density lipoprotein, phosphatidylcholine, glycerophosphate choline, hydroxybutyrate, alanine, and glutamate. QRHT could also decrease the levels of low-density lipoprotein/very low-density lipoprotein, lipids, N-acetyl glycoprotein, and O-acetyl glycoprotein. CONCLUSION: QRHT can effectively ameliorate metabolic disorders in YEH Patients with abundant phlegm-heat syndrome. 1H NMR-based metabolomic technology can provide an objective basis for the treatment of YEH patients with abundant phlegm-heat syndrome using QRHT.

[Quantitative metabolomics based on NMR].

Nuclear magnetic resonance (NMR) spectroscopy can be used to both identify and quantify chemicals from complex mixtures. Over the last several decades, significant technical and experimental advances have made quantitative nuclear magnetic resonance (qNMR) a valuable analytical tool for quantitative measurements of a wide variety of samples. This particular approach is now being exploited to characterize the metabolomes of many different biological samples and is called quantitative metabolomics or targeted metabolic profiling. In this review, some of the strengths, limitations of NMR-based quantitative metabolomics will be discussed as well as the practical considerations necessary for acquisition with an emphasis on their use for bioanalysis. Recent examples of the application of this particular approach to metabolomics studies will be also presented.

Evaluation of dose-related effects of 2', 3', 5'-tri-O-acetyl-N6-(3-hydroxylaniline)adenosine using NMR-based metabolomics.

2', 3', 5'-Tri-O-acetyl-N6-(3-hydroxylaniline)adenosine (WS070117) is a derivative compound of natural product cordycepin. It has significant lipids regulating activity and low toxicity which has been proved by in vitro and in vivo experiments. In this study, 1H NMR-based metabolomics was used to investigate the dose-related effects of WS070117 on hyperlipidemia of high-fat-fed hamsters. The hyperlipidemic hamsters were administrated with six different doses of WS070117, including 3, 12, 50, 100, 200 and 400 mg x kg(-1) x d(-1). 1H NMR spectra of hamster serum were visually and statistically analyzed using two multivariate analyses: principal component analysis (PCA) and orthogonal partial least squares-discriminant analysis (OPLS-DA). As a result, WS070117-treated groups showed dose-related regulation of metabolites associated with lipid metabolism, choline metabolism and glucose metabolism. The dose of 3 mg x kg(-1) x d(-1) of WS070117 only exhibited a little lipids regulating activity. However, the doses of 12 and 50 mg x kg(-1) x d(-1) of WS070117 both regulated the contents of metabolites to reverse significantly toward normal levels. When the dose of WS070117 reached 100 mg x kg(-1) x d(-1), it was more effective than positive control drugs. The work suggested that NMR-based metabolomics might be a valuable approach to evaluate dose-related effects of lipids regulating compounds.

Identification and Validation of the Glycolysis and Immune-related Gene Signature for Prognosis in Colorectal Cancer.

BACKGROUND/AIM: Glycolysis has a role in regulating the tumor immune microenvironment. However, the functions and clinical role for facilitating the prognosis prediction of colorectal cancer (CRC) based on glycolysis and immune-related genes remain to be identified. MATERIALS AND METHODS: Genes associated with glycolysis and immunity (GI) were identified from established databases (MSigDB and ImmPort). The TCGA (training cohort) and GSE39582 (validation cohort) datasets were used. Cox regression and least absolute shrinkage and selection operator (LASSO) Cox regression analyses were applied for model construction. The prognostic power of the GI signature was examined by multivariate Cox regression analysis. The correlations between the GI signature, immune cell infiltration and immune checkpoint blockade (ICB) genes were analyzed. To further validate the identified gene signature, quantitative RT-PCR was performed. Cell proliferation assays were conducted for CCK8 detection. RESULTS: A new GI model was constructed, and this signature may serve as an independent prognostic biomarker in CRC. The GI signature remained an effective tool for predicting prognosis among each clinical subgroup. This signature was related to immune cell infiltration of myeloid dendritic cells, cancer-associated fibroblasts (CAFs), CD4+ and CD8+ T cells and response to the ICB immunotherapy-related genes IDO1, BTLA, PD-L1 and PD-L2. In addition, our findings showed that PMM2, IL20RB, and NTF4 exhibited high expression levels in CRC. The upregulation of these genes resulted in the promotion of the proliferation of CRC cells. CONCLUSION: This novel prognostic signature contributed to CRC risk stratification and survival prediction based on glycolysis and immune status.

[A 1H NMR based metabonomics approach to progression of coronary atherosclerosis in a hamster model].

To obtain a better understanding of the progression of atherosclerosis and identify potential biomarkers, proton nuclear magnetic resonance spectroscopy (1H NMR)-based metabonomics was used to study the metabolic changes in the plasma of hamster fed with a high-fat/cholesterol diet. Plasma samples were collected at different time points during the progression of atherosclerosis and individual proton NMR spectra were visually and statistically assessed using multivariate analyses. NMR results for all samples showed a time-dependent development from physiological to pathophysiological status during atherosclerosis. Analysis of the identified biomarkers of atherosclerosis suggests that lipid and amino acid metabolisms are significantly disturbed, together with inflammation, oxidative stress, following cholesterol overloading. The results enriched our understanding of the mechanism of atherosclerosis and demonstrated the effectiveness of the NMR-based metabonomics approach to study such a complex disease.

Downregulation of ornithine decarboxylase by pcDNA-ODCr inhibits gastric cancer cell growth in vitro.

Ornithine decarboxylase (ODC), the first rate-limiting enzyme of polyamine biosynthesis, was found to be associated with cell growth, proliferation and transformation. ODC gene expression in gastric cancer was increased and its level was positively correlated with the degree of malignity of gastric mucosa and development of gastric lesions. In order to evaluate the therapeutic effects of antisense RNA of ODC on gastric cancer, an antisense RNA of ODC expressing plasmid pcDNA-ODCr which delivered a 120 bp fragment complementary to the initiation codon of ODC gene was constructed and transfected to gastric cancer cells SGC7901 and MGC803. Expression of ODC in gastric cancer cells was determined by western blot. Cell proliferation was assessed by MTS assay. Cell cycle was analyzed by flow cytometry and Matrigel assay was performed to assess the ability of gastric cancer cell invasiveness. The results showed that the ODC gene expression in gastric cancer cells transfected with the pcDNA-ODCr was downregulated efficiently. Tumor cell proliferation was suppressed significantly, and cell cycle was arrested at G1 phase. Gastric cancer cells had reduced invasiveness after gene transfer. Our study suggested that antisense RNA of ODC expressing plasmid pcDNA-ODCr had antitumor activity by inhibiting the expression of ODC, and downregulation of ODC expression using a gene therapy approach might be a novel therapeutic strategy for gastric cancer.

Long non-coding RNA PROX1-AS1 knockdown upregulates microRNA-519d-3p to promote chemosensitivity of retinoblastoma cells via targeting SOX2.

OBJECTIVE: Long non-coding RNAs (lncRNAs) and microRNAs (miRNAs) participate in tumor progression, while the role of PROX1-antisense RNA1 (PROX1-AS1) sponging miR-519d-3p in retinoblastoma (RB) remains largely unknown. We aim to explore the effect of the PROX1-AS1/miR-519d-3p/sex determining region Y-box 2 (SOX2) in chemosensitivity of RB cells. METHODS: Expression of PROX1-AS1, miR-519d-3p and SOX2 in RB tissues and cells was determined. The drug-resistant cell lines were established and respectively intervened with PROX1-AS1 or miR-519d-3p expression to explore their roles in drug resistance and malignant behaviors of the drug-resistant cells. The binding relationships between PROX1-AS1 and miR-519d-3p, and between miR-519d-3p and SOX2 were evaluated. RESULTS: PROX1-AS1 and SOX2 were upregulated while miR-519d-3p was downregulated in RB tissues and cells, especially in drug-resistant cells. The PROX1-AS1 inhibition or miR-519d-3p elevation suppressed the drug resistance, proliferation, migration and invasion, and promoted apoptosis of the drug-resistant RB cells. Moreover, PROX1-AS1 sponged miR-519d-3p and miR-519d-3p targeted SOX2. CONCLUSION: PROX1-AS1 knockdown upregulates miR-519d-3p to promote chemosensitivity of RB cells via targeting SOX2.

Effectiveness and Safety of Cryoablation in Patients With Atrial Fibrillation Episodes of <24 h Duration: A Propensity-Matched Analysis.

Background: Paroxysmal atrial fibrillation (AF) is closely related to pathophysiologic processes and clinical outcomes. However, it is uncertain whether cryoablation of pulmonary veins isolation is effective and safe for patients with symptomatic and drug refractory AF episodes of <24-h duration. Methods: The patients were designed into Group A (253 patients with paroxysmal AF episodes of <24-h duration) and Group B (253 patients with paroxysmal AF lasting for 24 h or longer) on a 1:1 basis by identical propensity scores. Mortality, stroke/transient ischemic attack (TIA), and complications relevant to the cryoablation procedure were compared, and recurrence of atrial tachyarrhythmia was analyzed for clinical independent predictors. Results: The rate of atrial tachyarrhythmia recurrence was 21.74% in Group A and 30.04% in Group B, respectively (P = 0.042). At 12-month follow-up from the procedure, lower incidences of stroke/TIA endpoint of the patients were observed in Group A compared with Group B by Kaplan-Meier analysis [HR 0.34 (0.13-0.87), P = 0.025]. No significant differences in mortality and complications relevant to the cryoablation procedure were observed between Group A and Group B. Moreover, adjusted multivariable Cox regression analysis showed that <24-h paroxysmal AF type (HR 0.644, 95% CI: 0.455-0.913, P = 0.014) and left atrium diameter (LAD) (>40 mm) (HR 1.696, 95% CI: 1.046-2.750, P = 0.032) were independently associated with the incidence of recurrence of atrial tachyarrhythmia in the study. Conclusion: Our findings indicated that <24-h paroxysmal AF type was obviously associated with an increased success rate of cryoablation and reduced incidence of stroke/TIA during the follow-up period. Therefore, there is superior effectiveness and similar safety in patients with AF episodes of <24-h duration compared with patients with longer paroxysmal AF duration.

A Novel Recombinant Virus-Like Particles Displaying B and T Cell Epitopes of Japanese Encephalitis Virus Offers Protective Immunity in Mice and Guinea Pigs.

Virus-like particles (VLPs) are non-replicative vectors for the delivery of heterologous epitopes and are considered one of the most potent inducers of cellular and humoral immune responses in mice and guinea pigs. In the present study, VLP-JEVe was constructed by the insertion of six Japanese encephalitis virus (JEV) envelope protein epitopes into different surface loop regions of PPV VP2 by the substitution of specific amino acid sequences without altering the assembly of the virus; subsequently, the protective efficacy of this VLP-JEVe was evaluated against JEV challenge in mice and guinea pigs. Mice immunized with the VLP-JEVe antigen developed high titers of neutralizing antibodies and 100% protection against lethal JEV challenge. The neutralizing and hemagglutination inhibition (HI) antibody responses were also induced in guinea pigs vaccinated with VLP-JEVe. In addition, immunization with VLP-JEVe in mice induced effective neutralizing antibodies and protective immunity against PPV (porcine parvovirus) challenge in guinea pigs. These studies suggest that VLP-JEVe produced as described here could be a potential candidate for vaccine development.

RNA interference-mediated silencing of UBCH10 gene inhibits colorectal cancer cell growth in vitro and in vivo.

1. UbcH10 is the cancer-related E2 ubiquitin-conjugating enzyme, and its overexpression has been demonstrated in a variety of malignancies. The aim of the present study is to silence UbcH10 gene by RNA interference (RNAi) and to observe its inhibitory effect on the colorectal cancer cell growth in vitro and in vivo. 2. We constructed the expression vector pGPU6/GFP/Neo/UbcH10-RNAi (pUbcH10-RNAi), which contained a UbcH10 short hairpin RNA expression cassette. Then the UbcH10 gene silencing cell lines LoVo/UbcH10-RNAi and HT-29/UbcH10-RNAi were established. Reverse transcription-polymerase chain reaction and western blot analysis were used to evaluate the expression of the UbcH10 gene. Cell Counting Kit-8 was used to assess properties of tumour cell growth in vitro. Flow cytometry was used to detect the effect of pUbcH10-RNAi on the cell cycle of colorectal cancer cells. Furthermore, the anti-tumour effects of pUbcH10-RNAi were evaluated in vivo in a nude mouse xenografts model. 3. Results demonstrated that UbcH10 gene expression was significantly decreased in pUbcH10-RNAi treated cells. Colorectal cancer cells growth was markedly suppressed in the pUbcH10-RNAi group compared with control conditions and colorectal cancer cells were arrested in the G2-M phase. In vivo, the downregulation of UbcH10 gene expression by pUbcH10-RNAi also inhibited tumour growth in a nude mice xenograft model. 4. Our study suggests that RNA interference-mediated silencing of UbcH10 gene has anti-tumour activity on colorectal cancer and might have therapeutic potential for the treatment of colorectal cancer.

STEAP4 Inhibits HIF-1α/PKM2 Signaling and Reduces High Glucose-Induced Apoptosis of Retinal Vascular Endothelial Cells.

BACKGROUND: Diabetic retinopathy (DR) is a vascular lesion induced by high glucose. STEAP4 is an indispensable membrane protein, which is closely related to hyperglycemic-induced cell inflammation and injury, while STEPT4 has not been studied in hyperglycemic-induced retinal vascular endothelial cell injury. METHODS: The expression of STEAP4 was detected by RT-qPCR and Western blot. CCK-8 was used to detect cell survival. STEAP4 was overexpressed by cell transfection. The expressions of cytokines TNF-α, IL-1, IL-6, ICAM-1, MDA, SOD and ROS were detected by ELISA. Cell apoptosis was detected by flow cytometry. The expressions of proteins associated with cell damage VEGF, KLF2, eNOS and apoptosis-related proteins Bax, cleaved caspase3 and Bcl2 were detected by Western blot. Finally, the expressions of HIFα and PKM2 were detected by immunofluorescence and Western blot. RESULTS: The expression of STEAP4 in hyperglycemic-induced retinal vascular endothelial cells (HRCECs) decreased gradually. Overexpression of STEAP4 reduced inflammation and apoptosis of HRCECs and improved dysfunction of them. Meanwhile, overexpression of steap4 inhibited the expression of HIF-1α/PKM2 signal. CONCLUSION: STEAP4 can be a potential therapeutic target for diabetic retinopathy by inhibiting HIF1/PKM2 signaling to reduce hyperglycemic-induced retinal cell apoptosis.

Combined effects of mulch film-derived microplastics and atrazine on oxidative stress and gene expression in earthworm (Eisenia fetida).

With the wide use of mulch film and pesticides, mulch film-derived microplastics are very likely to produce combined effects with pesticides in agricultural soil. However, little is known about their combined toxicity on terrestrial organisms. This study aimed to investigate the combined toxicity of unused or farmland residual transparent low-density polyethylene mulch film-derived microplastics (MPs and MPs-aged, respectively) (550-1000 µm) and atrazine (ATZ; 0.02 and 2.0 mg/kg) on the earthworm (Eisenia fetida). After single and combined exposure to ATZ and microplastics for 28 d, the results showed an accumulation of reactive oxygen species, a decrease in superoxide dismutase, catalase, and glutathione-S-transferase activities, an increase in the malondialdehyde and 8-hydroxydeoxyguanosine levels, and abnormal expression of annetocin, heat shock protein 70, translationally controlled tumor protein and calreticulin genes. Integrated biological response (IBR) values calculated at the biochemical level indicated that the combined exposure to ATZ and microplastics, particularly to high concentrations of ATZ, induced greater oxidative stress in E. fetida compared with that of exposure to ATZ or microplastics alone. In addition, the IBR values calculated at the gene level did not show regular changes after combined exposure to ATZ and microplastics compared with those of a single exposure. The oxidative stress and abnormal expression of genes in E. fetida induced by MPs-aged were higher than those induced by MPs; a similar trend was observed for oxidative stress induced by MPs/MPs-aged + ATZ2.0, whereas an opposite trend was observed for the abnormal expression of genes in E. fetida induced by MPs/MPs-aged + ATZ0.02/ATZ2.0. Our results suggest that mulch film-derived microplastics have the potential to enhance the toxicity of ATZ within the soil environment.