Upregulation of circ-IGF1R increased therapeutic effect of hypoxia-pretreated ADSC-derived extracellular vesicle by regulating miR-503-5p/HK2/VEGFA axis.

Diabetes mellitus is a major cause of blindness and chronic ulcers in the working-age population worldwide. Wound healing is deeply dependent on neovascularization to restore blood flow. Former research has found that differentially expressed circular RNAs (circRNAs) are associated with hyperglycaemia-induced endothelial cell damage, and hypoxia-pretreated adipose-derived stem cells (ADSCs)-extracellular vesicle (HEV) transplants have a more therapeutic effect to enhance wound healing in diabetic mice by delivery circRNA. The current investigation employed high-throughput sequencing to identify circRNAs that are abnormally expressed between EV and HEV. The regulatory mechanism and predicted targets of one differentially expressed circRNA, circ-IGF1R, were investigated utilizing bioinformatics analyses, luciferase reporter assays, angiogenic differentiation assays, flow cytometric apoptosis analysis and RT-qPCR. Circ-IGF1R expression increased in HEV, and downregulation of circ-IGF1R suppressed and reversed the promotion effect of HEV on angiogenesis in ulcerated tissue. Bioinformatics analyses and luciferase reporter assays confirmed that miR-503-5p was the downstream target of circ-IGF1R, and inhibiting miR-503-5p restored the promotion effect of HEV on angiogenesis after circ-IGF1R silence. The study also found that miR-503-5p can interact with 3'-UTR of both HK2 and VEGFA. Overexpression of HK2 or VEGFA restored the promotion effect of HExo on angiogenesis after circ-IGF1R silence. Overexpression miR-503-5p or silence HK2/VEGFA reversed the protective effect of circ-IGF1R to MLMECs angiogenic differentiation. Overexpression of circ-IGF1R increased the protective effect of HEV on the promotion of wound healing in mice with diabetes. Circ-IGF1R promotes HIF-1α expression through miR-503-5p sponging. Our data demonstrate that circ-IGF1R overexpression EVs from ADSCs suppress high glucose-induced endothelial cell damage by regulating miR-503-5p/HK2/VEGFA axis.

Exosomes derived from mmu_circ_0000250-modified adipose-derived mesenchymal stem cells promote wound healing in diabetic mice by inducing miR-128-3p/SIRT1-mediated autophagy.

More and more evidence advises that circular RNAs (circRNAs) function critically in regulating different disease microenvironments. Our previous study found that autotransplantation of adipose-derived mesenchymal stem cells (ADSCs) promotes diabetes wound healing. Exosomes derived in ADSCs play an important regulatory role. This study aimed to characterize if mmu_circ_0000250 played a role in ADSC-exosome-mediated full-thickness skin wound repair in diabetic rats. Endothelial progenitor cells (EPCs) were selected to study the therapeutic mechanism of exosomes in high-glucose (HG)-induced cell damage and dysfunction. Analysis and luciferase reporter assay were utilized to explore the interaction among mmu_circ_0000250, miRNA (miR)-128-3p, and sirtuin (SIRT)1. The diabetic rats were used to confirm the therapeutic effect of mmu_circ_0000250 against exosome-mediated wound healing. Exosomes containing a high concentration of mmu_circ_0000250 had a greater therapeutic effect on restoration of the function of EPCs by promotion autophagy activation under HG conditions. Expression of mmu_circ_0000250 promoted SIRT1 expression by miR-128-3p adsorption, which was confirmed via luciferase reporter assay and bioinformatics analysis. In vivo, exosomes containing a high concentration of mmu_circ_0000250 had a more therapeutic effect on wound healing when compared with wild-type exosomes from ADSCs. Immunohistochemistry and immunofluorescence detection showed that mmu_circ_0000250 increased angiopoiesis with exosome treatment in wound skin and suppressed apoptosis by autophagy activation. In conclusion, we verified that mmu_circ_0000250 enhanced the therapeutic effect of ADSC-exosomes to promote wound healing in diabetes by absorption of miR-128-3p and upregulation of SIRT1. Therefore, these findings advocate targeting the mmu_circ_0000250/miR-128-3p/SIRT1 axis as a candidate therapeutic option for diabetic ulcers.

Stem cell therapy with CRISPR/Cas9-mediated MALAT1 delivery modulates miR-142 and rescues wound healing in rats with age-associated diabetic foot ulcers.

BACKGROUND: Diabetic foot ulcer (DFU) is a serious diabetes complication, significantly impacting the quality of life, particularly in the elderly. Age-associated DFUs pose additional challenges due to impaired healing mechanisms. Our study aims to explore the role of metastasis-associated lung adenocarcinoma transcript 1 (MALAT1) as a miR-142 sponge in repairing diabetic rat foot ulcer tissue under age-associated diabetes, offering a new theoretical basis and therapeutic target for preventing and treating diabetic vascular disease in the elderly. METHODS: Using qPCR, we analyzed MALAT1 and miR-142 expression in EPCs and hUC-MSCs. Targetscan predicted potential interaction targets for MALAT1 and miR-142, confirmed by dual luciferase reporter gene assay. An age-associated diabetic rat model was established using Streptozotocin (STZ) injection. Hypoxia, apoptosis, and angiogenesis-related proteins were assessed through Western Blot. In vitro, miR-142 inhibition and MALAT1 overexpression promoted foot ulcer healing in diabetic rats. RESULTS: MALAT1 acted as a miR-142 sponge, downregulated in hUC-MSCs under high glucose, relevant to age-associated diabetic foot ulcers. MiR-142 negatively regulated SIRT1 and Nrf2. In vitro experiments demonstrated potential significance for age-related DFU treatment. CONCLUSIONS: MALAT1 in human umbilical cord mesenchymal stem cells expedited foot ulcer healing in diabetic rats, particularly in age-associated diabetes, through miR-142 sponge activity. These findings offer insights for novel therapeutic strategies targeting elderly diabetic foot ulcers, emphasizing exogenous stem cell transplantation's potential in effective DFU treatment for the elderly.

Molecular subtypes, prognostic and immunotherapeutic relevant gene signatures mediated by DNA methylation regulators in hepatocellular carcinoma.

Growing evidence has revealed the crucial role of epigenetics in tumor progression and immune response. However, the molecular subtypes and their microenvironment characterization mediated by DNA methylation regulators in hepatocellular carcinoma remain little known. In this study, we comprehensively integrated the transcriptome profiling of twenty DNA methylation regulators in hepatocellular carcinoma. Consensus clustering was used to identify distinct methylation regulator-related molecular subtypes. The prognostic DMS signature was constructed using principal components analysis. Most regulators experienced a low genomic variation, but we found a remarkably difference in mRNA expression of these regulators between normal and tumor tissues. Three distinct methylation regulator-related molecular subtypes were successfully identified according to the expression of 20 regulators, which had substantially different biological characteristics and prognosis. The classic carcinogenic pathways and stromal activity including TGF-beta, p53 and WNT signaling pathway were significantly activated in subtype B, leading to a survival inferiority in subtype B compared to other two subtypes. Further analysis demonstrated the constructed DMS signature was an independent predictive biomarker in patient prognosis. Two anti-checkpoint immunotherapy cohorts demonstrated patients with high DMS presented significantly improved treatment advantages and enhanced responses especially the survival prolonged. Generally, the high DMS groups improved more than 15% clinical response to immunotherapy than low DMS groups. In conclusion, this study identified three DNA methylation regulator-related subtypes with distinct clinical, molecular and biological characteristics, and constructed a prognostic and immunotherapeutic relevant gene signature. It might help to promote individualized immunotherapy for hepatocellular carcinoma from the perspective DNA methylation regulators.

Hypoxic ADSC-derived exosomes enhance wound healing in diabetic mice via delivery of circ-Snhg11 and induction of M2-like macrophage polarization.

Diabetes mellitus is a major cause of blindness and chronic ulcers in the working age population worldwide. Former research have found that differentially expressed circular RNAs (circRNAs) are associated with hyperglycemia (HG)-induced endothelial cell damage. And adipose-derived stem cells (ADSCs)-exosome transplant have more therapeutic effect to enhance wound healing in diabetic mice by delivery circRNA. The current investigation employed high-throughput sequencing to identify circRNAs that are abnormally expressed in endothelial progenitor cells (EPCs) under HG conditions. The regulatory mechanism and predicted targets of one differentially expressed circRNA, circ-Snhg11, were investigated utilizing bioinformatics analyses, luciferase reporter assays, angiogenic differentiation assays, flow cytometric apoptosis analysis, and RT-qPCR. The result show that circ-Snhg11 expression decreased in EPCs under HG conditions and that overexpression of circ-Snhg11 suppressed HG-induced endothelial cell damage and M1-like macrophage polarization. miR-144-3p and HIF-1α were identified as downstream targets of circ-Snhg11, which was further verified by luciferase reporter analysis. miR-144-3p overexpression or HIF-1α inhibition reversed circ-Snhg11 protective effect on HG-induced endothelial cell dysfunction, as evidenced by increased apoptosis, abnormal vascular differentiation, and secretion of inflammatory factors. In addition, miR-144-3p overexpression or inhibition of HIF-1α reversed protective effect regarding circ-Snhg11 on M2-like macrophage polarization under HG conditions. These findings suggest that circ-Snhg11 promotes HIF-1α expression through miR-144-3p sponging. Our data demonstrate that circ-Snhg11 overexpression exosome from ADSCs suppresses HG-induced endothelial cell damage and induces M2-like macrophage polarization via the miR-144-3p/HIF-1α axis.

A 3D multi-modal intelligent intervention system using electromagnetic navigation for real-time positioning and ultrasound images: a prospective randomized controlled trial.

Background: Anesthesia, nerve block, therapeutic injections, and biopsies all require an acupuncture intervention. However, traditional two-dimensional (2D) ultrasound-guided needle puncture is often challenging and therefore requires the use of three-dimensional (3D) ultrasound images to accurately identify and evaluate the patient's anatomical structure. Methods: In this study, a 3D multi-modal intelligent intervention system using electromagnetic navigation for real-time positioning and ultrasound images was described. A total of 190 cases requiring puncture were randomly divided into control (conventional 2D ultrasound instrument) and experimental (novel 3D ultrasound imedis9000) groups. The advantages and disadvantages of the two puncture methods were prospectively analyzed in the 190 cases, and the feasibility of electromagnetic navigation real-time positioning was compared to ultrasound imaging. Results: This study included 190 cases from two centers that required puncture treatment and were randomly assigned to the control (conventional 2D ultrasound instrument; n=95) or the experimental (novel 3D ultrasound imedis9000; n=95) groups. Percutaneous vascular puncture, percutaneous biopsy, percutaneous bile duct puncture, thoracic paravertebral nerve block, and sciatic nerve block operations were performed separately. The results indicated that the puncture time and number of trials in the experimental group were significantly lower than those in the control group. No significant difference was identified in the basic vital signs between the two groups before and after surgery. The success rate of the novel 3D ultrasound imedis9000 was 100%, and the success rate of the conventional 2D ultrasound instrument was 95.7%. Furthermore, the results also showed that the novel 3D ultrasound imedis9000 and the matching coaxial positioning channel puncture needle had low pain, good toughness and strength, and great convenience. Conclusions: The new 3D multi-modal intelligent intervention system using electromagnetic navigation real-time positioning and ultrasound images has significant advantages compared with conventional 2D ultrasound in terms of puncture time, number of trials, operation difficulty, and convenience, and is worthy of further promotion and use in clinics. Trial Registration: Beijing Municipal Drug Administration, 20190015.

High nuclear expression of APE1 correlates with unfavorable prognosis and promotes tumor growth in hepatocellular carcinoma.

APE1 is a multifunctional protein that plays important roles in cancer development. However, the association between APE1 expression and the clinicopathological parameters of HCC patients has not been fully characterized. In this study, bioinformatics analysis of APE1 was performed in several databases, including the TCGA, GeneCard, Human Protein Atlas and Ualcan databases. The relationship between APE1 mRNA expression and several attributes of liver cancer patients in TCGA was investigated. Then, the protein expression of APE1 was detected by IHC analysis in 95 HCC samples and the association between APE1 expression and the clinicopathological parameters of HCC patients was explored. GSEA-KEGG analysis was performed to predict the potential signaling pathways that associated with APE1 expression. Then the siRNA-mediated knockdown model of APE1 was constructed in HCC cell line to further detect the detailed function of APE1 in HCC development in vitro and in vivo. The results of the bioinformatics analysis showed that APE1 expression was primarily located in the cell nucleus. APE1 mRNA expression was substantially correlated with pathological grade and T status in TCGA database. Elevated APE1 expression was observed in HCC samples and was associated with unfavorable survival time in liver cancer patients. IHC data demonstrated that the nuclear expression of APE1 in HCC tissues was significantly higher than that in noncancerous tissues. The expression level of the APE1 protein in HCC was strongly associated with tumor diameter and overall survival. Survival analysis indicated that APE1 nuclear expression is an independent prognostic marker for the overall survival of HCC patients. GSEA-KEGG results confirmed that APE1 associated with the base excision repair signaling pathway. The data of phenotypic experiments indicated that APE1 remarkably promoted tumor growth both in HCC cells and xenografts. The findings firstly imply that nuclear expression of APE1 is a valuable prognostic marker for HCC. APE1 significantly facilitate HCC development and targeting APE1 may be a promising strategy for HCC treatment.

Epigenetic age acceleration of early stage hepatocellular carcinoma tightly associated with hepatitis B virus load, immunoactivation, and improved survival.

Properly stratifying high-risk individuals with early stage hepatocellular carcinoma (HCC) is essential to identify patients in which the potentially therapies can be offered. To this context, we systematically investigated the prognostic value of epigenetic clock with early stage HCC as well as the association with other molecular characteristics.We computed DNA methylation (DNAm) age of 256 early stage HCC patients and 50 normal samples from TCGA by Horvath clock model. The characteristics of two DNAm age subgroups were differentiated regarding HBV expression, pathway activity, epigenomic, and genomic alteration. Cox regression and restricted cubic spline (RCS) analysis were utilized to evaluate the prognostic value of epigenetic acceleration.DNAm age was significantly associated with chronological age in normal tissue but largely disrupted in tumors (P< .001), and showed significant negative correlation with HBV expression (P< .05). We identified two DNAm age groups (DNAmAge-ACC and DNAmAge-DEC), and the former presented with an immunoactive phenotype (all FDRs<0.05 in enrichment analysis), CpG island hypermethylation (P< .001), and lower mutation burden (P= .018). Every 10-year increase in DNAm age was associated with a 18% decrease in fatality after adjustment for major clinical variables; DNAmAge-ACC had 50% lower mortality risk than DNAmAge-DEC (HR: 0.50, 95% CI: 0.27-0.94, P= .03). RCS revealed the fatality risk significantly decreased as epigenetic age accelerated (P = .04). Conclusions.In summary, we highlighted the prognostic value of epigenetic age acceleration for early stage HCC; better prognosis, relatively lower HBV load, and higher enrichment of immune signatures were tightly associated with epigenetic age accelerated tumors.

Knockdown of Ror2 suppresses TNF­α­induced inflammation and apoptosis in vascular endothelial cells.

Wnt family member 5a (Wnt5a) is a noncanonical member of the Wnt family that is highly expressed in atherosclerosis. Studies have shown that Wnt5a/receptor tyrosine kinase­like orphan receptors 2 (Ror2) signaling can participate in the formation of foam cells; however, the role of Ror2 in vascular endothelial cells during atherosclerotic injury is unknown. Therefore, the present study aimed to investigate the role of Ror2 in tumor necrosis factor (TNF)­α­induced vascular endothelial cell injury and investigate whether it could be regulated by Wnt5a. Human umbilical vein endothelial cells were transfected with short hairpin RNA specific against Ror2 in the absence or presence of TNF­α. The alteration of inflammatory cytokine levels was detected, and the expression of adhesion molecules was assessed. Western blot and flow cytometry analyses were used to detect the activation of nuclear factor­κB (NF­κB) signaling and cell apoptosis. The interaction between Ror2 and Wnt5a was confirmed by immunoprecipitation. Ror2 was upregulated upon TNF­α stimulation. Knockdown of Ror2 inhibited the TNF­α­induced release of inflammatory cytokines, the expression of intercellular adhesion molecule­1 and vascular cell adhesion molecule­1 and the activation of NF­κB signaling. Furthermore, cell apoptosis induced by TNF­α was rescued by Ror2 silencing. In addition, Wnt5a expression was increased by TNF­α, and Ror2 could bind to Wnt5a, the knockdown of which could downregulate the levels of Ror2. In conclusion, it was demonstrated that Ror2 was upregulated upon TNF­α stimuli, and interference of Ror2 regulated by Wnt5a could suppress TNF­α­induced inflammation and apoptosis in vascular endothelial cells.

Control of Contaminant Transport Caused by Open-Air Heavy Metal Slag in Zhehai, Southwest China.

Slag heaps are formed by mining waste materials, and the improper treatment of leachate from such heaps can threaten nearby aquifers. The Zhehai slag heap in Yunnan Province, China, contains 2.7 million tons of zinc and cadmium slag, and is considered a heavy metal source threatening the local groundwater safety, however, the severity of contamination remains unknown. In this study, numerical modeling was used to predict the groundwater flow and contaminant transport in this area based on field data. The results show that the atmospheric precipitation infiltration recharge at the top of the heap is 81.8 m³/d, accounting for 93.76% of total infiltration. The south and east sides of the area are the main outflow channels for contaminants, accounting for 93.25% of the total discharge around the heap. To reduce aquifer contamination, an in situ system involving a "controlling the source, 'breaking' the path, and intercepting the flow" (CSBPIF) strategy is established. The results indicate that the system performs well because it not only decreases the flow velocity but also reduces the concentrations of contaminants adsorbed by clay media. Moreover, the equivalent bottom liner thicknesses of the clay layers were calculated to improve the applicability of the CSBPIF system. Compared with ex situ disposal, this scheme provides an economic and effective solution and can be used to prevent and control groundwater pollution in China.

Nanowire Array-Coated Flexible Substrate to Accommodate Lithium Plating for Stable Lithium-Metal Anodes and Flexible Lithium-Organic Batteries.

Although lithium metal is an ideal anode with high theoretical capacity, Li dendrite formation and volume change have limited its application. We report a vertical polyaniline nanowire-coated carbon nanotube (CNT/PANI) composite flexible electrode on which Li could be homogeneously deposited to obtain a CNT/PANI@Li anode. In the composite, CNT/PANI acted as a host matrix with well-distributed Li ion flux attributed to high electroactive surface area, thereby effectively suppressing the Li dendrite. Compared with the pure CNT electrode, the CNT/PANI electrode presented low overpotential and stable long-term cycling with much less fluctuant stripping/plating profiles. The potential application of CNT/PANI@Li in all-flexible full cells was demonstrated by combining flexible organic poly(2,5-dihydroxyl-1,4-benzoquinonyl sulfide)/carbon nanotube (PDHBQS/CNT) composite films, in which the cathode achieves an eminent performance of 120 mA h g-1 at 50 mA g-1. Furthermore, pouch batteries with good flexibility were tested successfully, which demonstrated a promising future for all-flexible and high-performance Li-metal batteries.

A carbon foam-supported high sulfur loading composite as a self-supported cathode for flexible lithium-sulfur batteries.

A binder-free, self-supported, flexible cathode is explored for application in flexible lithium-sulfur (Li-S) batteries. The cathode is constructed using nitrogen (N)-doped carbon foam/carbon nanotubes (CNTs) as the scaffold and filled with poly(3,4-ethylenedioxythiophene) (PEDOT)-encapsulated sulfur nanoparticles as the active material. The dense CNTs coated on the skeleton of the 3D N-doped foam enhance flexibility, and the highly conductive CNTs are crossed and twined together to create an interconnected skeleton for rapid electron transport. The conductive PEDOT shell of sulfur nanoparticles and the N-doping of the carbon foams restrain the dissolution of polysulfides through the enhanced chemisorption of lithium polysulfides. The best cathode with a sulfur loading of 2.6 mg cm-2 has an eminent capacity of 1395 mA h g-1 during the initial cycle at 0.1 C. Furthermore, freestanding cathodes are assembled into flexible Li-S batteries, which demonstrate significant achievement at various bending angles. The capacity fading rate is 0.16% per cycle at 30° after 120 cycles. Its high sulfur loading, high capacitance, and good flexibility make this cathode material a promising candidate for potential application in flexible electronics.

Evaluation of HCPTd1,d14-double passaged intervening chemotherapy protocol for hepatocellular carcinoma.

AIM: To establish a kind of standardization of the clinical chemotherapeutic prototypes for unresectable hepatocellular carcinomas (HCC). METHODS: 10-Hydroxycamptothecin (HCPT) was applied through transcatheter arterial embolization (TAE) to HCC patients who were categorized into three groups: (1) test group: treatment with HCPT twice (HCPT d1 and 14) through TAE and portal venous embolization. (2) Control I: treatment with anticancer drugs without HCPT. (3) Control II: treatment with HCPT as a major component in anticancer drugs once (HCPT d1). A set of comparisons between test groups and control I and II groups were performed before and after the treatment to study the effectiveness of each treatment, in terms of tumor volumes, dynamic variations in serum alpha-fetoprotein (AFP), gamma-glutamyl transferase hepatoma-specific band (GGT-II), patient survival and adverse events. RESULTS: The general effectiveness rate of the test group reached 62.1% (72/116), remarkably higher than that of control I (32.1%, 40/124) and control II (54.7%, 47/56), (P<0.01 and P<0.05, respectively). Especially, the reduction rate or disappearance of the portal vein tumor emboli was as high as 88.4% (61/69) in the test group, in contrast with 13.9% (10/72) in control I and 35.9% (18/51) in control II (P<0.01 and P<0.01, respectively). After treatment, AFP decreased or turned to negative levels at 52.3% (34/65) in control I, 67.3% (35/52) in control II, and 96.8% (60/62) in the test group. Also GGT-II declined or became negative at 37.8% (28/74) in control I, 69.5% (57/82) in control II, and 94.7% (89/94) in test group (P<0.01 and P<0.05, respectively). CONCLUSION: We have designed a good protocol (test group) to treat HCC with excellent advantages of high efficiency, low cost, low toxicity and low adverse events and easy application. It could be recommended as one of the standardizations for HCC treatment in clinical practice.

Responsiveness of neoadjuvant chemotherapy before surgery predicts favorable prognosis for cervical cancer patients: a meta-analysis.

BACKGROUND: Neoadjuvant chemotherapy (NAC) before surgery has already shown the therapy effectiveness inpatients with cervical cancer. The present meta-analysis was conducted to determine whether the response to NAC predicts for prognosis. METHODS: Systematic computerized searches of the Pub-Med and Web of Knowledge were performed. Prognosis outcomes included progression-free survival (PFS), and overall survival (OS). The pooled odd ratio (OR) was estimated by using fixed-effect model or random-effect model according to heterogeneity between studies. RESULTS: Eighteen studies with 1,785 patients were included. Cisplatin-based NAC treatments were most commonly used. The clinical response rate ranged from 48.4 to 93.0 %, and the pathological response rate ranged from 27.6 to 30.6 %. The pooled ORs estimating the association of PFS with NAC response were 5.707 (95 % CI3.564­9.137), 6.798 (95 % CI 4.716­9.799), 6.327 (95 %CI 4.398­9.102), and 5.214 (95 % CI 3.748­7.253) at 1-,2-, 3-, and 5-year follow-up, respectively, and the pooled ORs estimating the association of OS with NAC response were 6.179 (95 % CI 3.390­11.264), 9.155 (95 % CI5.759­14.555), 8.431 (95 % CI 5.667­12.543), and 5.785(95 % CI 4.124­8.115) at 1-, 2-, 3-, and 5-year follow-up,respectively. No obvious statistical heterogeneity was detected. Funnel plots and Egger's tests did not reveal publication bias. Sensitivity analysis showed the results of meta-analysis were robust. CONCLUSION: This meta-analysis confirms that response to NAC is an indicator for PFS and OS, and suggests that patients-achieving response of NAC before surgery predicts favorable prognosis for cervical cancer patients.