Exosome-mediated miR-106a-3p derived from ox-LDL exposed macrophages accelerated cell proliferation and repressed cell apoptosis of human vascular smooth muscle cells.

OBJECTIVE: Atherosclerosis (AS) is a leading disease with high mortality and morbidity in the world. It has been demonstrated that exosomes can transfer some miRNAs or proteins to regulate the biological functions of human vascular smooth muscle cells (VSMCs) and promote the progression of AS. In this study, we mainly aimed at exploring potential functions of exosomes derived from ox-LDL exposed macrophages and investigating the potential mechanisms of exosome-mediated miR-106a-3p in regulating VSMCs and promoting AS. MATERIALS AND METHODS: Ox-LDL was used to treat THP-1 macrophages, CCK-8 assay was performed to detect cell viability, and flow cytometric analysis was used to detect cell apoptosis. Exosomes were isolated and collected with centrifugation, and were determined by transmission electron microscopy and WB assay. RT-PCR was used to detect the expressions of miRNAs in exosomes and VSMCs, WB assay was used to detect protein expressions. MiR-106a-3p mimic was transfected into VSMCs to verify its functions and the Luciferase gene reporter assay was performed to prove the binding site of miR-106a-3p and CASP9. Finally, GW4869, an inhibitor for exosome secretion, was used to block exosome secretion by ox-LDL induced THP-1 and to confirm the effects of miR-106a-3p on cell proliferation and apoptosis in VSMCs. RESULTS: We found that ox-LDL induced THP-1 could promote cell proliferation and repress cell apoptosis of VSMCs, then, exosomes were successfully isolated, which could promote cell proliferation and repressed cell apoptosis of VSMCs after adding into VSMCs. Furthermore, we found that miR-106a-3p was significantly increased in exosomes from ox-LDL induced THP-1 and its expression was also increased in VSMCs after adding into VSMCs. Moreover, miR-106a-3p overexpression could promote cell viability and repress cell apoptosis, as well as regulate associated protein expressions. Additionally, the Luciferase gene reporter assay confirmed that miR-106a-3p could directly bind with CASP9 and regulate Caspase signaling in VSMCs. Finally, blocking exosomes from ox-LDL induced THP-1 reduced the cell viability and promoted cell apoptosis in VSMCs. CONCLUSIONS: Above all, this study demonstrated that miR-106a-3p was increased in exosomes from ox-LDL induced THP-1 and it could promote cell proliferation and repress cell apoptosis of VSMCs. We found that the exosomes-mediated miR-106a-3p could directly bind with CASP9 and repress Caspase signaling pathway in VSMCs, which might provide a potential target for treating AS.

RUNX3 protects against acute lung injury by inhibiting the JAK2/STAT3 pathway in rats with severe acute pancreatitis.

OBJECTIVE: Acute lung injury (ALI) is the most common complication of severe acute pancreatitis (SAP) in the early stage, which causes systemic inflammatory response and organ damage. Human runt-associated transcription factor 3 gene (RUNX3) has been reported to participate in various inflammatory diseases. However, the exact role of RUNX3 in SAP and its-related ALI remains unclear. MATERIALS AND METHODS: To establish the model of SAP, rats were retrogradely injected with 5% sodium taurocholate (1 mg/kg body weight) into the biliary-pancreatic duct. Cytokine level in serum was measured by ELISA, and the polymorphonuclear neutrophil (PMN) was isolated from rat's blood 12 h-post SAP induction. RESULTS: We found RUNX3 expression was significantly decreased with the progression of SAP. Both pancreas damages and cytokine production abilities were reduced in RUXN3-overexpressed SAP rats compared with control rats. Moreover, SAP-associated ALI was also improved upon RUNX3 overexpression in SAP rats. RUNX3 upregulation enhanced PMN apoptosis and inhibited Janus kinase 2/signal transducer and activator of transcription 3 (JAK2/STAT3) phosphorylation. CONCLUSIONS: Our study indicates that RUNX3 protects against SAP and SAP-associated ALI through controlling PMN apoptosis and regulating JAK2/STAT3 signaling pathway. RUNX3 could be regarded as a potent therapeutic target in SAP for future studies.

[Clinical study of FibroTouch and six serological models for assessing the degree of liver fibrosis in patients with chronic hepatitis B].

Objective: To evaluate the using value of FibroTouch and six serological models in detecting the degree of liver fibrosis in patients with chronic hepatitis B, in an attempt to provide reference for accurate diagnosis. Methods: Two hundred and fifty-eight cases with chronic hepatitis B admitted to Xixi Hospital of Hangzhou from September 1, 2015 to September 1, 2017 were selected. All patients underwent liver histopathological examination and FibroTouch measurement to determine liver stiffness (LSM). Serum biochemical parameters were detected and the scoring values of six serological models were calculated. SAS 9.4 statistical software was used for statistical analysis, and the correlation between FibroTouch and the six serological models was analyzed by Spearman correlation. The diagnostic value of FibroTouch and six serological models was analyzed by receiver operating characteristic curve (ROC) based on liver histopathological findings. Results: The median LSM of 258 cases with chronic hepatitis B was 9.4 (6.5-13.8) kPa. In the six serological models, the median value of aspartate transaminase to platelet ratio index (APRI), FIB-4 index, S-index, Forn's index, PRPindex, and FIB-5 were 0.42 (0.28-0.62), 1.27 (0.78-2.03), 0.11 (0.07-0.20), 6.95 (5.89-8.51), 0.000 8 (0.000 6-0.000 9),and 38.59 (36.28-40.97). FibroTouch had positive correlation with APRI, FIB-4, S-index, Forn's index, PRP, fibrosis stage (r= 0.73,P< 0.001) and inflammation grade, and had negative correlation with FIB-5, and both had statistical significance. The area under curve (AUC) of FT-LSM at S≥2, S≥3, S = 4 were 0.89, 0.90 and 0.85, respectively, which was significantly higher than serological models (P< 0.001). The AUC of S-index model at S≥2, S≥3, S = 4 were higher than other five serological models. Conclusion: The diagnostic performance of FibroTouch is significantly better than serological model. S-index model has the best diagnostic performance in the six serological models, and the combination of S-index and FT-LSM may better diagnose the grading of liver fibrosis, and thus can be applied and promoted in clinic.

Influence of aggregate pre-wetting and fly ash on mechanical properties of lightweight concrete.

This study has examined the mechanical properties of lightweight aggregate concrete with a density of 1800 kg/m3. The effects of the following parameters on the concrete properties have been analyzed: the pre-wetting time of the lightweight aggregate and the percentage of pulverized fly ash used as cementitious replacement material. The strength of the lightweight aggregate was found to be the primary factor controlling the strength of high-strength lightweight concrete. An increase in the cementitious content from 420 to 450 kg/m3 does not significantly increase the strength of lightweight aggregate concrete. The relationship between the flexural and compressive strength at 28 days can be represented by the equation fr=0.69/fck. The elastic modulus was found to be much lower than that of normal weight concrete, ranging from 15.0 to 20.3 GPa. The addition of PFA increases the slump and density of lightweight aggregate concrete.