Objective To compare the differences between exosomes derived from three-dimensional (3D) cultured human umbilical cord-derived mesenchymal stem cells (hUC-MSCs) and conventional (2D) cultured hUC-MSCs in promoting osteoblast differentiation, and to explore the application possibility of exosomes derived from 3D-cultured hUC-MSCs in treatment related to promoting bone formation. Methods hUC-MSCs were divided into 2D group and 3D group, and cultured respectively.The morphological characteristics of the both groups were observed under optical microscope, and the viability of 3D-cultured hUC-MSCs was detected by calcein AM/PI double staining of living and dead cells. Transcriptome sequencing was performed to screen the differentially expressed genes between 2D group and 3D group, and GO enrichment analysis was conducted. Exosomes of 2D group and 3D group were extracted and divided into 2D-Exo group and 3D-Exo group. Characterization of exosomes was conducted by transmission electron microscope, nanoparticle tracking analysis and Western blotting. The exosomes excreted by 2D group and 3D group were applied to C57BL/6J mouse calvarial osteoblasts, respectively, and osteogenic differentiation was induced in vitro. The effects of exosomes cultured in 2D and 3D on osteogenic differentiation were identified by alizarin red staining, alkaline phosphatase staining and RT-qPCR. Results Comparing to 2D group, cells in 3D group were equal in size and grew into spherical shape. 3D-cultured hUC-MSCs showed higher rate of viability verified by calcein AM/PI double staining of living and dead cells.Transcriptome sequencing results showed that the up-regulated genes in 3D group were enriched in bone mineralization, cartilage development, composition of extracellular matrix, osteoblast differentiation, angiogenesis, cell proliferation and gene expression.The down-regulated genes were enriched in negative regulation of cell proliferation, cell migration, and apoptotic process, etc.Transmission electron microscope observation showed that the exosome diameter in both 2D-Exo group and 3D-Exo group were around 100 nm and exhibited typical cup shape features, and expressed exosome related marker proteins CD9, CD63 and CD81.Staining results of osteoblasts in calvaria of newborn rats showed that the number of calcium nodules stained with alizarin red and the intensity of alkaline phosphatase staining increased significantly in 3D-Exo group than in 2D-Exo group. Results of RT-qPCR showed that the relative expression levels of osteogenic differentiation related genes Bglap, Runx2, Alp, Col1a1 and Spp1 mRNA increased significantly in 3D-Exo group than in 2D-Exo group with statistically significant differences (P<0.05). Conclusion Exosomes from 3D-cultured hUC-MSCs can up-regulate the expressions of osteogenic genes Bglap, Runx2, Alp, Col1a1 and Spp1,and have stronger capabilities to promote osteogenic differentiation compared with 2D-cultured exosomes.

Bullet vascular embolism is a serious complication of firearm injury, which is caused by military weapons in wartime and civil guns in peacetime. However, small caliber bullets and small mass fragments are mostly used in military weapons now, which makes it more likely that bullets/fragments will be left in the body, leading to a higher incidence of bullet vascular embolism. Therefore, the incidence of bullet vascular embolism in wartime in the future would be higher than it in the past. Bullet vascular embolism after firearm injury is more harmful to human body, and more emergency treatment is needed. This article reviews the mechanism, vulnerable vessels, embolization types, diagnostic methods and treatment points of bullet vascular embolism, so as to provide reference for clinical diagnosis and treatment of bullet vascular embolism.

Chronic hepatitis C virus (HCV) is a worldwide epidemic and the main cause of liver cirrhosis and hepatocellular carcinoma (HCC). The anti-HCV treatment has gone through two eras of pegylated interferon-α plus ribavirin (PR therapy) and direct-acting antiviral agent (DAA). Generally, achieving a sustained virological response (SVR) can reduce the incidence of HCC through antiviral treatment. In recent years, increasing researchers pay more attention to the issue whether DAA treatment might increase the risk of HCC occurrence or recurrence. This article aims to review the related studies on the risk of HCC after PR therapy and DAA treatment, summarize the risk factors, and explore the mechanism of HCC and its impact on the efficacy of DAA, in order to help clinicians to determine the timing of initiation of antiviral therapy and provide clinical evidence for individualized management.

Objective To explore the relativity of asymptomatic hyperuricemia(HUA) to lipoprotein associated phospholipase A2 (Lp-PLA2) in the middle-aged and elderly people. Methods From June 2019 to June 2020, 174 cases of epiphysically healthy middle-aged and elderly people were randomly screened from the Fuwai Hospital of the Chinese Academy of Medical Sciences for physical examination, and were divided into asymptomatic HUA group (n=58) and control group (n=116)according to the diagnostic criteria of HUA.The baseline clinical data of age, sex, body mass index (BMI) and laboratory data of blood routine, uric acid, creatinine, blood urea nitrogen, blood lipid, and Lp-PLA2 were retrospectively analyzed. Regression analysis was used to explore the risk factors of asymptomatic hyperuricemia. Results Among all the subjects, the incidence of HUA was significantly higher in women than in men (41.4% vs. 25.3%, P<0.05). The level of BMI, TG, LDL-C, Lp-PLA2 and the proportion of hypertension in asymptomatic HUA group was significantly higher than that in control group (P<0.05). Compared with the control group by sex, the incidence of HUA is higher in asymptomatic HUA group (P<0.05). The results of multivariate logistic regression analysis indicated that high level of Lp-PLA2 could be independent risk factors of high uric acid level. After divided into three quantile by the concentration of Lp-PLA2, compared with the lowest concentration, the OR value of the highest group increasing the risk of asymptomatic hyperuricemia was 4.61(95%CI 1.807-11.76, P<0.05). Conclusion Lipoprotein associated phospholipase A2 could be an independent risk factor of asymptomatic HUA in middle-aged and elderly adults.

Heart failure is the end stage of various cardiovascular diseases. Its pathological process is complex and regulated by many factors. With population aging intensifying, the prevalence rate of heart failure is increasing. MicroRNA (miRNA)participates in the development and growth of the body and plays an important role in the occurrence and development of heart failure. In addition, miRNA could be used as a marker and a target of treatment on heart failure. In this paper, CiteSpace was used to conduct bibliometric analysis on research on miRNA and heart failure, and keyword clustering and emergence was used to judge the research frontier in this field, that is, miRNA changes participate in the regulation of myocardial hypertrophy and myocardial fibrosis in the occurrence and development of heart failure. It is summarized research progress on traditional Chinese and Western medicine improving heart failure by regulating miRNA.

Objective To investigate the effect and its mechnism of ubiquitin ligase cullin3 (CUL3) on the proliferation, migration and invasion of gastric cancer cells. Methods The expression of CUL3 in gastric cancer and adjacent tissues and its relationship with clinical prognosis were analyzed based on TCGA database. HGC-27 and BGC-823 cells were transfected with CUL3 shRNA and empty lentivirus to construct a CUL3 knockdown gastric cancer cell line group (shCUL3 group) and an empty lentivirus transfection group (Vector group). CCK-8 assay, clone formation assay, EdU staining, scratch assay and Transwell assay were used to detect the changes of proliferation, migration and invasion ability of the two groups; Western blotting was used to detect the expression of epithelial-mesenchymal transition (EMT)-related proteins and PI3K/Akt/GSK3β pathway proteins in two groups;The changes of migration and invasion ability of shCUL3 group were detected after application of PI3K agonist 740 Y-P. Results In the TCGA database, CUL3 was highly expressed in gastric cancer, and was significantly associated with poor prognosis of gastric cancer patients. Compared with Vector group, CCK-8, clone formation assay and EdU staining showed that the proliferation of gastric cancer cells in shCUL3 group was inhibited (P<0.05), and CUL3 shRNA could significantly inhibit cell migration and invasion (P<0.05); In addition, compared with Vector group, the expression of E-cadherin protein in shCUL3 group increased, and the protein levels of vimentin, β-catenin, p-PI3K, p-Akt and p-GSK-3β significantly decreased (P<0.05); In contrast, application of the PI3K agonist 740 Y-P could partially reverse the inhibitory effect of CUL3 shRNA on gastric cancer cell migration and invasion. Conclusions Knockdown of CUL3 can inhibit the proliferation, migration and invasion of gastric cancer cells, and the mechanism of action of CUL3 in gastric cancer may be related to the activation of PI3K/Akt/GSK3β signaling pathway.

Objective To investigate the role of Toll-like receptor 4 (TLR4) in cardiac function injury of rats with heat stroke disease by regulating myocardial ferroptosis. Methods (1) 24 SD rats were randomly divided into control group and heat stroke disease (HS) group (12 in each group). During heat attack, anal temperature of rats was measured by anal thermometer, heart rate and mean arterial pressure of rats were measured by BL-420F biological function experiment system. (2) 24 SD rats were randomly divided into control group, ferroptosis inhibitor (Liproxstatin-1) group, HS group and HS+Liproxstatin-1 group (6 in each group). Thirty minutes before modeling, rats in Liproxstatin-1 group and HS+Liproxstatin-1 group were intraperitoneally injected with Liproxstatin-1 (10 mg/kg). The expression of solute carrier family 7 member 11 (SLC7A11) in myocardial tissue was detected by immunofluorescence staining, and the expression levels of SLC7A11 and glutathione peroxidase 4 (GPX4) were detected by Western blotting. (3) 48 SD rats were randomly divided into control group, TLR4 inhibitor (TAK-242) group, HS group and HS+TAK-242 group (12 in each group). Thirty minutes before modeling, rats in TAK-242 group and HS+TAK-242 group were intraperitoneally injected with TAK-242 (3 mg/kg). The expression of TLR4 in myocardial tissue was detected by immunofluorescence staining, cardiac hemodynamic indexes [left ventricular systolic pressure (LVSP), left ventricular end-diastolic pressure (LVEDP), maximum rate of increase (+dp/dt_max) and maximum rate of decrease (–dp/dt_max) of left ventricular pressure] were detected by BL-420F biological function experiment system, cardiac function indexes [cardiac output (CO), diastolic left ventricular posterior wall thickness (LVPWd) and systolic left ventricular posterior wall thickness (LVPWs)] were detected by cardiac ultrasound, the expression levels of TLR4/NF-κB-p53 signaling pathway protein [TLR4, nuclear factor kappa B (NF-κB), p53] and ferroptosis related protein (SLC7A11 and GPX4) were detected by Western blotting. Results Compared with control group, the core body temperature and heart rate of rats in HS group increased significantly after heat shock for 180 minutes (P<0.01), the mean arterial pressure decreased significantly (P<0.05). Immunofluorescence staining results showed that compared with control group, the expression of myocardial SLC7A11 in HS group significantly decreased (P<0.01); Compared with HS group, the expression of SLC7A11 in HS+TAK-242 group improved significantly (P<0.01). The results of Western blotting showed that compared with control group, the expression levels of SLC7A11 and GPX4 in myocardium of HS group decreased significantly (P<0.05); Compared with HS group, the expression levels of SLC7A11 and GPX4 in HS+Liproxstatin-1 group improved significantly (P<0.05). Immunofluorescence staining results showed that compared with control group, the expression level of myocardial TLR4 in HS group increased significantly (P<0.01); Compared with HS group, the expression level of TLR4 in HS+TAK-242 group improved significantly (P<0.01). The results of BL-420F biological function test system showed that compared with control group, LVEDP increased, LVSP decreased (P<0.01) in HS group. LVSP, LVEDP, +dp/dt_max and –dp/dt_max were improved in HS+TAK-242 group (P<0.05). Compared with control group, CO was significantly decreased, and LVPWd and LVPWs were increased in HS group (P<0.01). Compared with HS group, CO increased, and LVPWd and LVPWs decreased in HS+TAK-242 group (P<0.05). Western blotting results showed that compared with control group, the expression levels of TLR4, NF-κB and p53 in the HS group increased significantly (P<0.01), and of SLC7A11 and GPX4 decreased significantly (P<0.01); Compared with HS group, the expression levels of TLR4, NF-κB and p53 decreased significantly, and of SLC7A11 and GPX4 increased significantly (P<0.05) in HS+TAK-242 group. Conclusion Ferroptosis may exist in myocardial injury caused by heat stroke. Inhibition of TLR4 can improve cardiac function of rats with heatstroke, and the mechanism may be related to ferroptosis mediated by TLR4/NF-κB-p53/SLC7A11 signaling pathway.

Liver transplantation is currently an effective treatment for end-stage liver disease. Postoperative infection is the main important cause of death in patients after liver transplantation. Due to the lack of specificity of clinical symptoms of postoperative infections, further insight is needed into the incidence, risk factors, diagnosis and treatment of different infections, including bacterial, fungal and viral. In addition, it could reduce the perioperative mortality that understanding the characteristics of changes in immune function after liver transplantation and timely prevention and treatment of post-transplant complications of infection, which would ultimately improve the outcome of patients. Nowadays, the use of immune-supporting drugs such as thymosin α₁, intravenous immunoglobulin and ulinastatin have also shown some efficacy in patients with post-transplant infections.This article reviews the characteristics of infection, the changes of immune function and the feasibility of immunosupportive therapy after liver transplantation.

Chronic obstructive pulmonary disease (COPD) is one of the most common comorbidities in patients with atrial fibrillation, and the both diseases share a series of common risk factors. COPD promotes the occurrence and development of atrial fibrillation by variety of pathophysiological mechanisms, is the important factor affecting the prognosis of patient with atrial fibrillation, and results in an elevation of clinical adverse and cardiovascular death events. Atrial fibrillation also affects the treatment strategy and prognosis of COPD patients. Treatment plan for patients with both atrial fibrillation and COPD faced huge challenging.The research findings of home and abroad related to atrial fibrillation complicated with COPD were summarized in present paper from the aspects of epidemiology, pathophysiology mechanism, patient management, outcome and prognosis, in order to provide help for the integrated multidisciplinary management and individualized treatment for these patients.

Objective To investigate the targeting relationship of miR-149-5p and fibroblast growth factor 21 (FGF21), and the effect of miR-149-5p on hypoxia/reoxygenation (H/R) injury of rat`s cardiomyocytes. Methods Ten male C56BL/6J mice were used to construct the cardiac ischemia-reperfusion (I/R) models, and qRT-PCR was performed to detect the changes of miR-149-5p levels in the heart tissues of I/R mice. H9c2 cells cultured to logarithmic phase were divided into control group, H/R group, H/R+miR-149-5p inhibitor group, H/R+miR-149-5p NC group, H/R+miR-149-5p mimics group and H/R+miR-149-5p mimics+FGF21 group, and induced H/R model and transfection treatment. The activities of lactate dehydrogenase (LDH) and creatine kinase (CK) in H9c2 cardiomyocytes were detected by ELISA; Cell viability was detected by MTT assay; apoptosis was detected by flow cytometry; the level of miR-149-5p was detected by qRT-PCR; the expression levels of Bcl-2, Bax, cleaved caspase-3 and FGF21 proteins were detected by Western blotting; the targeting relationship between miR-149-5p and FGF21 were determined by dual luciferase assay. Results Compared with the sham operation group, the level of miR-149-5p in the heart tissue of I/R mice was significantly increased (P<0.05). Compared with those in control group, the apoptosis rate, LDH and CK activities and Bax, cleaved caspase-3 protein expression levels of H9c2 cells in H/R group were significantly increased (P<0.05), cell viability and FGF21, Bcl-2 protein expression levels decreased significantly (P<0.05). Compared with those in H/R group and H/R+miR-149-5p NC group, the apoptosis rate, LDH and CK activities and Bax, cleaved caspase-3 protein expression levels of H9c2 cells in H/R+miR-149-5p inhibitor group were significantly decreased (P<0.05), cell viability and FGF21, Bcl-2 protein expression levels were significantly increased (P<0.05). Compared with those in H/R group, the apoptosis rate, LDH and CK activities and Bax, cleaved caspase-3 protein expression levels of H9c2 cells in H/R+miR-149-5p mimics group were significantly increased (P<0.05), cell viability and FGF21, Bcl-2 protein expression levels decreased significantly (P<0.05). Compared with those in H/R+miR-149-5p mimics group, the apoptosis rate, and LDH and CK activities of H9c2 cells in miR-149-5p mimics+FGF21 group were significantly decreased (P<0.05), cell viability was significantly increased (P<0.05). The result of dual luciferase assay indicated the targeted regulatory relationship between miR-149-5p and FGF21. Conclusions MiR-149-5p is up-regulated significantly in the myocardial tissue of I/R mice, and negatively regulate the level of FGF21, leads to decreased cell viability and increased apoptosis of H/R-treated rat's cardiomyocytes.