Objective:To discuss the effect of knock-out MG53 on skeletal muscle,and to determine whether MG53 can protect skeletal muscle from injury during delayed onset muscle damage(DOMS).Method:Eighty-four eight-week old C57 mice were divided into two groups due to their gene (WT and KO),and each group were divided into 7 subgroups again.14 groups,6 mice in each.They were WC,WE0,WE2,WE12,WE24,WE48,WE72 and KC,KE0,KE2,KE12,KE24,KE48,KE72.At each time point after 3 days eccentric exercise,the mice were decapitated after drawing blood.The rectus femoris of left leg was used to detect the level of MG53 with Western blot,the other side were used to do electron microscope analysis.Blood CK were measured also.Result:The CK of K0 raised remarkably and decreased gradually until W 12.But the second peak appear at W24 and decreased again gradually since then.Ultrastructural changes of skeletal muscle after eccentric exercise at different time points is obvious.The injury is worsen gradually from Wo to W24,and improved gradually from W48 to W72.Comparing with W groups,the injury in K group were even worsen an each time point.The MG53 of W0 and W24 were 16％ and 11％ higher than those before exercise (p＜0.05)and restored to normal in 72 hours after exercise.Conclusion:The injury after severe exercise is more severity in MG53 knock-out mouse than those in wild mouse.and the MG53 protein increased significantly.It suggests that MG53 may significantly relieve the skeleton muscle injury in delayed muscle sore period.

Abstract: Objective To investigate the changes in expression of Toll-like receptor 3 (TLR3) and interferon-α (IFN-α) in patients with different clinical outcomes of hepatitis C virus (HCV) infection treated with direct-acting antiviral agents (DAAs), and to explore the relationship between the expression of TLR3 and IFN-α with the clinical outcomes of HCV infection. Methods A total of 149 HCV infected patients who received initial treatment were selected from Hainan General Hospital between September 2020 and August 2022. The patients were divided into two groups: chronic hepatitis C (CHC) group (n=129) and liver cirrhosis (LC) group (n=20). Additionally, 28 volunteers were selected as the control group during the same period. All patients with HCV infection were first treated with Sofosbuvir/Vipatavir tablets for 12 weeks. Blood samples were collected at 0, 4, 12, 24 and 48 weeks after treatment. Liver function indicators were detected by enzyme-linked immunosorbent assay (ELISA), while TLR3 mRNA were detected by real-time fluorescence quantitative polymerase chain reaction (qRCR), IFN-α was detected by Luminex multiplex cytokine assays. Measurement data subject to normal distribution were represented by x±s, and t test was used between groups. Compare differences between groups. Results TLR3 mRNA in CHC group was higher than that in LC group and control group at baseline (P<0.05). After 4 weeks of DAAs treatment, TLR3 mRNA in CHC and LC groups was significantly up-regulated (P<0.05). TLR3 mRNA in the CHC group was gradually down-regulated to the level of the control group at 12, 24, and 48 weeks. In addition, IFN-α expression gradually increased with prolonged treatment, while it decreased in the LC group. The liver inflammation indicators in both the CHC and LC groups partially recovered after treatment with DAAs. Conclusions TLR3 is involved in viral clearance and chronic inflammatory response. The expression difference of TLR3 in patients with different clinical outcomes of HCV infection after DAAs treatment may be related to the severity of the disease.

This study investigated the potential role of ERK1/2-cyclinE1 signaling pathway in rat pulmonary artery smooth muscle cells (rPASMCs) proliferation and pulmonary vascular remodeling induced by cigarette smoke exposure. A total of 24 male Wistar rats were randomly divided into 4 groups: control group (C group), S-1M, S-3M and S-6M groups (animals in the groups were exposed to smoke for 1, 3, and 6 months, respectively). HE staining and anti-α-smooth muscle actin antibody staining were performed to observe the degree of pulmonary vascular remodeling. Immunohistochemistry and Western blotting were performed to evaluate ERK1/2 and cyclinE1 expression in pulmonary vessels. Primary cultured rat pulmonary artery smooth muscle cells (rPASMCs) were exposed to cigarette smoke extract (CSE). ERK inhibitor (PD98059) and cyclinE1 siRNA were used to verify the role of ERK1/2 and cyclinE1 in CSE-induced rPASMCs proliferation. Cell proliferation was assessed by cell counting and 5-bromo-2-deoxyuridine (BrdU) incorporation. Our results showed that abnormal pulmonary vascular remodeling was found in cigarette smoked rats. Compared to C group, activated ERK1/2 and cyclinE1 expression was significantly increased in smoke-exposure groups. This up-regulated expression was positively correlated with the severity of pulmonary vascular remodeling, and there was positive correlation between the expression of ERK1/2 and cyclinE1. PD98059 and cyclinE1 siRNA inhibited the proliferation of rPASMCs. The expression of cyclinE1 could be down-regulated by PD98059. Our data demonstrated that increased expression of ERK1/2 and cyclinE1 might be involved in the pathogenesis of abnormal rPASMCs proliferation and rat pulmonary vascular remodelling induced by cigarette smoke exposure.
Animals ; Cells, Cultured ; Cyclins ; metabolism ; MAP Kinase Signaling System ; Male ; Myocytes, Smooth Muscle ; metabolism ; pathology ; Pulmonary Artery ; metabolism ; pathology ; Rats ; Rats, Wistar ; Smoking ; metabolism ; pathology ; Up-Regulation

Objective:To investigate the influence of cognitive-behavior therapy on the psychological status of family members of terminal cancer patients. Method:A total of 60 families of terminal cancer patients were selected and randomly divided into observation group (30 cases) and control group (30 cases). The observation group was treated with cognitive-behavior therapy, while the control group was given general supportive psychological care. The Hamilton Depression Scale ( HAMD) , Hamilton Anxiety Scale ( HAMA) and Pittsburgh Sleep Quality Index ( PSQL) were used to evaluate the family members of the two groups of patients before and after the intervention. Results: Before the intervention, there was no statistical significance difference in the scores of HAMA, HAMD and PSQI between the two groups (P>0. 05). After the intervention, the scores of HAMA, HAMD and PSQI in the observation group were significantly lower than those before the intervention ( P <0 . 05 ); and the scores of HAMA, HAMD and PSQI in the observation group were significantly lower than those in the control group ( P<0 . 05 ) . Conclusion:Cognitive-behavior therapy can significantly improve the negative emotions of depression, anxiety and sleep disorder among family members of terminal tumor patients.

Objective: Remuzzi scoring system is utilized for assessing the degree of renal tissue damage in donors with hypertensive cerebral hemorrhage and donors with brain trauma after cardiac death. To explore the prognosis of hypertensive cerebral hemorrhage donor kidney in renal transplant recipients. Methods: The kidney donated by DCD between January 1, 2016 to June 1, 2018 were retrospectively reviewed. Pathological biopsy was performed before transplantation and hematoxylin-eosin (HE) staining after sectioning. The degree of renal tissue lesions was evaluated by Remuzzi scoring system. According to the source of donor kidney, they were divided into two groups of donors with heart failure due to hypertensive cerebral hemorrhage (HCH) and those with brain trauma (BT). Both groups of donor kidneys were preserved by low-temperature machine perfusion. The immunosuppressive regimen was identical in both groups. The prognosis of two groups was compared by serum creatinine (Scr) at Month 1/6/12 post-operation and cumulative graft survival rate over a follow-up period of 12-36 months. Results: The renal Remuzzi score of HCH donors was significantly higher than that of BT donors. The maximal creatinine clearance rate was significantly lower than that of BT donors [(86.8±27.8) vs (115.4±23.2) ml/min, P<0.05]. At 1/6/12 months post-transplantation, serum creatinine levels were (76.1±18.5), (72.4±16.2) and (71.4±16.8) μmol/L in BT group and (160.3±33.4), (154.3±32.6) and (146.4±29.1) μmol/L in HCH group. The SCr in BT group at 1/6/12 months was lower than that in HCH group (P<0.05). Kaplan-meier analysis showed no significant inter-group difference in graft survival between two groups over a follow-up period of 12 to 36 months (Log-Rank test, P=0.485). Conclusions No significant difference exists in short-term survival rate of kidneys from HCH and BT donors. The recipients of HCH donor's kidney have higher serum creatinine levels than those of BT donors. Selective use of kidney transplants in patients with cardiac death caused by HCH may greatly reduce the waste of donor kidney and improve the quality-of-life of patients with end-stage renal disease.

BACKGROUND: Standard two-dimensional (2D) culture has confirmed the mechanism of mast cells (MCs) in the pathogenesis of inflammatory bowel disease (IBD), but the regulation of signaling responses of MCs may well differ in three-dimensional (3D) microenvironments. The aim of the study was to develop a 3D culture model based on decellularized intestinal scaffolds (DIS) and verify how MCs influenced fibroblasts phenotype in the 3D model. METHODS: DIS were achieved using the detergent technique and extracellular matrix (ECM) components were verified by histologic analysis, quantification and scanning electron microscope. After human colon fibroblasts recellularized into the scaffolds and activated by MCs tryptase and TGFb1, the changes in genes and signaling pathways during fibroblasts activation in 3D were studied and compared with the changes in 2D cell culture on plastic plates. RESULTS: Decellularization process effectively removed native cell debris while retaining natural ECM components and structure. The engrafted fibroblasts could penetrate into the scaffolds and maintain its phenotype. No matter whether fibroblasts were cultured in 2D or 3D, MCs tryptase and transforming growth factor b1 (TGF-b1) could promote the differentiation of fibroblasts into fibrotic-phenotype myofibroblasts through Akt and Smad2/3 signaling pathways. Furthermore, the pro-collagen1a1 and fibronectin synthesis of myofibroblasts in 3D was higher than in 2D culture. CONCLUSION Our results demonstrated that the DIS can be used as a bioactive microenvironment for the study of intestinal fibrosis, providing an innovative platform for future intestinal disease modeling and screening of genes and signaling pathways.