BACKGROUND:Cel co-culture can maximize the simulation ofin vivomicroenvironment. Cel scratch test and interleukin-1β can destroy the balance between matrix metaloproteinases (MMPs) and matrix metaloproteinase inhibitors (TIMPs), resulting in extracelular matrix degradation of the articular cartilage, functional disorders of chondrocytes and articular cartilage degeneration. OBJECTIVE:To study the effect of interleukin-1β on migration, MMP and TIMP expression of chondrocytes co-cultured with osteoblast supernatantin vitro. METHODS:There were three groups: chondrocyte monoculture group, osteoblast+chondrocyte group (co-culture group), osteoblast+chondrocyte+interleukin-1β group (interleukin-1β group). Cel scratch test was conducted to observe the migration of chondrocytes within 24 hours. Semi-quantitative PCR test was used to detect the changes in expressions of MMP-1, MMP-2, MMP-3, MMP-9, TIMP-1, TIMP-2, TIMP-3, TIMP-9 in chondrocytes within 24 hours. RESULTS AND CONCLUSION:Compared with the monoculture group, cel migration rate of the other two groups were increased significantly (P< 0.01). Compared with the monoculture group, the gene expressions of MMP-1, MMP-2, MMP-3 and MMP-9 were increased significantly in the coculture group (P < 0. 05); the gene expressions of MMP-1, MMP-3, MMP-9 were increased significantly in the interleukin-1β group (P< 0. 01). Compared with monoculture group, the gene expression of TIMP-1 was increased significantly (P < 0. 01), but the gene expressions of TIMP-3 and TIMP-4 were declined significantly (P < 0. 05) in the other two groups. These findings indicate that co-culture of chondrocytes with osteoblasts can promote chondrocytes migration, enhance gene expression of chondrocytes MMP-1, MMP-2, MMP-3, MMP-9 and regulate gene expression of TIMPs family. Interleukin-1β inhibitsthe migration of chondrocytes co-cultured with osteoblasts and gene expression of TIMPs family.

Objective To investigate the safety and therapeutic effects ot a novel multi-layer flat-plate bioartificial liver (BAL) for patients with liver failure.Methods Thirty-eight patients with liver failure from Dec.2010 to Dec.2011 were treated with a novel BAL based on multi-layer flatplate bioreactor and the co-cultured cells of the porcine hepatocytes and mesenchymal stem cells.A total of 48 treatments was performed,4 h each time.The clinical signs and symptoms,liver function,ammonia,coagulation function and complete blood count were evaluated before,during and after the treatment.DNA in the collected PBMCs was extracted for PCR with PERV specific primers and the porcine specific primer Sus scrofa cytochrome B.The RT activity was detected as well.Levels of xenoantibodies (IgG,IgM) were determined by using ELISA kit. Titers of complement were quantified by CH50 kit.Results All treatment procedures were completed successfully without any adverse reaction. All samples presented negative PERV DNA and RT activity. The levels of antibodies were similar before and after treatment.Treatment was associated with a temporary decline in levels of complement,and then the levels were recovered quickly.The clinical symptoms such as acratia,anorexia and abdominal distension were improved.The stage of hepatic encephalopathy in 16 patients was decreased. The liver function and ammonia was reduced disproportionately. Seven patients in all were bridged to liver transplantation,2 patients died and 2 patients gave up the treatment,and the others were turned better.After the outcome judgment according to the standard developed by the Artificial Liver Group,and Chinese Association of Infectious and Parasitic Diseases,there were 9 patients with clinical healing,25 patients with improvement and 4 patients with no effect,and the cure-improvement rate was 89.5％.Conclusion The novel multi-layer flat-plate BAL could be used as a safe and effective therapy for patients with liver failure.

Objective To investigate the therapeutic effect of bone marrow mesenchymal stromal cell (BMSC) transplantation on D-galactosamine-induced acute liver failure in Sprague-Dawley (SD) rats,as well as the mechanism of neutrophils in this process.Methods A total of 39 male SD rats were divided into control group (8 rats,intraperitoneal injection of isotonic saline),model group (10 rats,intraperitoneal injection of D-galactosamine),solvent group (9 rats,tail vein injection of isotonic saline at 2 hours after intraperitoneal injection of D-galactosamine),and treatment group (12 rats,tail vein injection of MSCs at 2 hours after intraperitoneal injection of D-galactosamine).The rats were sacrificed at 24 hours after the model of D-galactosamine-induced acute liver failure was established,and the blood and liver tissue were harvested.The serum levels of alanine aminotransferase (ALT) and aspartate aminotransferase (AST) and total bilirubin (TBil) were measured,and blood analysis was performed to measure the number and percentage of neutrophils in peripheral blood.Immunofluorescence assay was used to measure the expression of the neutrophil marker Ly6g in the liver,the myeloperoxidase (MPO) kit was used to measure the activity of MPO in liver,and RT-PCR was used to measure the mRNA expression of inflammatory cytokines and chemokines in the liver,i.e.,tumor necrosis factor-α (TNF-α),interleukin-1β (IL-1β),interferon-γ (IFN-γ),interleukin-10 (IL-10),CXC chemokine ligand 1 (CXCL1),and CXC chemokine ligand 2 (CXCL2).Another 64 male SD rats were randomly divided into groups,and the survival rates of rats in each group were observed for 7 days.The independent samples t-test was used for comparison between any two groups (Levene homogeneity test of variance,and the corrected t-test was used for a P value of ＜ 0.05),and the log-rank test was used for comparison of survival rates between any two groups.Results At 24 hours after acute liver failure was induced by D-galactosamine in the SD rats,there were significant increases in the liver function parameters (ALT:2884.1±541.0 U/L vs 45.4±11.0 U/L,P ＜ 0.001;AST:3634.9±755.9 U/L vs 143.9±23.7 U/L,P ＜ 0.001;TBil:44.4±8.4 μmmol/L vs 0.9±0.2 μmmol/L,P ＜ 0.001) and the number and percentage of peripheral blood neutrophils [number:(4.7±1.1)×109 vs (1.4±0.4)× 109,P ＜0.001;percentage:44.9％±8.0％ vs 18.3％±4.4％,P ＜ 0.001].A large number of neutrophils aggregated in the liver tissue,and there were significant increases in the MPO activity (4.72±1.09 U/g vs 1.13±0.24 U/g,P ＜ 0.001),inflammatory cytokines,and chemokines.Compared with the model group,the treatment group showed significant improvements in liver function (ALT:1 823.9a389.2 U/L vs 2 884.1±541.0 U/L,P ＜ 0.001;AST:2173.0±567.3U/L vs 3634.9±755.9 U/L,P ＜ 0.001;TBil:30.9±6.5 μmmol/L vs 44.4±8.4 μmmol/L,P ＜ 0.001) and survival rate (50％ vs 12.5％,P=0.023).Meanwhile,the treatment group also showed significant reductions in the number and percentage of peripheral blood neutrophils [number:(3.5±1.0)× 109 vs (4.7±1.1)×109,P =0.012;percentage:35.9％±8.9％ vs 44.9％±8.0％,P =0.021],number of neutrophils in the liver,and MPO activity (3.52±1.03 U/g vs 4.72±1.09 U/g,P =0.040),as well as significantly inhibited expression of inflammatory cytokines and chemokines (TNF-α:2.458±0.762 vs 3.778±1.046,P =0.005;IL-1β:2.498±0.547 vs 4.065 ± 0.953,P =0.002;IFN-γ:3.977±1.039 vs 5.418±1.255,P =0.025;IL-10:6.056±1.542 vs 3.368±0.952,P=0.001;CXCL1:7.988±1.911 vs 10.366±1.239,P =0.010;CXCL2:3.441±1.005 vs 4.847±1.113,P=0.019).Conclusion BMSC transplantation has a therapeutic effect on D-galactosamine-induced acute liver failure in rats,and this process is accompanied by reduced aggregation and activity of neutrophils in peripheral blood and liver.Inflammatory cytokines and chemokines may be involved in the mechanism of regulation of these two aspects.

Objective:To analyze the current status of salary system reform in public hospitals in Mianyang City, Sichuan Province, and provide reference for the deepening of salary system reform in public hospitals.Methods:From April to May 2023, a self-designed survey questionnaire was used to conduct a survey of medical staff in five municipal tertiary public hospitals through the Wenjuanxing platform. The survey questionnaire mainly included the basic information of the survey subjects, the implementation status of the salary system, and employee satisfaction with salary. In May 2023, semi-structured interviews were conducted with relevant personnel from government departments and public hospitals involved in the reform. The interview content included the reform measures taken and their impact, changes in medical staff salaries after the reform, and the current difficulties faced by the reform. Descriptive analysis was conducted on the data, and Chi-square test was used to compare the count data. Kendall′s tau-b test was used to analyze the correlation between salary level and basic information of the survey subjects.Results:In the reform of the salary system in public hospitals in Mianyang City, measures such as system construction, total salary quota determination and dynamic control, and salary management for non-establishment employees have been mainly adopted. 826 valid questionnaires were collected, with an effective response rate of 94.5%. The annual income level was positively correlated with age, education level, years of work experience, and professional title level ( r=0.420, P<0.001; r=0.245, P<0.001; r=0.364, P<0.001; r=0.504, P<0.001). Compared with the personnel in the same positions at the same level of hospitals in other cities and states in Sichuan Province, 274 people (33.2%) were quite satisfied or very satisfied with the current salary level; 350 people (42.3%) were quite satisfied or very satisfied with the salary distribution mechanism, and 441 people (53.4%) were quite satisfied or very satisfied with non-economic salaries. The proportion of survey respondents who believe that the existing salary structure and performance evaluation methods could reflect their actual work value was 464 people (56.2%) and 423 people (51.2%), respectively. A total of 10 people were interviewed, with 4 individuals believing that the salary level has increased after the reform, but the change was not significant compared to before the reform; 8 individuals believed that the current reform does not involve the reform of the salary structure; 6 individuals believed that the internal performance based salary distribution mechanism in hospitals was not sound; 3 individuals believed that there was insufficient attention to the reform of non-economic compensation; 7 individuals believe that the difficulties in the salary system reform were mainly concentrated on the unified understanding of policies, how to determine the increase in the total salary quota, and how to reasonably distribute it internally. Conclusions:Mianyang City has scientifically determined the total salary and implemented dynamic control, enhancing the public welfare nature of public hospitals, while also taking into account the salary management of employees both inside and outside the establishment, promoting equal pay for equal work. However, there were also issues such as disagreements on the dynamic adjustment of the total salary, an imperfect internal distribution mechanism within hospitals, and an incomplete salary system reform. It is necessary to further strengthen top-level design, reasonably increase salary levels, improve the internal distribution mechanism, and optimize the salary structure to continue deepening the reform of the salary system in public hospital.