BACKGROUND:Forskolin,a diterpenoid natural compound extracted from Coleus forskohlii,has a crucial regulatory role in skeletal muscle repair.However,the regulatory role of forskolin on myogenic differentiation of C2C12 skeletal muscle cells has not been fully explored.OBJECTIVE:To explore the effects of forskolin on the differentiation of C2C12 myoblast cell line and probe into the underlying molecular mechanisms.METHODS:C2C12 cells were treated with 0,0.1,0.25,0.5,1,5,10 and 20 μmol/L forskolin during growth,and cell proliferation was detected by cell counting kit-8 and qRT-PCR.C2C12 cells were treated with 0,0.25,0.5 and 1 μmol/L forskolin during the induction of myogenic differentiation.Immunofluorescence staining and qRT-PCR were used to detect C2C12 cells differentiation.Western blot was used to detect the expression level of myogenic differentiation-related signaling pathway proteins.RESULTS AND CONCLUSION:(1)The viability of C2C12 cells was decreased and cell proliferation was inhibited after treatment with high concentrations(＞1 μmol/L)of forskolin.(2)The qRT-PCR results showed that forskolin up-regulated the expression of Myh2,Myh4,Myomaker,but down-regulated the expression of Myh7 compared with the 0 μmol/L group,when C2C12 cells were differentiated for 4 days.Immunofluorescence staining results showed that the fusion index and myotube diameter of C2C12 cells were increased after forskolin treatment,and the number of myotubes was also increased.(3)Western blot results showed that the phosphorylated extracellular signal-regulated kinase 1/2 expression was inhibited;however,the phosphorylated protein kinase B was promoted after treatment with forskolin.The protein expression level of the myogenic differentiation transcription factor Myogenin was significantly up-regulated after treatment with forskolin.The above results demonstrate that forskolin may promote myogenic differentiation of C2C12 skeletal muscle cells through the extracellular signal-regulated kinase 1/2 and protein kinase B signaling pathway.

BACKGROUND:Forskolin,a diterpenoid natural compound extracted from Coleus forskohlii,has a crucial regulatory role in skeletal muscle repair.However,the regulatory role of forskolin on myogenic differentiation of C2C12 skeletal muscle cells has not been fully explored.OBJECTIVE:To explore the effects of forskolin on the differentiation of C2C12 myoblast cell line and probe into the underlying molecular mechanisms.METHODS:C2C12 cells were treated with 0,0.1,0.25,0.5,1,5,10 and 20 μmol/L forskolin during growth,and cell proliferation was detected by cell counting kit-8 and qRT-PCR.C2C12 cells were treated with 0,0.25,0.5 and 1 μmol/L forskolin during the induction of myogenic differentiation.Immunofluorescence staining and qRT-PCR were used to detect C2C12 cells differentiation.Western blot was used to detect the expression level of myogenic differentiation-related signaling pathway proteins.RESULTS AND CONCLUSION:(1)The viability of C2C12 cells was decreased and cell proliferation was inhibited after treatment with high concentrations(＞1 μmol/L)of forskolin.(2)The qRT-PCR results showed that forskolin up-regulated the expression of Myh2,Myh4,Myomaker,but down-regulated the expression of Myh7 compared with the 0 μmol/L group,when C2C12 cells were differentiated for 4 days.Immunofluorescence staining results showed that the fusion index and myotube diameter of C2C12 cells were increased after forskolin treatment,and the number of myotubes was also increased.(3)Western blot results showed that the phosphorylated extracellular signal-regulated kinase 1/2 expression was inhibited;however,the phosphorylated protein kinase B was promoted after treatment with forskolin.The protein expression level of the myogenic differentiation transcription factor Myogenin was significantly up-regulated after treatment with forskolin.The above results demonstrate that forskolin may promote myogenic differentiation of C2C12 skeletal muscle cells through the extracellular signal-regulated kinase 1/2 and protein kinase B signaling pathway.

BACKGROUND:It has been proved clinically that adhesion of fibrous scar with the dura mater or nerve root after lumbar operation is an important factor for postoperative symptoms,such as postoperative pain and numbness. OBJECTIVE:To verify the inhibitory effect of autologous ligamentum flavum on the formation of epidural fibrous scar after lumbar surgery and explore the possible molecular biological mechanism. METHODS:Forty-eight Japanese white rabbits(6-8 months old)were randomly divided into three groups:a ligamentum flavum preservation group,a ligamentum flavum non-preservation group,and an autologous fat reposition group.A lumbar laminectomy model was established in all the three groups of rabbits,and rabbit epidural tissues were collected at 3 and 6 weeks after modeling.Hematoxylin-eosin staining was used to observe histological changes and the number and density of fibroblasts,VG staining was used to observe the percentage of collagen fiber area,and immunohistochemistry was used to observe the expression of transforming growth factor β1 and Smad3 proteins. RESULTS AND CONCLUSION:Hematoxylin-eosin staining results revealed that fibroblasts in the ligamentum flavum preservation group were few and loosely arranged,while the cells in the ligamentum flavum non-preservation and autologous fat reposition groups were more numerous and closely arranged.The number density of fibroblasts in the ligamentum flavum preservation group was lower than that in the ligamentum flavum non-preservation and autologous fat reposition groups at 3 and 6 weeks after surgery(P＜0.05);however,there was no significant difference between the latter two groups.VG staining results showed that the collagen fibers in the ligamentum flavum preservation group were sparse and distributed unevenly,while a lot of red collagen fibers were gathered in the ligamentum flavum non-preservation and autologous fat reposition groups.The area percentage of collagen fibers in the ligamentum flavum preservation group was lower than that in the ligamentum flavum non-preservation and autologous fat reposition groups at 3 and 6 weeks after surgery(P＜0.05),but there was no significant difference between the latter two groups.The results of immunohistochemistry showed that the degree of positive staining of retained histone the ligamentum flavum preservation group was significantly lower than that of the other two groups.The absorbance value of transforming growth factor β1 and Smad3 in the ligamentum flavum preservation group was significantly lower than that in the other two groups at 3 and 6 weeks after surgery(P＜0.05),but there was no significant difference between the latter two groups.To conclude,there are different degrees of epidural fibrous scar formation after lumbar surgery.If the ligamentum flavum is preserved,it can help to reduce the number of epidural fibroblasts as well as the formation of collagen fibers,thus reducing the adhesion of the fibrous scar tissue to the dural sac and nerve root.The mechanism is not only a purely mechanical blockade,but also to reduce the formation of epidural fibrous scar by interfering with the transforming growth factor β1/Smad3 signaling pathway.

Objective To evaluate the application of rabbit liver cancer model in teaching interventional medicine for graduate students.Methods A total of 10 first-year master graduate students majoring in Radiological Imaging(Interventional Medicine).who were studying at Zhengzhou University of China,were enrolled in this study.The rabbit liver cancer model was used as the experimental teaching materials.The teaching contents included the establishment of rabbit liver cancer model,the interventional operation of rabbit liver cancer,the method of scientific research and teaching,the evaluation of the teaching effect,and the survey of student satisfaction.Results Under the guidance of teaching tutor,the success rate of VX2 rabbit liver cancer modeling performed by the 10 master graduate students majoring in interventional medicine was 100％,and the mean operational quality assessment score was(11.5±2.0)points.During the operation of interventional surgery,the success rate of femoral artery puncture was also 100％,and the mean score for each interventional operation was(11.8±2.3)points.The students'experimental designs were evaluated by the expert group,the results were as follows:2 cases were rated as excellent,7 cases were rated as good,and one case was rated as moderate.The degree of students'satisfaction with experimental teaching method was high,the specific scores of each item are as follows:the understanding of the rabbit liver cancer model was(4.80±0.40)points,the command of interventional technology was(4.60±0.49)points,and the quality and practicability of teaching materials was(4.90±0.30)points.Conclusion This teaching method of using rabbit liver cancer model experiment can improve the animal experiment ability,interventional operation ability and scientific research innovation ability of graduate students.Animal model teaching method is an innovation of teaching mode for graduate students majoring in interventional medicine.

Fungal infection is an increasingly serious clinical problem,especially in immunocompromised patients.Fungi,such as Candida albicans,Aspergillus,and Cryptococcus,can form biofilm,which enhance their drug re-sistance and immune evasion ability,thus become major obstacles to the treatment of infection.Biofilm formation is not limited to human body,it can also form on the surface of medical devices,leading to chronic and recurrent infec-tion.This article elaborates the biofilm formation mechanisms and hazards caused by different fungi such as Candi-da albicans and Aspergillus,emphasizes immune suppression,chronic diseases,and medical devices as high-risk factors for biofilm formation.Fungi can form biofilm through multiple stages such as adhesion,proliferation,extra-cellular matrix(ECM)construction,and cell dispersion,and enhance drug resistance based on physical barriers and gene regulation.Although existing antifungal agents are effective in infection caused by planktonic fungi,their treatment efficacy on fungal biofilm is limited.Therefore,the article explores new treatment strategies,including non-pharmacological therapies such as photodynamic therapy and electric field therapy,as well as targeted gene edi-ting and the application of new biomaterials.It is necessary to strengthen the combination of basic research and clini-cal applications,develop efficient and low toxicity treatment scheme to improve the successful treatment rate for patients.

Objectives:Financial pressure could have severe consequences for research hospitals.Studying the financial statement information and management measures of"top"hospitals in the US.can provide references for research hospitals in China to ensure long-term financial sustainability and improve competitiveness.Methods:The latest audited financial statements of Mayo Clinic,Cleveland Clinic,The Johns Hopkins Health System,and Mass General Brigham in the United States were collected,descriptive statistical analysis was employed.Results:The"top"four medical institution have a good asset composition,moderate debt ratios,large investment,with net assets continue to increase.They have diversified sources of revenue,with specialists being its key asset,human resource expense accounting for 57％～58％of total expenses,pursuing technological leadership.When facing financial pressure,the first choice is to improve operational efficiency and continue to reinvest in core missions.Conclusion:The practice of four"top"research medical institutions in the US.has provide new perspective for research hospitals in China to ensure long-term financial sustainability and improve market competitiveness.

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OBJECTIVE To explore the efficacy of platelet-rich plasma （PRP） intrauterine infusion combined with Gushen antai pills in the treatment of recurrent abortion （RSA） and its impacts on endometrial receptivity and hormone levels in patients. METHODS A total of 108 patients with RSA treated in our hospital from January 2021 to January 2024 were selected and evenly divided into control group and study group using the random number table method， with 54 cases in each group. On the basis of conventional treatment， patients in the control group were administered with Gushen antai pills， while patients in the study group received PRP intrauterine infusion combined with Gushen antai pills. The clinical efficacy， TCM syndrome scores before and after treatment， endometrial receptivity [endometrial thickness （EST）， spiral artery resistance index （RI）， and pulsatility index （PI）]， hormone [progesterone （P）， estradiol （E2）， and human chorionic gonadotropin （HCG）] levels， as well as pregnancy outcomes， were compared between the two groups. Additionally， the occurrence of adverse reactions in both groups was recorded. RESULTS The total effective rate （91.44% vs. 81.48%） and infant live birth rate （96.30% vs. 83.33%） of the study group were significantly higher than those of the control group （P＜0.05）. Following treatment， various TCM syndrome scores and total score， spiral artery RI and PI levels in both groups were markedly lower than those in the same groups before treatment， with the study group showing significantly lower levels than the control group（P＜0.05）. Conversely， the P， E2， HCG and EST levels in both groups were significantly higher than those in the same groups before treatment， and the study group exhibited notably higher levels than the control group （P＜0.05）. There was no statistically significant difference in the total incidence of adverse reactions between the two groups （P＞0.05）. CONCLUSIONS PRP intrauterine infusion combined with Gushen antai pills has good clinical efficacy in the treatment of RSA. It can improve TCM syndromes， enhance endometrial receptivity， regulate hormone levels and improve pregnancy outcomes， and it is highly safe.

Objective: To investigate the characteristics of malignant tumor incidence and mortality in cancer registration areas of Sichuan Province in 2021. Methods: Cancer registration data from 142 registries in Sichuan Province in 2021 were collected via the China Cancer Registry Platform. Crude incidence and crude mortality were calculated. The Chinese population-standardized incidence and world population-standardized incidence were standardized using the age structure of the standard population from the Fifth National Population Census in 2000 and Segi's world standard population. Descriptive analyses examined the distribution of rates by genders, urban/rural areas, and ages, and the ranking of leading cancer sites. Results: In 2021, there were 248 600 new malignant tumor cases reported in Sichuan Province, with a crude incidence of 296.37/100 000. The Chinese population-standardized incidence and world population-standardized incidence were 164.67/100 000 and 160.47/100 000, respectively. There were 158 673 malignant tumor deaths, with a crude mortality of 189.16/100 000. The Chinese population-standardized mortality and world population-standardized mortality were 92.47/100 000 and 92.00/100 000, respectively. The Chinese population-standardized incidence and mortality in males were higher than in females (179.56/100 000 vs. 151.62/100 000, 125.09/100 000 vs. 60.35/100 000). The Chinese population-standardized incidence and mortality in urban areas were higher than in rural areas (175.74/100 000 vs. 157.54/100 000, 93.63/100 000 vs. 91.82/100 000). Both the crude incidence and crude mortality increased with age. The top ten malignant tumors by crude incidence were lung cancer, colorectal cancer, liver cancer, breast cancer, esophageal cancer, gastric cancer, cervical cancer, prostate cancer, thyroid cancer, and corpus uteri cancer, accounting for 76.33% of all new cases. The top ten by crude mortality were lung cancer, liver cancer, esophageal cancer, colorectal cancer, gastric cancer, breast cancer, pancreatic cancer, cervical cancer, prostate cancer, and brain tumors, accounting for 82.39% of all cancer deaths. Conclusions In registration areas of Sichuan Province, the incidence and mortality of malignant tumors are relatively low. Key populations such as males, urban residents, and the elderly require focused prevention and control efforts. Comprehensive measures should be prioritized for malignant tumors including lung cancer, liver cancer, esophageal cancer, gastric cancer, colorectal cancer, and breast cancer.

BACKGROUND: Macrophage polarization anomalies and dysfunction play a crucial role in the pathogenesis of immune thrombocytopenia (ITP). Itaconate is a Krebs cycle-derived immunometabolite synthesized by myeloid cells to modulate cellular metabolism and inflammatory responses. This study aimed to evaluate the immunoregulatory effects of an itaconate derivative on macrophages in patients with ITP. METHODS: Peripheral blood-derived macrophages from patients with ITP and healthy controls were treated with 4-octyl itaconate (4-OI), a derivative of itaconate that can penetrate the cell membrane. Macrophage polarization, antigen-presenting functions, and phagocytic capability were measured via flow cytometry and enzyme-linked immunosorbent assay (ELISA). Macrophage glycolysis in patients with ITP and the metabolic regulatory effect of 4-OI were detected using a Seahorse XFe96 Analyzer. An active murine model of ITP was used to evaluate the therapeutic effects of 4-OI in vivo . RESULTS: 4-OI reduced the levels of CD80 and CD86 in M1 macrophages and suppressed the release of tumor necrosis factor-α (TNF-α) and interleukin (IL)-6 pro-inflammatory cytokines, suggesting that 4-OI could hinder the polarization of macrophages toward an M1 phenotype. We found that 4-OI pretreated M1 macrophages reduced the proliferation of CD4 + T cells and promoted the differentiation of regulatory T cells. In addition, after 4-OI treatment, the phagocytic capacity of M1 macrophages toward antibody-coated platelets decreased significantly in patients with ITP. In addition, the glycolytic function of M1 macrophages was elevated in individuals with ITP compared to those in healthy controls. 4-OI treatment downregulated glycolysis in M1 macrophages. The glycolysis inhibitor 2-deoxy-d-glucose (2-DG) also inhibited the polarization of M1 macrophages and restored their functions. In vivo , 4-OI treatment significantly increased platelet counts in the active ITP murine model. CONCLUSIONS Itaconate derivative 4-OI inhibited M1 macrophage polarization and restored impaired functions through metabolic reprogramming. This study provides a novel therapeutic option for ITP.
Macrophages/metabolism* ; Humans ; Animals ; Succinates/pharmacology* ; Mice ; Male ; Female ; Adult ; Middle Aged ; Flow Cytometry ; Tumor Necrosis Factor-alpha/metabolism* ; Enzyme-Linked Immunosorbent Assay ; Purpura, Thrombocytopenic, Idiopathic/metabolism* ; Glycolysis/drug effects* ; Metabolic Reprogramming