Background/Aims: Liver transplantation (LT) following total hepatectomy is a life-saving treatment for hepatocellular carcinoma (HCC). The HCC recurrence after LT hinders the effectiveness of the procedure. The objective of this study is to develop a pre-operative risk stratification model based on a liquid biopsy. Methods: We conducted a comprehensive multi-omics study of 260 HCC patients from three centers, including clinical data, low-coverage whole-genome sequencing of cell-free DNA (cfDNA) from plasma, as well as whole-exome, single-nucleus RNA, and spatial transcriptomics from matched tumor and non-tumor tissues. Results: We identified cfDNA-derived copy number alteration (CNA) signatures associated with post-transplant recurrence. By integrating cfDNA-derived CNA profiles with single-cell transcriptomic data, we traced recurrence-associated cfDNA to a distinct subpopulation of malignant cells within the primary tumor. These cells were embedded in a pro-metastatic microenvironment of specialized endothelial subtypes and cancer-associated fibroblasts. Notably, most recurrence-associated lesions were detectable in cfDNA prior to liver transplantation (LT). Building on these insights, we developed the ZJU Criteria based on CNA fragments and tumor markers, a pre-LT risk prediction tool that integrates conventional clinical factors with cfDNA-derived CNA signatures, and validated it using internal and independent external cohorts. Conclusion Our findings suggest that post-transplant recurrence commonly originates from advanced subclones that emerge late during tumor evolution. The ZJU Criteria provides an accurate, non-invasive strategy that significantly improves pre-LT risk stratification and clinical decision-making for patients with HCC.

In recent years, China’s capacity for new drug research and development has been significantly strengthened, with major breakthroughs across multiple therapeutic areas, including cancer, autoimmune diseases, and metabolic disorders. An increasing number of domestically developed innovative drugs have been approved for marketing, gradually establishing a competitive position against imported drugs and expanding into global markets. This article systematically reviews the latest research progress of Class 1 innovative drugs in China from 2023 to 2024, categorizes representative approved drugs by therapeutic area, summarizes the clinical advantages and characteristics of representative drugs approved, analyzes in depth the current development trends of and challenges for China’s innovative pharmaceutical industry, and explores future directions, aiming to provide some reference for innovative drug research, industrial development, and policy-making in China.

Numerous studies have shown that depression is main-ly associated with the abnormal expression of connexin 43(Cx43)in astrocytes(Astro)and its mediated dysfunction of gap junction(GJ).However,the molecular mechanism of post-translational modifications targeting Cx43 to regulate neuroin-flammation-associated depression is still unclear.Post-transla-tional modifications of Cx43 mainly include phosphorylation of specific amino acid sites by PKC,PKA,PKG,MAPK and PTK,and protein degradation of Cx43 through the K48/K63 polyubiq-uitylation and deubiquitination pathways,which ultimately lead to protein degradation through K48/K63 polyubiquitination and deubiquitination.These modifications are ultimately involved in the regulation of neuroinflammatory responses through the associ-ation of GJ function.In this paper,we systematically review the role of Cx43 post-translational modifications in neuroinflamma-tion,with the aim of further exploring the potential application of targeting these modifications to modulate the inflammatory re-sponse mechanism in improving depressive symptoms.

Aim To investigate the effect of p-AMPK activity on autophagy in different subtypes of MDA-MB-231(triple-negative breast cancer cells)and MCF-7(estrogen receptor-positive cells)and its regulatory mechanism.Methods MDA-MB-231 cells were trea-ted with EBSS,Baf-A1,and EBSS+Baf-A1 for four hours,and MCF-7 cells for eight hours.The effects of autophagy on cell proliferation and apoptosis were ob-served,mitochondrial morphology was examined,and the expression of autophagy markers LC3B,P62,LAMP1,TOM20,AMPK,p-AMPK,ULK1,and Bec-lin1/VPS34 proteins was detected.The autophagy pathway was validated by inhibiting AMPK activity.Results Breast cancer cells underwent autophagy af-ter starvation induction(EBSS),with inconsistent au-tophagy processes observed in different subtypes of breast cancer cells.Autophagy inhibited cell prolifera-tion.In MDA-MB-231 cells,autophagy led to an in-crease in p-AMPK levels and a decrease in ULK1 lev-els,initiating autophagy through p-AMPK activation of ULK1.In MCF-7 cells,both p-AMPK and ULK1 levels decreased after autophagy,suggesting that autophagy might not be mediated by p-AMPK activation.Conclu-sions MDA-MB-231 cells primarily initiate autophagy by directly activating ULK1 by p-AMPK,independent of the MTOR pathway.In MCF-7 cells autophagy might be triggered by inhibiting MTOR through AMPK activity or directly activating MTOR through other up-stream factors.Regulating p-AMPK activity based on the autophagy pathways in different cell subtypes could enable more precise targeting and treatment of different types of breast cancer.

BACKGROUND:Exosomes derived from human umbilical cord mesenchymal stem cells are widely used for bone repair and reconstruction,significantly enhancing osteogenesis and promoting angiogenesis.OBJECTIVE:To explore the mechanisms of exosomes derived from human umbilical cord mesenchymal stem cells in the treatment of osteoporotic fractures.METHODS:H uman umbilical cord mesenchymal stem cells were extracted using tissue block culture method.Exosomes derived from human umbilical cord mesenchymal stem cells were extracted using ultracentrifugation method for identification.Thirty 12-week-old female SD rats were randomly divided into sham-operated group(n=6)and ovariectomized group(n=24).Osteoporosis model was established by castration in the ovariectomized group.12 weeks after modeling,6 rats in the ovariectomized group and 6 rats in the sham-operated group were randomly selected for Micro-CT and hematoxylin-eosin staining to verify the models.After verification,the remaining 18 rats in the ovariectomized group were randomly assigned to three groups to establish osteoporotic fracture models.The fracture end was separately injected with PBS(PBS group),exosomes at 1.5×1011 particles/mL(low-concentration exosome group),and 3×1011 particles/mL(high-concentration exosome group).Four weeks after operation,fracture healing and bone angiogenesis were evaluated by imaging and histological observations.RESULTS AND CONCLUSION:(1)In the gross specimens,compared with the PBS group,the exosome group had faster fracture healing and more callus formation.(2)The X-ray score of fracture healing in the exosome group was significantly higher than that in the PBS group,and the difference was significant(P＜0.05).(3)Micro-CT three-dimensional imaging:Compared with the PBS group,the fracture healing in the exosome group was accelerated and the callus formation was significantly increased;the bone volume fraction in the exosome group was significantly higher than that in the PBS group,and the difference was significant(P＜0.01).(4)Hematoxylin-eosin staining and Masson's trichrome staining showed that bone trabeculae and the new bone tissue in the exosome group were more than those in the PBS group.(5)Immunohistochemical staining showed that the expression of CD31 and osteocalcin in the exosome group was significantly higher than that in the PBS group.The high-concentration exosome group had a higher density of new blood vessels,more callus formation,and faster fracture healing than the low-concentration exosome group,showing a concentration-dependent manner.The results show that exosomes derived from human umbilical cord mesenchymal stem cells can promote the repair of osteoporotic fracture by enhancing the angiogenesis and osteogenesis.The mechanism may be related to the increased expression of CD31 and osteocalcin.

Sepsis, a life-threatening syndrome characterized by acute organ dysfunction due to a dysregulated host response to infection, remains a significant global health challenge. Despite the identification of numerous biomarkers, their clinical translation remains challenging due to various limitations. In this article, we review the historical evolution of sepsis biomarker research, tracing the progression from early inflammatory marker research to the advent of multi-omics technologies and artificial intelligence-driven precision medicine. We summarize the current clinical applications of biomarkers in early diagnosis, pathogen identification, and precision treatment. Furthermore, we discuss key barriers including the complex pathophysiology of sepsis, insufficient real-world studies, and the lack of standardized evaluation frameworks in advancing biomarker research and utilization. Looking ahead, leveraging advanced approaches such as dynamic monitoring and precision stratification, combined with multidisciplinary collaboration, is expected to facilitate biomarker clinical translation and support the development of precision medicine for sepsis.

Numerous studies have shown that depression is main-ly associated with the abnormal expression of connexin 43(Cx43)in astrocytes(Astro)and its mediated dysfunction of gap junction(GJ).However,the molecular mechanism of post-translational modifications targeting Cx43 to regulate neuroin-flammation-associated depression is still unclear.Post-transla-tional modifications of Cx43 mainly include phosphorylation of specific amino acid sites by PKC,PKA,PKG,MAPK and PTK,and protein degradation of Cx43 through the K48/K63 polyubiq-uitylation and deubiquitination pathways,which ultimately lead to protein degradation through K48/K63 polyubiquitination and deubiquitination.These modifications are ultimately involved in the regulation of neuroinflammatory responses through the associ-ation of GJ function.In this paper,we systematically review the role of Cx43 post-translational modifications in neuroinflamma-tion,with the aim of further exploring the potential application of targeting these modifications to modulate the inflammatory re-sponse mechanism in improving depressive symptoms.

Sepsis, a life-threatening syndrome characterized by acute organ dysfunction due to a dysregulated host response to infection, remains a significant global health challenge. Despite the identification of numerous biomarkers, their clinical translation remains challenging due to various limitations. In this article, we review the historical evolution of sepsis biomarker research, tracing the progression from early inflammatory marker research to the advent of multi-omics technologies and artificial intelligence-driven precision medicine. We summarize the current clinical applications of biomarkers in early diagnosis, pathogen identification, and precision treatment. Furthermore, we discuss key barriers including the complex pathophysiology of sepsis, insufficient real-world studies, and the lack of standardized evaluation frameworks in advancing biomarker research and utilization. Looking ahead, leveraging advanced approaches such as dynamic monitoring and precision stratification, combined with multidisciplinary collaboration, is expected to facilitate biomarker clinical translation and support the development of precision medicine for sepsis.

Aim To investigate the effect of p-AMPK activity on autophagy in different subtypes of MDA-MB-231(triple-negative breast cancer cells)and MCF-7(estrogen receptor-positive cells)and its regulatory mechanism.Methods MDA-MB-231 cells were trea-ted with EBSS,Baf-A1,and EBSS+Baf-A1 for four hours,and MCF-7 cells for eight hours.The effects of autophagy on cell proliferation and apoptosis were ob-served,mitochondrial morphology was examined,and the expression of autophagy markers LC3B,P62,LAMP1,TOM20,AMPK,p-AMPK,ULK1,and Bec-lin1/VPS34 proteins was detected.The autophagy pathway was validated by inhibiting AMPK activity.Results Breast cancer cells underwent autophagy af-ter starvation induction(EBSS),with inconsistent au-tophagy processes observed in different subtypes of breast cancer cells.Autophagy inhibited cell prolifera-tion.In MDA-MB-231 cells,autophagy led to an in-crease in p-AMPK levels and a decrease in ULK1 lev-els,initiating autophagy through p-AMPK activation of ULK1.In MCF-7 cells,both p-AMPK and ULK1 levels decreased after autophagy,suggesting that autophagy might not be mediated by p-AMPK activation.Conclu-sions MDA-MB-231 cells primarily initiate autophagy by directly activating ULK1 by p-AMPK,independent of the MTOR pathway.In MCF-7 cells autophagy might be triggered by inhibiting MTOR through AMPK activity or directly activating MTOR through other up-stream factors.Regulating p-AMPK activity based on the autophagy pathways in different cell subtypes could enable more precise targeting and treatment of different types of breast cancer.

This paper reviews the current situation of massive open online course (MOOC) construction both domestically and internationally, highlighting the similarities, differences, and limitations of MOOC construction across nations. Based on the full-cycle MOOC construction of the "clinical epidemiology" course at Southern Medical University, including course design, resource integration, online deployment, and teaching evaluation, this study explored the significance, implementation path, and challenges of MOOC construction. This paper also reflects on the activation of teaching content, teacher-student interaction, and teaching mode, aiming to provide a reference for the construction and continuous enhancement of MOOC in China.