Most patients with hepatocellular carcinoma(HCC)are diagnosed at an intermediate or advanced stage,losing the opportunity for surgical resection.Conversion therapy,which uses non-surgical approaches to render initially unresectable tumors resectable,has gradually become part of routine clinical practice and a research focus.However,multiple challenges remain,including the lack of clear criteria for identifying"potentially resectable"cases,difficulty in selecting individualized systemic regimens from multiple approved options and determining whether to combine them with locoregional therapy,controversy over the necessity of surgery in patients achieving radiological complete response,the need for optimization of perioperative assessment and management,uncertainty in determining the optimal timing of surgery,and the absence of consensus on postoperative sequential systemic therapy regimens and duration.Addressing these issues requires multidisciplinary collaboration and high-quality evidence from multicenter randomized controlled trials.With the accumulation of clinical experience,growing evidence,and advances in treatment,more patients with initially unresectable HCC are expected to gain surgical opportunities and achieve long-term disease-free survival.

To construct a subgenomic replication subsystem of foot-and-mouth disease virus(FM-DV)carrying fluorescent protein(UnaG),the full-length cDNA plasmid of FMDV was employed as the template.A few of structural and non-structural protein genes of FMDV were removed by double-enzyme digestion.By substituting reporter gene sequences expressing the green fluorescent protein UnaG for specific structural protein sequences of FMDV,FMDV-UnaG replicators were successfully created.After PCR and sequencing,linearized FMDV-UnaG replicons were transfected into BSR/T7 cells expressing T7 RNA polymerase,and the fluorescence signal was observed through fluorescence microscopy and laser confocal technique.The results demonstrated that the constructed FMDV-UnaG replicons could effectively express UnaG protein,and the protein colo-calized with FMDV 3A protein.Additionally,Western blot and RT-qPCR also detected that the replicator RNA could express the non-structural proteins of the virus and replicate autonomously in BSR/T7 cells,respectively.In conclusion,the successful construction of FMDV-UnaG sub-genomic replicators offers a favorable tool for further research on the replication and translation mechanism of FMDV and the development of vaccine vectors.

Purpose To investigate the differential expression of proteins secreted by diffuse-type gastric cancer-associated fibroblasts(DGC-CAFs)and intestinal-type gastric cancer-associated fibroblasts(IGC-CAFs)according to Lauren classification of gastric cancer.Methods Fresh surgery samples were acquired to extract primary cancer-asso-ciated fibroblasts(CAFs)to obtain the conditional medium,including three cases of diffuse-type gastric cancer(DGC)and three cases of intestinal-type gastric cancer(IGC).Then high performance liquid chromatography-tandem mass spectrometry(HPLC-MS/MS)was used to detect the differences in secreted factors between DGC-CAFs and IGC-CAFs.Additionally,a total of 1 174 cases were collected from the GEO and TCGA databases,and the distribution of stromal cells was assessed via ESTIMATE to derive stromal scores for both DGC and GC.Furthermore,Kaplan-Meier survival analysis was performed to evaluate the relationship between high stromal scores,elevated CAFs proportions,and patient prognosis.Subsequently,GO enrichment analysis was performed to investigate associated genes and their bi-ological functions.Results HPLC-MS/MS analysis showed that only 10 proteins were identified to be expressed in both DGC-CAFs and IGC-CAFs,namely SPARC,COL6A1,COL1A2,COL1A1,COL3A1,FN1,DCN,ACTG1,TIMP1 and SHROOM3.There were 8 proteins expressed higher in DGC-CAFs than IGC-CAFs in all cases with ratio＞2,namely COL3A1(5.28),COL1A2(3.98),DCN(3.71),TIMP1(3.40),COL6A1(3.35),ACTG1(3.13),COL1A1(2.84)and SHROOM3(2.50).Two other proteins,TUBB and BASP1,were identified in all three cases of DGC-CAFs.The former was only identified in one case of IGC-CAFs,while the latter was not identified in all three cases of IGC-CAFs.The results of bioinformatics analysis showed that the stromal score of DGC was higher than IGC,which correlated with poor prognosis.Analysis of cell components revealed that the related genes were more enriched in ECM,collagen-containing extracellular matrix and cell-cell junction.Also,the biological process and molecular function was based on the components,which was consistent with HPLC-MS/MS analysis results.Conclusion There are differ-ences in secreted proteins of DGC-CAFs and IGC-CAFs,both on the types and the content.The differential proteins are mostly enriched in ECM-related signaling pathways.Presumably,the high content of CAFs in the stroma affects the prognosis of gastric cancer patients by influencing the gene expression and the function of receptor pathway of ECM.

Most patients with hepatocellular carcinoma(HCC)are diagnosed at an intermediate or advanced stage,losing the opportunity for surgical resection.Conversion therapy,which uses non-surgical approaches to render initially unresectable tumors resectable,has gradually become part of routine clinical practice and a research focus.However,multiple challenges remain,including the lack of clear criteria for identifying"potentially resectable"cases,difficulty in selecting individualized systemic regimens from multiple approved options and determining whether to combine them with locoregional therapy,controversy over the necessity of surgery in patients achieving radiological complete response,the need for optimization of perioperative assessment and management,uncertainty in determining the optimal timing of surgery,and the absence of consensus on postoperative sequential systemic therapy regimens and duration.Addressing these issues requires multidisciplinary collaboration and high-quality evidence from multicenter randomized controlled trials.With the accumulation of clinical experience,growing evidence,and advances in treatment,more patients with initially unresectable HCC are expected to gain surgical opportunities and achieve long-term disease-free survival.

To construct a subgenomic replication subsystem of foot-and-mouth disease virus(FM-DV)carrying fluorescent protein(UnaG),the full-length cDNA plasmid of FMDV was employed as the template.A few of structural and non-structural protein genes of FMDV were removed by double-enzyme digestion.By substituting reporter gene sequences expressing the green fluorescent protein UnaG for specific structural protein sequences of FMDV,FMDV-UnaG replicators were successfully created.After PCR and sequencing,linearized FMDV-UnaG replicons were transfected into BSR/T7 cells expressing T7 RNA polymerase,and the fluorescence signal was observed through fluorescence microscopy and laser confocal technique.The results demonstrated that the constructed FMDV-UnaG replicons could effectively express UnaG protein,and the protein colo-calized with FMDV 3A protein.Additionally,Western blot and RT-qPCR also detected that the replicator RNA could express the non-structural proteins of the virus and replicate autonomously in BSR/T7 cells,respectively.In conclusion,the successful construction of FMDV-UnaG sub-genomic replicators offers a favorable tool for further research on the replication and translation mechanism of FMDV and the development of vaccine vectors.

Purpose To investigate the differential expression of proteins secreted by diffuse-type gastric cancer-associated fibroblasts(DGC-CAFs)and intestinal-type gastric cancer-associated fibroblasts(IGC-CAFs)according to Lauren classification of gastric cancer.Methods Fresh surgery samples were acquired to extract primary cancer-asso-ciated fibroblasts(CAFs)to obtain the conditional medium,including three cases of diffuse-type gastric cancer(DGC)and three cases of intestinal-type gastric cancer(IGC).Then high performance liquid chromatography-tandem mass spectrometry(HPLC-MS/MS)was used to detect the differences in secreted factors between DGC-CAFs and IGC-CAFs.Additionally,a total of 1 174 cases were collected from the GEO and TCGA databases,and the distribution of stromal cells was assessed via ESTIMATE to derive stromal scores for both DGC and GC.Furthermore,Kaplan-Meier survival analysis was performed to evaluate the relationship between high stromal scores,elevated CAFs proportions,and patient prognosis.Subsequently,GO enrichment analysis was performed to investigate associated genes and their bi-ological functions.Results HPLC-MS/MS analysis showed that only 10 proteins were identified to be expressed in both DGC-CAFs and IGC-CAFs,namely SPARC,COL6A1,COL1A2,COL1A1,COL3A1,FN1,DCN,ACTG1,TIMP1 and SHROOM3.There were 8 proteins expressed higher in DGC-CAFs than IGC-CAFs in all cases with ratio＞2,namely COL3A1(5.28),COL1A2(3.98),DCN(3.71),TIMP1(3.40),COL6A1(3.35),ACTG1(3.13),COL1A1(2.84)and SHROOM3(2.50).Two other proteins,TUBB and BASP1,were identified in all three cases of DGC-CAFs.The former was only identified in one case of IGC-CAFs,while the latter was not identified in all three cases of IGC-CAFs.The results of bioinformatics analysis showed that the stromal score of DGC was higher than IGC,which correlated with poor prognosis.Analysis of cell components revealed that the related genes were more enriched in ECM,collagen-containing extracellular matrix and cell-cell junction.Also,the biological process and molecular function was based on the components,which was consistent with HPLC-MS/MS analysis results.Conclusion There are differ-ences in secreted proteins of DGC-CAFs and IGC-CAFs,both on the types and the content.The differential proteins are mostly enriched in ECM-related signaling pathways.Presumably,the high content of CAFs in the stroma affects the prognosis of gastric cancer patients by influencing the gene expression and the function of receptor pathway of ECM.

Cancer incidence in China has been rising steadily,with a particularly heavy burden from several high-prevalence malignancies.Medical examination for cancer plays a critical role in the early detection of cancer,precancerous lesions,and precursor conditions,thereby facilitating timely diagnosis and intervention.Such examination also addresses the growing demand for person-alized cancer screening services among diverse population groups.The development of evidence-based,context-specific cancer screening guidelines is essential to enhance the standardization,quality,and equity of preventive screening practices across the country,ultimately improving out-comes in early cancer detection and treatment.Guided by the Department of Medical Emergency Response of the National Health Commission,the Guidelines for Medical Examination for Cancer in Health Examination Agency(2025 Edition)were developed under the leadership of the National Cancer Center.A multidisciplinary panel of experts formulated the guidelines in accordance with the principles and methodology of the World Health Organization Handbook for Guideline Deve-lopment.The guidelines provide evidence-based recommendations on key clinical domains:target cancers and populations,overall screening workflow,screening protocols,diagnostic technolo-gies,result interpretation,follow-up procedures,and quality control.The primary objective is to standardize cancer screening practices in health examination agency and strengthen China's ca-pacity for prevention and control of high-burden cancers.

Objective To investigate the impact of ultrasound intervention combined with tripterygium for ovarian endometriosis cyst(OEC)on the ovarian function of patients.Methods The clinical data of 107 patients with clinically-conformed OEC,who received treatment at the Yuyao Municipal People's Hospital of China from December 2022 to June 2024,were retrospectively analyzed.According to different treatment methods,the patients were divided into control group(n=53)and observation group(n=54).The patients of control group were treated with laparoscopic cystectomy,while the patients of observation group were treated with ultrasound intervention combined with tripterygium therapy.The uterine artery blood flow parameters,sex hormone levels,tumor marker levels,ovarian reserve function were compared between the two groups.Results After treatment,uterine artery blood flow parameters were decreased in both groups,the levels of uterine artery blood flow parameters in the observation group were significantly lower than those in the control group(P＜0.05);the levels of sex hormones were elevated in both groups,the levels of sex hormones in the observation group were obviously higher than those in the control group(P＜0.05).After treatment,the tumor-related indicators generally decreased,especially in the observation group receiving combination therapy,the levels of tumor-related indicators were significantly lower than those in the control group receiving simple surgical treatment(P＜0.05),indicating that the therapeutic effect of the observation group was better than that of the control group.After treatment,the ovarian reserve function of both groups were increased when compared with their preoperative values,and the ovarian reserve function of the observation group was higher than that of the control group(P＜0.05).Conclusion In treating OEC patients,ultrasound intervention combined with tripterygium can regulate the levels of sex hormones,reduce tumor marker levels,improve local uterine blood flow status,and protect ovarian reserve function.This therapy is worth promoting and applying in clinical practice.

It is believed that the basic mechanism of cough associated with interstitial lung disease is collaterals deficiency with latent wind： the deficiency of the lung organs and lung collaterals is the basis of its pathogenesis， and latent wind in lung collaterals is the key mechanism of the cough which is difficult to cure. Treatment is based on the principle of supplementing deficiency and treating wind， dispelling the pathogens and unblocking the collaterals. Supplementing deficiency should supplement lungs， boost kidneys， and strengthen spleens to consolidate the root and banking up the origin， and regulate and tonify the lung collaterals； dispelling the pathogens should treat the internal and external winds at the same time， and taking into account the combined pathogens of phlegm， stasis and dampness to clear the stagnation of the lung collaterals.

Foot-and-mouth disease (FMD) is one of the major animal infectious diseases in the world. All cloven-hoofed animals are susceptible to FMD. Vaccination is still the first choice for the prevention and control of FMD. mRNA vaccines can be rapidly designed, synthesized, and produced on a large scale in vitro, and they can induce effective protective immune responses, demonstrating the advantages of rapid development, easy preparation, and low biosafety risks. The design of untranslated regions is a key to enhancing the expression and efficacy of mRNA vaccines. In order to generate an efficient FMD mRNA vaccine, we designed three FMD P12A3C expression vectors with different untranslated regions and synthesized corresponding mRNAs. By comparing expression efficiency of these vectors and their mRNAs at different time points and in different cell lines, we found that the mRNA P12A3C-UTR3 had the best expression and universality. This study laid a foundation for the development of mRNA vaccines against FMD and provided a theoretical basis for the optimal sequence design of efficient mRNA.
Foot-and-Mouth Disease Virus/genetics* ; Animals ; RNA, Messenger/biosynthesis* ; Foot-and-Mouth Disease/immunology* ; Antigens, Viral/biosynthesis* ; Viral Vaccines/biosynthesis* ; Genetic Vectors/genetics* ; Cell Line ; Vaccines, DNA/immunology*