Extracellular vesicles(EVs)are nanoscale membrane vesicles released by cells that carry bioactive molecules such as proteins and nucleic acids. They play a key role in intercellular communication. In recent years, they have shown significant potential in the diagnosis and treatment of corneal and ocular surface diseases. As diagnostic biomarkers, EVs derived from tears or other sources can reflect disease-specific characteristics, providing a basis for the early disease detection. In terms of therapy, mesenchymal stem cells(MSCs)-derived EVs deliver miRNAs to modulate inflammatory pathways, promote corneal epithelial repair, and inhibit fibrosis. Furthermore, as natural drug vehicles, EVs enable precise delivery of therapeutic agents and promote corneal wound healing. This review systematically summarizes the recent research advances in the application of EVs in the field.

Objective To evaluate the clinical efficacy and safety of Huoxue Jiedu Prescription in the treatment of polycythemia vera with heat toxicity and blood stasis syndrome.Methods Totally 155 patients of polycythemia vera with heat toxicity and blood stasis syndrome from 5 hospitals including Langfang Traditional Chinese Medicine Hospital from October 2022 to March 2024 were collected.Patients were divided into an observation group(79 cases)and control group(76 cases)using a random number table method.Both groups received conventional Western medicine treatment.The observation group was given Huoxue Jiedu Prescription,one dosage per day,taken orally twice a day;both groups received one treatment course of one month,and three treatment courses were observed.The efficacy of Western medicine and TCM syndromes was observed,and the total symptom assessment scale of myeloproliferative neoplasms(MPN-10)scores,hematological indicators,coagulation function before and after treatment were compared.The safety indicators of the two groups were monitored.Results The control group and observation group lost 2 and 4 cases,respectively.The total effective rate of Western medicine in the observation group was 90.67%(68/75),while the control group was 67.57%(50/74),with statistical significance(P<0.01).The total effective rate of TCM syndromes in the observation group was 94.67%(71/75),while in the control group was 71.62%(53/74),with statistical significance(P<0.01).Compared with before treatment,the total score of MPN-10 in both groups significantly decreased(P<0.05);after treatment,the total score of MPN-10 in the observation group was lower than that in the control group(P<0.05).Compared with before treatment,both groups showed significant reductions in hemoglobin,white blood cell count,hematocrit and platelet count after treatment(P<0.05);after treatment,the above hematological indicators in the observation group were better than those in the control group(P<0.05).Compared with before treatment,the levels of D-dimer and fibrinogen in both groups significantly decreased after treatment,and the activated partial thromboplastin time and prothrombin time were significantly shortened(P<0.05);after treatment,the observation group showed better improvement in the coagulation function indicators compared to the control group(P<0.05).There were no significant adverse reactions in the two groups.Conclusion Huoxue Jiedu Prescription can improve clinical efficacy of polycythemia vera with heat toxicity and blood stasis syndrome,improve hematological indexes,reduce coagulation indexes,and has good safety.

Methodological quality assessment is a pivotal link between primary studies and reliable evidence-based practice,and an essential pathway for operationalizing the core principles of the Guide on Methodological Standards in Pharmacoepidemiology(2nd edition).A prevalent challenge in practice,however,is the conflation of appraising methodological robustness(risk of bias assessment)with verifying reporting transparency(adherence to reporting guidelines).This paper systematically addresses this fundamental challenge,beginning with a clear distinction between the essence and boundaries of these two concepts.On this basis,the article provides a comprehensive review of mainstream quality assessment tools,covering the methodological features and evolutionary trajectory of numerous instruments for interventional(e.g.,RoB 2,ROBINS-I),observational(e.g.,NOS,the JBI/SIGN/NIH series),secondary(e.g.,AMSTAR 2),and other specific types of studies such as health economic evaluations.Furthermore,a complete case study is used to illustrate the practical application of the ROBINS-I tool.The paper's central thesis advocates for an"appraisal-informed design"philosophy,urging a conceptual shift from the retrospective critique of existing literature to the prospective quality control of new research by internalizing appraisal standards as design principles,while also exploring the emerging paradigm of artificial intelligence in assisting assessment.This paper provides a comprehensive methodological reference for researchers and practitioners to prudently select appropriate assessment tools and to conduct rigorous critical appraisals of pharmacoepidemiological evidence.

Methodological quality assessment is a pivotal link between primary studies and reliable evidence-based practice,and an essential pathway for operationalizing the core principles of the Guide on Methodological Standards in Pharmacoepidemiology(2nd edition).A prevalent challenge in practice,however,is the conflation of appraising methodological robustness(risk of bias assessment)with verifying reporting transparency(adherence to reporting guidelines).This paper systematically addresses this fundamental challenge,beginning with a clear distinction between the essence and boundaries of these two concepts.On this basis,the article provides a comprehensive review of mainstream quality assessment tools,covering the methodological features and evolutionary trajectory of numerous instruments for interventional(e.g.,RoB 2,ROBINS-I),observational(e.g.,NOS,the JBI/SIGN/NIH series),secondary(e.g.,AMSTAR 2),and other specific types of studies such as health economic evaluations.Furthermore,a complete case study is used to illustrate the practical application of the ROBINS-I tool.The paper's central thesis advocates for an"appraisal-informed design"philosophy,urging a conceptual shift from the retrospective critique of existing literature to the prospective quality control of new research by internalizing appraisal standards as design principles,while also exploring the emerging paradigm of artificial intelligence in assisting assessment.This paper provides a comprehensive methodological reference for researchers and practitioners to prudently select appropriate assessment tools and to conduct rigorous critical appraisals of pharmacoepidemiological evidence.

Recently, the World Health Organization (WHO) R&D Blueprint for Epidemics released the 2024 priority pathogen list. Over 200 scientists around the world evaluated the evidences of transmissions, virulences of 1 652 pathogens and the availability of medical countermeasures in two years. Finally, 33 pathogens that could potentially cause the next pandemic were included in the priority pathogen list. This list introduces the scientific framework of "family-priority pathogen-prototype pathogen", emphasizing the importance of studying entire pathogen family rather than specific pathogen. The list can be used to guide and coordinate the research and development of medical countermeasures, including vaccine, blood product, antibody, antimicrobial or antiviral drug, diagnostic test, and personal protective equipment, thereby improving the global rapid response to unknown infectious disease threats. This article summarizes the scientific background and main contents of the WHO priority pathogen list and the enlightenment for pandemic preparedness in the development of medical countermeasures in China to provide reference for the disease prevention and control in China.

Objective To investigate the expression of Cyclin O(CCNO)in cervical cancer and its molecular mechanism in the progression of cervical cancer.Methods The pathological sections of 60 patients with cervical cancer in the Department of Oncology and Gynecology of the First Affiliated Hospital of Bengbu Medical University from January 2018 to December 2023 were collected,and the expression of CCNO in cervical cancer was detected by immunohistochemistry.The cells were divided into Vector group,CCNO group,siNC group and siCCNO group by transfection technology.CCK-8 assay and colony formation assay were used to evaluate the ability of CCNO to affect cell proliferation.Transwell assay and wound healing assay were used to evaluate the effect of CCNO on cell invasion and migration.Glycolysis assay and Western blot assay were used to evaluate the effect and mechanism of CCNO on glycolysis of cervical cancer cells.Results The results of immunohistochemistry and WB showed that CCNO was highly expressed in cervical cancer(P<0.001).Overexpression of CCNO promoted the proliferation,invasion,migration and glycolysis of cervical cancer cells(P<0.05),whereas the opposite effect inhibited the proliferation,invasion,migration and glycolysis of cervical cancer cells(P<0.05).Bioinformatics analysis and WB results showed that CCNO overexpression may regulate the occurrence of cervical cancer by activating the PI3K/AKT signaling pathway(P<0.05).Conclusion Cyclin O may promote the proliferation and metastasis of cervical cancer cells by regulating glycolysis.

Objective To explore the relationship between the oral health status with the severity and outcome of pediatric bronchopneumonia.Methods A total of 150 children patients with bronchopneumonia admitted and treated in this hospital from January 2020 to August 2022 were selected as the observation group(n=150).According to the severity of the illness,the children patients were divided into the mild group(n=97)and severe group(n=53).In addition,150 children undergoing physical examination during the same pe-riod were selected as the control group(n=150).The changes in the decay missing filling surface(DMFS),simplified oral hygiene index(OHI-s)and plaque index(PLI)were analyzed.Spearman was used to analyze the relationship between the oral health status and the severity of bronchopneumonia.After one month of treatment,the changes in oral health status were analyzed under different outcome conditions.The receiver op-erating characteristic(ROC)curve was drawn to assess the predictive effect of the oral health status on the outcome of pediatric bronchopneumonia.Results The DMFS,OHI-s scores and PLI scores in the observation group were higher than those in the control group(P＜0.05).The DMFS,OHI-s score and PLIscores in the severe group were higher than those in the mild group(P＜0.05).Spearman analysis found that DMFS,OHI-s and PLI were positively correlated with the severity(r=0.441,0.253,0.328,P＜0.05).The levels of DMFS,OHI-s and PLI in the poor prognosis group were higher than those in the good prognosis group(P＜0.05).The area under the curve(AUC)of DMFS,OHI-s and PLI combination in predicting the outcome of bronchopneumonia was 0.851(0.703～0.999),with a sensitivity of 83.78％and a specificity of 92.92％.Con-clusion The oral health status is closely related to the severity of pediatric bronchopneumonia,which can be used to assess the outcome of the children patients.

Objective To explore the potential role and underlying mechanism of the functionally uncharacterized gene Gm49394 on regulating β-cell apoptosis under diabetic conditions.Methods The expression and translational activity of Gm49394 in pancreatic β-cell lines and non-β-cell lines were validated using RNA fluorescence in situ hybridization(RNA-FISH),quantitative real-time PCR(qPCR),Western blotting,and immunofluorescence(IF)assay.The β-cell lines(NIT-1/Min6)with Gm49394 overexpression or knockdown were constructed.The proliferation,apoptosis,mitochondrial function,as well as oxidative stress and endoplasmic reticulum stress markers in these β-cell lines under physiological homeostasis or pathological stress conditions,such as high glucose(30 mmol/L),inflammation(10 ng/mL IFN-γ alone or combined with 10 ng/mL IL-6),and hydrogen peroxide(100 μmol/L H2O2)were detected by flow cytometry and Western blotting.Results RNA-FISH and qPCR indicated that Gm49394 was specifically expressed in pancreatic β-cell lines and up-regulated under high glucose or inflammatory stimulation.IF assay and Western blotting showed that Gm49394 had protein-coding activity.Flow cytometry and Western blotting identified that Gm49394 overexpression did not affect β-cell proliferation,but promoted β-cell apoptosis and increased reactive oxygen species(ROS)and mitochondrial superoxide(MitoSOX)levels in β cells under physiological homeostasis or pathological stress conditions(P<0.05).Under physiological conditions,Gm49394 knockdown failed to induce significant alterations on β-cell apoptosis,ROS,or MitoSOX levels.Under pathological stress conditions,Gm49394 knockdown significantly suppressed β-cell proliferation,apoptosis,as well as oxidative and endoplasmic reticulum stress(P<0.05).Conclusion Gm49394 may promote β-cell apoptosis via oxidative stress and endoplasmic reticulum stress.

Recently, the World Health Organization (WHO) R&D Blueprint for Epidemics released the 2024 priority pathogen list. Over 200 scientists around the world evaluated the evidences of transmissions, virulences of 1 652 pathogens and the availability of medical countermeasures in two years. Finally, 33 pathogens that could potentially cause the next pandemic were included in the priority pathogen list. This list introduces the scientific framework of "family-priority pathogen-prototype pathogen", emphasizing the importance of studying entire pathogen family rather than specific pathogen. The list can be used to guide and coordinate the research and development of medical countermeasures, including vaccine, blood product, antibody, antimicrobial or antiviral drug, diagnostic test, and personal protective equipment, thereby improving the global rapid response to unknown infectious disease threats. This article summarizes the scientific background and main contents of the WHO priority pathogen list and the enlightenment for pandemic preparedness in the development of medical countermeasures in China to provide reference for the disease prevention and control in China.

BACKGROUND:Early diagnosis of pulmonary fibrosis is the foundation for timely antifibrotic drug therapy.Therefore,exploring and discovering ideal biomarkers that can be effectively used for the early diagnosis of pulmonary fibrosis is crucial for the treatment of the disease.OBJECTIVE:To conduct an in-depth analysis of key autophagy-related genes involved in the process of pulmonary fibrosis by means of bioinformatics and machine learning techniques,in order to investigate whether autophagy-related core genes of pulmonary fibrosis can be used as reliable biomarkers in the assessment of the progression of pulmonary fibrosis.METHODS:Two datasets of pulmonary fibrosis,GSE24206 and GSE110147,were downloaded from the Gene Expression Omnibus(GEO)database(a public database developed and maintained by the U.S.National Center for Biotechnology Information to store and share bioinformatics data),and the gene expression matrices of these two datasets were normalized by using the"limma"package in R software.The autophagy-related genes were extracted from GeneCards database(a database created by the U.S.National Center for Biotechnology Information,which automatically integrates gene-centric data from about 200 Web sources,including genomic,transcriptomic,proteomic,genetic,clinical,and functional information).Differential gene analysis was performed on the pulmonary fibrosis dataset,and the common genes were extracted by cross-comparing the differential genes with the autophagy genes,so as to identify autophagy genes that may play a role in the process of pulmonary fibrosis.The intersecting genes were analyzed for functional enrichment and cellular immune infiltration by gene ontology and Kyoto Encyclopedia of Genes and Genomes.Core genes of pulmonary fibrosis associated with autophagy were screened by protein-protein interactions and machine learning,and core genes were subjected to the enrichment analysis.Diagnostic models were constructed from the identified core genes.Calibration curves were used to assess the predictive ability of the line graph model.An external dataset,GSE21369,was used to perform a receiver operating characteristic curve analysis to validate the expression profiles of pulmonary fibrosis genes associated with autophagy,as well as to predict Chinese herbs associated with the genes IL6 and COL1A2 via the Coremine database.Finally,human embryonic lung fibroblasts were cultured and modelled by transforming growth factor-β1 treatment,and the relative expression of genes in the model cells was verified using qRT-PCR.RESULTS AND CONCLUSION:(1)A total of 51 pulmonary fibrosis differential genes and 25 genes intersecting with autophagy genes were obtained.Gene ontology analysis showed that the 25 intersecting genes were related to extracellular matrix tissue,collagen metabolism,collagen pro-fibroblasts,and growth factor binding,etc.The results of Kyoto Encyclopedia of Genes and Genomes enrichment analysis indicated that they were mainly related to the Phosphatidylinositol 3-kinase/protein kinase B signaling pathway and the signaling pathway of the extracellular matrix-receptor interactions.(2)Immunoinfiltration analysis revealed that the expression of activated memory CD4+T cells,M0 macrophages,and resting dendritic cells was significantly elevated in the pulmonary fibrosis group(P＜0.05),showing a strong correlation.(3)Two autophagy signature genes involved in the progression of pulmonary fibrosis were identified:COL1A2 and IL6.The column-line diagram model showed that the two core genes predicted the onset of pulmonary fibrosis more accurately,and the receiver operating characteristic curve analysis showed that the two characteristic genes had diagnostic significance.COL1A2 and IL6 were related to the cell-cycle pathway,mitogen-activated protein kinase signaling pathway,Janus kinase-signal transduction and activator of transcription signaling pathway and cytokine-cytokine receptor interactions.A total of 20 Chinese herbs were predicted to be related to COL1A2 and IL6 genes,and their efficacies were mainly to clear away heat and detoxify toxins and to invigorate blood and move qi.COL1A2 and IL6 were verified to be highly expressed in pulmonary fibrosis.To conclude,COL1A2 and IL6 may be potential diagnostic biomarkers for pulmonary fibrosis,but its specificity to pulmonary fibrosis needs to be further investigated.