BACKGROUND:Exosomes are nanoscale extracellular vesicles secreted by various types of cells,with advantages such as high bioavailability,low toxicity,low immunogenicity,and good biocompatibility.However,natural exosomes have certain limitations in clinical therapy.By using bioengineering techniques to modify and engineer exosomes,the engineered exosomes not only improve their original therapeutic effects but also exhibit excellent biostability and targeting specificity,showing great potential for application in the field of tissue repair.OBJECTIVE:To summarize the various strategies for engineering exosomes,including functional loading and surface modification,outline the research progress of engineered exosomes in different tissue repairs,and explore the therapeutic potential of engineered exosomes in tissue repair.METHODS:PubMed database was searched for relevant literature published between 2010 and 2024 using the search terms"engineered exosomes,tissue repair,biomaterials,tissue engineering,wound healing,parenchyma,bone regeneration,cartilage,neural,myocardial,hepatic."Studies that were not closely related to the article's theme,of poor quality,repetitive,or outdated were excluded.A total of 115 articles met the inclusion criteria.RESULTS AND CONCLUSION:(1)Functional loading is used to combine therapeutic molecules with exosomes to obtain additional properties or to enhance the original physiological function of the exosome,among which ultrasonication and extrusion are simple to operate and can obtain higher drug loading capacity at the same time.(2)Surface modification can make exosomes express desired proteins or enhance their targeting,including genetic engineering and chemical modification.Genetic engineering is complicated,poorly reproducible,and the end product is poorly controllable.Chemical modification,on the other hand,is relatively simple and versatile,and is more suitable for designing highly targeted and functionally specific engineered exosomes.(3)Among the techniques for pre-treating cells to obtain engineered exosomes,hypoxic pre-treatment is more widely used because of its simplicity and clearer mechanism,which can activate glycolysis to promote cell proliferation,and regulate the vascular endothelial growth factor receptor signaling pathway through the generation of hypoxia-inducible factors to promote angiogenesis.(4)The function of exosomes is affected by various factors such as cell source,cell state,synthesis process,and extracellular environment.If the engineering strategy is complicated,it is more difficult to ensure the functional consistency of the final engineered exosomes,so the relatively simple and reliable engineering strategy is more suitable for its clinical application.(5)Engineered exosomes combined with biomaterials or scaffolds can be used to treat complex wounds of skin soft tissue,such as infected wounds and diabetic ulcers.This approach enhances exosome delivery and controls their release,promotes tissue repair,controls infection,and regulates the local microenvironment of the wound.(6)A single mechanism of engineered exosomes is often ineffective due to the specificity of the bone tissue fracture,so dual or even multi-functional engineered exosomes are needed to promote fracture repair while anti-inflammatory or remodeling the vascular system.(7)The source of exosomes has a significant impact on neural tissue repair.Exosomes derived from different neural cells promote neural repair through different effects.In addition,the combination of stents and engineered exosomes for traumatic brain injury has obvious advantages,the stent itself provides hemostasis and support,combined with the engineered exosomes itself to promote the repair effect,can obtain better therapeutic effect.(8)In cardiac and hepatic tissue repair,it is needed to develop anti-fibrotic engineered exosomes to resist the abnormal repair of cardiac and hepatic tissues themselves,which will require further research in the future.

BACKGROUND:Exosomes are nanoscale extracellular vesicles secreted by various types of cells,with advantages such as high bioavailability,low toxicity,low immunogenicity,and good biocompatibility.However,natural exosomes have certain limitations in clinical therapy.By using bioengineering techniques to modify and engineer exosomes,the engineered exosomes not only improve their original therapeutic effects but also exhibit excellent biostability and targeting specificity,showing great potential for application in the field of tissue repair.OBJECTIVE:To summarize the various strategies for engineering exosomes,including functional loading and surface modification,outline the research progress of engineered exosomes in different tissue repairs,and explore the therapeutic potential of engineered exosomes in tissue repair.METHODS:PubMed database was searched for relevant literature published between 2010 and 2024 using the search terms"engineered exosomes,tissue repair,biomaterials,tissue engineering,wound healing,parenchyma,bone regeneration,cartilage,neural,myocardial,hepatic."Studies that were not closely related to the article's theme,of poor quality,repetitive,or outdated were excluded.A total of 115 articles met the inclusion criteria.RESULTS AND CONCLUSION:(1)Functional loading is used to combine therapeutic molecules with exosomes to obtain additional properties or to enhance the original physiological function of the exosome,among which ultrasonication and extrusion are simple to operate and can obtain higher drug loading capacity at the same time.(2)Surface modification can make exosomes express desired proteins or enhance their targeting,including genetic engineering and chemical modification.Genetic engineering is complicated,poorly reproducible,and the end product is poorly controllable.Chemical modification,on the other hand,is relatively simple and versatile,and is more suitable for designing highly targeted and functionally specific engineered exosomes.(3)Among the techniques for pre-treating cells to obtain engineered exosomes,hypoxic pre-treatment is more widely used because of its simplicity and clearer mechanism,which can activate glycolysis to promote cell proliferation,and regulate the vascular endothelial growth factor receptor signaling pathway through the generation of hypoxia-inducible factors to promote angiogenesis.(4)The function of exosomes is affected by various factors such as cell source,cell state,synthesis process,and extracellular environment.If the engineering strategy is complicated,it is more difficult to ensure the functional consistency of the final engineered exosomes,so the relatively simple and reliable engineering strategy is more suitable for its clinical application.(5)Engineered exosomes combined with biomaterials or scaffolds can be used to treat complex wounds of skin soft tissue,such as infected wounds and diabetic ulcers.This approach enhances exosome delivery and controls their release,promotes tissue repair,controls infection,and regulates the local microenvironment of the wound.(6)A single mechanism of engineered exosomes is often ineffective due to the specificity of the bone tissue fracture,so dual or even multi-functional engineered exosomes are needed to promote fracture repair while anti-inflammatory or remodeling the vascular system.(7)The source of exosomes has a significant impact on neural tissue repair.Exosomes derived from different neural cells promote neural repair through different effects.In addition,the combination of stents and engineered exosomes for traumatic brain injury has obvious advantages,the stent itself provides hemostasis and support,combined with the engineered exosomes itself to promote the repair effect,can obtain better therapeutic effect.(8)In cardiac and hepatic tissue repair,it is needed to develop anti-fibrotic engineered exosomes to resist the abnormal repair of cardiac and hepatic tissues themselves,which will require further research in the future.

Hemoglobin A1c (HbA1c) has a unique structure that makes monoclonal antibody (mAb) preparation challenging. This study aims to develop a method for preparing HbA1c mAbs and establish a fluorescent immunochromatographic assay (FICA) for rapid detection of HbA1c. Three glycosylated peptides were synthesized and used to prepare complete antigens, which were identified by dot enzyme-linked immunosorbent assay (Dot-ELISA) and ultraviolet absorption spectroscopy. The complete antigens and natural HbA1c were used for cross-immunization of mice, and the optimal complete antigen was selected. The mouse with the highest serum titer was chosen for mAb preparation. The purity and specificity of the mAbs were verified, and a FICA method was developed. The optimal complete antigen, with a titer of 1:512 000, was successfully prepared and selected. Fusion with splenocytes resulted in four specific HbA1c antibodies (purity > 90%). The best antibody exhibited a binding constant (Ka) of 1.67×10¹⁰ L/mol with the antigen. Based on this antibody, a FICA method was successfully established, capable of producing results within 15 min. The method demonstrated a good linear range (3%-13% HbA1c, y=0.071 3x+0.005 6, R²=0.993 7), recovery rates of 98%-102%, precision < 10.00%, and no nonspecific reactions. Clinical testing of 210 samples showed positive agreement of 96.36%, negative agreement of 97.00%, and overall agreement of 96.68%. The receiver operating characteristic (ROC) curve analysis yielded an area under curve (AUC) of 0.980 9 [95% confidence interval (CI): 0.961 0-1.000 0], with high consistency verified in multicenter studies. We successfully developed a key technique for preparing HbA1c monoclonal antibodies and established a FICA method for rapid detection of HbA1c. It will provide an efficient and convenient detection method for the early diagnosis and long-term management of diabetes and its complications.
Antibodies, Monoclonal/biosynthesis* ; Animals ; Mice ; Glycated Hemoglobin/immunology* ; Mice, Inbred BALB C ; Humans ; Antibody Specificity ; Chromatography, Affinity/methods* ; Enzyme-Linked Immunosorbent Assay/methods* ; Female

Objective To observe the expression levels of base excision repair(BER)pathway-related proteins in small cell lung cancer(SCLC)tissues,and analyze their relationship with the prognosis and tumor immune microenvironment.Methods A retrospective cohort study was conducted on 74 patients with limited-stage SCLC undergoing surgical treatment in our medical center from December 2018 to June 2023.Immunohistochemical staining was performed to analyze the protein expression of BER pathway components,apurinic/apyrimidinic endonuclease 1(APE1),8-oxoguanine DNA glycosylase 1(OGG1),DNA polymerase β(POLβ),X-ray repair cross-complementing protein 1(XRCC1),ATP-dependent DNA ligase I(LIGⅠ),and immune cell infiltration markers of CD3? T cells,CD8? T cells,CD68? macrophages in SCLC tissues.Chi-square test was applied to analyze the relationship of BER protein expression and clinicopathological features;Kaplan-Meier survival curve was plotted to evaluate the impacts of BER protein expression and immune cells on disease-free survival(DFS)and overall survival(OS),multivariate Cox regression analysis was utilized to identify DFS prognostic factors,and Spearman correlation analysis was performed to analyze the correlation of BER-immune cell infiltration.In in vitro experiments,transient transfection was applied in H196 cells to overexpress APE1/POLβ/LIGⅠ,respectively.Thus,the cells were divided into negative control(NC,empty vector)and overexpression(OE,target plasmids)groups.CCK-8 and TUNEL assays were employed to determine the effects of OEAPE1,OEPOLβ and OELIGⅠon cell sensitivity to cisplatin.In in vivo experiments,nude mice bearing xenograft tumors were grouped into WT,E3330(APE1 inhibitor),cisplatin,and cisplatin+E3330 groups to determine the effects of the combination therapy on tumor growth.Results There were no significant correlations of the expression levels of key BER pathway proteins with clinicopathological characteristics,including gender,age,smoking history,tumor location,Ki67 index,or TNM stage(all P>0.05).The patients with low expression of APE1,POLβ,and LIGⅠ had obviously higher DFS rates than those with high expression(P<0.05),and the patients with larger proportion of CD3+T cells also had higher DFS rates than those with smaller proportion(P=0.043).Multivariate Cox regression analysis indicated that tumor TNM stage(HR=2.465)and APE1 expression(HR=2.730)were independent risk factors for the prognosis of SCLC patients(P<0.05).Spearman correlation analysis demonstrated a positive correlation between APE1 and CD8+T cell proportion in the SCLC patients(r=0.27,P<0.05).In vitro experiments showed that the overexpression(OE)cells(OEAPE1 and OELIGⅠ)exhibited reduced sensitivity to cisplatin than the NC group(P<0.05).Animal experiments indicated that cisplatin+E3330 significant inhibited xenograft tumor growth,indicating enhanced therapeutic efficacy(P<0.01).Conclusion High expression of APE1,POLβ,and LIGⅠ in the BER pathway indicates poor prognosis and low DFS rate in SCLC patients.High expression of APE1 is positively correlated with CD8+T cells,and can be used as an auxiliary marker for SCLC immunotherapy.

Objective:It aimed to provide a review on health insurance literacy,including its definition,measurement tools,influencing factors,and its effects on insurance enrollment and the healthcare utilization.Methods:A literature review was conducted by searching for the keywords"health insurance literacy"(Chinese/English)in the CNKI and PubMed databases.Results:The findings indicated that international research had developed a relatively mature definition of health insurance literacy and measurement tools,while domestic research remained in early stage.Health insurance literacy was significantly effected by age,gender,education level,and income.Conclusion:It is suggested to enhance health insurance literacy,develop localized measurement tools,implement targeted educational programs,and promote broader public understanding of health insurance.

To prepare a recombinant adeno-associated virus(rAAV9-ZsGreen1-EsASP)capable of expressing the aspartic protease protein of Eimeria stiedai(E.stiedai),and explore its in vitro ac-tivity.The EsASP gene was amplified by PCR,and recombinant adeno-associated viral vector pAAV-IRES-ZsGreen1 was combined with the EsASP gene using homologous recombination to construct the pAAV-ZsGreen1-EsASP expression plasmid;pAAV-ZsGreen1-EsASP expression plasmid,pHelper,and pAAV-RC9 plasmids were cotransfected into HEK-293T cells by liposomal transfection to package and produce recombinant adeno-associated virus rAAV9-ZsGreen1-EsASP capable of expressing EsASP protein.rAAV9-ZsGreen1-EsASP was purified using chloroform treatment-PEG/NaCl precipitation-chloroform extraction method,the purity of the virus was iden-tified by silver staining,the virus morphology was observed by TEM,and virus titer was detected by qRT-PCR;the purified recombinant virus was further infected into HEK-293T cells,and EsASP expression was detected by observing green fluorescent protein ZsGreen1 and Western blot method.The results indicated that double enzyme digestion and DNA sequencing confirmed that the EsASP gene had been successfully constructed into the pAAV-IRES-ZsGreen1 expression plas-mid.The expression of green fluorescent protein in HEK-293T cells suggested that co-transfection was successful.Western blot results of cell protein preparation showed that EsASP protein was successfully expressed;The purified recombinant viral capsid proteins show three distinct bands(VP1-3).The purified rAAV9-ZsGreen1-EsASP was uniform in size,around 20 nm,and had a titer higher than 1.0 × 1012 vg/mL;Green fluorescent protein expression was observed after infection of HEK-293T cells with the recombinant virus,and EsASP expression was detected by Western blot.The results suggest that rAAV9-ZsGreen1-EsASP with in vitro infectious activity was suc-cessfully obtained,providing a material basis for the development of a novel vaccine against Eimer-ia stiedai.

Objective To investigate the predicting value of cardiac magnetic resonance feature tracking(CMR-FT)for adverse left ventricular remodeling(ALVR)in patients with acute anterior wall ST-segment elevation myocardial infarction(STEMI).Methods The clinical data and cardiac magnetic resonance(CMR)images of 161 acute anterior wall STEMI patients within 1 week and 6 months after emergency percutaneous coronary intervention(PCI)were retrospectively analyzed.ALVR was defined as an increase of left ventricular end-diastolic volume(LVEDV)over 20％at the second CMR examination compared to the baseline.The CMR parame-ters were analyzed by CVI42 post-processing software.The logistic regression analysis was used to screen the independent predictors of ALVR,and the receiver operating characteristic(ROC)curve was used to evaluate the predictive efficiency of ALVR.Results The incidence of ALVR at 6 months was 21.7％(35/161).The logistic regression analysis showed that the left ventricular global circumferential strain(LVGCS)and right ventricular global longitudinal strain(RVGLS)at baseline were independent predictors for ALVR(P＜0.001).When LVGCS was-13.89％and RVGLS was-15.07％at baseline,the sensitivity of predicting ALVR was 0.714 and 0.743,the specificity was 0.833 and 0.810,and the area under the curve(AUC)was 0.806 and 0.835,respectively.The sensitivity of LVGCS combined with RVGLS in predicting ALVR was 0.802,the specificity was 0.952,and the AUC was 0.888.The DeLong test showed that the AUC of LVGCS com-bined with RVGLS in predicting ALVR was significantly higher than that of individuals,and the difference was statistically significant(P＜0.05).Conclusion The LVGCS and RVGLS at baseline are independent predictors for ALVR in patients with acute anterior wall STEMI,their combination can significantly improve the pre-dictive efficiency of ALVR in these patients.

To prepare a recombinant adeno-associated virus(rAAV9-ZsGreen1-EsASP)capable of expressing the aspartic protease protein of Eimeria stiedai(E.stiedai),and explore its in vitro ac-tivity.The EsASP gene was amplified by PCR,and recombinant adeno-associated viral vector pAAV-IRES-ZsGreen1 was combined with the EsASP gene using homologous recombination to construct the pAAV-ZsGreen1-EsASP expression plasmid;pAAV-ZsGreen1-EsASP expression plasmid,pHelper,and pAAV-RC9 plasmids were cotransfected into HEK-293T cells by liposomal transfection to package and produce recombinant adeno-associated virus rAAV9-ZsGreen1-EsASP capable of expressing EsASP protein.rAAV9-ZsGreen1-EsASP was purified using chloroform treatment-PEG/NaCl precipitation-chloroform extraction method,the purity of the virus was iden-tified by silver staining,the virus morphology was observed by TEM,and virus titer was detected by qRT-PCR;the purified recombinant virus was further infected into HEK-293T cells,and EsASP expression was detected by observing green fluorescent protein ZsGreen1 and Western blot method.The results indicated that double enzyme digestion and DNA sequencing confirmed that the EsASP gene had been successfully constructed into the pAAV-IRES-ZsGreen1 expression plas-mid.The expression of green fluorescent protein in HEK-293T cells suggested that co-transfection was successful.Western blot results of cell protein preparation showed that EsASP protein was successfully expressed;The purified recombinant viral capsid proteins show three distinct bands(VP1-3).The purified rAAV9-ZsGreen1-EsASP was uniform in size,around 20 nm,and had a titer higher than 1.0 × 1012 vg/mL;Green fluorescent protein expression was observed after infection of HEK-293T cells with the recombinant virus,and EsASP expression was detected by Western blot.The results suggest that rAAV9-ZsGreen1-EsASP with in vitro infectious activity was suc-cessfully obtained,providing a material basis for the development of a novel vaccine against Eimer-ia stiedai.

Objective:It aimed to provide a review on health insurance literacy,including its definition,measurement tools,influencing factors,and its effects on insurance enrollment and the healthcare utilization.Methods:A literature review was conducted by searching for the keywords"health insurance literacy"(Chinese/English)in the CNKI and PubMed databases.Results:The findings indicated that international research had developed a relatively mature definition of health insurance literacy and measurement tools,while domestic research remained in early stage.Health insurance literacy was significantly effected by age,gender,education level,and income.Conclusion:It is suggested to enhance health insurance literacy,develop localized measurement tools,implement targeted educational programs,and promote broader public understanding of health insurance.