Extracellular vesicles (EVs) are crucial for facilitating intercellular communication, promoting cell migration, and orchestrating the immune response. Recently, EVs can diagnose and treat tumors. EVs can be measured as biomarkers to provide information about the type of disease and therapeutic efficacy. Furthermore, EVs with lower immunogenicity and better biocompatibility are natural carriers of chemicals and gene drugs. Herein, we review the molecular composition, biogenesis, and separation methods of EVs. We also highlight the important role of EVs from different origins as biomarkers and drug delivery systems in tumor therapy. Finally, we provide deep insights into how EVs play a role in reversing the immunosuppressive microenvironment.

Plant-derived extracellular vesicles (PDEVs), describe a group of nanoparticles released by plants. These particles are characterized by a lipid bilayer structure containing various proteins, lipids, nucleic acids, and unique metabolites. Although the study on PDEVs is relatively new, having only been around for ten years, they have shown promising development prospects in both basic research and clinical transformation areas. Evidence suggests that PDEVs have excellent application prospects in regulating inflammation and treating tumors. Their distinctive, vesicle-mimicking architecture and stellar biocompatibility render them prime candidates for ferrying various anti-cancer agents, including RNA, proteins, and conventional chemotherapy drugs. Increasingly, studies have shown that PDEVs can be engineered as an innovative platform for combination cancer immunotherapy. Consequently, this paper provides an extensive summary of current developments in engineering methods and strategies for PDEVs in cancer treatment and combined cancer immune therapeutics. The essential characteristics of PDEVs, including the biogenesis process and components, as well as their anti-tumor activity and mechanism, are summarized. Finally, the in vivo safety of PDEVs as delivery vectors and the challenges of scale-up production and clinical transformation are discussed.

OBJECTIVES: To evaluate the protective effect of Lycium barbarum polysaccharides (LBP) against cisplatin-induced ovarian granulosa cell injury and investigate its possible mechanisms. METHODS: Human granulosa-like tumor cell line (KGN) were treated with 2.5 µg/mL cisplatin for 24 h, followed by treatment with 100, 500, and 1000 mg/L LBP, and the changes in cell viability, apoptosis, level of anti-Müllerian hormone (AMH), and cell ultrastructure were detected with CCK-8 assay, flow cytometry, ELISA and transmission electron microscopy. The cellular expressions of Bax, caspase-3, Bcl-2, and the PI3K/AKT pathway proteins were analyzed using Western blotting, and the expression of miR-23a was detected with RT-qPCR. KGN cell models with lentivirus-mediated miR-23a overexpression or knockdown were used to verify the therapeutic mechanism of LBP. RESULTS: Cisplatin treatment significantly inhibited cell viability, induced apoptosis, decreased AMH level, caused ultrastructural abnormalities, increased Bax and caspase-3 expression, and lowered Bcl-2 expression in KGN cells. Cisplatin also suppressed the activation of the PI3K/AKT signaling pathway and upregulated miR-23a expression in the cells. LBP intervention obviously alleviated cisplatin-induced injuries in KGN cells, and in particular, LBP treatment at the medium dose for 24 h significantly improved KGN cell viability, reduced apoptosis, enhanced their endocrine function, and ameliorated ultrastructural abnormalities. Mechanistically, medium-dose LBP obviously activated the PI3K/AKT pathway by downregulating miR-23a in cisplatin-treated cells, subsequently inhibiting Bax and caspase-3 while upregulating Bcl-2. Overexpression of miR-23a weakened while knockdown of miR-23a significantly enhanced the protective effects of LBP. CONCLUSIONS LBP alleviates cisplatin-induced apoptosis in KGN cells by inhibiting miR-23a expression and activating the PI3K/AKT pathway, suggesting a potential therapeutic strategy for ovarian function preservation.
Humans ; Cisplatin/adverse effects* ; MicroRNAs/genetics* ; Female ; Granulosa Cells/cytology* ; Apoptosis/drug effects* ; Drugs, Chinese Herbal/pharmacology* ; Down-Regulation ; Signal Transduction/drug effects* ; Proto-Oncogene Proteins c-akt/metabolism* ; Phosphatidylinositol 3-Kinases/metabolism* ; Cell Line, Tumor ; Cell Survival/drug effects*

Objective To evaluate the protective effect of Lycium barbarum polysaccharides(LBP)against cisplatin-induced ovarian granulosa cell injury and investigate its possible mechanisms.Methods Human granulosa-like tumor cell line(KGN)were treated with 2.5 μg/mL cisplatin for 24 h,followed by treatment with 100,500,and 1000 mg/L LBP,and the changes in cell viability,apoptosis,level of anti-Müllerian hormone(AMH),and cell ultrastructure were detected with CCK-8 assay,flow cytometry,ELISA and transmission electron microscopy.The cellular expressions of Bax,caspase-3,Bcl-2,and the PI3K/AKT pathway proteins were analyzed using Western blotting,and the expression of miR-23a was detected with RT-qPCR.KGN cell models with lentivirus-mediated miR-23a overexpression or knockdown were used to verify the therapeutic mechanism of LBP.Results Cisplatin treatment significantly inhibited cell viability,induced apoptosis,decreased AMH level,caused ultrastructural abnormalities,increased Bax and caspase-3 expression,and lowered Bcl-2 expression in KGN cells.Cisplatin also suppressed the activation of the PI3K/AKT signaling pathway and upregulated miR-23a expression in the cells.LBP intervention obviously alleviated cisplatin-induced injuries in KGN cells,and in particular,LBP treatment at the medium dose for 24 h significantly improved KGN cell viability,reduced apoptosis,enhanced their endocrine function,and ameliorated ultrastructural abnormalities.Mechanistically,medium-dose LBP obviously activated the PI3K/AKT pathway by downregulating miR-23a in cisplatin-treated cells,subsequently inhibiting Bax and caspase-3 while upregulating Bcl-2.Overexpression of miR-23a weakened while knockdown of miR-23a significantly enhanced the protective effects of LBP.Conclusion LBP alleviates cisplatin-induced apoptosis in KGN cells by inhibiting miR-23a expression and activating the PI3K/AKT pathway,suggesting a potential therapeutic strategy for ovarian function preservation.

OBJECTIVE To investigate the in vitro antitumor activity and in vivo targeting of nanopolymers co-loaded with arse-nic trioxide(ATO),an active ingredient of traditional Chinese medicine arsenic,and photosensitizer(IR780)to activate the patient's own immune system in the treatment of brain glioma in synergistically with chemotherapy and phototherapy.METHODS PLGA/AI containing ATO and IR780 was prepared by volatilization with multiple emulsion solvent.The particle size,potential and polydispersity index(PDI)were determined by dynamic light scatterometer(DLS)at different feed ratios.Transmission electron microscopy(TEM)was used to detect the morphology of PLGA/AI.Fluorescence spectrophotometer was used to investigate the spectroscopic characteris-tics.UV-vis spectrophotometer was used to determine the drug loading and encapsulation rate of IR780.Under laser irradiation,the physiological release of ATO was measured by dialysis bag method.The uptake of PLGA/AI in GL261 cells was photographed by confo-cal microscopy(CLSM).The cell uptake mechanism of PLGA/AI was investigated by flow cytometry(FCM).3D tumor spheroid mod-el was constructed to simulate the deep penetration of PLGA/AI in solid tumors.The synergistic anti-tumor effects of PLGA/AI in vitro were studied by MTT assay and dying staining.DCFH-DA staining and FCM determination of the co-expression of CD80 CD86 co-stimulatory molecules verified the immune activation induced by PLGA/AI in vitro photochemotherapy.The in vivo targeting of PLGA/AI and the tissue distribution of the main organs were investigated by means of in vivo imaging of small animals after tail vein injection.RESULTS The optimal ratio of ATO to IR780 was 1∶10,its particle size was(134.11±2.19)nm,Zeta potential was(-7.02±0.649)mV,PDI was 0.254±0.059,and it was a uniform spherical shape under TEM.The fluorescence spectra showed that IR780 was successfully loaded onto PLGA,and the drug loading and encapsulation rate of IR780 in PLGA/AI were(2.53±0.02)%and(71.26±0.38)%respectively.The results of in vitro release experiments showed that ATO could be released in response to laser light by IR780-mediated photo-dynamics.Cell uptake experiments showed that the nano-polymer could effectively enter tumor cells under clathrin-mediated endocytosis.In vitro investigation of cell viability,ROS detection and dendritic cell maturation experiment showed that it could effectively kill tumor cells by inducing a large amount of ROS to generate and activate immune cells.In vivo targe-ting and biological distribution study confirmed that PLGA/AI could effectively penetrate BBB into tumor sites.CONCLUSION The active ingredients of traditional Chinese medicine arsenic,ATO and IR780,use PLGA as carriers to form nano-polymers through the volatilization of multiple emulsion solvents,which can cross the BBB and effectively accumulate at the tumor site.Based on"strengthe-ning the healthy qi and eliminating pathogenic factors"and combined with chemotherapy and photo-immune activation,it has the po-tential for long-term anti-glioblastoma treatment.

Remarkable achievements have been made in the reform and development of higher medical education in China, as evidenced by the 315 awards for medical-related projects in the last six sessions of National Educational Achievement Awards, accounting for 8.99% of the total awards. However, several challenges persist, including an unbalanced regional distribution (e.g., the eastern region consistently accounted for more than 50% of the awards), insufficient investment in ordinary universities, imbalanced talent structure, and underdeveloped collaborative education mechanisms among universities, governments, enterprises. In response, higher medical education should fully implement the fundamental task of fostering virtue and cultivating talent, promote balanced development of medical education nationwide, increase support for ordinary universities, enhance the training of talents in fields with shortages such as public health, and improve the coordination between universities and hospitals as well as multi-party collaboration in talent education. These measures are essential to maximize the practical benefits and guiding values of the award-winning projects.

OBJECTIVE To investigate the in vitro antitumor activity and in vivo targeting of nanopolymers co-loaded with arse-nic trioxide(ATO),an active ingredient of traditional Chinese medicine arsenic,and photosensitizer(IR780)to activate the patient's own immune system in the treatment of brain glioma in synergistically with chemotherapy and phototherapy.METHODS PLGA/AI containing ATO and IR780 was prepared by volatilization with multiple emulsion solvent.The particle size,potential and polydispersity index(PDI)were determined by dynamic light scatterometer(DLS)at different feed ratios.Transmission electron microscopy(TEM)was used to detect the morphology of PLGA/AI.Fluorescence spectrophotometer was used to investigate the spectroscopic characteris-tics.UV-vis spectrophotometer was used to determine the drug loading and encapsulation rate of IR780.Under laser irradiation,the physiological release of ATO was measured by dialysis bag method.The uptake of PLGA/AI in GL261 cells was photographed by confo-cal microscopy(CLSM).The cell uptake mechanism of PLGA/AI was investigated by flow cytometry(FCM).3D tumor spheroid mod-el was constructed to simulate the deep penetration of PLGA/AI in solid tumors.The synergistic anti-tumor effects of PLGA/AI in vitro were studied by MTT assay and dying staining.DCFH-DA staining and FCM determination of the co-expression of CD80 CD86 co-stimulatory molecules verified the immune activation induced by PLGA/AI in vitro photochemotherapy.The in vivo targeting of PLGA/AI and the tissue distribution of the main organs were investigated by means of in vivo imaging of small animals after tail vein injection.RESULTS The optimal ratio of ATO to IR780 was 1∶10,its particle size was(134.11±2.19)nm,Zeta potential was(-7.02±0.649)mV,PDI was 0.254±0.059,and it was a uniform spherical shape under TEM.The fluorescence spectra showed that IR780 was successfully loaded onto PLGA,and the drug loading and encapsulation rate of IR780 in PLGA/AI were(2.53±0.02)%and(71.26±0.38)%respectively.The results of in vitro release experiments showed that ATO could be released in response to laser light by IR780-mediated photo-dynamics.Cell uptake experiments showed that the nano-polymer could effectively enter tumor cells under clathrin-mediated endocytosis.In vitro investigation of cell viability,ROS detection and dendritic cell maturation experiment showed that it could effectively kill tumor cells by inducing a large amount of ROS to generate and activate immune cells.In vivo targe-ting and biological distribution study confirmed that PLGA/AI could effectively penetrate BBB into tumor sites.CONCLUSION The active ingredients of traditional Chinese medicine arsenic,ATO and IR780,use PLGA as carriers to form nano-polymers through the volatilization of multiple emulsion solvents,which can cross the BBB and effectively accumulate at the tumor site.Based on"strengthe-ning the healthy qi and eliminating pathogenic factors"and combined with chemotherapy and photo-immune activation,it has the po-tential for long-term anti-glioblastoma treatment.

Remarkable achievements have been made in the reform and development of higher medical education in China, as evidenced by the 315 awards for medical-related projects in the last six sessions of National Educational Achievement Awards, accounting for 8.99% of the total awards. However, several challenges persist, including an unbalanced regional distribution (e.g., the eastern region consistently accounted for more than 50% of the awards), insufficient investment in ordinary universities, imbalanced talent structure, and underdeveloped collaborative education mechanisms among universities, governments, enterprises. In response, higher medical education should fully implement the fundamental task of fostering virtue and cultivating talent, promote balanced development of medical education nationwide, increase support for ordinary universities, enhance the training of talents in fields with shortages such as public health, and improve the coordination between universities and hospitals as well as multi-party collaboration in talent education. These measures are essential to maximize the practical benefits and guiding values of the award-winning projects.

Objective:To observe the in vivo microscopic histological changes during the treatment of melasma using reflectance confocal microscopy (RCM) combined with optical coherence tomography (OCT), and to explore its application value in evaluating therapeutic efficacy. Methods:Clinical data were collected from 30 melasma patients treated at the Department of Dermatology, Wuhan Hospital of Traditional Chinese and Western Medicine, Tongji Medical College, Huazhong University of Science and Technology, from January to October 2023. Retrospective analyses were performed on clinical photographs taken before treatment and at weeks 4, 8, 12, and 16 after the start of treatment, and the efficacy was evaluated by using the melasma area and severity index (MASI) scores. Meanwhile, changes in RCM imaging indicators, such as pigment distribution patterns, melanocyte morphology and spatial distribution patterns, and inflammatory infiltration in the superficial dermis in melasma lesions, were retrospectively analyzed. For clear elucidation of concepts and subsequent quantitative analysis, characteristic microscopic histological changes in melasma lesions before treatment were classified into several patterns based on RCM findings and previous literature reports. Additionally, optical coherence signals collected by OCT were used to asses characteristic vessel parameters in the lesions before and after treatment. Finally, correlations between various microscopic histological changes and clinical efficacy were analyzed using Spearman's correlation analysis.Results:With the increase in treatment duration, the proportions of pigment distribution patterns Ⅱ and Ⅲ observed by RCM gradually decreased, while the proportion of the nearly normal pattern Ⅰ markedly increased; the proportions of the 4 heterogeneous morphological patterns of melanocytes all decreased, while the pattern Ⅱ melanocytes were still focally distributed in the lesions at the last time of observation; meanwhile, the proportions of melanocyte spatial distribution patterns Ⅱ and Ⅲ decreased, which gradually restored to the normal pattern Ⅰ; the infiltration of melanophages and inflammatory cells in the superficial dermis diminished, and the proportion of the pattern Ⅲ inflammatory infiltration markedly decreased at week 4, but it lasted for a long time. The decrease rate of MASI scores was significantly correlated with the decrease rate of the proportions of pigment distribution patterns Ⅱ and Ⅲ ( r = 0.82, 0.97, respectively, both P < 0.05), melanocyte morphological patterns Ⅰ, Ⅲ, and Ⅳ ( r = 0.84, 0.83, 0.83, respectively, all P < 0.05), and inflammatory infiltration patterns Ⅰ and Ⅲ ( r = 0.85, 0.81, respectively, both P < 0.05), as well as the increase rate of the proportion of melanocyte spatial distribution pattern Ⅰ ( r = 0.89, P < 0.05), indicating that these indicators can be sensitive evaluation indicators for clinical efficacy. Moreover, OCT revealed a decrease in the average diameter and density of vessels in the lesions after treatment, and the decrease rates were correlated with the decrease rate of MASI scores ( r = 0.76, 0.78, respectively, both P < 0.05) ; there was no significant difference between the pre- and post-treatment average depth of vascular plexuses. Conclusion:During the treatment of melasma, RCM combined with OCT could serve as a non-invasive approach with objective and robust evidence for evaluating the therapeutic efficacy.

Objective To analyze differential metabolites (DMs) in the urine of workers with occupational nickel exposure using non-targeted metabolomics, and to screen differential metabolic pathways. Methods A total of 30 nickel exposed workers were selected as the exposure group, and 30 administrative staff from the same factory were selected as the control group using the judgment sampling method. Urine samples of the individuals from the two groups were collected. The ultra-high performance liquid chromatography with quadrupole time-of-flight mass spectrometry and non-targeted metabolomics were used to detect and identify metabolites. The differential metabolic profiles were compared between workers of the two groups, and key differential metabolic pathways and potential biomarkers were screened. The association of DMs and urinary nickel level were evaluated by Spearman correlation coefficients. The sensitivity and specificity of biomarkers were assessed by receiver operating characteristic (ROC) curve analysis. Results A total of 418 metabolites were identified in the urine of worker in the exposure and control groups. The result of principal component analysis and orthogonal partial least squares analysis showed that there were 128 DMs in the urine of workers in the exposure group compared with the control group. These DMs were mainly enriched in glutathione metabolism, carnitine synthesis, and amino acid and nucleotide metabolism pathways, including glycine and serine metabolism. The result of correlation analysis and ROC curve analysis revealed that 4-methylcatechol, 4-vinylphenol sulfate, 2-hydroxyphenylacetone sulfate, 2-dodecylbenzenesulfonic acid, and decylbenzenesulfonic acid could be the potential biomarkers for nickel exposure (all area under the ROC curve >0.800). Conclusion There were significant differences in the urinary metabolic profiles of workers with occupational nickel exposure. The five DMs including 4-methylcatechol, 4-vinylphenol sulfate, 2-hydroxyphenylacetone sulfate, 2-dodecylbenzenesulfonic acid, and decylbenzenesulfonic acid. These DMs could be potential biomarkers of occupational nickel exposure.