Objectives miR-207 has been identified as being expressed in natural killer (NK) cell exosomes that play a role in disease progression; however, to date, there are no studies specifically linking miR-207 to tuberculosis (TB). Methods Bioinformatics methods employed for prediction, followed by a dual luciferase reporter assay to determine whether lysosome-associated membrane protein 2 (LAMP2) is targeted by miR-207. The experiments were divided into four groups using the liposome transfection method (OP-LAMP2 group: co-transfected with miR-207 mimics and LAMP2 overexpression plasmid; EP group: co-transfected with mimics NC and null-loaded plasmid; siLAMP2 group: transfected with siLAMP2; and siLAMP2-NC group: transfected with siLAMP2-NC). TB infection was modeled using H37Ra-infected Ana-1 cells. The impact of LAMP2 on intracellular mycobacterial load and clearance of extracellular residual mycobacteria were assessed by tuberculosis colony-forming unit counting. Flow cytometry was used to assess the total apoptosis rate. Real-time fluorescent quantitative PCR was conducted to determine the relative expression of LAMP2, apoptosis genes, pyroptosis genes, and autophagy genes. Western blot analysis was performed to measure the relative expression of LAMP2 proteins, apoptosis proteins, pyroptosis proteins, and autophagy proteins. Results Dual luciferase reporter assay test showed that there was a targeting relationship between LAMP2 and miR-207. The transfection model was successfully constructed under real-time fluorescent quantitative PCR and Western blot statistical analysis, and microscopic observation. The infection model was successfully established under microscopic observation. Colony forming unit counting revealed that the number of colonies in the OP-LAMP2 group was lower than that in the EP group, while the number of colonies in the siLAMP2 group was higher than that in the siLAMP2-NC group. Flow cytometry assay revealed that the total apoptosis in OP-LAMP2 group was lower than that in EP group, and the total apoptosis in siLAMP2 group was higher than that in siLAMP2-NC group. Real-time fluorescence quantitative PCR and Western blot analysis revealed that the relative expression of apoptosis and pyroptosis-related proteins and genes in the control group was lower in the OP-LAMP2 group compared to the EP group, and higher in the siLAMP2 group compared to the siLAMP2-NC group. Real-time fluorescence quantitative PCR detected that the relative expression of autophagy positively regulated genes Microtubule-associated protein 1 light chain 3(LC3)and Beclin1 in the OP-LAMP2 group was higher in the OP-LAMP2 group compared to the EP group, and lower in the siLAMP2 group compared to the siLAMP2-NC group, while the relative expression of negatively regulated autophagy genes followed the opposite trend to that of autophagy positively regulated genes. The relative expression of autophagy-related proteins was consistent with the trend of autophagy genes. Conclusions miR-207 enhances macrophage apoptosis, cellular pyroptosis and inhibits autophagy, promoting survival of Mycobacterium tuberculosis by targeting the autophagy-related protein LAMP2, thus offering a novel therapeutic direction for tuberculosis.
Lysosomal-Associated Membrane Protein 2/metabolism* ; MicroRNAs/metabolism* ; Mycobacterium tuberculosis/physiology* ; Autophagy/genetics* ; Humans ; Macrophages/metabolism* ; Apoptosis/genetics* ; Tuberculosis/metabolism* ; Cell Line ; Pyroptosis/genetics*

Pulmonary diseases, as a prevalent category of respiratory system disorders, have become a significant global public health concern. The increasing incidence of these diseases, caused by environmental pollution and occupational hazards, poses a substantial threat to human health and the overall quality of life. Mesenchymal stem cells (MSCs) are known for their remarkable immunomodulatory, anti-bacterial, and anti-apoptotic capabilities. Exosomes derived from MSCs, carrying a diverse array of proteins, lipids, nucleic acids, and other bio-active molecules, have demonstrated considerable therapeutic potential in treating pulmonary diseases, and have come to the forefront of medical research. This review summarized the therapeutic role of exosomes derived from various sources of mesenchymal stem cells in the context of pulmonary diseases, aiming to provide a robust foundation for their clinical application in diagnosis and treatment.
Exosomes/transplantation* ; Humans ; Mesenchymal Stem Cells/metabolism* ; Lung Diseases/therapy* ; Animals

Objective To screen the key N6-methyladenosine(m6A)methylation related genes in renal clear cell carcinoma(ccRCC),and to study their expression and relationship with the prognosis,migration and invasion of renal clear cell carcinoma.Methods The RNA sequencing data and clinical data of ccRCC and ad-jacent tissues were downloaded from the Cancer Genome Atlas(TCGA)and GTEx(Genotype-Tissue Expres-sion).The expression profile and prognosis were analyzed with R 4.1.1,and the key genes were screened.Clinical specimens of 10 patients with ccRCC were collected.The mRNA and protein expressions were detec-ted by RT-qPCR and immunohistochemistry,respectively.In human ccRCC cell line RCC23,siRNA was used to knock down key genes,and CCK-8 was used to detect the survival rate of cells.Scratch test and Trans well test were used to detect the migration and invasion of cells,respectively.Results Among the 19 m6A methyl-ation related genes,only insulin-like growth factor 2 mRNA binding protein 3(IGF2BP3)was highly ex-pressed in cancer tissues,and the high expression was significantly positively correlated with poor prognosis.The high expression of IGF2BP3 was verified in clinical specimens by RT-qPCR and immunohistochemistry.After knockdown of IGF2BP3 by siRNA,the survival rate of RCC23 cells decreased significantly,and the mi-gration and invasion ability of cut cells decreased.Conclusion These results suggest that IGF2BP3 may be an effective biomarker and potential drug target for predicting the prognosis of patients with ccRCC.

Objective To prepare a thermosensitive hydrogel scaffold loaded with bone marrow mesenchymal stem cells（BMSCs） and resveratrol liposomes （RSV-LIP） to form a therapeutic unit and evaluate its treatment efficacy for traumatic brain injury （TBI）. Methods BMSCs were extracted from rats, and RSV-LIP was prepared and characterized. Cell models were constructed to investigate the pharmacological effects of BMSCs combined with RSV-LIP. BMSCs and RSV-LIP were then loaded into the hydrogel, and a TBI mouse model was established to evaluate the therapeutic effects of the hydrogel. Results The RSV-LIP had a particle size of 127.8 nm, a Zeta potential of −4.9 mV, an encapsulation efficiency of 78.50%, and a drug loading content of 2.37%. Live-dead staining indicated good biocompatibility of the hydrogel. The combination of BMSCs and RSV-LIP significantly inhibited TNF-α and reduced ROS levels, promoting cell migration in scratch assays. Compared to the control group, the hydrogel group showed significantly lower mNSS scores （P<0.01）, higher hanging scores （P<0.001）, and reduced stepping errors （P<0.001）. Conclusion The combination of BMSCs and RSV-LIP exhibited antioxidative stress, anti-inflammatory, and neurogenic cell migration-promoting effects. When loaded into a hydrogel scaffold and locally implanted, it could improve the motor and sensory functions in TBI mice.

Objective To investigate the in vitro metabolites and metabolic pathways of phenethylamine new psychoactive substances 2C-B-FLY and 25B-NBOH in human liver microsomes.Methods The human liver microsome incubation model was established,and the samples were centrifuged,blow-dried and re-dissolved after 3 h of co-incubation in a water bath at 37℃.The samples were detected by ultra-high performance liquid chromatography Q Exactive mass spectrometry in ESI+mode,and the parent drug and its metabolites were examined by applying Full scan-ddMS2 data-dependent acquisition mode.Results After in vitro incubation with human liver microsomes,2C-B-FLY was metabolised to produce a total of four phase I metabolites and one acetylated phase II metabolite,which underwent metabolic reactions such as hydroxylation,debromination,and N-acetylation.25B-NBOH was metabolised to produce a total of eight phase I metabolites and two glucuronidated phase II metabolites,which comprised the major biotransformation reactions of O-demethylation,hydroxylation,amine dehydrogenation,N-dealkylation and glucuronidation.Conclusion This study describes the metabolites and metabolic pathways of 2C-B-FLY and 25B-NBOH in a human liver microsomal incubation system,providing a scientific basis for the in vivo detection of new psychoactive substance of the phenethylamine group.

Ferroptosis is a newly discovered mode of programmed cell death characterized by the accumulation of intracellular iron-dependent lipid peroxidation.Current research has mainly focused on the role of ferroptosis in the field of cancer,but increasing evidence shows that ferroptosis is also related to the occurrence of infectious diseases.Ferroptosis has accordingly been detected in cases of COVID-19,tuberculosis,and cryptococcal meningitis,as well as other diseases.This article reviews the role of ferroptosis in infectious diseases,to provide new ideas for the prevention and treatment of ferroptosis-related infectious diseases.

Objective:To discuss the effect of NOD-like receptor protein 3(NLRP3)inflammasome on the renal interstitial fibrosis in the unilateral ureteral obstruction(UUO)model rats,and to clarify its potential mechanism.Methods:Thirty healthy male Wistar rats were randomly divided into sham operation group(n=6)and UUO group(n=24).The rats in sham operation group underwent the dissection of the ureter without ligation,while the rats in UUO group were sacrificed on the 3rd,7th,and 14th days after operation,and based on the treatment duration,the rats were divided into UUO 3 d group(n=8),UUO 7 d group(n=8),and UUO 14 d group(n=8).HE staining and Masson staining were used to observe the pathomorphology of kidney tissue of the rats in various groups;reagent kits were used to detect the levels of malondialdehyde(MDA),activities of superoxide dismutase(SOD),and levels of hydroxyproline(HYP)in kidney tissue of the rats in various groups;immunohistochemistry method was used to detect the expression levels of α-smooth muscle actin(α-SMA)and transforming growth factor-β1(TGF-β1)proteins in kidney tissue of the rats in various groups;Western blotting method was used to detect the expression levels of NLRP3 protein in kidney tissue of the rats in various groups.Results:The HE staining results showed significant tubular dilation,interstitial edema,and widening,with increased infiltration of inflammatory cells,and shedding of epithelial cells was seen in parts of the tubular lumina of the rats in UUO group.Compared with sham operation group,the interstitial fibrosis scores of the rats in UUO 3 d,UUO 7 d,and UUO 14 d groups were significantly increased(P<0.05);compared with UUO 3 d group and UUO 7 d group,the interstitial fibrosis score of the rats in UUO 14 d group was significantly decreased(P<0.05).The Masson staining results showed that in UUO group,there was evident infiltration of inflammatory cells in the renal interstitium and a noticeable increase in fibrotic tissue proliferation;with the increasing of duration of UUO,some tubular structures disappeared,and the interstitial widened further with gradually increasing collagen deposition,particularly at the corticomedullary junction.Compared with sham operation group,the interstitial fibrosis scores of the rats in UUO 3 d,UUO 7 d,and UUO 14 d groups were significantly increased(P<0.05);and compared with UUO 3 d and UUO 7 d groups,the interstitial fibrosis score of the rats in UUO 14 d group was significantly decreased(P<0.05).Compared with sham operation group,the levels of MDA in obstructed kidney tissue of the rats in UUO 3 d,UUO 7 d,and UUO 14 d groups were significantly increased(P<0.05),and the SOD activities were significantly decreased(P<0.05).Compared with sham operation group,the levels of HYP in obstructed kidney tissue of the rats in UUO 3 d,UUO 7 d,and UUO 14 d groups were also significantly increased(P<0.05);compared with UUO 3 d group,the level of HYP in obstructed kidney tissue of the rats in UUO 14 d group was significantly increased(P<0.05).The immunohistochemistry results showed that compared with sham operation group,the expression levels of α-SMA protein in kidney tissue of the rats in UUO 3 d,UUO 7 d,and UUO 14 d groups were significant increased(P<0.05);compared with UUO 3 d and UUO 7 d groups,the expression levels of α-SMA protein in kidney tissue of the rats in UUO 14 d group was significantly increased(P<0.05);compared with sham operation group,the expression levels of TGF-β1 protein in renal tubular epithelial cells and renal tubule interstitial tissue of the rats in UUO 3 d,UUO 7 d,and UUO 14 d groups were also significantly increased(P<0.05);compared with UUO 3 d group,the expression levels of TGF-β1 protein in the tubular epithelial cells and renal tubule interstitial tissue of the rats in UUO 14 d group were significantly decreased(P<0.05).The Western blotting results showed that compared with sham operation group,the expression levels of NLRP3 protein in kidney tissue of the rats in UUO 7 d and UUO 14 d groups were significantly increased(P<0.05).Conclusion:The NLRP3 inflammasome plays a critical role in renal fibrosis of the UUO rats,and its mechanism may be related to the increasing of oxidative stress and the increasing of expression level of TGF-β1 protein.

Objective miR-207 is differentially expressed in many diseases.We investigated the mechanism by which miR-207 overexpression regulates the survival of Mycobacterium tuberculosis(H37Ra)in macrophages,to provide a theoretical basis for the targeted therapy of tuberculosis.Methods Macrophages were divided into four groups:blank(Ana-1 cells),control(cells infected with H37Ra),mi(infected with H37Ra and transfected with miRNA-207 mimics),and mi-NC(infected with H37Ra and transfected with NC mimics)groups.A model of tuberculosis infection was established using H37Ra-infected Ana-1 cells,and miRNA-207 and NC mimics were transfected into Ana-1 cells using the liposome transfection method.Tuberculosis colony-forming units were counted to assess the effect of miR-207 on intracellular mycobacterial load and clearance of extracellular residual mycobacteria.The total apoptosis rate was detected by flow cytometry.The relative expression levels of miR-207 and apoptosis,pyroptosis,inflammation,and autophagy genes were measured by quantitative real-time polymerase chain reaction(qPCR).Relative expression levels of apoptosis,pyroptosis,and autophagy proteins were detected by Western blot.Fluorescence microscopy and multifunctional enzyme labeling were used to detect the fluorescence intensity of intracellular reactive oxygen species(ROS)and lactate dehydrogenase(LDH).Results Successful establishment of the infection model was observed under the microscope.qPCR showed that miR-207 expression was lower in the control compared with the blank group(P＜0.01),indicating differential expression between these two groups.miR-207 expression was significantly higher in the mi compared with the mi-NC group(P＜0.0001),indicating successful establishment of the transfection model.The number of colonies and total apoptosis were both higher in the mi group compared with the mi-NC and control groups(P＜0.001).qPCR and Western blot showed that the relative expression levels of apoptotic genes and proteins were higher in the control group than in the blank group(P＜0.05),and higher in the mi group than in the mi-NC group(P＜0.05).The relative expression levels of inflammatory genes were higher in the control than in the blank group(P＜0.001).The relative expression levels of inflammatory genes were higher in the mi group than in the mi-NC group(P＜0.05),and the relative expression levels of pyroptosis genes and proteins were higher in the control group compared with the blank group(P＜0.01)and higher in the mi group compared with the mi-NC group(P＜0.05).The relative expression levels of the autophagy positively-regulated genes LC3 and Beclin1 were higher in the control compared with the blank group(P＜0.0001),and lower in the mi than in the mi-NC group(P＜0.05),while negatively-regulated autophagy genes showed the opposite trend.Autophagy-related proteins showed similar trends to the autophagy genes.ROS fluorescence intensity was higher in the control compared with the blank group(P＜0.05),and higher in the mi compared with the mi-NC group(P＜0.001).LDH content was higher in the control than in the blank group(P＜0.01),but there was no significant difference between the mi and mi-NC groups(P＞0.05).Conclusions miR-207 overexpression promotes apoptosis,cellular pyroptosis,and inflammation,inhibits autophagy,and favors H37Ra survival.These result provide a potential new direction for the treatment of tuberculosis.

Objective miR-207 is differentially expressed in many diseases.We investigated the mechanism by which miR-207 overexpression regulates the survival of Mycobacterium tuberculosis(H37Ra)in macrophages,to provide a theoretical basis for the targeted therapy of tuberculosis.Methods Macrophages were divided into four groups:blank(Ana-1 cells),control(cells infected with H37Ra),mi(infected with H37Ra and transfected with miRNA-207 mimics),and mi-NC(infected with H37Ra and transfected with NC mimics)groups.A model of tuberculosis infection was established using H37Ra-infected Ana-1 cells,and miRNA-207 and NC mimics were transfected into Ana-1 cells using the liposome transfection method.Tuberculosis colony-forming units were counted to assess the effect of miR-207 on intracellular mycobacterial load and clearance of extracellular residual mycobacteria.The total apoptosis rate was detected by flow cytometry.The relative expression levels of miR-207 and apoptosis,pyroptosis,inflammation,and autophagy genes were measured by quantitative real-time polymerase chain reaction(qPCR).Relative expression levels of apoptosis,pyroptosis,and autophagy proteins were detected by Western blot.Fluorescence microscopy and multifunctional enzyme labeling were used to detect the fluorescence intensity of intracellular reactive oxygen species(ROS)and lactate dehydrogenase(LDH).Results Successful establishment of the infection model was observed under the microscope.qPCR showed that miR-207 expression was lower in the control compared with the blank group(P＜0.01),indicating differential expression between these two groups.miR-207 expression was significantly higher in the mi compared with the mi-NC group(P＜0.0001),indicating successful establishment of the transfection model.The number of colonies and total apoptosis were both higher in the mi group compared with the mi-NC and control groups(P＜0.001).qPCR and Western blot showed that the relative expression levels of apoptotic genes and proteins were higher in the control group than in the blank group(P＜0.05),and higher in the mi group than in the mi-NC group(P＜0.05).The relative expression levels of inflammatory genes were higher in the control than in the blank group(P＜0.001).The relative expression levels of inflammatory genes were higher in the mi group than in the mi-NC group(P＜0.05),and the relative expression levels of pyroptosis genes and proteins were higher in the control group compared with the blank group(P＜0.01)and higher in the mi group compared with the mi-NC group(P＜0.05).The relative expression levels of the autophagy positively-regulated genes LC3 and Beclin1 were higher in the control compared with the blank group(P＜0.0001),and lower in the mi than in the mi-NC group(P＜0.05),while negatively-regulated autophagy genes showed the opposite trend.Autophagy-related proteins showed similar trends to the autophagy genes.ROS fluorescence intensity was higher in the control compared with the blank group(P＜0.05),and higher in the mi compared with the mi-NC group(P＜0.001).LDH content was higher in the control than in the blank group(P＜0.01),but there was no significant difference between the mi and mi-NC groups(P＞0.05).Conclusions miR-207 overexpression promotes apoptosis,cellular pyroptosis,and inflammation,inhibits autophagy,and favors H37Ra survival.These result provide a potential new direction for the treatment of tuberculosis.

Objective To evaluate the clinical efficacy of comprehensive treatment for synchronous primary advanced gastric and esophageal cancer by propensity score matching(PSM).Methods A total of 2 551 patients with advanced gastric cancer admitted to Jiangsu Province Hospital of Chinese Medicine from January 2013 to December 2022 were retrospectively analyzed.Among them,45 patients with synchronous primary esophageal cancer were distributed to the observation group,and 2 506 patients without esophageal cancer were distributed to the control group.Through the PSM method,the control group was matched with the observation group and the equilibrium samples of covariates between two groups were obtained.The overall survival(OS)between the two groups were compared.Results Both observation and control group contained 45 patients in this study.According to the treatment regimen,the patients in the observation group was divided into radical resection treatment subgroup(n=22)and chemoradiotherapy(CRT)subgroup(n=23).In the radical resection subgroup,4 patients underwent the simultaneous surgical resection of gastric and esophageal tumors through proximal gastrectomy with the Ivor Lewis operation.Eighteen patients underwent endoscopic submucosal dissection(ESD)of their esophageal tumors and gastric cancer radical resection.Radical resection of gastric cancer combined with preoperative chemoradiotherapy of esophageal cancer was performed in the CRT subgroup.Survival analysis showed that OS in the observation group was significantly shorter than that in the control group(P=0.042)and there was no significant difference in OS between the radical resection subgroup and the control group(P=0.799).The 1-,3-,and 5-year survival rates of the patients in the CRT subgroup were significantly lower than those of the control group(P=0.003).While the 1-,3-,and 5-year survival rates of the patients in the radical resection subgroup were not statistically significant,compared to those of the CRT subgroup(P=0.071).Conclusions Multidisciplinary and comprehensive treatment can significantly improve the prognosis of patients with synchronous primary advanced gastric and esophageal cancer.Radical resection of gastric cancer combined with ESD of esophageal cancer is an optional treatment for patients with gastric cancer complicated with early esophageal cancer.Radical resection of gastric cancer combined with CRT of esophageal cancer can improve the prognosis of patients with advanced gastric cancer complicated with unresectable esophageal cancer.