AIM: To identify the primary active components and underlying mechanisms of Yijing Decoction(YJD)in treating early diabetic retinopathy(DR)based on liquid chromatography-mass spectrometry and network pharmacology.METHODS: Active components of YJD were characterized through LC-MS. Components with optimal ADME(absorption, distribution, metabolism, excretion)properties were selected as key bioactive candidates. Network pharmacology approaches were employed to predict YJD-DR therapeutic targets. Protein-protein interaction(PPI)networks, gene ontology(GO)enrichment analysis, and Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway analysis were subsequently conducted to predict core targets and networks. Critical targets and pathways were experimentally validated through Western blot.RESULTS: Ten core therapeutic targets were identified, including TNF, Alb, EGFR, STAT3, PTGS2, ESR1, PPAR, MMP9, TLR4, and MAPK. YJD was related to cancer-related signaling, fluid shear stress and atherosclerosis, and neurodegenerative diseases, encompassing key biological processes such as inflammatory response regulation, programmed cell death activation, and enhanced cell migration. Furthermore, Western blot analysis confirmed that YJD significantly inhibited high glucose-induced phosphorylation of STAT3(P-STAT3/STAT3)and ERK(P-ERK/ERK)in rat retinal microvascular endothelial cells.CONCLUSION: This study revealed YJD's pharmacodynamical basis and its multi-component, multi-target, and multi-paths pharmacology. YJD exerts therapeutic effects on DR by coordinately regulating critical signaling pathways and alleviating intraocular inflammation, thus preserving retinal vascular endothelial cells, maintaining blood-retinal barrier integrity, and facilitating retinal neurovascular repair.

Objective:To investigate whether exosomes derived from atorvastatin (ATV)-pretreated human umbilical cord mesenchymal stem cells (ATV-MSC-EXO) alleviate high glucose-induced injury in human retinal vascular endothelial cells (HREC) via the protein kinase B (AKT)/endothelial nitric oxide synthase (eNOS) signaling pathway.Methods:The optimal pretreatment concentration of ATV was determined using the cell counting Kit-8 (CCK-8) assay. Exosomes derived from mesenchymal stem cells (MSC-EXO) and ATV-pretreated MSC (ATV-MSC-EXO) were isolated and extracted, and their morphology and surface markers were characterized by transmission electron microscopy, nanoparticle tracking analysis, and Western blotting (WB). The uptake capacity of exosomes by human retinal vascular endothelial cells (HREC) was evaluated using a fluorescence labeling assay. In vitro cultured HREC were divided into the following groups: normal control group (NC group), high glucose group (HG group), high glucose+MSC-EXO group (MSC-EXO group), high glucose+ATV-MSC-EXO group (ATV-MSC-EXO group), high glucose+ATV-MSC-EXO+AKT inhibitor group (ATV-MSC-EXO-MK-2206-2HCL group), and high glucose+ATV-MSC-EXO+eNOS inhibitor group (ATV-MSC-EXO-L-NAME group). Cell proliferation and apoptosis were detected using CCK-8 and flow cytometry, respectively. The protein expression levels of B-cell lymphoma/leukemia-2 (Bcl-2), Bcl-2-associated protein (Bax), and Caspase-3 were measured by WB. In addition, the regulatory effects of ATV-MSC-EXO on the AKT/eNOS signaling pathway and its downstream functional molecules were analyzed by detecting the phosphorylation levels of AKT (P-AKT/AKT) and eNOS (P-eNOS/eNOS) via WB, the mRNA expression levels of AKT and eNOS by quantitative real-time polymerase chain reaction, and the concentrations of nitric oxide (NO) and endothelin-1 (ET-1) using commercial NO and ET-1 assay kits.Results:The optimal pretreatment concentration of ATV was 1 μmol/L. ATV-MSC-EXO exhibited similar morphology and particle size to MSC-EXO and were efficiently taken up by HREC. Under high glucose conditions, ATV-MSC-EXO significantly enhanced the viability of HREC ( F=83.24, P<0.000 1) and inhibited apoptosis ( F=77.39, P<0.000 1). WB analysis further confirmed that ATV-MSC-EXO upregulated the expression of the anti-apoptotic protein Bcl-2 ( F=53.17), while downregulating the pro-apoptotic proteins Bax ( F=36.49) and Caspase-3 ( F=60.75) ( P<0.001). In addition, ATV-MSC-EXO markedly increased the protein levels of P-AKT/AKT ( F=107.60) and P-eNOS/eNOS ( F=38.59), as well as the relative mRNA expression of AKT, eNOS ( F=203.60, 315.00; P<0.000 1). Furthermore, ATV-MSC-EXO promoted NO production ( F=407.40) and suppressed the relative expression of ET-1 ( F=49.76) ( P<0.000 1). Conclusion:ATV-MSC-EXO enhances the viability and inhibits apoptosis of HREC under high glucose conditions by activating the AKT/eNOS signaling pathway.

BACKGROUND:Studies have shown that mitochondrial oxidative stress has an important role in the development of knee osteoarthritis,and Hemin can regulate the expression of mitochondria-related proteins. OBJECTIVE:To study the regulatory effect of Hemin on oxidative stress in mouse chondrocytes and its interventional effect and mechanism in knee osteoarthritis. METHODS:(1)In vitro cell experiment:Primary chondrocytes from C57BL/6 mice were extracted and induced with 10 ng/mL interleukin-1β to construct an in vitro chondrocyte model of osteoarthritis.The optimal concentration of Hemin(0,1,10,20,40,80,and 160 μmol/L)for the intervention in mouse chondrocytes was determined by cell counting kit-8 method.Chondrocytes were randomly divided into control group,model group(interleukin-1β)and Hemin group(interleukin-1β+Hemin).Reactive oxygen species,mitochondrial membrane potential and apoptosis of chondrocytes in each group were detected.(2)In vivo experiment:Adult C57BL/6 mice were randomly divided into normal group,model group(osteoarthritis)and Hemin group(osteoarthritis+Hemin),with eight mice in each group.After 4 weeks of Hemin treatment,the behavioral test and histopathological observation of the knee joint were performed in each group.Changes in extracellular matrix-related protein expression and apoptosis in chondrocytes and the expression level of Nrf2/HO-1 protein in cartilage tissue were detected. RESULTS AND CONCLUSION:In vitro experiment:the optimal concentration of Hemin on primary chondrocytes was 40 μmol/L.Compared with the model group,the level of reactive oxygen species was significantly reduced,the mitochondrial membrane potential was significantly improved,and the apoptosis of chondrocytes was reduced in the hemin-treated interleukin-1β-induced chondrocytes.In vivo experiment:After 4 weeks of treatment,compared with the model group,the lower limb function of mice in the Hemin group was significantly improved,the histopathological score was significantly improved,and the apoptosis of knee chondrocytes was significantly reduced.All these findings indicate that Hemin can alleviate oxidative stress,restore mitochondrial function and reduce apoptosis in mouse chondrocytes induced by interleukin-1β.Hemin can improve extracellular matrix degradation,promote chondrocyte anabolism,reduce catabolism and reduce chondrocyte apoptosis in knee osteoarthritis.It may act by activating the chondrocyte Nrf2/HO-1 signaling pathway in the inflammatory environment.

Endometrial cancer is one of the common malignant tumors of the female reproductive system.Its incidence rate in China is second only to cervical cancer,posing a serious threat to women's life and health,and the onset shows a trend of affecting younger individuals.Currently,the primary treatment for endometrial cancer is surgical intervention,and some postoperative patients and those with postoperative recurrence require adjuvant radiotherapy.Pelvic external beam radiotherapy(EBRT)and vaginal brachytherapy(VBT)are two modalities of postoperative radiotherapy.The appropriate postoperative radiotherapy approach should be selected based on individual conditions and guideline recommendations.Most patients can achieve satisfactory treatment outcomes through surgery and postoperative adjuvant radiotherapy.However,the specific indications for postoperative radiotherapy in endometrial cancer patients and the radiotherapy principles for those with postoperative recurrence are relatively complex,and recommendations for radiotherapy modalities in postoperative patients vary slightly across different guidelines.This article reviews the treatment principles for endometrial cancer patients in recent years,as well as the indications for postoperative and post operative recurrent radiotherapy and the radiotherapy techniques,aiming to provide a reference for radiotherapy in the endometrial cancer patients.

Objective:To investigate whether exosomes derived from atorvastatin (ATV)-pretreated human umbilical cord mesenchymal stem cells (ATV-MSC-EXO) alleviate high glucose-induced injury in human retinal vascular endothelial cells (HREC) via the protein kinase B (AKT)/endothelial nitric oxide synthase (eNOS) signaling pathway.Methods:The optimal pretreatment concentration of ATV was determined using the cell counting Kit-8 (CCK-8) assay. Exosomes derived from mesenchymal stem cells (MSC-EXO) and ATV-pretreated MSC (ATV-MSC-EXO) were isolated and extracted, and their morphology and surface markers were characterized by transmission electron microscopy, nanoparticle tracking analysis, and Western blotting (WB). The uptake capacity of exosomes by human retinal vascular endothelial cells (HREC) was evaluated using a fluorescence labeling assay. In vitro cultured HREC were divided into the following groups: normal control group (NC group), high glucose group (HG group), high glucose+MSC-EXO group (MSC-EXO group), high glucose+ATV-MSC-EXO group (ATV-MSC-EXO group), high glucose+ATV-MSC-EXO+AKT inhibitor group (ATV-MSC-EXO-MK-2206-2HCL group), and high glucose+ATV-MSC-EXO+eNOS inhibitor group (ATV-MSC-EXO-L-NAME group). Cell proliferation and apoptosis were detected using CCK-8 and flow cytometry, respectively. The protein expression levels of B-cell lymphoma/leukemia-2 (Bcl-2), Bcl-2-associated protein (Bax), and Caspase-3 were measured by WB. In addition, the regulatory effects of ATV-MSC-EXO on the AKT/eNOS signaling pathway and its downstream functional molecules were analyzed by detecting the phosphorylation levels of AKT (P-AKT/AKT) and eNOS (P-eNOS/eNOS) via WB, the mRNA expression levels of AKT and eNOS by quantitative real-time polymerase chain reaction, and the concentrations of nitric oxide (NO) and endothelin-1 (ET-1) using commercial NO and ET-1 assay kits.Results:The optimal pretreatment concentration of ATV was 1 μmol/L. ATV-MSC-EXO exhibited similar morphology and particle size to MSC-EXO and were efficiently taken up by HREC. Under high glucose conditions, ATV-MSC-EXO significantly enhanced the viability of HREC ( F=83.24, P<0.000 1) and inhibited apoptosis ( F=77.39, P<0.000 1). WB analysis further confirmed that ATV-MSC-EXO upregulated the expression of the anti-apoptotic protein Bcl-2 ( F=53.17), while downregulating the pro-apoptotic proteins Bax ( F=36.49) and Caspase-3 ( F=60.75) ( P<0.001). In addition, ATV-MSC-EXO markedly increased the protein levels of P-AKT/AKT ( F=107.60) and P-eNOS/eNOS ( F=38.59), as well as the relative mRNA expression of AKT, eNOS ( F=203.60, 315.00; P<0.000 1). Furthermore, ATV-MSC-EXO promoted NO production ( F=407.40) and suppressed the relative expression of ET-1 ( F=49.76) ( P<0.000 1). Conclusion:ATV-MSC-EXO enhances the viability and inhibits apoptosis of HREC under high glucose conditions by activating the AKT/eNOS signaling pathway.

Objective:To explore the involvement of miR-126 and the role of mammalian target of rapamycin (mTOR)/hypoxia-induced factor 1 α (HIF-1 α) pathway in regulating human umbilical cord mesenchymal stem cells (hUCMSCs) exosomes (Exo) on vascular endothelial growth factor (VEGF)-A levels in high glucose-induced human retinal vascular endothelial cells (HRECs).Methods:The hREC was cultured in EGM-2-MV endothelial cell culture medium with 30 mmol/L glucose and placed in hypoxic cell incubator by 1% oxygen concentration. The cell model of high glucose and low oxygen was established. After modeling, divided HRECs into Exo group, phosphate buffer saline (PBS) group, PBS+anti-miR126 group, Exo+anti-miR126 group, PBS+anti-mTOR group, and PBS+anti-HIF-1 α group. High-glucose and hypoxia-induced hREC in the PBS and Exo groups were respectively co-cultured with PBS and 100 μg/ml hUCMSC Exo. PBS+anti-mTOR group, PBS+anti-HIF-1 α group: 500 nmol/L mTOR inhibitor ADZ2014, 25 μmol/L HIF-1 α inhibitor YC-1 pretreatment for hREC 12 h, and then co-culture with PBS after High-glucose and hypoxia-induced. PBS+anti-miR126 group, Exo+anti-miR126 group: miR-126 LNA power inhibitor probe was transfected with high glucose, and co-cultured with PBS and hUCMSC Exo 6 h after transfection. Real-time polymerase chain reaction (qPCR) measured miRNA-126 expression levels in PBS, and Exo groups for 0, 8, 16 and 24 h. After 24 h of co-culture, the levels of mTOR and HIF-1 α in the cells of PBS and Exo groups were detected by immunofluorescence, Western blot and qPCR, respectively. Western blot, qPCR detection of VEGF-A expression levels in cells of the PBS+anti-mTOR and PBS+anti-HIF-1 α groups. The expression of VE GF-A, mTOR, and HIF-1 α mRNA was measured in cells of PBS+anti-miR126 group and Exo+anti-miR126 group by qPCR. Comparison between two groups was performed by t-test; one-way ANOVA was used for comparison between multiple groups. Results:At 0, 8, 16 and 24 h, the relative mRNA expression of miR-126 gradually increased in the Exo group ( F=95.900, P<0.05). Compared with the PBS group, The mTOR, HIF-1 α protein ( t=3.466, 6.804), mRNA in HRECs in the Exo group, VEGF-A mRNA expression ( t=8.642, 7.897, 6.099) were all downregulated, the difference was statistically significant ( P<0.05). The relative expression level of VEGF-A protein ( t=3.337, 7.380) and mRNA ( t=8.515, 10.400) was decreased in HRECs of the anti-mTOR+PBS group and anti-HIF-1 α+PBS group, differences were statistically significant ( P<0.05). The relative expression of VEGF-A, mTOR, and HIF-1 α mRNA was significantly increased in the cells of the Exo+anti-miR126 group, the differences were all statistically significant ( t=4.664, 6.136, 6.247; P<0.05). Conclusions:miR-126 plays a role in regulating the effect of hUCMSCs exosomes on VEGF-A levels in high glucose-induced HRECs via mTOR-HIF-1 α pathway.

Objective:To evaluate the effect of urethral stent tube irrigation in preventing postoperative complications in adolescents with hypospadias.Methods:A retrospective analysis was conducted using the clinical data of adolescents with hypospadias treated at the Department of Urology, Children’s Hospital of Chongqing Medical University, from April 2016 to July 2023. Patients were divided into control group and study group according to whether urethral stent tube was inserted in urethra during operation. All surgeries were performed by the same surgeon. For two days before surgery, patients were instructed to cleanse the surgical site daily with body wash and a 5% povidone-iodine solution. In the control group, patients received postoperative care with an indwelling urinary catheter. In the study group, in addition to the urinary catheter, a urethral stent tube was placed in the posterior urethra, and postoperative urethral irrigation with saline was performed. Postoperative complications, including wound infection, urethral fistula, urethral stricture, and urethral diverticulum, were monitored. Categorical data were expressed as frequencies and percentages and were analyzed using the χ2 test or Monte Carlo-based χ2 test. Continuous data with normal distribution are presented as Mean±SD and were analyzed using the t-test. Non-normally distributed continuous data were expressed as M ( Q1, Q3) and were analyzed using the Wilcoxon rank-sum test. P<0.05 was considered statistically significant. Results:A total of 76 patients were enrolled in the study, with 43 patients in the control group [median age 12.9(12.2, 13.7) years, reconstructed urethral length (3.88±1.86) cm] and 33 patients in the study group [median age 12.5(11.3, 14.1) years, reconstructed urethral length (3.30±1.45)cm]. Postoperatively, 32 patients (74.4%) in the control group experienced complications, including 24 cases (55.8%) of wound infection, 16 cases (37.2%) of urethral fistula, 6 cases (14.0%) of urethral stricture, and 2 cases (4.7%) of diverticulum. In the study group, only 1 patient (3.0%) developed both wound infection and urethral fistula. The incidence of wound infection, urethral fistula, and overall complications was significantly lower in the study group than in the control group (all P<0.01). Conclusion:Postoperative urethral stent irrigation in adolescents with hypospadias significantly reduces the incidence of wound infection and urethral fistula.

Objective To investigate the effect and mechanism of long non-coding RNA(lncRNA)maternally expressed gene 3(MEG3)on the alleviation of myocardial ischemia-reperfusion(IR)injury by sponging miR-21 in rats after ovariectomy(OVX).Methods A total of 108 female SD rats were subjected,and 48 of them were randomly divided into sham group,OVX group,IR group and combined group 1(OVX+IR),with 12 rats in each group.And the remaining 60 rats were given an injection with blank adeno-associated virus(AAV,negative control),lncRNA MEG3 AAV and miR-21 AAV,respectively through the tail vein before OVX and myocardial IR modeling,and then consequently assigned into negative sham group,negative model group,lncRNA MEG3 group,miR-21 group,and combined group 2(lncRNA MEG3+miR-21),with 12 rats in each group.Myocardial infarction size,left ventricular ejection fraction(LVEF),left ven-tricular fractional shortening(LVFS),serum lactate dehydrogenase(LDH),creatine kinase(CK),creatine kinase isoenzyme(CK-MB)content,cardiomyocyte apoptotic rate and expression level of cleaved Caspase-3 were determined.Dual luciferase reporter gene assay was used to detect lncRNA MEG3 targeting miR-21 in H9c2 cardiomyocytes.Results Compared with the sham group,the expression of lncRNA MEG3 was decreased and that of miR-21 was increased in the OVX group,IR group and combined group 1(P＜0.05).The combination group 1 had significant-ly lower lncRNA MEG3 expression and higher miR-21 expression than the OVX group and IR group(P＜0.05).Compared with the negative model group,the myocardial infarction size,serum LDH,CK,CK-MB,cardiomyocyte apoptotic rate,and cleaved Caspase-3 expression were de-creased,while LVFS and LVEF were increased in the lncRNA MEG3 group(P＜0.05).Compared with the lncRNA MEG3 group,myocardial infarction size,serum LDH,CK,CK-MB content,car-diomyocyte apoptotic rate,Cleaved Caspase-3 expression were increased,while LVFS and LVEF were decreased in the combined group 2[(43.58±3.32)％vs(50.37±4.29)％,(57.12±4.28)％vs(68.47±5.61)％,P＜0.05].Conclusion Overexpression of lncRNA MEG3 alleviates the myo-cardium IR injury in OVX rats by sponging miR-21.

Objective:Study on the correlation between the active components of Salviea Miltiorrhizae Radix et Rhizoma screened by high-throughput sequencing and the regulation of lncRNA-mRNA in human lung adenocarcinoma A549 cells.Methods:A549 cells were cultured, and the IC 50 dose of cryptotanshinone and tanshinone ⅡA was confirmed according to the cell proliferation experiment. A549 cells were randomly divided into blank control group, cryptotanshinone group, and tanshinone IIA group using a random number table method. After 24 hours of intervention, the cell cycle was detected by flow cytometry. High-throughput sequencing technique was used to detect the expressions of lncRNA and mRNA in A549 cells in intervention group and non-intervention group. By analyzing the expression profiles of differential genes related to cryptotanshinone and tanshinone ⅡA, the obtained differential genes were analyzed by GO and KEGG. Results:The cell cycle results showed that the proportion of G0/G1 phase cells in cryptotanshinone and Tanshinone ⅡA was increased ( P<0.01), the proportion of S phase cells was decreased ( P<0.01), and the proportion of G2/M phase cells in cryptotanshinone was decreased ( P<0.01). The results of high-throughput screening showed that cryptotanshinone could up-regulate 4 698 lncRNA, down-regulate 1 557 lncRNA, up-regulate 4 810 mRNA and down-regulate 5 644 mRNA. Tanshinone ⅡA could up-regulate 1 348 lncRNA, down-regulate 1 299 lncRNA, up-regulate 4646 mRNA and down-regulate 4 741 mRNA. The function and pathway enrichment analysis of differential lncRNA and mRNA showed that the differentially expressed genes of cryptotanshinone and tanshinone ⅡA were mainly related to cell cycle, autophagy, AMPK signaling pathway, FoxO signaling pathway and EGFR signaling pathway. GAS5 may be one of the targets for the inhibitory effects of cryptotanshinone and tanshinone ⅡA. Conclusion:Cryptotanshinone and tanshinone ⅡA have certain inhibitory effects on A549 cells, and there are differentially expressed genes of lncRNA-mRNA, which are closely related to cell cycle and signal pathway.

Objective:To investigate the effectiveness and safety of the coaxial needle technique in percutaneous liver biopsy for patients with coagulation function abnormalities.Methods:Clinical data of 210 patients who underwent percutaneous liver biopsy using the coaxial needle technique under ultrasound guidance from December 2018 to May 2021 in 3 centers were collected. A retrospective analysis was conducted to compare the puncture success rate, number of samples obtained, pathology qualification rate, intraoperative and postoperative bleeding rates between the group with coagulation function abnormalities and the group with normal coagulation function.Results:After propensity score matching, there were 105 patients in each group, with a puncture success rate of 100% in both groups. The pathology qualification rate was 100% for all samples.Intraoperative bleeding occurred in 78 cases (74.3%, 78/105) in the coagulation function abnormalities group and in 64 cases (61.0%, 64/105) in the normal coagulation function group, with a statistically significant difference between the two groups ( P=0.006). Postoperative bleeding occurred in 3 cases (2.9%, 3/105) in the coagulation function abnormalities group and in 0 case in the normal coagulation function group, with no statistically significant difference between the two groups ( P=0.081). Conclusions:The use of the coaxial needle technique for percutaneous liver biopsy in patients with coagulation function abnormalities not only allows for obtaining an adequate tissue sample but also demonstrates good safety.