Objective To investigate the effects of Bushen Shengjing Tiaohe Qixue Prescription on rats with diminished ovarian reserve(DOR)based on Wnt/β-catenin signaling pathway.Methods Totally 60 8-week-old SD female rats were randomly divided into control group,model group,estradiol valerate group and TCM low-,medium-and high-dosage groups,with 10 rats in each group.Except for the control group,the rats in the other groups were given a single intraperitoneal injection of cyclophosphamide 75 mg/kg to replicate the DOR rat model.From the next day of modeling,the corresponding drugs was administered for medication groups by gavage for 3 weeks.The general conditions of the rats were observed,ELISA was used to detect the contents of serum follicle-stimulating hormone(FSH),luteinizing hormone(LH),estradiol(E2),anti-mullerian hormone(AMH)and inhibin B(INHB),HE staining was used to observe the morphologic changes in ovarian tissue,Western blot was used to detect GSK3β,β-catenin,CyclinD1 and c-MYC protein expressions in ovarian tissues,immunohistochemical staining was used to observe β-catenin expression in ovarian tissue,RT-qPCR was used to detect Wnt1,Wnt2,Wnt4,GSK3β,β-catenin,CyclinD1 and c-MYC mRNA expression in ovarian tissue,transmission electron microscopy was used to observe ultrapathologic structure of ovarian granulosa cells.Results Compared with the control group,the rats in the model group had reduced intake of food and water,slow or even reduced body mass gain,and disturbed estrous cycle,the serum contents of FSH and LH increased(P<0.01),the contents of E2,AMH and INHB decreased(P<0.01);the number of follicles of all levels in ovarian tissue was reduced,and there was increased number of atretic follicles with disturbed arrangement of the granulosa cells;the expression of GSK3β protein in ovarian tissue increased(P<0.01),the expression of β-catenin,CyclinD1 and c-MYC protein decreased(P<0.05,P<0.01),and β-catenin positive expression decreased(P<0.01),the expression of Wnt1,Wnt2,Wnt4,β-catenin,CyclinD1 and c-MYC mRNA in ovarian tissue decreased(P<0.01),GSK3β mRNA expression was increased(P<0,01);organelles such as mitochondria and nuclei of ovarian granulosa cells were highly swollen and partially lysed.Compared with the model group,the rats in the administered group all had different degrees of body mass increase,partial restoration of the motility cycle,serum FSH and LH contents decreased,and serum E2,AMH and INHB contents increased(P<0.01,P<0.05);the number of follicles at all levels in ovarian tissue increased to varying degrees,and the atretic follicles were reduced;the expression of GSK3β protein in ovarian tissue decreased,β-catenin,CyclinD1 and c-MYC protein expressions increased,β-catenin positive expression increased,the expression of Wnt1,Wnt2,Wnt4,β-catenin,CyclinD1 and c-MYC mRNA in ovarian tissue increased,and GSK3β mRNA expression decreased,with statistical significance in estradiol valerate group and TCM high-dosage group(P<0.05,P<0.01);mitochondria,endoplasmic reticulum and other organelles of ovarian granulosa cells were mildly to moderately swollen.Conclusion Bushen Shengjing Tiaohe Qixue Prescription can improve ovarian function and regulate serum sex hormone levels in DOR model rats,and its mechanism might be related to the activation of Wnt/β-catenin signaling pathway.

Objective To analyze the clinical value of plasma insulin-like factor 6(INSL6)in predicting the risk of acute aortic syndromes(AAS)and adverse outcomes.Methods A retrospective case-control trial was conducted on 194 AAS patients admitted to Department of Cardiovascular Diseases of Second Affiliated Hospital of Army Medical University between April 2021 and June 2023.Another 194 sex-,age-and BMI-matched individuals without aortic diseases were recruited from the health examination center between December 2021 and January 2024.Their plasma INSL6 level was measured with ELISA,and the general clinical data and results of some laboratory tests were collected and compared between the 2 groups.Spearman correlation analysis was used to assess the relationship between plasma INSL6 and other variables,receiver operating characteristic(ROC)curve was plotted to evaluate the diagnostic efficacy of INSL6 for AAS occurrence,multivariate conditional logistic regression model was utilized to analyze the association between plasma INSL6 and AAS onset,and Kaplan-Meier survival curve and multivariate Cox proportional hazards regression model was applied to analyze the relationship between acute-phase plasma INSL6 level and adverse prognosis in AAS patients.Results The plasma INSL6 level was significantly higher in AAS patients at acute phase than the control individuals[704.40(481.32～1 152.62)vs 141.24(107.60～163.72)pg/mL,P<0.001],but no statistical difference was observed in the level among the patients with different AAS subtypes(aortic dissection,intramural hematoma,and penetrating aortic ulcer).Spearman correlation analysis showed that the plasma INSL6 level was positively correlated with platelet count(r=0.325,P<0.001)and hemoglobin concentration(r=0.186,P=0.009),and negatively with IL-6(r=-0.182,P=0.011),INF-γ(r=-0.283,P<0.001),and D-dimer levels(r=-0.195,P=0.006).Multivariate conditional logistic regression analysis revealed that plasma INSL6 level was independently associated with the occurrence of AAS(OR=28.634,95%CI:7.267～112.820,P<0.001).ROC curve analysis further confirmed that the optimal cutoff value of plasma INSL6 in predicting AAS was 259.425 pg/mL,with a sensitivity of 95.9%and a specificity of 98.5%at this threshold.Kaplan-Meier curve analysis demonstrated that AAS patients with low INSL6 level had significantly lower cumulative survival rates(P=0.020)and event-free survival rates(P=0.004)than those with high INSL6 level(P<0.05).Multivariate COX regression analysis revealed that,after adjusting for sex,age,systolic blood pressure,ST classification,and surgical treatment,acute-phase INSL6 level was independently associated with all-cause mortality(HR:0.999,95%CI:0.999～1.000,P=0.023),AAS-related mortality(HR:0.999,95%CI:0.998～1.000,P=0.012),and composite endpoint events(HR:0.999,95%CI:0.999～1.000,P=0.026)in AAS patients during follow-up.Conclusion Plasma INSL6 level is closely associated with the occurrence and adverse prognosis of AAS,and the indicator is expected to be an effective biomarker for diagnosing AAS and predicting its prognosis.

Corticotomy is a clinical procedure to accelerate orthodontic tooth movement characterized by the regional acceleratory phenomenon (RAP). Despite its therapeutic effects, the surgical risk and unclear mechanism hamper the clinical application. Numerous evidences support macrophages as the key immune cells during bone remodeling. Our study discovered that the monocyte-derived macrophages primarily exhibited a pro-inflammatory phenotype that dominated bone remodeling in corticotomy by CX3CR1^CreERT2; R26^GFP lineage tracing system. Fluorescence staining, flow cytometry analysis, and western blot determined the significantly enhanced expression of binding immunoglobulin protein (BiP) and emphasized the activation of sensor activating transcription factor 6 (ATF6) in macrophages. Then, we verified that macrophage specific ATF6 deletion (ATF6^f/f; CX3CR1^CreERT2 mice) decreased the proportion of pro-inflammatory macrophages and therefore blocked the acceleration effect of corticotomy. In contrast, macrophage ATF6 overexpression exaggerated the acceleration of orthodontic tooth movement. In vitro experiments also proved that higher proportion of pro-inflammatory macrophages was positively correlated with higher expression of ATF6. At the mechanism level, RNA-seq and CUT&Tag analysis demonstrated that ATF6 modulated the macrophage-orchestrated inflammation through interacting with Tnfα promotor and augmenting its transcription. Additionally, molecular docking simulation and dual-luciferase reporter system indicated the possible binding sites outside of the traditional endoplasmic reticulum-stress response element (ERSE). Taken together, ATF6 may aggravate orthodontic bone remodeling by promoting Tnfα transcription in macrophages, suggesting that ATF6 may represent a promising therapeutic target for non-invasive accelerated orthodontics.
Animals ; Mice ; Macrophages/metabolism* ; Tumor Necrosis Factor-alpha/genetics* ; Tooth Movement Techniques/methods* ; Activating Transcription Factor 6/metabolism* ; Bone Remodeling ; Flow Cytometry ; Blotting, Western

Objective To investigate the effects of Bushen Shengjing Tiaohe Qixue Prescription on rats with diminished ovarian reserve(DOR)based on Wnt/β-catenin signaling pathway.Methods Totally 60 8-week-old SD female rats were randomly divided into control group,model group,estradiol valerate group and TCM low-,medium-and high-dosage groups,with 10 rats in each group.Except for the control group,the rats in the other groups were given a single intraperitoneal injection of cyclophosphamide 75 mg/kg to replicate the DOR rat model.From the next day of modeling,the corresponding drugs was administered for medication groups by gavage for 3 weeks.The general conditions of the rats were observed,ELISA was used to detect the contents of serum follicle-stimulating hormone(FSH),luteinizing hormone(LH),estradiol(E2),anti-mullerian hormone(AMH)and inhibin B(INHB),HE staining was used to observe the morphologic changes in ovarian tissue,Western blot was used to detect GSK3β,β-catenin,CyclinD1 and c-MYC protein expressions in ovarian tissues,immunohistochemical staining was used to observe β-catenin expression in ovarian tissue,RT-qPCR was used to detect Wnt1,Wnt2,Wnt4,GSK3β,β-catenin,CyclinD1 and c-MYC mRNA expression in ovarian tissue,transmission electron microscopy was used to observe ultrapathologic structure of ovarian granulosa cells.Results Compared with the control group,the rats in the model group had reduced intake of food and water,slow or even reduced body mass gain,and disturbed estrous cycle,the serum contents of FSH and LH increased(P<0.01),the contents of E2,AMH and INHB decreased(P<0.01);the number of follicles of all levels in ovarian tissue was reduced,and there was increased number of atretic follicles with disturbed arrangement of the granulosa cells;the expression of GSK3β protein in ovarian tissue increased(P<0.01),the expression of β-catenin,CyclinD1 and c-MYC protein decreased(P<0.05,P<0.01),and β-catenin positive expression decreased(P<0.01),the expression of Wnt1,Wnt2,Wnt4,β-catenin,CyclinD1 and c-MYC mRNA in ovarian tissue decreased(P<0.01),GSK3β mRNA expression was increased(P<0,01);organelles such as mitochondria and nuclei of ovarian granulosa cells were highly swollen and partially lysed.Compared with the model group,the rats in the administered group all had different degrees of body mass increase,partial restoration of the motility cycle,serum FSH and LH contents decreased,and serum E2,AMH and INHB contents increased(P<0.01,P<0.05);the number of follicles at all levels in ovarian tissue increased to varying degrees,and the atretic follicles were reduced;the expression of GSK3β protein in ovarian tissue decreased,β-catenin,CyclinD1 and c-MYC protein expressions increased,β-catenin positive expression increased,the expression of Wnt1,Wnt2,Wnt4,β-catenin,CyclinD1 and c-MYC mRNA in ovarian tissue increased,and GSK3β mRNA expression decreased,with statistical significance in estradiol valerate group and TCM high-dosage group(P<0.05,P<0.01);mitochondria,endoplasmic reticulum and other organelles of ovarian granulosa cells were mildly to moderately swollen.Conclusion Bushen Shengjing Tiaohe Qixue Prescription can improve ovarian function and regulate serum sex hormone levels in DOR model rats,and its mechanism might be related to the activation of Wnt/β-catenin signaling pathway.

Objective:This study aimed to evaluate the prognostic factors of clinical and histopathological parameters,including age,gender,tumor stage,tumor grade,tumor differentiation,lymph node metastasis(LNM),tumor frequency,and tumor count,in patients undergoing radical cystectomy(RC)combined with Mainz Pouch Ⅱ bladder reconstruction.Methods:A total of 237 bladder cancer patients(198 male and 39 female)who underwent RC combined with Mainz Pouch Ⅱ bladder reconstruction without chemotherapy or radiotherapy,from January 2004 to January 2023,were included in this study.Kaplan-Meier and Cox regression analyses were performed to assess the impact of age,tumor grade,tumor stage,tumor differentiation,LNM,tumor frequency,and tumor count on 5-year overall survival(OS)and 5-year cancer-specific survival(CSS).Results:The mean age at diagnosis was 59.8 years,with 198 male and 39 female patients.The mean follow-up duration was 47.8 months.In univariate analysis,patients younger than 65 years had significantly higher 5-year OS and 5-year CSS compared to those aged≥65 years.Patients with urothelial carcinoma showed better 5-year OS than those with non-urothelial carcinoma.Additionally,tumor stage,tumor grade,and LNM were negatively associated with 5-year OS and 5-year CSS.On multivariate analysis,only tumor grade and LNM remained statistically significant(P<0.05).Conclusions:Tumor grade and LNM were identified as independent prognostic risk factors for 5-year OS and 5-year CSS following RC combined with Mainz PouchⅡ bladder reconstruction.Moreover,the application of RC combined with Mainz Pouch Ⅱ bladder reconstruction should consider the patient's preferences and physical condition.

ObjectiveTo investigate the mechanism of the modified of Bazhentang combined with Guishentang in improving pregnancy outcomes in mouse models of embryo implantation dysfunction by regulating T helper 1/T helper 2 （Th1/Th2） immune balance. MethodsEighty ICR female mice were randomly divided into four groups （n=20 per group） on gestational day 1 （GD1）： control， model， western medicine， and traditional Chinese medicine （TCM） groups. Except for the control group， all mice received mifepristone solution （0.2 mg/mouse） via oral gavage on GD4 to induce embryo implantation dysfunction. The TCM group received a water decoction of the modified of Bazhentang combined with Guishentang （20.8 g·kg^-1）， with the western medicine group administered dydrogesterone （3.9 mg·kg^-1）， and the control/model groups given equal volumes of saline. All treatments were administered once daily from GD1 until one day before sample collection. Outcomes included implantation site counts （macroscopic observation）， pregnancy rates， body weight， endometrial histopathology （hematoxylin-eosin staining）， uterine expression of T-box expressed in T cells （T-bet）， GATA-binding protein 3 （GATA3）， interferon gamma （IFN-γ）， and interleukin-4 （IL-4） at protein （Western blot） and mRNA （real-time polymerase chain reaction， Real-time PCR） levels， serum IFN-γ and IL-4 levels （enzyme-linked immunosorbent assay， ELISA）， and Th1/Th2 immune balance evaluated by calculating T-bet/GATA3 and IFN-γ/IL-4 ratios. ResultsCompared to the control group， the model group showed no significant change in pregnancy rate but exhibited a marked reduction in average implantation sites and body weight （P<0.01）. Histopathological analysis revealed endometrial abnormalities， including decreased glandular density， stromal compaction， and absence of nucleolar vacuoles. At the molecular level， uterine tissue in the model group demonstrated significantly upregulated expression of T-bet and IFN-γ （P<0.05， P<0.01）， alongside markedly downregulated GATA3 and IL-4 expression （P<0.05， P<0.01）. Serum analysis confirmed markedly elevated IFN-γ （P<0.01） and reduced IL-4 levels （P<0.01）， resulting in significantly increased T-bet/GATA3 and IFN-γ/IL-4 ratios （P<0.01）. Compared to the model group， pregnancy rates in all treatment groups showed no significant change. Implantation sites and body weight increased substantially （P<0.01）， with restored endometrial morphology characterized by enhanced glandular density， stromal edema， and reappearance of nucleolar vacuoles. Significant downregulation of T-bet and IFN-γ （P<0.01） and upregulation of GATA3 and IL-4 （P<0.05， P<0.01） in uterine tissue were observed. Serum IFN-γ levels were significantly reduced （P<0.05， P<0.01）， while IL-4 levels were significantly elevated （P<0.05）. The Th1/Th2 ratios were significantly decreased （P<0.01）. ConclusionThe modified of Bazhentang combined with Guishentang significantly enhances the number of embryo implantation sites in mice with embryo implantation dysfunction， potentially through modulating T-bet/GATA3 expression， restoring Th1/Th2 immune balance， and improving endometrial receptivity.

Objective:To explore the prognosis of patients with acute aortic syndrome (AAS) in the real world and to examine the risk factors associated with poor outcomes in AAS.Methods:This is a single-center retrospective study. Patients diagnosed with AAS at Xinqiao Hospital from January 2021 to July 2023 were included. The primary endpoints were all-cause mortality and aorta-related mortality, while the secondary endpoints included stroke, myocardial infarction, secondary interventions, and readmission for any cause. Survival analysis was performed using Kaplan-Meier curves, and risk factors for primary endpoint events were analyzed using multivariate Cox regression.Results:A total of 254 AAS patients, aged (58.9±13.2) years were included in this study. There were 178 cases of aortic dissection, 69 cases of aortic intramural hematoma, and 7 cases of aortic penetrating ulcer. The median follow-up time was 545 days. Seventy-three all-cause deaths occurred among patients with AAS, including 61 aorta-related deaths; 3 strokes, 1 myocardial infarction, 9 secondary surgeries, and 35 readmissions for any cause were observed. Kaplan-Meier curve analysis demonstrated significant differences in all-cause mortality rates based on the Stanford classification, AAS disease classification, eGFR, and albumin levels (all P<0.05), and similar results were also observed in aorta-related death (all P<0.05). Multivariate Cox regression suggested that albumin<35 g/L ( HR=2.372, 95% CI 1.337-4.210, P=0.003), eGFR<90 ml·min -1·1.73 m -2 ( HR=2.457, 95% CI 1.261-4.786, P=0.008), and Stanford type A AAS ( HR=3.420, 95% CI 1.998-5.856, P<0.001) were independent risk factors for all-cause mortality in AAS patients; albumin<35 g/L( HR=2.432, 95% CI 1.295-4.570, P=0.006), eGFR<90 ml·min -1·1.73 m -2( HR=2.523,95% CI 1.243-5.122, P=0.010), and Stanford type A AAS ( HR=3.455,95% CI 1.819-6.564, P<0.001) were independent risk factors for aorta-related mortality in AAS patients. Conclusions:In the real world, the prognosis of patients with AAS remains pessimistic. Patients with type A AAS, renal dysfunction, hypoproteinemia may have a higher risk of poor prognosis.

Objective:This study aimed to evaluate the prognostic factors of clinical and histopathological parameters,including age,gender,tumor stage,tumor grade,tumor differentiation,lymph node metastasis(LNM),tumor frequency,and tumor count,in patients undergoing radical cystectomy(RC)combined with Mainz Pouch Ⅱ bladder reconstruction.Methods:A total of 237 bladder cancer patients(198 male and 39 female)who underwent RC combined with Mainz Pouch Ⅱ bladder reconstruction without chemotherapy or radiotherapy,from January 2004 to January 2023,were included in this study.Kaplan-Meier and Cox regression analyses were performed to assess the impact of age,tumor grade,tumor stage,tumor differentiation,LNM,tumor frequency,and tumor count on 5-year overall survival(OS)and 5-year cancer-specific survival(CSS).Results:The mean age at diagnosis was 59.8 years,with 198 male and 39 female patients.The mean follow-up duration was 47.8 months.In univariate analysis,patients younger than 65 years had significantly higher 5-year OS and 5-year CSS compared to those aged≥65 years.Patients with urothelial carcinoma showed better 5-year OS than those with non-urothelial carcinoma.Additionally,tumor stage,tumor grade,and LNM were negatively associated with 5-year OS and 5-year CSS.On multivariate analysis,only tumor grade and LNM remained statistically significant(P<0.05).Conclusions:Tumor grade and LNM were identified as independent prognostic risk factors for 5-year OS and 5-year CSS following RC combined with Mainz PouchⅡ bladder reconstruction.Moreover,the application of RC combined with Mainz Pouch Ⅱ bladder reconstruction should consider the patient's preferences and physical condition.

Objective:To explore the prognosis of patients with acute aortic syndrome (AAS) in the real world and to examine the risk factors associated with poor outcomes in AAS.Methods:This is a single-center retrospective study. Patients diagnosed with AAS at Xinqiao Hospital from January 2021 to July 2023 were included. The primary endpoints were all-cause mortality and aorta-related mortality, while the secondary endpoints included stroke, myocardial infarction, secondary interventions, and readmission for any cause. Survival analysis was performed using Kaplan-Meier curves, and risk factors for primary endpoint events were analyzed using multivariate Cox regression.Results:A total of 254 AAS patients, aged (58.9±13.2) years were included in this study. There were 178 cases of aortic dissection, 69 cases of aortic intramural hematoma, and 7 cases of aortic penetrating ulcer. The median follow-up time was 545 days. Seventy-three all-cause deaths occurred among patients with AAS, including 61 aorta-related deaths; 3 strokes, 1 myocardial infarction, 9 secondary surgeries, and 35 readmissions for any cause were observed. Kaplan-Meier curve analysis demonstrated significant differences in all-cause mortality rates based on the Stanford classification, AAS disease classification, eGFR, and albumin levels (all P<0.05), and similar results were also observed in aorta-related death (all P<0.05). Multivariate Cox regression suggested that albumin<35 g/L ( HR=2.372, 95% CI 1.337-4.210, P=0.003), eGFR<90 ml·min -1·1.73 m -2 ( HR=2.457, 95% CI 1.261-4.786, P=0.008), and Stanford type A AAS ( HR=3.420, 95% CI 1.998-5.856, P<0.001) were independent risk factors for all-cause mortality in AAS patients; albumin<35 g/L( HR=2.432, 95% CI 1.295-4.570, P=0.006), eGFR<90 ml·min -1·1.73 m -2( HR=2.523,95% CI 1.243-5.122, P=0.010), and Stanford type A AAS ( HR=3.455,95% CI 1.819-6.564, P<0.001) were independent risk factors for aorta-related mortality in AAS patients. Conclusions:In the real world, the prognosis of patients with AAS remains pessimistic. Patients with type A AAS, renal dysfunction, hypoproteinemia may have a higher risk of poor prognosis.

Bronchopulmonary dysplasia (BPD), characterized by impaired alveolarization and dysregulated vascularization, is a severe health burden for neonates worldwide. Hyperoxia induced acute lung injury is a major contributor to the progression and deterioration of BPD. An increasing number of animal studies have revealed that human umbilical cord blood derived mononuclear cells (hUCB-MNCs) infusion significantly attenuated the hyperoxia-induced acute lung injury through regeneration capacity. Currently, clinical application requires determination of the optimal dose and adjustment to good manufacturing practices. In this work, we comprehensively investigated the optimal dose of hUCB-MNCs in alleviating hyperoxia-induced lung injury in neonatal C57BL6/J mice. Mice with hyperoxia exposure were implanted with low (3×104cells/kg, Dl), middle (3×105 cells/kg, Dm) and high (3×106 cells/kg, Dh) dose of hUCB-MNCs at postnatal day 7. Three weeks after graft, characteristics exhibited in lungs including morphology, function and cytokine expression were thoroughly analyzed. Implantation of hUCB-MNCs sharply reverted the impaired lung architecture induced by hyperoxia exposure dose dependently as evidenced by indicated parameters. Attenuated expression of IL-1β concomitant with enhanced expression of IL-10 and IL-2 were shown in Dh inoculated groups, where Dl and Dm failed to restore the level of IL-10, IL-1β and IL-2. Significantly re-escalated marker of angiogenic marker VEGFA, CD31 in lung tissue were uniquely observed in Dh group. Mechanistically, our study revealed the appropriate dose of intravenous infusion of hUCB-MNCs in alleviating hyperoxia-induced lung injury through modulating reactive oxygen species response in neonatal mice.Therefore, a tight control of hUCB-MNCs density or levels of CB-MNC related products is of great significance.