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.

Objective To identify genetic regulators of ionizing radiation(IR)sensitivity through clustered regularly interspaced short palindromic repeats(CRISPR)/CRISPR-associated nuclease 9(Cas9)genome-wide screening.Methods A specialized single guide RNA(sgRNA)library was developed targeting 987 stress-response regulatory genes identified through Kyoto Encyclopedia of Genes and Genomes(KEGG),Reactome and gene ontology(GO)databases,followed by construction of sgRNA plasmids and packaging into a lentiviral library.Using colon adenocarcinoma Caco-2 cells as a model system,the Cas9-stable transgenic line was established via lentiviral transduction and antibiotic selection.Library-transduced cells underwent puromycin selection,followed by γ-irradiation(dose optimized via preliminary experiments).Post-irradiation phenotypic selection was conducted after 14 days,with subsequent next-generation sequencing of surviving cell populations to identify enriched/depleted sgRNAs.Candidate genes were functionally validated through orthogonal assays:cell counting kit-8(CCK-8)proliferation analysis,clonogenic survival assays and flow cytometric quantification of reactive oxygen species(ROS)and apoptotic markers.Results The optimized screening platform identified 281 radiation response genes(165 radiosensitive and 116 radioprotective candidates).Functional validation revealed Bcl2-associated athanogene 3(BAG3)knockdown significantly enhanced radioresistance.Proliferation was increased 1.2-1.5 fold,clonogenic survival improved 1.5-2.0 fold,and ROS was reduced by 13％-25％coupled with 32％-73％apoptosis attenuation compared to control groups.Conclusion The integrated CRISPR/Cas9 screening platform effectively identifies novel radiation response regulators,as evidenced by the discovery of BAG3 as a critical radiosensitivity determinant.This high-throughput functional genomics approach provides a robust methodology for systematically mapping molecular determinants of cellular radiation response,with potential applications in both mechanistic studies and therapeutic target discovery.

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 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 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 To investigate the effect of erythroid differentiation associated gene(EDAG)on hematopoietic stem/progenitor cells(HSPCs)under chronic inflammation.Methods In this study,EDAG-/-and wild type(WT)mice were divided into the experiment group and control group.An infectious chronic inflammation model was established via multiple intraperitoneal injections of Listeria monocytogenes(LM),while a sterile chronic inflammation model was generated via multiple intraperitoneal injections of polyinosinic-polycytidylic acid[Poly(I∶C)].The effect of EDAG on HSPCs was explored under chronic inflammation conditions.Results In the LM repeated infection model,EDAG deletion led to a decrease in HSPCs and long-term hematopoietic stem cells(LT-HSCs)in mice as well as a significant bias towards myeloid differentiation in peripheral blood.Similarly,EDAG knockout also resulted in reduced numbers of HSPCs and decreased colony-forming ability in aseptic chronic inflammation models.Conclusion EDAG deficiency accelerates HSPC depletion in young mice under chronic inflammation,indicating strong protection of EDAG against HSPC damage induced by chronic inflammation.

Objective:To investigate the difference in the radiation sensitivity of hematopoietic stem and progenitor cells (HSPCs) derived from fetal liver and bone marrow.Methods:HSPCs from fetal liver of 14.5 d embryo or bone marrow of 8 week-old mice were isolated to receive a single dose of 5 or 10 Gy irradiation in vitro using a 60Co irradiator. Twelve hours later, the cell apoptosis, mitochondrial reactive oxygen species (ROS) level, colony formation ability and DNA damage in HSPCs were detected. Freshly isolated HSPCs were injected into lethally irradiated CD45.1 + C57BL/6J mice (4.5 Gy+ 5 Gy with an interval of 30 min) Chimerism rate, lineage constitution, and cell cycle were analyzed 12 weeks after transplantation. Results:Compared with bone marrow HSPCs after irradiation, the percentage of apoptosis in fetal liver HSPCs was significantly higher ( t=16.21, 12.27, P<0.05), the level of ROS was dramatically elevated ( t=68.72, 18.89, P<0.05). At 10 Gy, fetal liver HSPCs could not form colonies at all ( t=12.41, 15.67, 9.46, P<0.05). γ-H2AX immunofluorescence staining showed that the DNA damage of fetal liver HSPCs was more severe after irradiation, and the number of Foci formed was significantly higher than that of bone marrow HSPCs ( t=2.27, 2.03, P< 0.05), which indicated that fetal liver HSPCs were more sensitive to radiation. The chimerism rate of transplanted fetal liver HSPCs was lower than that of bone marrow cells ( t=5.84, P<0.05) with a higher proportion of myeloid lineage, suggesting that fetal liver HSPCs had lower in vivo reconstitution capacity than bone marrow HSPCs and were more prone to myeloid differentiation. The cell cycle of bone marrow HSPCs from transplanted chimeric mice was examined, and the proportion of S-phase was significantly higher in the fetal liver group than that in the bone marrow group ( t=2.89, P<0.05). Mitochondrial stress results showed that fetal liver HSPCs had higher basal respiratory capacity ( t=39.19, P<0.05), proton leakage ( t=6.64, P<0.05), ATP production ( t=9.33, P<0.05), and coupling efficiency ( t=7.10, P<0.05) than bone marrow c-Kit + cells, while respiratory reserve capacity ( t=5.53, P< 0.05) was lower than that of bone marrow c-Kit + cells. Conclusions:HSPCs derived from fetal liver display higher radiosensitivty compared with bone marrow HSPCs, laying the foundation for an in-depth illustration of the effects of radiation on hematopoietic stem cells at different developmental stages.

Objective:To study the effect of different doses of 60Co γ-ray ionizing radiation on mitochondrial function in mouse hematopoietic stem and progenitor cells (HSPCs). Methods:C57BL/6 mice were divided into control group, 1 Gy irradiation group and 4.5 Gy irradiation group. The mitochondrial functions were detected at 12 h and 24 h after irradiation, including ROS level, membrane potential, mitochondrial structure, and mitochondrial stress. Bone marrow c-Kit + cells received a single 15 Gy irradiation in vitro, after 24 h, mitochondrial function was detected. Results:It was found that mice leukocytes ( t=12.41, 18.31, 16.48, 14.16, 19.08, 20.25, P<0.05), red blood cells ( t=4.81, 6.62, P<0.05) and platelets ( t=4.33, 6.68, P<0.05) were significantly reduced. The numbers of bone marrow colony formation unit ( t=16.27, 55.66, 17.06, 43.75, P<0.05), and HSPCs ( t=5.16, 11.55, P<0.05) were decreased dose-dependently post-irradiation. Under 1 Gy irradiation, the mitochondrial function and mitochondrial basal metabolic index of HSPCs ( t= 7.36, 3.68, 4.58, 3.15, 3.15, P<0.05) were enhanced at 24 h post-irradiation. Under 4.5 Gy irradiation, mitochondrial number, mitochondrial membrane potential ( t=12.29, 10.46, P<0.05), maximal respiration and spare respiratory capacity were decreased ( t=7.81, 5.78, 6.70, 5.83, P<0.05), ROS level was increased ( t=4.63, 4.12, P<0.05). The basal respiration and oxidative phosphorylated ATP production were reduced at 12 h after irradiation ( t=8.48, 3.80, P<0.05); and the proton leakage was increased ( t=6.57, P<0.05) and coupling efficiency was reduced ( t=11.43, P<0.05) at 24 h after irradiation. In cultured c-Kit + cells, the level of ROS ( t=11.30, P<0.05) and the maximum respiration and spare respiratory capacity were increased ( t=4.25, 3.44, P<0.05) while the mitochondrial membrane potential was decreased ( t=34.92, P<0.05) significantly. Conclusions:A method for systematically assessing mitochondrial function in HSPCs was established, and the effect of ionizing radiation on mitochondrial function of HSPCs was clarified, laying a foundation for further revealing the mechanism of ionizing radiation-induced mitochondrial damage in HSPCs.

To report a 4-year-old boy with severed right index finger amputations in 2 segments. There were severe contusion on the 2 amputated sections of finger. According to the prevention and control requirement for the novel coronavirus disease(COVID-19), the patient was firstly checked to exclude the COVID-19. Then the replantation surgery was successfully carried out under the strict protective measures. The replanted index finger survived well at 2 weeks after surgery.