Objective To develop an erythrocyte-based delivery system for butyrylcholinesterase(BChE)that is capable of prophylaxis against organophosphorus nerve agents.Methods Recombinant BChE was produced and analyzed for oligomerization via polyacrylamide gel electrophoresis(PAGE)and Western blotting.A modified hypotonic preswelling method was employed to prepare BChE-loaded erythrocytes.The drug loading capacity and encapsulation efficiency were quantified using enzyme-linked immunosorbent assay(ELISA).Catalytic activity was assessed in vitro with an activity detection kit.The system was characterized via scanning electron microscopy(SEM),flow cytometry and a hematology analyzer.Results Recombinant BChE predominantly existed as dimers(85％dimer,15％monomer).The optimized volume ratio of erythrocytes to hypotonic solution was determined as 1:7.Compared with native and empty erythrocytes,BChE-loaded erythrocytes exhibited significantly higher catalytic activity(P＜0.001).The mean corpuscular volume of BChE-loaded erythrocytes increased(P＜0.001),while the mean content of corpuscular hemoglobin and hemoglobin in erythrocytes per 100 mL decreased(P＜0.001).SEM revealed no morphological differences(biconcave disc shape).Hypotonic preswelling moderately increased erythrocyte apoptosis(P＜0.001),but no statistical difference was observed between BChE-loaded and hypotonic-treated erythrocytes(P＞0.05).CD47 expression remained unchanged compared to native erythrocytes(P＞0.05).Conclusion The modified hypotonic preswelling method can generate BChE-loaded erythrocytes that retain the characteristics of native erythrocytes while conferring catalytic activity,offering a novel strategy for clinical intervention against organophosphorus poisoning.

Objective To observe the value of whole-brain CT perfusion(CTP)parameters at different time and clinical data for predicting delayed cerebral ischemia(DCI)and 3-month poor prognosis in patients with aneurysmal subarachnoid hemorrhage(aSAH).Methods Totally 127 aSAH patients were retrospectively enrolled.Clinical and CTP data within 24 h of symptom onset and during DCI time window(DCITW)were collected.The patients were divided into DCI group(n=34)and non-DCI group(n=93)based on DCI occurred or not during hospitalization,also into poor outcome group(modified Rankin scale[mRS]≥3,n=36)and good outcome group(mRS≤2)based on 3-month's follow-up.Multivariate logistic regression was performed to select independent predictive factors among variates being significantly different between groups.Then receiver operating characteristic curve was drawn,and the area under the curve(AUC)was calculated to evaluate the predictive performance of logistic regression model.Results Patients'age,modified Fisher score(mFS),subarachnoid hemorrhage early brain edema score(SEBES)and mean flow extraction product(mFEP)within 24 h of onset were all identified as independent predictive factors of DCI,and the AUC of their combination for predicting DCI during hospitalization was 0.817.Patients' age and mFS within 24 h of onset,alternatively,World Federation of Neurosurgical Societies(WFNS)grade and mFEP during DCITW were all independent predictive predictors of 3 months' prognosis,and the combination of the latter two showed better predictive performance(AUC=0.922)tahn the former two(AUC=0.822,P＜0.05).Conclusion Whole-brain CTP parameters combined with clinical data within 24 h of onset of aSAH could be used to predict the occurrence of DCI during hospitalization,whole-brain CTP parameters during DCITW could be used to predict 3 months'poor prognosis.

Objective:To explore the application value of real-time virtual sonography (RVS) combined with intraductal biliary contrast-enhanced ultrasound (IB-CEUS) in percutaneous transhepatic cholangial drainage (PTCD).Methods:This retrospective cohort study included data from 71 patients who underwent PTCD at the Department of Hepatobiliary and Pancreatic Surgery in the Second Affiliated Hospital of Nanchang University between May 2021 and August 2022. There were 36 male and 35 female patients,aged 35 to 94 years. Based on the guidance modality used,patients were divided into two groups: the RVS combined with IB-CEUS group ( n=36) and the digital subtraction angiography (DSA) group ( n=35). PTCD was performed under the guidance of RVS combined with IB-CEUS in the RVS+IB-CEUS group,and under conventional DSA fluoroscopic guidance in the DSA group. Two clinicians classified the biliary conditions as either simple or complex based on preoperative ultrasound and CT (or MRI) imaging. Statistical analyses were conducted using independent sample t-tests,rank-sum tests, χ2 tests,or Fisher′s exact tests,as appropriate. Results:Significant differences were observed between the RVS+IB-CEUS group and the DSA group in terms of the number of punctures (1.0±0.2 vs. 2.2±1.4, t=-5.148, P<0.01) and postoperative complication rate(2.8% (1/35) vs. 17.1% (6/36), P=0.049). There were 9 patients with complex biliary conditions in the DSA group and 12 in the RVS+IB-CEUS group. The number of punctures in both the simple and complex subgroups of the RVS+IB-CEUS group(1.0±0.2 and 1.0±0.0) remained lower than that in the corresponding DSA subgroups(2.2±1.6 and 2.4±0.4) ( t=-3.606, P<0.01; t=-3.959, P=0.002). Moreover,the complication rate in the simple biliary subgroup of the RVS+IB-CEUS group was significantly lower than that of the DSA group(0 (0/24) vs. 19.2% (5/26), P=0.031),whereas no significant difference was found in the complex biliary subgroup (1/12 vs. 1/9, P=0.686). Conclusion:Guided by RVS and IB-CEUS, PTCD can help reduce the number of punctures during surgery and postoperative complications, and patients with complex bile duct conditions can still benefit from PTCD.

Objective To investigate the biological function of the proprotein convertase subtilisin/kexin type 9(PCSK9)in prostate cancer(PCa)and its effect on ferroptosis sensitivity Methods Bioinformatics was used to analyze the relationship between the expression of PCSK9 and the prognosis of prostate cancer.The expression of PCSK9 in PCa cell lines were detected using RT-qPCR.PCa cells with PCSK9 knockdown were constructed using siRNA,and the The effect of PCSK9 on cell proliferation,migration,and invasion were detected using CCK-8 assays and Transwell assays.The Cancer Therapeutics Response Portal(CTRP)was employed to investigate the correlation between PCSK9 and ferroptosis drug sensitivity,and PCa cells with PCSK9 knockdown were treated with the ferroptosis inducer(RSL3)to detect the sensitivity to ferroptosis.Results Bioinformatics showed low expression of PCSK9 had longer disease specific survival(P<0.05).The results of the in vitro experiments showed that PCSK9 knockdown significantly inhibited the proliferation,migration,and invasion of PCa cells(P<0.001).Furthermore,CTRP analysis showed that cellular sensitivity to ferroptosis inducers correlated with the expression level of PCSK9.PCSK9 knockdown cells exhibited higher sensitivity to the ferroptosis inducer RSL3.Conclusion Knockdown of PCSK9 inhibits the proliferation,migration,and invasion of PCa cells,and increases the sensitivity of cells to ferroptosis inducers.PCSK9 may provide new insights for the treatment of PCa.

Objective To investigate the biological function of the proprotein convertase subtilisin/kexin type 9(PCSK9)in prostate cancer(PCa)and its effect on ferroptosis sensitivity Methods Bioinformatics was used to analyze the relationship between the expression of PCSK9 and the prognosis of prostate cancer.The expression of PCSK9 in PCa cell lines were detected using RT-qPCR.PCa cells with PCSK9 knockdown were constructed using siRNA,and the The effect of PCSK9 on cell proliferation,migration,and invasion were detected using CCK-8 assays and Transwell assays.The Cancer Therapeutics Response Portal(CTRP)was employed to investigate the correlation between PCSK9 and ferroptosis drug sensitivity,and PCa cells with PCSK9 knockdown were treated with the ferroptosis inducer(RSL3)to detect the sensitivity to ferroptosis.Results Bioinformatics showed low expression of PCSK9 had longer disease specific survival(P<0.05).The results of the in vitro experiments showed that PCSK9 knockdown significantly inhibited the proliferation,migration,and invasion of PCa cells(P<0.001).Furthermore,CTRP analysis showed that cellular sensitivity to ferroptosis inducers correlated with the expression level of PCSK9.PCSK9 knockdown cells exhibited higher sensitivity to the ferroptosis inducer RSL3.Conclusion Knockdown of PCSK9 inhibits the proliferation,migration,and invasion of PCa cells,and increases the sensitivity of cells to ferroptosis inducers.PCSK9 may provide new insights for the treatment of PCa.

The immune microenvironment plays a pivotal role in maintaining ovarian homeostasis. Polycystic ovary syndrome (PCOS), a common endocrine and metabolic disorder, is closely associated with immune microenvironment imbalance. This review systematically describes the dysregulation of innate immune cells (e.g., macrophages, natural killer cells and dendritic cells) and adaptive immune cells (e.g., Th1, Th2, Treg and Th17) in PCOS, highlighting their impacts on ovarian function, insulin resistance, and hyperandrogenemia. These findings underscore the central role of immune microenvironment disturbances in PCOS pathogenesis. Additionally, the association between gut microbiota dysbiosis and PCOS is explored, emphasizing how gut microbiota influences metabolic byproducts and hormonal levels to contribute to PCOS development. Furthermore, therapeutic strategies targeting immune microenvironment imbalance such as modulating macrophage polarization, restoring Th1/Th2 and Th17/Treg balance, and ameliorating gut microbiota dysbiosis are discussed, offering novel insights for PCOS immunotherapy. In conclusion, this review comprehensively analyzes the pathogenesis of PCOS from the perspective of the immune microenvironment, aiming to provide a theoretical foundation and reference for future research and clinical practice.

The immune microenvironment plays a pivotal role in maintaining ovarian homeostasis. Polycystic ovary syndrome (PCOS), a common endocrine and metabolic disorder, is closely associated with immune microenvironment imbalance. This review systematically describes the dysregulation of innate immune cells (e.g., macrophages, natural killer cells and dendritic cells) and adaptive immune cells (e.g., Th1, Th2, Treg and Th17) in PCOS, highlighting their impacts on ovarian function, insulin resistance, and hyperandrogenemia. These findings underscore the central role of immune microenvironment disturbances in PCOS pathogenesis. Additionally, the association between gut microbiota dysbiosis and PCOS is explored, emphasizing how gut microbiota influences metabolic byproducts and hormonal levels to contribute to PCOS development. Furthermore, therapeutic strategies targeting immune microenvironment imbalance such as modulating macrophage polarization, restoring Th1/Th2 and Th17/Treg balance, and ameliorating gut microbiota dysbiosis are discussed, offering novel insights for PCOS immunotherapy. In conclusion, this review comprehensively analyzes the pathogenesis of PCOS from the perspective of the immune microenvironment, aiming to provide a theoretical foundation and reference for future research and clinical practice.

Objective To observe the value of whole-brain CT perfusion(CTP)parameters at different time and clinical data for predicting delayed cerebral ischemia(DCI)and 3-month poor prognosis in patients with aneurysmal subarachnoid hemorrhage(aSAH).Methods Totally 127 aSAH patients were retrospectively enrolled.Clinical and CTP data within 24 h of symptom onset and during DCI time window(DCITW)were collected.The patients were divided into DCI group(n=34)and non-DCI group(n=93)based on DCI occurred or not during hospitalization,also into poor outcome group(modified Rankin scale[mRS]≥3,n=36)and good outcome group(mRS≤2)based on 3-month's follow-up.Multivariate logistic regression was performed to select independent predictive factors among variates being significantly different between groups.Then receiver operating characteristic curve was drawn,and the area under the curve(AUC)was calculated to evaluate the predictive performance of logistic regression model.Results Patients'age,modified Fisher score(mFS),subarachnoid hemorrhage early brain edema score(SEBES)and mean flow extraction product(mFEP)within 24 h of onset were all identified as independent predictive factors of DCI,and the AUC of their combination for predicting DCI during hospitalization was 0.817.Patients' age and mFS within 24 h of onset,alternatively,World Federation of Neurosurgical Societies(WFNS)grade and mFEP during DCITW were all independent predictive predictors of 3 months' prognosis,and the combination of the latter two showed better predictive performance(AUC=0.922)tahn the former two(AUC=0.822,P＜0.05).Conclusion Whole-brain CTP parameters combined with clinical data within 24 h of onset of aSAH could be used to predict the occurrence of DCI during hospitalization,whole-brain CTP parameters during DCITW could be used to predict 3 months'poor prognosis.

Objective:To explore the application value of real-time virtual sonography (RVS) combined with intraductal biliary contrast-enhanced ultrasound (IB-CEUS) in percutaneous transhepatic cholangial drainage (PTCD).Methods:This retrospective cohort study included data from 71 patients who underwent PTCD at the Department of Hepatobiliary and Pancreatic Surgery in the Second Affiliated Hospital of Nanchang University between May 2021 and August 2022. There were 36 male and 35 female patients,aged 35 to 94 years. Based on the guidance modality used,patients were divided into two groups: the RVS combined with IB-CEUS group ( n=36) and the digital subtraction angiography (DSA) group ( n=35). PTCD was performed under the guidance of RVS combined with IB-CEUS in the RVS+IB-CEUS group,and under conventional DSA fluoroscopic guidance in the DSA group. Two clinicians classified the biliary conditions as either simple or complex based on preoperative ultrasound and CT (or MRI) imaging. Statistical analyses were conducted using independent sample t-tests,rank-sum tests, χ2 tests,or Fisher′s exact tests,as appropriate. Results:Significant differences were observed between the RVS+IB-CEUS group and the DSA group in terms of the number of punctures (1.0±0.2 vs. 2.2±1.4, t=-5.148, P<0.01) and postoperative complication rate(2.8% (1/35) vs. 17.1% (6/36), P=0.049). There were 9 patients with complex biliary conditions in the DSA group and 12 in the RVS+IB-CEUS group. The number of punctures in both the simple and complex subgroups of the RVS+IB-CEUS group(1.0±0.2 and 1.0±0.0) remained lower than that in the corresponding DSA subgroups(2.2±1.6 and 2.4±0.4) ( t=-3.606, P<0.01; t=-3.959, P=0.002). Moreover,the complication rate in the simple biliary subgroup of the RVS+IB-CEUS group was significantly lower than that of the DSA group(0 (0/24) vs. 19.2% (5/26), P=0.031),whereas no significant difference was found in the complex biliary subgroup (1/12 vs. 1/9, P=0.686). Conclusion:Guided by RVS and IB-CEUS, PTCD can help reduce the number of punctures during surgery and postoperative complications, and patients with complex bile duct conditions can still benefit from PTCD.

Objective To investigate the performance of different methods in common autoantibody detec-tion,and to provide basis for the detection strategy of autoantibody.Methods A total of 2 657 cases were in-cluded in this study,whose serum antinuclear antibody(ANA)by indirect immunofluorescence method(IIF)and antinuclear antibody spectrum(ANAs,a total of 15 antibodies)by immunoblot method(IB)were tested simultaneously,and anti-double strand DNA antibody(anti-dsDNA antibody)were tested by IIF,enzyme linked immunosorbent assay(ELISA)and IB.ANCA and ANCA spectrum[anti-myeloperoxidase antibody(anti-MPO antibody),anti-protease 3 antibody(anti-PR3 antibody)]were tested by IIF and ELISA respec-tively in 2 348 cases.Results Firstly,the detection rate of ANA was significantly higher than that of ANAs(57.77％vs.30.64％,P＜0.001).Among the specimens detected with ANA,40.78％detected at least one specific antibody positive.Among the specimens detected with ANAs positive,ANA was not detected in 23.10％of cases.Secondly,the ANA positive cases were mainly of lower titers,and cases with titer of ≤1∶320 accounted for 84.95％of the total detected cases.But the higher the titer,the higher the detection rate of ANAs specific antibodies(29.63％in 1∶100,48.56％in 1∶320,77.78％in 1∶1 000,92.42％in≥1∶3 200).Thirdly,among the specimens detected ANAs positive,the composition ratios of anti-Scl-70,anti-PM-Scl and anti-PCNA antibodies in ANA positive cases was significantly lower than those in ANA negative cases(P＜0.001).Fourthly,the detection rate of anti-dsDNA antibody by ELISA was the highest(4.14％),which was significantly higher than those by IIF or IB(both P＜0.001),and the difference in detection efficacy of the latter two methods was also statistically significant(P＜0.001).Even so,the IIF and IB for the detection of anti-dsDNA antibody still had a certain complementary effect on ELISA.Fifthly,the positive rate of ANCA by IIF was significantly higher than that of ANCA spectrum(9.16％vs.2.43％),but among those who were detected anti-MPO antibodies and/or anti-PR3 antibodies positive,47.37％were ANCA negative.Conclusion Different methods for detecting autoantibodies have significantly different detection efficacy,and it is partic-ularly necessary to adopt correct detection strategies based on evidence-based medicine.