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.

Objectives:To investigate the clinical value of cardiac magnetic resonance imaging(CMR)feature-tracking strain analysis in risk stratification of diabetic heart failure with preserved ejection fraction(HFpEF).Methods:In this retrospective study,a total of 215 patients with diabetic HFpEF who underwent CMR at Chinese Academy of Medical Sciences Fuwai Hospital from January 2012 to December 2018 were included.Myocardial strain parameters were calculated using CMR feature-tracking technology.Patients were followed up by medical records or telephone calls.Composite endpoint event,all-cause death or heart failure hospitalization during follow-up were recorded.Patients were divided into event group and event-free group.Univariable and multivariable Cox proportional hazard regression analyses were performed to determine the risk factors for the outcomes in diabetic HFpEF.The effects of hypertension and obesity on the prognosis of diabetic HFpEF patients and whether they affect the prognostic value of CMR feature-tracking strain analysis were also analyzed.Results:During a follow-up of(7.1±1.8)years,93(43.3%)patients had endpoint events(event group),including 28 all-cause deaths and 65 heart failure hospitalization.Compared with the event-free group(n=122),patients in the event group had significantly lower left ventricular ejection fraction,higher prevalence and extent of late gadolinium enhancement,and significantly reduced global longitudinal strain(GLS),global circumferential strain,global radial strain,and global systolic longitudinal strain rate(all P<0.05).The absolute GLS value was significantly lower in event group than in event-free group,regardless of the presence of hypertension and obesity.Multivariate Cox regression analysis showed that estimated glomerular filtration rate(HR=0.983,95%CI:0.972-0.993,P=0.001),left atrial volume index(HR=1.015,95%CI:1.005-1.026,P=0.004),and GLS(HR=1.142,95%CI:1.060-1.231,P<0.001)were independent risk factors for adverse cardiovascular events in diabetic HFpEF patients.However,adjusted N-terminal pro-brain natriuretic peptide was not an independent prognostic factor.The cut-offvalue of GLS to predict outcome was-14.09%from ROC curve analysis.The Kaplan-Meier curve showed that in patients with and without hypertension and obesity,patients with the GLS>-14.09%had lower event-free survival compared to patients with GLS≤-14.09%(all P<0.05),and the ability of GLS to predict adverse outcomes was not affected by hypertension and obesity.Conclusions:GLS obtained by CMR feature-tracking strain analysis is an independent predictor of adverse outcomes in diabetic HFpEF,and its ability to predict adverse outcomes is independent of hypertension and obesity.

Objectives:To investigate the clinical value of cardiac magnetic resonance imaging(CMR)feature-tracking strain analysis in risk stratification of diabetic heart failure with preserved ejection fraction(HFpEF).Methods:In this retrospective study,a total of 215 patients with diabetic HFpEF who underwent CMR at Chinese Academy of Medical Sciences Fuwai Hospital from January 2012 to December 2018 were included.Myocardial strain parameters were calculated using CMR feature-tracking technology.Patients were followed up by medical records or telephone calls.Composite endpoint event,all-cause death or heart failure hospitalization during follow-up were recorded.Patients were divided into event group and event-free group.Univariable and multivariable Cox proportional hazard regression analyses were performed to determine the risk factors for the outcomes in diabetic HFpEF.The effects of hypertension and obesity on the prognosis of diabetic HFpEF patients and whether they affect the prognostic value of CMR feature-tracking strain analysis were also analyzed.Results:During a follow-up of(7.1±1.8)years,93(43.3%)patients had endpoint events(event group),including 28 all-cause deaths and 65 heart failure hospitalization.Compared with the event-free group(n=122),patients in the event group had significantly lower left ventricular ejection fraction,higher prevalence and extent of late gadolinium enhancement,and significantly reduced global longitudinal strain(GLS),global circumferential strain,global radial strain,and global systolic longitudinal strain rate(all P<0.05).The absolute GLS value was significantly lower in event group than in event-free group,regardless of the presence of hypertension and obesity.Multivariate Cox regression analysis showed that estimated glomerular filtration rate(HR=0.983,95%CI:0.972-0.993,P=0.001),left atrial volume index(HR=1.015,95%CI:1.005-1.026,P=0.004),and GLS(HR=1.142,95%CI:1.060-1.231,P<0.001)were independent risk factors for adverse cardiovascular events in diabetic HFpEF patients.However,adjusted N-terminal pro-brain natriuretic peptide was not an independent prognostic factor.The cut-offvalue of GLS to predict outcome was-14.09%from ROC curve analysis.The Kaplan-Meier curve showed that in patients with and without hypertension and obesity,patients with the GLS>-14.09%had lower event-free survival compared to patients with GLS≤-14.09%(all P<0.05),and the ability of GLS to predict adverse outcomes was not affected by hypertension and obesity.Conclusions:GLS obtained by CMR feature-tracking strain analysis is an independent predictor of adverse outcomes in diabetic HFpEF,and its ability to predict adverse outcomes is independent of hypertension and obesity.

Objective To explore the clinical efficacy of the regimen based on sacubitril/valsartan and empagliflozin in the treatment of elderly hypertension patients with type 2 diabetes(T2DM)and determine the impact on left ventricular remodeling.Methods A prospective randomized trial was conducted on 100 elderly patients with essential hypertension combined with T2DM admitted in our hospital from January 2022 to January 2023.All of them had left ventricular hypertrophy(LVH).They were randomly divided into an observation group(n=50,empagliflozin combined with sacubitril/valsartan)and a control group(n=50,empagliflozin combined with valsartan).Baseline data,and SBP,DBP,left ventricular ejection fraction,left ventricular mass(LVM),interventricular septal thickness(IVST),left ventricular posterior wall thickness(LVPWT),and LVM index(LVMI)before and at 3 and 6 months after treatment were recorded and compared between the two groups.The incidence of adverse events was also compared between them.Results Repeated measures ANOVA showed that the values of SBP,DBP,IVST,LVPWT,LVM and LVMI were gradually reduced in the two groups with the extension of treatment time,and reached the lowest levels at 6 months of treatment(Ftime=4.829,10.037,121.202,65.784,214.796,162.801,P＜0.05,P＜0.01).With the elapse of treatment time,the observation group obtained more significant declines in terms of SBP,DBP,IVST,LVPWT,LVM and LVMI(Ftreatment×time=5.252,5.685,4.574,6.239,8.292,11.660,P＜0.05,P＜0.01).The inter-group effect was statistically significant.After 6 months of treatment,the values of above indicators were notably lower in the observation group than the control group(Ftreatment=79.151,97.673,4.250,8.080,9.598,20.091,P＜0.05,P＜0.01).SBP(129.80±7.67 mm Hg vs 138.48±12.35 mm Hg.P＜0.01),DBP(79.76±5.96 mm Hg vs 87.46±9.57 mm Hg,P＜0.01),IVST,LVPWT,LVM and LVMI were obviously lower in the observation group than the control group.Both valsartan+empagliflozin and sacubitril/valsartan+empagliflozin showed good safety.Conclusion Sacubitril/valsartan can effectively improve blood pressure,cardiac function,and left ventricular remodeling in elderly hypertensive patients with T2DM and LVH,with good safety.

Objective To explore the clinical efficacy of the regimen based on sacubitril/valsartan and empagliflozin in the treatment of elderly hypertension patients with type 2 diabetes(T2DM)and determine the impact on left ventricular remodeling.Methods A prospective randomized trial was conducted on 100 elderly patients with essential hypertension combined with T2DM admitted in our hospital from January 2022 to January 2023.All of them had left ventricular hypertrophy(LVH).They were randomly divided into an observation group(n=50,empagliflozin combined with sacubitril/valsartan)and a control group(n=50,empagliflozin combined with valsartan).Baseline data,and SBP,DBP,left ventricular ejection fraction,left ventricular mass(LVM),interventricular septal thickness(IVST),left ventricular posterior wall thickness(LVPWT),and LVM index(LVMI)before and at 3 and 6 months after treatment were recorded and compared between the two groups.The incidence of adverse events was also compared between them.Results Repeated measures ANOVA showed that the values of SBP,DBP,IVST,LVPWT,LVM and LVMI were gradually reduced in the two groups with the extension of treatment time,and reached the lowest levels at 6 months of treatment(Ftime=4.829,10.037,121.202,65.784,214.796,162.801,P＜0.05,P＜0.01).With the elapse of treatment time,the observation group obtained more significant declines in terms of SBP,DBP,IVST,LVPWT,LVM and LVMI(Ftreatment×time=5.252,5.685,4.574,6.239,8.292,11.660,P＜0.05,P＜0.01).The inter-group effect was statistically significant.After 6 months of treatment,the values of above indicators were notably lower in the observation group than the control group(Ftreatment=79.151,97.673,4.250,8.080,9.598,20.091,P＜0.05,P＜0.01).SBP(129.80±7.67 mm Hg vs 138.48±12.35 mm Hg.P＜0.01),DBP(79.76±5.96 mm Hg vs 87.46±9.57 mm Hg,P＜0.01),IVST,LVPWT,LVM and LVMI were obviously lower in the observation group than the control group.Both valsartan+empagliflozin and sacubitril/valsartan+empagliflozin showed good safety.Conclusion Sacubitril/valsartan can effectively improve blood pressure,cardiac function,and left ventricular remodeling in elderly hypertensive patients with T2DM and LVH,with good safety.

Objectives:To analyze the consistency of evaluating left ventricular hemodynamics (HDF) based on single plane and multi plane cine sequences of magnetic resonance mitral valve orifice.Methods:A prospective study was conducted on 48 healthy adults, and two methods were used to measure the mitral valve diameter and calculate HDF parameters. The first method was to measure the diameter of the mitral valve opening in the left ventricular three chamber cine sequence; The second method is to measure the mitral valve diameter using cine sequences of two chamber, three chamber, and four chamber hearts, and then take the average value. Paired t-tests were used to compare the differences in HDF measured by two methods, and Pearson correlation coefficient ( r), intra group correlation coefficient ( ICC), and Bland-Altman analysis were used to test the consistency and reproducibility of the two methods. Results:The root mean square (RMS) of longitudinal HDF calculated using single plane and multi plane mitral valve diameters were [(17.28±4.41)% vs (17.21±4.61)%] ( P=0.379) for the entire cardiac cycle, [(21.45±5.54)% vs (21.49±5.68)%] ( P=0.646) for systolic phase, and [(12.78±4.10)% vs (12.54±4.24)%] ( P=0.106) for diastolic phase, respectively. The difference in the calculation results of HDF parameters related to ventricular function was not statistically significant (all P>0.05), and there was good consistency ( r=0.924-0.996, ICC=0.924-0.995). The two HDF parameters related to atrial function were sensitive to the measurement method of mitral valve orifice diameter [RMS of longitudinal HDF during active atrial emptying: (3.26±1.51)% vs (3.32±1.55)%, P=0.006; longitudinal HDF pulse during active atrial emptying: (-2.60±1.28)% vs (-2.76±1.30)%, P<0.001]. Conclusions:The ventricular function related HDF parameters obtained from the analysis of mitral valve orifice diameter using single plane and multi plane methods have good consistency, and can be evaluated using relatively simple single plane methods for left ventricular HDF.

Objective: To determine the active components of Eupolyphaga sinensis Walker (Tu Bie Chong) and explore the mechanisms underlying its fracture-healing ability. Methods: A modified Einhorn method was used to develop a rat tibial fracture model. Progression of bone healing was assessed using radiological methods. Safranin O/fast green and CD31 immunohistochemical staining were performed to evaluate the growth of bone cells and angiogenesis at the fracture site. Methylthiazoletetrazolium blue and wound healing assays were used to analyze cell viability and migration. The Transwell assay was used to explore the invasion capacity of the cells. Tubule formation assays were used to assess the angiogenesis capacity of human vascular endothelial cells (HUVECs). qRT-PCR was used to evaluate the changes in gene transcription levels. Results: Tu Bie Chong fraction 3 (TF3) significantly shortened the fracture healing time in model rats. X-ray results showed that on day 14, fracture healing in the TF3 treatment group was significantly better than that in the control group (P = .0086). Tissue staining showed that cartilage growth and the number of H-shaped blood vessels at the fracture site of the TF3 treatment group were better than those of the control group. In vitro, TF3 significantly promoted the proliferation and wound healing of MC3T3-E1s and HUVECs (all P < .01). Transwell assays showed that TF3 promoted the migration of HUVECs, but inhibited the migration of MC3T3-E1 cells. Tubule formation experiments confirmed that TF3 markedly promoted the ability of vascular endothelial cells to form microtubules. Gene expression analysis revealed that TF3 significantly promoted the expression of VEGFA, SPOCD1, NGF, and NGFR in HUVECs. In MC3T3-E1 cells, the transcript levels of RUNX2 and COL2A1 were significantly elevated following TF3 treatment. Conclusion TF3 promotes fracture healing by promoting bone regeneration associated with the RUNX2 pathway and angiogenesis associated with the VEGFA pathway.

Objective To investigate the pathological damage to and inflammation of different intestinal segments in a rat model of severe hemorrhage,and to explore the effect of polarization of intestinal macrophage on the pathophysiology of intestinal inflammation.Methods Male Wistar rats were randomly divided into two groups:the sham operation group and hemorrhage group.In the hemorrhage group,40％of the total blood volume was lost in 25-30 minutes,while in the sham operation group,only the femoral artery and vein were intubated without bleeding.The rats were killed at 0,3,6,12 and 24 hours.The entire intestine was isolated quickly,and sections of the intestine were cut at the duodenum,jejunum,ileocecal junction,colon and rectum for histopathological evaluation.ELISA was adopted to determine related inflammation factors while multi-color immunohistochemistry was used to calculate macrophage surface markers.The data was statistically analyzed.Results(1)Compared with the sham group,there was no significant difference in colon histology at 3 h and 6 h,but significant difference was detected in rectum scores only at 24 h.The scores of other intestinal segments were significantly different at each time point.The severity of ileocecal and colonic lesions after bleeding increased with time.The duodenum,jejunum and ileocecum were more critically injured at 3 h than the rectum at 6 h.The injury to the duodenum,jejunum,ileum and colon was much more pronounced than to the rectum at 12 h.(2)The expressions of TNF-α and IL-1β in the rectum were increased significantly at 12 h post operation.The expressions of IL-1β,TNF-α in the jejunum increased obviously at 3 h and 6 h,respectively.(3)Three hours after severe bleeding,the level of macrophages in the jejunum and ileocececal area increased significantly,and the percentage of M1 macrophages was higher.After 6 hours,the proportion of M2 macrophages in the jejunum and M1 macrophages decreased significantly.After 3 hours,the percentage of M1 macrophages in the colon decreased,but that of M2 macrophages increased.The proportion of M2 polarized macrophages in the duodenum and rectum increased at 3 h after severe bleeding but decreased at 6 h.Conclusion Pathological damage to intestinal sections after bleeding varies depending on the time,and is correlated with the inflammatory level of macrophages.

Objective To analyze the relationship between fragmented QRS(fQRS)and microalbuminuria(MAU)and serum uric acid(SUA)in patients with type 2 diabetes mellitus(T2DM).Methods From October 2022 to June 2023,245 T2DM patients diagnosed by The Department of Endocrinology,Jinhua Hospital Affiliated to Zhejiang University School of Medicine were selected and divided into fQRS group(fQRS,n=111)and T2DM group(n=134)according to ECG results.The general data,biochemical indexes,left ventricular end-diastolic diameter(LVEDd),left ventricular end-systolic diameter(LVESd),interventricular septum(IVS),left ventricular posterior wall(LVPW),left ventricular ejection fraction(LVEF),left atrial diameter(LAd),peak value ratio of mitral orifice blood flow velocity in early and late diastole(E/A)and mitral orifice blood flow velocity in early diastole were compared between the two groups.Pearson correlation analysis was used to analyze the correlation between fQRS wave and other indexes in T2DM patients,and Logistic regression analysis was used to analyze the influencing factors of fQRS.Results The prevalence rates of UACR,SUA,HbA1c and MAU in fQRS group were higher than in T2DM group(P＜0.05 or P＜0.01).LVEDd,LAd,E,E/e'and E/A>1.5 in QRS group were higher than in T2DM group(P＜0.05 or P＜0.01).Pearson correlation analysis showed that fQRS wave was positively correlated with smoking history,BMI,HbA1c,UACR,MAU,SUA,LVEDd,LAd,E/A and E/e'(P＜0.05).Logistic regression analysis showed that MAU was the influencing factor of fQRS wave in T2DM patients.Conclusions The fQRS in ECG in patients with T2DM is associated with MAU and SUA and may be an important evidence of myocardial fibrosis associated with subclinical cardiac diastolic dysfunction.

Objective To explore the effects of prucalopride(PRUC)on the intestinal function during the perioperative period of robot-assisted laparoscopic radical cystectomy(RARC)and urinary diversion.Methods A total of 75 patients undertaking RARC with urinary diversion(orthotopic neobladder or ileal bladder)in Nanjing Drum Hospital during Jan.and Dec.2021 were divided into PRUC group(n=28)and control group(n=47)according to whether they took PRUC or not.Postoperative intestinal ventilation time and defecation time,drainage tube retention time,tolerance time for first intake of semi-flow food,postoperative hospital stay,and incidence of complications were observed and recorded in the two groups.Postoperative C-reactive protein(CRP)and neutrophil/lymphocyte ratio(NLR)were compared.Results The PRUC group had shorter intestinal ventilation time and defecation time[(47.14±16.31)h vs.(74.04±35.33)h,P＜0.01;(86.14±30.47)h vs.(123.57±79.12)h,P=0.02],smaller change of ΔCRP and ΔNLR[(79.99±29.71)mg/L vs.(127.75±56.98)mg/L;(9.24±6.43)vs.(16.11±9.90),P＜0.01].All complications were minor,the incidence of intestinal obstruction in PRUC group tended to decrease within 90 days after operation(P=0.38),and there was no significant difference in other complications between the two groups(P＞0.05).Conclusion The perioperative use of PRUC in RARC with urinary diversion is safe and effective,which can promote the recovery of intestinal function after operation.