Objective To investigate the changes in eosinophil counts and the activation of microglial cells in the brain tissues of mice at different stages of Angiostrongylus cantonensis infection, and to examine the role of microglia in regulating the progression of angiostrongyliasis and unravel the possible molecular mechanisms. Methods Fifty BALB/c mice were randomly divided into the control group and the 7-d, 14-d, 21-day and 25-d infection groups, of 10 mice in each group. All mice in infection groups were infected with 30 stage III A. cantonensis larvae by gavage, and animals in the control group was given an equal amount of physiological saline. Five mice were collected from each of infection groups on days 7, 14, 21 d and 25 d post-infection, and 5 mice were collected from the control group on the day of oral gavage. The general and focal functional impairment was scored using the Clark scoring method to assess the degree of mouse neurological impairment. Five mice from each of infection groups were sacrificed on days 7, 14, 21 d and 25 d post-infection, and 5 mice from the control group were sacrificed on the day of oral gavage. Mouse brain tissues were sampled, and the pathological changes of brain tissues were dynamically observed using hematoxylin and eosin (HE) staining. Immunofluorescence staining with eosinophilic cationic protein (ECP) and ionized calcium binding adaptor molecule 1 (Iba1) was used to assess the degree of eosinophil infiltration and the counts of microglial cells in mouse brain tissues in each group, and the morphological parameters of microglial cells (skeleton analysis and fractal analysis) were quantified by using Image J software to determine the morphological changes of microglial cells. In addition, the expression of M1 microglia markers Fcγ receptor III (Fcgr3), Fcγ receptor IIb (Fcgr2b) and CD86 antigen (Cd86), M2 microglia markers Arginase 1 (Arg1), macrophage mannose receptor C-type 1 (Mrc1), chitinase-like 3 (Chil3), and phagocytosis genes myeloid cell triggering receptor expressed on myeloid cells 2 (Trem2), CD68 antigen (Cd68), and apolipoprotein E (Apoe) was quantified using real-time quantitative reverse transcription PCR (RT-qPCR) assay in the mouse cerebral cortex of mice post-infection. Results A large number of A. cantonensis larvae were seen on the mouse meninges surface post-infection, and many neuronal nuclei were crumpled and deeply stained, with a large number of bleeding points in the meninges. The median Clark scores of mouse general functional impairment were 0 (interquartile range, 0), 0 (interquartile range, 0.5), 6 (interquartile range, 1.0), 14 (interquartile range, 8.5) points and 20 (interquartile range, 9.0) points in the control group and the 7-d, 14-d, 21-d and 25-d groups, respectively (H = 22.45, P < 0.01), and the median Clark scores of mouse focal functional impairment were 0 (interquartile range, 0), 2 (interquartile range, 2.5), 7 (interquartile range, 3.0), 18 (interquartile range, 5.0) points and 25 (interquartile range, 6.5) points in the control group and the 7-d, 14-d, 21-d and 25-d groups, respectively (H = 22.72, P < 0.01). The mean scores of mice general and focal functional impairment were all higher in the infection groups than in the control group (all P values < 0.05). Immunofluorescence staining showed a significant difference in the eosinophil counts in mouse brain tissues among the five groups (F = 40.05, P < 0.000 1), and the eosinophil counts were significantly higher in mouse brain tissues in the 14-d (3.08 ± 0.78) and 21-d infection groups (5.97 ± 1.37) than in the control group (1.00 ± 0.28) (both P values < 0.05). Semi-quantitative analysis of microglia immunofluorescence showed a significant difference in the counts of microglial cells among the five groups (F = 17.66, P < 0.000 1), and higher Iba1 levels were detected in mouse brain tissues in 14-d (5.75 ± 1.28), 21-d (6.23 ± 1.89) and 25-d infection groups (3.70 ± 1.30) than in the control group (1.00 ± 0.30) (all P values < 0.05). Skeleton and fractal analyses showed that the branch length [(162.04 ± 34.10) μm vs. (395.37 ± 64.11) μm; t = 5.566, P < 0.05] and fractal dimension of microglial cells (1.30 ± 0.01 vs. 1.41 ± 0.03; t = 5.266, P < 0.05) were reduced in mouse brain tissues in the 21-d infection group relative to the control group. In addition, there were significant differences among the 5 groups in terms of M1 and M2 microglia markers Fcgr3 (F = 48.34, P < 0.05), Fcgr2b (F = 55.46, P < 0.05), Cd86 (F = 24.44, P < 0.05), Arg1 (F = 31.18, P < 0.05), Mrc1 (F = 15.42, P < 0.05) and Chil3 (F = 24.41, P < 0.05), as well as phagocytosis markers Trem2 (F = 21.19, P < 0.05), Cd68 (F = 43.95, P < 0.05) and Apoe (F = 7.12, P < 0.05) in mice brain tissues. Conclusions A. cantonensis infections may induce severe pathological injuries in mouse brain tissues that are characterized by massive eosinophil infiltration and persistent activation of microglia cells, thereby resulting in progressive deterioration of neurological functions.

OBJECTIVE To study the efficacy of Tianjiang xueshuantong pills in the treatment of coronary heart disease. METHODS In accordance with the common pathogenesis of coronary heart disease， acute myocardial ischemia model， hyperlipidemia model， blood stasis model， and carotid artery thrombosis model were established using Wistar rats or SD rats as the experimental subjects. The effects of Tianjiang xueshuantong pills administered at high， medium， and low doses （0.6， 1.2 and 2.4 g/kg） on hemodynamic parameters and myocardial enzyme markers [lactate dehydrogenase （LDH）， creatine kinase-MB （CK- MB）]， oxidative stress factors [superoxide dismutase （SOD）， malondialdehyde （MDA）， glutathione （GSH）]， inflammatory cytokines [tumor necrosis factor-α （TNF-α）， interleukin-6 （IL-6）， IL-1β， monocyte chemotactic protein-1 （MCP-1）， intercellular adhesion molecule-1 （ICAM-1）]， myocardial infarction percentage， serum lipid indexes [total cholesterol （TC）， triglycerides （TG）， low-density lipoprotein （LDL）， high-density lipoprotein （HDL）]， platelet aggregation function 话：022-84845240。E-mail：jiangx@tjipr.com [maximum aggregation rate （MAR）]， and thrombus formation indexes [thrombosis time， thrombus mass， thrombus protein content， plasminogen activator inhibitor-1 （PAI-1）， and tissue-type plasminogen activator （t-PA）] were evaluated in the rat models. RESULTS In myocardial ischemia tests， Tianjiang xueshuantong pills significantly reduced the percentage of myocardial infarction and the levels of CK-MB， LDH， MDA， GSH， IL-6， TNF-α， IL- 1β， and MCP-1 in serum （P＜0.05 or P＜0.01）. In hyperlipidemia tests， high dose of Tianjiang xueshuantong pills significantly reduced the serum levels of TC， LDL and significantly increased the level of HDL in rats after 2 weeks and 4 weeks of administration. In blood stasis tests， different doses of Tianjiang xueshuantong pills significantly reduced MAR of rats （P＜0.01）. In artery thrombosis tests， high dose of Tianjiang xueshuantong pills significantly prolonged the time of thrombosis formation （P＜ 0.01）， significantly reduced the weight and protein content of thrombus and the level of PAI-1 in serum （P＜0.01）. CONCLUSIONS Tianjiang xueshuantong pills exert therapeutic effects on coronary heart disease through multi-dimensional synergistic actions， including anti-myocardial ischemia， lipid-lowering， and anti-thrombotic effects.

Objective To evaluate the effectiveness of screening-intervention management program for high risk population of stroke in community.Methods Participants aged≥60 years old in Tairi Community,Fengxian District,Shanghai from May 2019 to July 2022 were selected as screening and intervention objects.The first round of stroke high-risk group screening was conducted in 2019 and 2020 in two years respectively,and the second round of stroke high-risk group screening(re-screening)was conducted in 2021 and 2022 respectively for the 2019 and 2020 screening groups,and the groups who had received stroke high-risk screening in both rounds of screening(overlapping groups)were selected as the observation objects of this study.The cerebrovascular function score was used to screen the high-risk individuals of stroke,75-100 was classified as non-high-risk,＜75 were classified as high risk,among which 50-74,25-49,0-24 were light,medium and severe risk,in turn.Baseline and follow-up data were collected for all screening groups,including systolic blood pressure,diastolic blood pressure,overweight or obesity,fasting blood glucose,glycated hemoglobin,triglyceride,total cholesterol,low-density lipoprotein cholesterol,high-density lipoprotein cholesterol,and blood uric acid.After the completion of screening,the test report interpretation and first diagnosis intervention were carried out on the screening site,and the screening results were recorded into the health examination file.The first intervention includes lifestyle intervention,risk factor intervention and therapeutic intervention for high-risk individuals.Lifestyle intervention and risk factor intervention were conducted through the distribution of popular science handbook for stroke prevention and individualized face-to-face guidance.Therapeutic intervention for high-risk individuals was guided by anti-platelet aggregation drug therapy,statin therapy,and further examination and treatment of cerebral vessels according to stroke risk assessment results and the incidence of related chronic diseases.Prior to the implementation of the project,the incidence of stroke in the community in 2018 was retrospectively investigated to compare annual changes in stroke screening-intervention.In the process of implementation of intervention management,stroke incidence monitoring of the whole community registered population was carried out,and the monitoring method was to conduct stroke incidence registration once a year,and cooperate with the disease control and community police station to obtain the community stroke incidence monitoring data and death registration information provided by the police station from 2018 to 2022.Results A total of 5 188 subjects who completed both the initial screening and the follow-up screening and met the inclusion and exclusion criteria were identified,of whom 2 269 were male and 2 923 were female.The age of participants ranged from 60 to 93 years at the time of the first round of screening,with a mean age of(68±6)years.The proportions of subjects in the age groups of 60-64,65-69,70-74,and≥75 years were 30.3％,34.7％,21.1％,and 14.0％,respectively.(1)After screening and intervention,the proportion of individuals with increased systolic blood pressure,diastolic blood pressure,fasting blood glucose,triglyceride,and low-density lipoprotein cholesterol all decreased(respectively 49.4％vs.57.3％,26.6％vs.28.7％,9.6％vs.10.9％,14.7％vs.17.0％,2.4％vs.3.3％;all P＜0.05),but the proportion of individuals with hyperuricemia increased(15.8％vs.13.1％,P＜0.01),with statistically significant differences.(2)Before the implementation of the screening-intervention program in 2018,the stroke incidence rate in the community was 332.1/100 000.The stroke incidence rates in the community during the period from 2019 to 2022 after the implementation of the screening-intervention program were 335.0/100 000,270.8/100 000,235.0/100 000,and 193.6/100 000,respectively.The incidence rates of ischemic stroke(x2trend=8.350,P=0.004)and stroke(x2trend=9.910,P=0.002)decreased during the period from 2019 to 2022,while the incidence rate of hemorrhagic stroke did not show a decreasing trend(x2trend=1.636,P=0.201).(3)The median baseline and follow-up cerebrovascular function scores for the 5 188 elderly individuals undergoing residual stroke risk screening were 82.50(52.50,98.75)and 88.5(59.00,100.00),respectively,with stroke risk rates of 39.8％and 35.6％before and after intervention,respectively.After intervention,the follow-up cerebrovascular function scores increased compared to the baseline,and the stroke risk rate decreased.The distribution of stroke risk levels before and after screening-intervention had statistically significant differences(P＜0.01).Conclusion Implementing a stroke high-risk population screening-intervention management program for the elderly in the community,combined with health examinations and family doctor team services,can significantly reduce the incidence,high-risk rate,and exposure level of risk factors for stroke in the community.

This study was aimed at investigating the prevalence and characteristics of Angiostrongylus cantonensis infection in snails for sale in select regions of Yunnan Province between April and September in 2019-2023.A total of 1 584 snails were collected from 13 regions,and third stage larvae with A.cantonensis infection were identified with the digestion method.The A.cantonensis infection rate in snails was 3.66％.The infection rate of Pomacea canaliculata was highest(6.41％),that of Cipangopaludina cathayensis was 2.74％,and that of Bellamya aeruginosa was 2.56％.The A.cantonensis infection rate in each snail species decreased each year,except in P.canaliculata.The A.cantonensis infection rate was highest in July(7.87％).Snails infected with A.cantonensis were found in regions except Chuxiong.The highest infection rate in these 12 re-gions occurred in Honghe(9.52％).In conclusion,A.cantonensis infection was identified in snails for sale in select areas of Yunnan province,thus posing hidden food safety risks.Strengthened supervision and management of snail food products,in-creased education,environmental improvement,and deratization deratize are necessary.Moreover,molecular biological detec-tion provides a sensitive tool for the prevention and control of A.cantonensis.

Objective:To investigate the value of immunoglobulin heavy chain(IgH)gene rearrangement in monitoring minimal residual disease(MRD)in multiple myeloma(MM)received autologous hematopoietic stem cell transplantation(auto-HSCT).Methods:The clinical data of 26 MM patients who received auto-HSCT in the Department of Hematology,Wuhan First Hospital from 2018 to 2022 were collected.IgH rearrangement was detected by multiplex PCR combined with capillary electrophoresis fragment analysis to evaluate minimal residual disease(MRD),and the outcome of the disease was analyzed statistically.Results:Among the 26 MM patients,18 were males and 8 were females,with a median age of 59(41-70)years.The median follow-up time after transplantation was 33(7-52)months.Compared with the IgH rearrangement negative group(n=17),the proportion of CR and sCR of patients with IgH rearrangement positive in bone marrow samples before auto-HSCT at 3 months after transplantation was lower(1/9 vs 14/17),and the duration of remission(DOR)after transplantation was shorter(10.78±4.35 vs 15.88±5.22 months),with statistically significant difference in DOR between the two groups(P＜0.05).Compared with IgH rearrangement negative group(n=21),the proportion of CR and sCR of patients with positive IgH rearrangement results from peripheral blood stem cell collection at 3 months after transplantation was lower(0/5 vs 15/21),the duration of remission(DOR)after transplantation was shorter(9.60±4.83 vs 15.19±5.11 months),and the difference in DOR between the two groups was statistically significant(P＜0.05).During the follow-up period,5 patients(5/9)with positive IgH rearrangement results in bone marrow specimens died,and all patients with negative IgH rearrangement results survived.Four patients(4/5)with positive IgH rearrangement results by peripheral blood stem cell samples died,while one patient(1/21)with negative IgH rearrangement results died.In both bone marrow and peripheral blood stem cell samples,the survival time of IgH rearrangement-positive patients after transplantation was shorter than that of IgH rearrangement-negative patients(P＜0.05).Logistic regression analysis showed that gender,disease stage,the proportion of bone marrow smear plasma cells at initial diagnosis,stem cell mobilization plan,efficacy evaluation before transplantation(≥ CR and＜CR),and CD34+cell count had no effect on IgH rearrangement results of stem cell collection(P＞0.05).Conclusion:By detecting IgH rearrangement of MM patients receiving auto-HSCT,the depth of MRD can be further evaluated,which has a certain guiding significance for the efficacy and prognosis of the disease.

Objective:To investigate the reversal effect and mechanism of asiatic acid (AA)on multidrug resistance in human adriamycin (ADR)chronic myeloid leukemia K562/ADR cells.Methods:CCK-8 assay was used to detect the resistance of K562 cells and K562/ADR cells to ADR.CCK-8 assay was used to detect the effect of AA on K562/ADR cell viability and adriamycin sensitization.After K562/ADR cells were treated with non-toxic doses of AA(10,20μmol/L),the average fluorescence intensity of ADR was detected by flow cytometry.Real-time quantitative PCR was used to detect the expression levels of MRP1,P-gp,β-catenin,C-myc and cyclinD1 mRNA.Western blot was used to detect the expression levels of MRP1,P-gp,β-catenin,C-myc and cyclinD1 proteins.Western blot assay was used to detect the expression levels of MRP1,P-gp,β-catenin,C-myc and cyclinD1 proteins in K562/ADR cells treated with 20μmol/L AA and Wnt/β-catenin pathway agonist WAY-262611 (5 μmol/L).Results:The CCK-8 assay showed that the drug resistance of K562/ADR cells was 56.57 times that of K562 cells,showing stable drug resistance,and the difference was statistically significant (P<0.05 ).AA inhibited the proliferative activity of K562/ADR cells in a concentration-dependent manner(r=0.9666).Compared with 0 μmol/L AA group,the 10 and 20 μmol/L AA groups could significantly enhance the average fluorescence intensity of intracellular ADR (P<0.05 ),and reverse the cell resistance to ADR (P<0.05).The mRNA and protein expressions of MRP1,P-gp,β-catenin,C-myc and cyclinD1 in cells were down-regulated (P<0.05).Compared with 20μmol/L AA group,the expression levels of MRP1,P-gp,β-catenin,C-myc and cyclinD1 protein in 20 μmol/L AA+WAY group were significantly increased (P<0.05 ). Conclusion:AA inhibits K562/ADR cell proliferation in a concentration-dependent manner and reverse their resistance to ADR,the reversal mechanism may be related to the down-regulation of MRP1 and P-gp expression after inhibiting Wnt/β-catenin signaling pathway.

Objectives:Present study aimed to use quantitative flow ratio based on Murray's law to analyze the risk factors of impaired side branches perfusion without naked eye visible slowing of blood flow in branches after interventional treatment of true bifurcation lesions of the coronary arteries. Methods:A total of 211 patients with non-left main coronary artery true bifurcation coronary artery disease who underwent percutaneous coronary intervention(PCI)in Hunan Provincial People's Hospital from June 2022 to September 2023 were continuously enrolled,with a total of 234 bifurcation lesions.The general clinical indicators,anatomical characteristics of coronary artery bifurcation lesions,branch protection methods,postoperative branch TIMI blood flow and other data were collected,and quantitative flow ratio(μQFR)was measured for postoperative branch blood vessels.Post-PCI μQFR<0.8 was considered as impaired branch perfusion and was included in the postoperative impaired branch perfusion group(n=51,53 branch lesions).Patients with μQFR≥0.8 were included in the postoperative normal branch perfusion group(n=160,181 branch lesions).Multivariate Logistic regression analysis was used to evaluate the effects of various clinical and anatomical factors on branch perfusion after PCI. Results:The post-PCI branch flow grading of all patients was TIMI grade Ⅲ.The postoperative branch μQFR of 53 vessels(22.6％)in the group with impaired postoperative branch perfusion was 0.70±0.10,and 0.93±0.05 in the group with normal postoperative branch perfusion,and the difference between the two groups was statistically significant(P<0.001).Compared with the postoperative group with normal branch perfusion,the postoperative group with impaired branch perfusion was featured with an elevated branch lesion length,branch reference diameter,postoperative branch opening diameter stenosis rate,postoperative branch narrowest lumen diameter stenosis rate,and a lower main branch-to-branch diameter ratio,preoperative branch narrowest lumen diameter stenosis rate,and preoperative main branch μQFR,all of which were statistically significant(all P<0.05).The postoperative branch opening diameter stenosis rate(r=-0.490,P<0.001),postoperative branch narrowest lumen diameter stenosis rate(r=-0.788,P<0.001),preoperative branch narrowest lumen diameter stenosis rate(r=-0.280,P<0.001),branch narrowest lumen diameter(r=-0.469,P<0.001),branch lesion length(r=-0.157,P=0.016)were negatively correlated with postoperative branch μQFR,and branch reference diameter(r=0.173,P=0.008),main branch/side branch diameter ratio(r=0.194,P=0.003),and branch opening diameter(r=0.328,P<0.001)were positively correlated with postoperative branch μQFR,and none of them were significantly correlated with clinical baseline data(all P>0.05).Multifactorial logistic regression analysis showed that following four factors were independent risk factors for impaired branch perfusion:postoperative stenosis of the narrowest branch lumen diameter(OR=1.228,95％CI:1.144-1.318,P<0.001),postoperative stenosis of the branch opening diameter(OR=1.110,95％CI:1.055-1.168,P<0.001),postoperative stenosis of the narrowest lumen diameter of the main branch(OR=1.115,95％CI:1.042-1.192,P=0.001),and length of the branch lesion(OR=1.121,95％CI:1.021-1.231,P=0016). Conclusions:Some of the patients whose branch flow reached TIMI grade Ⅲ after PCI are still faced the risk of hemodynamical impairment and should be functionally evaluated after PCI.The postoperative stenosis rate of the narrowest branch lumen diameter,postoperative stenosis rate of the branch opening diameter,postoperative stenosis rate of the narrowest lumen diameter of the main branch,and branch lesion length are the risk factors of branch perfusion impairment after PCI for coronary bifurcation lesions.

Prepubertal-type testicular neuroendocrine tumor is a rare neoplasm of low malignant potential, which is classified as germ cell tumors unrelated to germ cell neoplasia in situ, and needs to be differentiated from metastatic neuroendocrine tumor, postpubertal-type testicular neuroendocrine tumor, and testicular seminoma. The clinicopathological and molecular features of a case of prepubertal-type testicular neuroendocrine tumor were reported. The tumour cells were uniform in size and arranged in nested and insular pattern. The tumor was positive for CgA and Syn, and the Ki-67 index was less than 2% by immunostaining. Next-generation sequencing identified no variants of pathogenicity, potential pathogenicity or uncertain significance. The patient was followed without evidence of recurrence and metastasis 56 months after surgery.

Objective:To reduce peri-mitral atrial flutter/tachycardia(AFL/AT), this study modified the ablation method of the mitral isthmus(MI) by adopting bipolar radiofrequency ablation (RFA) combined with "cut-and-sew" technique.Methods:This was a clinical retrospective study, involving a total of 138 consecutive patients with AF who underwent a mitral valve surgery and concomitant maze Ⅳ procedure. According to methods of MI linear lesion, patients were divided into the modified group and the RFA group. The primary end points are maintenance of sinus rhythm (SR) rate and the incidence of AFL/AT.Results:During the perioperative period, one patient in the modified group died of suspected cardiogenic shock and septic shock. Follow up for two years, no deaths or stroke events occurred. The proportion of 6-, 12-, and 24-month maintenance of SR off class Ⅰ/Ⅲ antiarrhythmic drugs after surgery were 91.0%, 89.7%, and 88.5% in the modified group and 89.7%, 84.5%, and 81.0% in the RF group, respectively ( P>0.05). The incidence of postoperative AFL/AT was 5.1% in the modified group and 15.3% in the RF group ( P=0.044). The healthcare facility use expense in the RF group was 1.5 times that in the modified group. Early recurrence of AF was an independent risk factor for AF recurrence. Conclusion:In the mitral valve surgery and concomitant maze Ⅳ procedure, the modified method of bipolar RF ablation combined with "cut-and-sew" technique for the MI line is safe and feasible. Its incidence of postoperative AFL/AT is lower than that of the RF method, which reduces existing medical costs and could be accepted by more patients and regions.

Objective:To retrospectively analyze the surgical treatment of isolated pulmonary mucormycosis, explore the timing and effect of operation.Methods:Clinical data of patients with isolated pulmonary mucormycosis who underwent surgical treatment from April 2019 to April 2024 were collected and retrospectively analyzed, combined with literature review.Results:The clinical data of 13 patients with pulmonary mucormycosis who underwent operation were collected, including 9 males and 4 females, aged 22~64(45.4±15.4)years old. The basic diseases included diabetes mellitus in 10 cases and hematological malignancy in 3 cases. Seven patients had hemoptysis during the course of the disease. All 13 patients received adequate antifungal therapy before surgery, and 12 patients received bronchoscopic interventional therapy. Ten patients underwent lobectomy, two underwent pneumonectomy, and one underwent a right sleeve pneumonectomy. All patients were cured except one patient with bronchopleural fistula after operation. All patients achieved complete elimination of mucor and also discontinuation of antifungal drug.Conclusion:For isolated pulmonary mucormycosis, on the basis of systemic antifungal treatment, actively seek the opportunity of surgical intervention, can shorten the course of the disease and achieve good therapeutic results.