Objective To establish an immortalized tree shrew corneal stromal cells(CSCs)line and to study its response to virus infection.Methods Primary tree shrew CSCs were isolated and cultured by the tissue block adhesion method.CSCs were then transfected with a lentivirus carrying the SV40T gene and monoclonal cells were selected for passage culture.The characteristics of the CSCs were investigated by morphological observation and compared with 40 generations until the 50 generations or more,immunofluorescence identification of vimentin and SV40T genes,karyotype examination,and cell proliferation curve.The CSCs were infected with herpes simplex virus-1(HSV-1)(McKrae strain),Zika virus(ZIKV,GZ01 strain),Dengue virus typeⅡ,and H1N1(PR8).Results The immortalized tree shrew CSCs after＞50 passages appeared spindle-shaped with good cell morphology and structure compared with 40 generations.Positive immunofluorescence expression of vimentin and SV40T genes.The cell growth curve showed that the cells were in logarithmic-phase growth on days 4～5 and grew vigorously.The number of chromosomes in the primary cells was stable at 62,while immortalized CSCs had 64 chromosomes at P21 and P56.The virus titer results showed that the immortalized tree shrew CSCs were sensitive to HSV-1(McKrae strain),ZIKV(GZ01 strain),Dengue virus typeⅡ,and H1N1(PR8),with virus titers of 1.32×105,5.62×106,2.69×107,and 7.76×104 CCID50/mL,respectively.Conclusions The immortalized tree shrew CSCs were established successfully,suggesting that this cell line is suitable for studies of the mechanisms of HSV,ZIKV,Dengue virus,and influenza A virus infection in relation to corneal diseases and antiviral drugs.

Objective:To evaluate an effective manner to replace the drainage tube due to drainage complications during the irrigation treatment of abdominal infection after pancreaticoduodenectomy (PD).Methods:Clinical data of 39 patients with abdominal infection after PD due to drainage complications who were successfully treated by replacement of the drainage tubes with continued flushing in the Department of Hepatobiliary Surgery, Yuncheng Central Hospital Affiliated to Shanxi Medical University from August 2020 to August 2023 were retrospectively analyzed, including 23 males and 16 females, aged (54.8±9.6) years. According to the fashion of tube replacement, patients were divided into the observation group ( n=21), in which a flushing tube was placed through the original abdominal drainage tube sinus tract combined with external negative pressure suction; and the control group ( n=18), in which two parallel drainage tubes were placed through the original abdominal drainage tube sinus tract for flushing and drainage. The two groups were compared in terms of indicators before tube replacement, including the primary tumor classification, incidence of pancreatic fistula and biliary fistula after PD, complications of abdominal drainage, time from the surgery to tube replacement; and indicators after tube replacement, including total hospital stay, hospitalization cost, continuous abdominal lavage time, fever and elevated white blood cell count, number of dressing changes, etc. Results:There were no significant difference in the primary tumor classification, incidence of pancreatic fistula and biliary fistula after PD, complications of abdominal drainage, and time from PD to tube replacement between the two groups before tube replacement (all P>0.05). After tube replacement, the total hospitalization time(32.7±1.9 vs 44.7±14.5, d), hospitalization cost (67 604±16 052 vs 91 845±19 826, yuan), continuous abdominal lavage time [4.0 (3.0, 5.0) vs 9.0 (8.0, 10.8), d], fever [23.8% (5/21) vs 55.6% (10/18)] and leukocytosis rate [28.6% (6/21) vs 66.7% (12/18)], and times of dressing change times [10.0 (7.0, 13.0) vs 22.0 (18.2, 27.0)] of the observation group were lower than those of the control group (all P<0.05). Conclusion:Inserting a flushing tube through the sinus tract of original abdominal drainage tube combined with external negative pressure drainage is an effective way to manage the abdominal infection after PD surgery due to drainage complications, featuring good irrigation and drainage, short irrigation time, and early control of the abdominal infection.

Objective:To investigate the protective effects and related mechanisms of Astragaloside Ⅳ(ASⅣ)alleviating Angiotensin II-induced cardiomyocyte hypertrophy.Methods:H9c2 cardiomyocytes were divided into six groups: normal control group, ASⅣ group(ASⅣ 100 μmol/L), AngⅡ group(AngⅡ 1 μmol/L), and three ASⅣ dose experiments(AngⅡ 1 μmol/L + ASⅣ 25 μmol/l group, AngⅡ 1 μmol/L+ ASⅣ 50 μmol/l group, AngⅡ1 μmol/L+ ASⅣ 100 μmol/L group), and simultaneously cultured for 24 hours.Cardiomyocyte viability was assessed by CCK8 assay, and surface area of culturedcardiomyocytes in each group was assessed by immunofluorescence assay.Atrial natriuretic peptide(ANP)mRNA expression was assessed by fluorescence real-time quantitative RT-PCR.And LC3 protein expression, an autophagy related protein, was assessed by Western blotting as well as immunofluorescence.Results:(1)AngⅡ decreased cardiomyocyte H9c2 viability in a dose-dependent manner( P<0.05). ASⅣ could inhibit the decrease of cardiomyocyte H9c2 viability in response to AngⅡ in a dose-dependent manner( P<0.05). (2)H9c2 cardiomyocytes induced by AngⅡ showed a significantly larger cell area and significantly higher ANP mRNA and ANP protein expression compared with controls.Different concentrations of ASⅣ intervention could reverse the increase of cardiomyocyte H9c2 area induced by AngⅡ and also decreased the expression of ANP protein induced by AngⅡ in a dose-dependent manner(all P<0.05). (3)Compared with the control group, the autophagy level and the expression of autophagy marker LC3II/I of H9c2 cardiomyocytes induced by AngⅡ were significantly increased(all P<0.05). ASⅣ could inhibit AngⅡ-activated autophagy, and the difference was statistically significant( P<0.05). ASⅣ inhibited the expression of LC3II/I in H9c2 cardiomyocytes stimulated by AngⅡ, and the difference was statistically significant( P<0.05). Conclusions:ASⅣ inhibits AngⅡ-induced cardiac hypertrophy by inhibiting autophagy of cardiomyocytes.

Background: and Purpose Despite administration of evidence-based therapies, residual risk of stroke recurrence persists. This study aimed to evaluate the residual risk of recurrent stroke in acute ischemic stroke or transient ischemic attack (TIA) with adherence to guideline-based secondary stroke prevention and identify the risk factors of the residual risk. Methods: Patients with acute ischemic stroke or TIA within 7 hours were enrolled from 169 hospitals in Third China National Stroke Registry (CNSR-III) in China. Adherence to guideline-based secondary stroke prevention was defined as persistently receiving all of the five secondary prevention medications (antithrombotic, antidiabetic and antihypertensive agents, statin and anticoagulants) during hospitalization, at discharge, at 3, 6, and 12 months if eligible. The primary outcome was a new stroke at 12 months. Results: Among 9,022 included patients (median age 63.0 years and 31.7% female), 3,146 (34.9%) were identified as adherence to guideline-based secondary prevention. Of all, 864 (9.6%) patients had recurrent stroke at 12 months, and the residual risk in patients with adherence to guidelinebased secondary prevention was 8.3%. Compared with those without adherence, patients with adherence to guideline-based secondary prevention had lower rate of recurrent stroke (hazard ratio, 0.85; 95% confidence interval, 0.74 to 0.99; P=0.04) at 12 months. Female, history of stroke, interleukin-6 ≥5.63 ng/L, and relevant intracranial artery stenosis were independent risk factors of the residual risk. Conclusions There was still a substantial residual risk of 12-month recurrent stroke even in patients with persistent adherence to guideline-based secondary stroke prevention. Future research should focus on efforts to reduce the residual risk.

Background and Objectives: Galectin-3 promotes fibroblast-to-myofibroblast differentiation and facilitates injury repair. Previous studies have shown that exosomes derived from human umbilical cord mesenchymal stem cells (hucMSC-ex) promote the differentiation of myocardial fibroblasts into myofibroblasts under inflammatory environment. Whether hucMSC-ex derived Galectin-3 (hucMSC-ex-Galectin-3) plays an important role in fibroblast-to-myofibroblast differentiation is the focus of this study. Methods: and Results: Galectin-3 was knocked-down by siRNA in hucMSCs, and then exosomes were extracted. Fibroblasts were treated with LPS, LPS＋hucMSC-ex, LPS＋negative control-siRNA-ex (NC-ex), or LPS＋ Galectin-3-siRNA-ex (si-ex) in vitro. The coronary artery of the left anterior descending (LAD) branch was permanently ligated, followed by intramyocardial injection with phosphate buffered saline(PBS), hucMSC-ex, hucMSC-NC-ex, or hucMSC-si-ex in vivo. Western blot, RT-PCR, and immunohistochemistry were used to detect the expression of markers related to fibroblast-to-myofibroblast differentiation and inflammatory factors. Migration and contraction functions of fibroblasts were evaluated using Transwell migration and collagen contraction assays, respectively. β-catenin expression was detected by western blot and immunofluorescence. The results showed that hucMSC-ex increased the protein expression of myofibroblast markers, anti-inflammatory factors, and β-catenin. HucMSC-ex also reduced the migration and promoted the contractility of fibroblasts. However, hucMSC-si-ex did not show these activities. Conclusions HucMSC-ex-Galectin-3 promoted the differentiation of cardiac fibroblasts into myofibroblasts in an inflammatory environment, which was associated with increased β-catenin levels.

Humans ; Ischemic Attack, Transient ; Recurrence ; Risk Factors ; Stroke/epidemiology*

Background and Objectives: Galectin-3 promotes fibroblast-to-myofibroblast differentiation and facilitates injury repair. Previous studies have shown that exosomes derived from human umbilical cord mesenchymal stem cells (hucMSC-ex) promote the differentiation of myocardial fibroblasts into myofibroblasts under inflammatory environment. Whether hucMSC-ex derived Galectin-3 (hucMSC-ex-Galectin-3) plays an important role in fibroblast-to-myofibroblast differentiation is the focus of this study. Methods: and Results: Galectin-3 was knocked-down by siRNA in hucMSCs, and then exosomes were extracted. Fibroblasts were treated with LPS, LPS＋hucMSC-ex, LPS＋negative control-siRNA-ex (NC-ex), or LPS＋ Galectin-3-siRNA-ex (si-ex) in vitro. The coronary artery of the left anterior descending (LAD) branch was permanently ligated, followed by intramyocardial injection with phosphate buffered saline(PBS), hucMSC-ex, hucMSC-NC-ex, or hucMSC-si-ex in vivo. Western blot, RT-PCR, and immunohistochemistry were used to detect the expression of markers related to fibroblast-to-myofibroblast differentiation and inflammatory factors. Migration and contraction functions of fibroblasts were evaluated using Transwell migration and collagen contraction assays, respectively. β-catenin expression was detected by western blot and immunofluorescence. The results showed that hucMSC-ex increased the protein expression of myofibroblast markers, anti-inflammatory factors, and β-catenin. HucMSC-ex also reduced the migration and promoted the contractility of fibroblasts. However, hucMSC-si-ex did not show these activities. Conclusions HucMSC-ex-Galectin-3 promoted the differentiation of cardiac fibroblasts into myofibroblasts in an inflammatory environment, which was associated with increased β-catenin levels.

Objective:To explore the clinical features and possible pathogenesis of spontaneous remission of acute myeloid leukemia (AML) .Methods:We retrospectively analyzed the clinical data of a patient with spontaneous remission of AML, MLL-AF9 rearrangement, and abnormal liver function in the First Affiliated Hospital of Zhengzhou University, and the relevant pieces of literature were summarized.Results:The patient experienced lung infection, fever, and liver dysfunction and was treated with anti-infection and blood transfusion. After complete response (CR) , the patient remained in CR with mild, indirect bilirubin elevation at 35 months of follow-up. Additionally, 56 cases of adult AML (non-acute promyelocytic leukemia) were reported in the literature from 1990 to June 2021. The cases were checked by bone marrow aspiration, and our patients were summarized and analyzed. Furthermore, 57 patients, including 37 males and 20 females, with a median age of 51 (20-83) years and a median remission time of five months; 52 patients achieved complete remission. In addition, there were five cases with long-term remission and a chromosomal record, with no recurrence so far, three with normal karyotype and two with t (9;11) (q21;q23) .Conclusion:The spontaneous remission of leukemia is rare and may be related to immunosuppression and genes.

Objective:To develop the Fear of Disease Progression Scale for Patients with Acute Myocardial Infarction, and test its reliability and validity.Methods:The first draft of the scale was formed through literature review, qualitative interviews, and Delphi expert consultations. From March to July 2020, convenience sampling method was adopted to conduct a formal survey of 200 patients with acute myocardial infarction admitted to the Department of Cardiology in three ClassⅢ Grade A hospitals in Jiangsu Province. The formal scale was formed through the project analysis and exploratory factor analysis, and its reliability and validity were determined. 200 questionnaires were distributed, and 178 valid questionnaires were recovered, with a valid recovery rate of 89.00%.Results:The final scale contained 17 items, and exploratory factor analysis extracted 3 common factors, and the cumulative contribution rate of variance was 62.671%. The item-level content validity index ( I- CVI) was from 0.83 to 1.00, the scale-level content validity index ( S- CVI) was 0.82, and the average S- CVI was 0.97. The Cronbach's α coefficient of the total scale was 0.892, the half-reliability was 0.764, and the test-retest reliability was 0.964. Conclusions:The developed Fear of Disease Progression Scale for Patients with Acute Myocardial Infarction has good reliability and validity, which can be used to assess the current status of fear of disease progression in patients with acute myocardial infarction.

Objective:To explore the status quo of meaning of life in patients undergoing radical mastectomy and analyze the influencing factors.Methods:The convenient sampling method was used to select 302 patients with breast cancer who were treated in 2 Class Ⅲ Grade A hospitals in Zhenjiang from June 2019 to February 2020 as research objects. The general data questionnaire, Chinese version of Meaning in Life Questionnaire (C-MLQ) , Health-related Hardiness Scale (HRHS) and Cognitive Reassessment Strategy Scale were used to investigate them.Results:A total of 315 questionnaires were issued and 302 valid questionnaires were collected.The total score of C-MLQ of 302 patients undergoing radical mastectomy was (31.65±6.71) , the total score of HRHS was (92.59±5.68) and the total score of Cognitive Reassessment Strategy Scale was (16.23±5.04) .The results of multiple linear regression analysis showed that cognitive reassessment, postoperative time, health-related hardiness and its control dimension, family monthly income, gender, radiotherapy and chemotherapy were the influencing factors of meaning of life in patients undergoing radical mastectomy ( P<0.05) , which explained 85.8% of the variation in meaning of life. Conclusions:The meaning of life of patients undergoing radical mastectomy is at a medium low level. Clinical medical staff should take effective intervention measures to improve cognitive reassessment ability of patients, so that they can have correct cognition of the disease. They should also encourage them to face the disease actively and bravely and strengthen their own health-related hardinessin order to improve the meaning of life.