Objective To establish a method for rapid detection of tramadol in urine by liquid-liquid extraction（LLE）-surface-enhanced Raman spectroscopy （SERS）. Methods Tramadol was extracted from urine with chloroform∶isopropyl alcohol （9∶1） extractant and detected in urine samples by enhanced Raman spectroscopy （wavelength 785 nm）. Results The quantitative curve of tramadol was Y=204.35 X−465.62, r=0.9952, and the linear range was 1-100 μg/ml. The detection limit of tramadol by this method （S/N=3） was 0.53 μg/ml. The sensitivity of SERS was higher than that of conventional methods, and it had reasonable reproducibility. Conclusion This method is simple, efficient and economical, and can be used for qualitative and quantitative analysis of tramadol personalized medicine.

Background/Aims: Metabolic dysfunction–associated steatotic liver disease (MASLD) is a chronic liver disease characterized by hepatic steatosis. Ubiquitin-specific protease 29 (USP29) plays pivotal roles in hepatic ischemiareperfusion injury and hepatocellular carcinoma, but its role in MASLD remains unexplored. Therefore, the aim of this study was to reveal the effects and underlying mechanisms of USP29 in MASLD progression. Methods: USP29 expression was assessed in liver samples from MASLD patients and mice. The role and molecular mechanism of USP29 in MASLD were assessed in high-fat diet-fed and high-fat/high-cholesterol diet-fed mice and palmitic acid and oleic acid treated hepatocytes. Results: USP29 protein levels were significantly reduced in mice and humans with MASLD. Hepatic steatosis, inflammation and fibrosis were significantly exacerbated by USP29 deletion and relieved by USP29 overexpression. Mechanistically, USP29 significantly activated the expression of genes related to fatty acid β-oxidation (FAO) under metabolic stimulation, directly interacted with long-chain acyl-CoA synthase 5 (ACSL5) and repressed ACSL5 degradation by increasing ACSL5 K48-linked deubiquitination. Moreover, the effect of USP29 on hepatocyte lipid accumulation and MASLD was dependent on ACSL5. Conclusions USP29 functions as a novel negative regulator of MASLD by stabilizing ACSL5 to promote FAO. The activation of the USP29-ACSL5 axis may represent a potential therapeutic strategy for MASLD.

Background/Aims: Metabolic dysfunction–associated steatotic liver disease (MASLD) is a chronic liver disease characterized by hepatic steatosis. Ubiquitin-specific protease 29 (USP29) plays pivotal roles in hepatic ischemiareperfusion injury and hepatocellular carcinoma, but its role in MASLD remains unexplored. Therefore, the aim of this study was to reveal the effects and underlying mechanisms of USP29 in MASLD progression. Methods: USP29 expression was assessed in liver samples from MASLD patients and mice. The role and molecular mechanism of USP29 in MASLD were assessed in high-fat diet-fed and high-fat/high-cholesterol diet-fed mice and palmitic acid and oleic acid treated hepatocytes. Results: USP29 protein levels were significantly reduced in mice and humans with MASLD. Hepatic steatosis, inflammation and fibrosis were significantly exacerbated by USP29 deletion and relieved by USP29 overexpression. Mechanistically, USP29 significantly activated the expression of genes related to fatty acid β-oxidation (FAO) under metabolic stimulation, directly interacted with long-chain acyl-CoA synthase 5 (ACSL5) and repressed ACSL5 degradation by increasing ACSL5 K48-linked deubiquitination. Moreover, the effect of USP29 on hepatocyte lipid accumulation and MASLD was dependent on ACSL5. Conclusions USP29 functions as a novel negative regulator of MASLD by stabilizing ACSL5 to promote FAO. The activation of the USP29-ACSL5 axis may represent a potential therapeutic strategy for MASLD.

Background/Aims: Metabolic dysfunction–associated steatotic liver disease (MASLD) is a chronic liver disease characterized by hepatic steatosis. Ubiquitin-specific protease 29 (USP29) plays pivotal roles in hepatic ischemiareperfusion injury and hepatocellular carcinoma, but its role in MASLD remains unexplored. Therefore, the aim of this study was to reveal the effects and underlying mechanisms of USP29 in MASLD progression. Methods: USP29 expression was assessed in liver samples from MASLD patients and mice. The role and molecular mechanism of USP29 in MASLD were assessed in high-fat diet-fed and high-fat/high-cholesterol diet-fed mice and palmitic acid and oleic acid treated hepatocytes. Results: USP29 protein levels were significantly reduced in mice and humans with MASLD. Hepatic steatosis, inflammation and fibrosis were significantly exacerbated by USP29 deletion and relieved by USP29 overexpression. Mechanistically, USP29 significantly activated the expression of genes related to fatty acid β-oxidation (FAO) under metabolic stimulation, directly interacted with long-chain acyl-CoA synthase 5 (ACSL5) and repressed ACSL5 degradation by increasing ACSL5 K48-linked deubiquitination. Moreover, the effect of USP29 on hepatocyte lipid accumulation and MASLD was dependent on ACSL5. Conclusions USP29 functions as a novel negative regulator of MASLD by stabilizing ACSL5 to promote FAO. The activation of the USP29-ACSL5 axis may represent a potential therapeutic strategy for MASLD.

【Objective】 To explore the risk factors for the production of anti-HLA antibodies in patients with hematological diseases before hematopoietic stemcell transplantation. 【Methods】 The results and clinical data of 1 008 patients with hematological diseases in our hospital who underwent anti-HLA antibody testing were collected by using Luminex technology platform before transplantation from 2016 to 2018 for statistical analysis. 【Results】 The total positive rate of anti-HLA antibodies in 1 008 patients was 24.08%. Multivariate analysis showed that independent risk factors associated with the production of anti-HLA antibodies included age≥30 years old(P=0.046, OR1.467, 95%CI1.007-2.136), time from disease diagnosis to antibody testing≥41 days(P=0.000, OR1.830, 95%CI1.306-2.565), initial platelet count<20×10⁹/L(P=0.020, OR1.543, 95%CI1.072-2.220), prior pregnancy(P=0.000, OR5.187, 95%CI3.689-7.293), transfusions before admission(P=0.001, OR1.762, 95%CI1.257-2.470)and total platelet transfusion volumes after admission≥30 U(P=0.000, OR2.352, 95%CI1.638-3.376). Age ≥30 years old(P=0.023, OR=1.839, 95%CI1.088-3.108)and prior pregnancy(P=0.042, OR=5.258, 95%CI1.062-26.038)are associated with the production of anti-HLA class Ⅰ and class Ⅱ antibodies, respectively. The time from disease diagnosis to antibody testing≥41 days(P=0.000, OR=2.873, 95%CI1.612-5.119), initial platelet count<20×10⁹/L(P=0.008, OR=2.164, 95%CI1.225-3.822), prior pregnancy(P=0.002, OR=6.734, 95%CI1.993-22.751), transfusions before admission(P=0.001, OR=2.746, 95%CI1.531-4.925)and total platelet transfusion volumes after admission>30 U(P=0.006, OR=3.459, 95%CI1.416-8.451)are associated with the production of anti-HLA class Ⅰ and Ⅱ antibodies. 【Conclusion】 Older age, longer course of disease, lower PLT count, history of pregnancy and blood transfusion, and higher total amount of PLT transfusion are risk factors which affect the production of anti-HLA antibodies.Therefore, it is advisable to test for anti-HLA antibodies according to the situation before transplantation, which is of great value in guiding donor selection, monitoring antibody changes and improving transplant prognosis.

Objective To systematically evaluate and analyze the qualitative research on the experience of maintenance hemodialysis patients,so as to provide references for the implementation of exercise interventions in maintenance hemodialysis patients.Methods We searched databases including PubMed,Embase,Web of Science,Cochrane Library,PsycINFO,CINAHL,CBM,CNKI,WanFang Database and VIP from inception to September 30,2022,to collect qualitative studies about exercise experience of maintenance hemodialysis patients.The quality of included studies was evaluated according to JBI Critical Appraisal Tool for qualitative studies in Australia.The results were integrated by integrating methods.Results A total of 14 studies were included.47 completed findings were grouped according to their similarities to form 12 categories.These categories resulted in 4 synthesized findings.Integration results 1:recognizing the positive effects and potential risks of intradialytic exercise.Integration results 2:barriers and facilitators of intradialytic exercise.Integration results 3:needs of intradialytic exercise.Integration results 4:exercise experience on non-dialysis day.Conclusion The experience and needs of maintenance hemodialysis patients should be considered to ensure patient compliance and continuity when exercise programs were designed.

Objective:To analyze the fail mode of neoadjuvant therapy combined with surgery for locally advanced esophageal squamous cell carcinoma (ESCC) after long-term follow-up.Methods:Clinical data of consecutive 238 patients with locally advanced resectable ESCC who underwent neoadjuvant therapy combined with surgery in Zhejiang Cancer Hospital from September 2012 to October 2019 were retrospectively analyzed. The failure mode in the whole cohort was analyzed after long-term follow-up. The overall survival (OS) and disease free survival (DFS) rates were analyzed by Kaplan-Meier method. Survival differences were determined by log-rank test.Results:The pathological complete response (pCR) rate was 42.0% in 238 patients. After a median follow-up of 46.1 months, tumor progression occurred in 96 patients (40.3%), including 25 patients (10.5%) with local recurrence, 61 patients (25.6%) with distant metastases, and 10 patients (4.2%) with simultaneous local recurrence and distant metastases. The median OS and DFS were 64.7 months and 49.9 months. And the 3-, 5-, and 7-year OS and DFS rates were 70.0%, 52.8%, 36.4% and 63.5%, 42.5%, and 30.0%, respectively. The 3-, 5-, and 7-year locoregional recurrence-free survival rates and distant metastasis-free survival rates were 86.0%, 71.4%, 61.2% and 70.6%, 55.9%, 43.0%. Compared with non-pCR patients, the overall progression rate and distant metastasis rate of pCR patients were lower (26.0% vs. 50.7%, 16.0% vs. 32.6%, both P<0.05). And the 3-, 5-, and 7-year OS (83.0% vs. 60.2%, 69.7% vs. 41.7%, 50.4% vs. 27.7%, all P<0.001) and DFS rates (80.4% vs. 51.4%, 63.9% vs. 31.2%, 45.9% vs. 20.3%, all P<0.001) were significantly better in pCR patients. Conclusions:Distant metastasis is the main failure mode of patients with locally advanced ESCC after neoadjuvant therapy. Patients with postoperative pCR can achieve better long-term survival.

OBJECTIVE To investigate the mechanism of Jianpi Yishen Huazhuo Recipe in regulating ovarian function in rats with polycystic ovary syndrome(PCOS) through the Bax/Bcl-2/Caspase-3 pathway. METHODS SD rats were fed with a high-fat diet combined with letrozole to establish a model of PCOS. The successfully modeled rats were randomly divided into blank control group, model group, high-, medium-, and low-dose Jianpi Yishen Huazhuo Recipe groups, as well as positive drug metformin group. ELISA was used to detect the serum levels of LH, T, and LH/FSH in each group of rats. HE staining was used to observe the morphology of ovarian tissue. Immunohistochemistry and Western blotting was used to detect the protein expression of Bax, Bcl-2, and Caspase-3 in ovarian tissues of rats. RESULTS ELISA results showed that the levels of LH, T, and LH/FSH in each dose group of Jianpi Yishen Huazhuo Recipe significantly decreased, while the level of FSH significantly increased. HE staining results showed that ovarian tissue in each dose group of Jianpi Yishen Huazhuo Recipe was significantly improved. Immunohistochemistry results showed that Bax expression increased in granulosa cells, interstitial cells, and luteal cells, while Bcl-2 expression increased in oocytes, luteal cells, and follicular fluid. Caspase-3 expression increased in granulosa cells, oocyte membranes, and interstitial cells. Western blotting results showed that the relative expression of Bax and Caspase-3 proteins decreased in each dose group of Jianpi Yishen Huazhuo Recipe, while the relative expression of Bcl-2 protein increased, and the ratio of Bax/Bcl-2 decreased. CONCLUSION Jianpi Yishen Huazhuo Recipe can improve ovarian ovulation function by downregulating the expression of Bax and Caspase-3 proteins in ovarian tissue of PCOS rats, reducing the ratio of Bax/Bcl-2, upregulating the expression of Bcl-2 protein, adjusting the process of ovarian cell proliferation and apoptosis, improving the ovarian microenvironment, regulating endocrine levels, and promoting follicular development and maturation.

Objective:To explore the current status of readiness for young and middle-aged maintenance hemodialysis (MHD) patients to return to work and analyze its influencing factors, with the aim of providing reference for the evaluation and intervention of patients returning to work.Methods:From October to December 2022, convenience sampling was used to select 425 patients from six hospitals in the urban area of Yangzhou as the subject. A cross-sectional survey was conducted using the General Information Questionnaire, Readiness for Return-To-Work Scale (RRTWS), Distress Disclosure Index (DDI) and Perceived Social Support Scale (PSSS). Binary Logistic regression was used to analyze the influencing factors of returning to work.Results:Among 425 young and middle-aged patients undergoing MHD, 105 (24.7%) returned to work, of which 79 (75.2%) were in the uncertain maintenance stage and 26 (24.8%) were in the active maintenance stage. 320 did not return to work, including 148 (46.3%) in the pre-intention stage, 86 (26.9%) in the intention stage, 42 (13.1%) in the action preparation self-evaluation stage, and 44 (13.8%) in the action preparation behavior stage. Age, per capita monthly income of the family, number of comorbidities, level of self-disclosure, and level of perceived social support were factors that affected patients' readiness to return to work.Conclusions:The rate of young and middle-aged MHD patients returning to work needs to be improved. The return of patients to work is influenced by multiple factors. Medical and nursing staff should focus on patients who are old, have low per capita monthly income of the family, and have a large number of comorbidities. Targeted interventions and guidance should be provided to patients, such as self-disclosure training and improving their perceived social support, in order to increase the rate of patients returning to work rate.