Objective To explore the the detection of MMP-2,-7,-9,and-12 enzymatic activity using the CEACAM1-derived fluorescent peptide substrate Site 84,investigating the application of substrate Site 84 to distinguishing between MMP-2 and MMP-9 in the gelatinase spectrum of MMPs.Methods The fluorescent enzymatic method was employed to observe the detection of MMP-2,-7,-9,and-12 enzymatic activity using substrate Site 84;further observations were made on the sensitivity and specificity of substrate Site 84 to enzymatic activity of MMP-2 and MMP-9 within the gelatinase spectrum;the kinetic parameters(Km and Kcat)of the enzymatic reaction between substrate Site 84 and MMP-2 were obtained.Results Using Site 84 as a substrate,enzymic kinetics curves for MMP-12,-7,-2 were obtained,but no enzymatic activity curve for MMP-9 was observed.Furthermore,Site 84 specifically detected the enzymatic activity of MMP-2 within the gelatinase spectrum,capable of detecting low concentration(0.6 μM)of MMP-2 enzymatic activity,but no obvious enzymatic reaction was observed for high concentration(6 μM)of MMP-9;the kinetics parameters for the enzymatic reaction between Site 84 and MMP-2 were Km=315 μM,Kcat/Km=2 565/MS.Conclusion The CEACAM1-derived substrate Site 84 serves as a novel fluorescent peptide substrate,enabling the acquisition of enzymatic activity curves for MMP-12,-7 and-2,and specifically detecting the enzymatic activity of MMP-2 within the MMP gelatinase spectrum.

BACKGROUND:Allogeneic hematopoietic stem cell transplantation is an effective and even the only way to cure various hematological diseases,but the short-term mortality rate is relatively high after transplantation. OBJECTIVE:To investigate the risk factors affecting the overall survival of patients with hematological diseases in the short term(within 100 days)after allogeneic hematopoietic stem cell transplantation,so as to reduce mortality and effectively prevent related risks in the short term(within 100 days)after allogeneic hematopoietic stem cell transplantation. METHODS:Clinical data of 585 patients with hematological diseases who underwent allogeneic hematopoietic stem cell transplantation at the Hematopoietic Stem Cell Transplantation Center of First Affiliated Hospital of Zhengzhou University from January 1,2018 to June 30,2021 were retrospectively analyzed.The risk factors that affected overall survival within 100 days after allogeneic hematopoietic stem cell transplantation were explored. RESULTS AND CONCLUSION:A total of 585 patients with hematologic diseases underwent allogeneic hematopoietic stem cell transplantation.92 patients died within 100 days after transplantation,with a mortality rate of 15.7%(92/585).The median age of death cases was 26.5 years old(1-56 years),and the median survival time of death cases was 48 days(0-97 days).Univariate analysis exhibited that age≥14 years old,acute graft-versus-host disease,grade IV acute graft-versus-host disease,bacterial bloodstream infection,as well as carbapenem-resistant organism bloodstream infection,were risk factors for overall survival within 100 days after allogeneic hematopoietic stem cell transplantation(P<0.05).Multivariate regression analysis showed that age≥14 years old,grades Ⅲ-Ⅳ acute graft-versus-host disease,bacterial bloodstream infection,and carbapenem-resistant organism bloodstream infections were independent risk factors for overall survival(within 100 days)in patients after allogeneic hematopoietic stem cell transplantation.Hazard ratios were 1.77(95%CI 1.047-2.991),7.926(95%CI 3.763-16.695),2.039(95%CI 1.117-3.722),and 3.389(95%CI 1.563-7.347),respectively.In conclusion,all-cause mortality rate after allogeneic hematopoietic stem cell transplantation is relatively high in the short term.A timely diagnosis and effective treatment of bacterial bloodstream infection and acute graft-versus-host disease are essential to improving allogeneic hematopoietic stem cell transplantation outcomes.

Objective:To investigate the teaching effects of blended teaching based on massive open online courses (MOOC) combined with team-based learning (TBL) for the general elective course First Aid for Burns for non-medical college students. Methods:We assigned 73 non-medical college students choosing the Common Home Health Issues and Care course in the spring semester of 2023 to receive blended teaching based on MOOC and TBL (experimental group), and 113 non-medical college students choosing the same course in the spring semester of 2024 to receive traditional lecture-based learning (control group). A self-evaluation questionnaire was used to evaluate the teaching effects and satisfaction with the course of the two groups. SPSS 27.0 was used to perform the t-test and chi-square test. Results:The experimental group showed significantly higher proportions of students who had a complete mastery of knowledge of first aid for burns (54.8% vs. 33.6%, P<0.01) and students who provided positive feedback on improving autonomous learning ability (98.6% vs. 78.8%, P<0.001), improving the ability of teamwork (100.0% vs. 68.1%, P<0.001), improving communication ability (98.6% vs. 72.6%, P<0.001), and overall satisfaction with the course (97.3% vs. 76.6%, P<0.001) compared with the control group. Conclusions:The blended teaching model based on MOOC and TBL can improve the teaching effects of First Aid for Burns for non-medical college students, enhancing their autonomous learning, teamwork, and communication abilities. This method is applicable to medical general education courses, which is worthy of promotion and application.

Objective:To study the reproductive toxicity of lithium chloride (LiCl) in male mice and to explore the molecular mechanism of the protective effect of quercetin on testosterone production dysfunction.Methods:Twenty-five male C57BL/6 mice aged 4-5 weeks were randomly divided into five groups according to the random number table method: control group, LiCl infected group [38.4 mg/(kg·d) LiCl+corn oil, noted as LiCl group], quercetin control group [50 mg/(kg·d) quercetin, noted as High-Quer group], low-dose quercetin combined with LiCl infection group [38.4 mg/(kg·d) LiCl+10 mg/(kg·d) quercetin, noted as Low-Quer+LiCl group] and high dose quercetin combined with LiCl infected group [38.4 mg/(kg·d) LiCl+50 mg/(kg·d) quercetin, noted as High-Quer+LiCl group]. The structure of testicular tissue, semen parameters, and the ultrastructure of Leydig cells in mice were detected by HE staining, computer-aided sperm analysis system (CASA), and transmission electron microscopy, respectively. TM3 mouse Leydig cells were treated with 0 mmol/L, 5 mmol/L, 10 mmol/L, 20 mmol/L LiCl for 24 h. The activities of superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px), mitochondrial membrane potential (MMP), lipid peroxidation levels, and expression levels of testosterone-related protein were measured by SOD and GSH-Px kits, TMRE probe, Image-iT TM lipid peroxidation probe, and Western blotting, respectively. Intracellular Fe 2+ concentration was detected by Fe 2+ detection kit and FerroOrange probe. The levels of testosterone, progesterone, and estradiol were measured using enzyme-linked immunosorbent assay kits. Results:In the LiCl group, the total sperm count [(36.78±1.81)×10 6], sperm concentration [(18.39±0.90)×10 6/mL], sperm motility [(25.70±3.32)%] and serum testosterone level [(7.26±0.29) μg/L] were lower than those of control group [(51.60±4.96)×10 6, P=0.002; (25.80±2.48)×10 6/mL, P=0.002; (41.47±2.83)%, P=0.001; (7.87±0.29) μg/L, P=0.013], the expression levels of Cyp11a1, StAR and Cyp17a1, Leydig cell biomarkers (3β-HSD1, 17β-HSD3) and ferroptosis regulatory proteins (GPX4, SLC7A11 and Nrf2) in testis were significantly down-regulated compared with control group (all P<0.001). LiCl also induced mitochondrial vacuoles and swelling of Leydig cells. However, low-dose quercetin intervention could significantly ameliorate the above LiCl-induced damage (total sperm count: P<0.001, sperm concentration: P<0.001, sperm motility: P=0.015, serum testosterone level: P=0.026, Cyp11a1, StAR, Cyp17a1, 3β-HSD1, 17β-HSD3, GPX4, SLC7A11, and Nrf2: all P<0.001). In 20 mmol/L LiCl group, the testosterone level [(7.28±0.24) μg/L] in TM3 cells was lower than that of 0 mmol/L LiCl group [(12.50±0.38) μg/L, P<0.001], the protein levels of Cyp11a1, StAR and Cyp17a1 were significantly down-regulated compared with 0 mmol/L LiCl group (all P<0.001), the activities of SOD [(2.42±0.11) U/mg], GSH-Px [(1.29±0.03) mU/mg] and MMP [(57.24±1.69)%] were lower than those of 0 mmol/L LiCl group [(3.11±0.09) U/mg, (1.54±0.01) mU/mg, (100.00±0)%, all P<0.001], the level of lipid peroxidation [(211.18±3.60)%] and the concentration of Fe 2+ [(26.44±0.94) μmol/L] were higher than those of 0 mmol/L LiCl group [(100.00±0)%, (7.12±0.29) μmol/L, all P<0.001], and the expression of ferroptosis regulatory proteins was significantly down-regulated compared with 0 mmol/L LiCl group (all P<0.001). Compared with 20 mmol/L LiCl group cells [lipid peroxidation: (194.46±3.16)%, (194.70±3.93)%; MMP: (78.74±0.52)%, (75.32±1.29)%], ferroptosis inhibitors Fer-1 and quercetin significantly decreased the level of lipid peroxidation [(181.71±3.80)%, P=0.004; (166.88±3.22)%, P<0.001], increased the level of MMP [(86.26±0.79)%, P=0.040; (81.09±1.32)%, P=0.001], and significantly up-regulated the expression of key enzymes in testosterone synthesis and ferroptosis regulatory proteins (all P<0.05). Conclusion:Quercetin may protect LiCl-induced Leydig cells injury and testosterone production dysfunction by inhibiting cell ferroptosis.

Objective:To study the reproductive toxicity of lithium chloride (LiCl) in male mice and to explore the molecular mechanism of the protective effect of quercetin on testosterone production dysfunction.Methods:Twenty-five male C57BL/6 mice aged 4-5 weeks were randomly divided into five groups according to the random number table method: control group, LiCl infected group [38.4 mg/(kg·d) LiCl+corn oil, noted as LiCl group], quercetin control group [50 mg/(kg·d) quercetin, noted as High-Quer group], low-dose quercetin combined with LiCl infection group [38.4 mg/(kg·d) LiCl+10 mg/(kg·d) quercetin, noted as Low-Quer+LiCl group] and high dose quercetin combined with LiCl infected group [38.4 mg/(kg·d) LiCl+50 mg/(kg·d) quercetin, noted as High-Quer+LiCl group]. The structure of testicular tissue, semen parameters, and the ultrastructure of Leydig cells in mice were detected by HE staining, computer-aided sperm analysis system (CASA), and transmission electron microscopy, respectively. TM3 mouse Leydig cells were treated with 0 mmol/L, 5 mmol/L, 10 mmol/L, 20 mmol/L LiCl for 24 h. The activities of superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px), mitochondrial membrane potential (MMP), lipid peroxidation levels, and expression levels of testosterone-related protein were measured by SOD and GSH-Px kits, TMRE probe, Image-iT TM lipid peroxidation probe, and Western blotting, respectively. Intracellular Fe 2+ concentration was detected by Fe 2+ detection kit and FerroOrange probe. The levels of testosterone, progesterone, and estradiol were measured using enzyme-linked immunosorbent assay kits. Results:In the LiCl group, the total sperm count [(36.78±1.81)×10 6], sperm concentration [(18.39±0.90)×10 6/mL], sperm motility [(25.70±3.32)%] and serum testosterone level [(7.26±0.29) μg/L] were lower than those of control group [(51.60±4.96)×10 6, P=0.002; (25.80±2.48)×10 6/mL, P=0.002; (41.47±2.83)%, P=0.001; (7.87±0.29) μg/L, P=0.013], the expression levels of Cyp11a1, StAR and Cyp17a1, Leydig cell biomarkers (3β-HSD1, 17β-HSD3) and ferroptosis regulatory proteins (GPX4, SLC7A11 and Nrf2) in testis were significantly down-regulated compared with control group (all P<0.001). LiCl also induced mitochondrial vacuoles and swelling of Leydig cells. However, low-dose quercetin intervention could significantly ameliorate the above LiCl-induced damage (total sperm count: P<0.001, sperm concentration: P<0.001, sperm motility: P=0.015, serum testosterone level: P=0.026, Cyp11a1, StAR, Cyp17a1, 3β-HSD1, 17β-HSD3, GPX4, SLC7A11, and Nrf2: all P<0.001). In 20 mmol/L LiCl group, the testosterone level [(7.28±0.24) μg/L] in TM3 cells was lower than that of 0 mmol/L LiCl group [(12.50±0.38) μg/L, P<0.001], the protein levels of Cyp11a1, StAR and Cyp17a1 were significantly down-regulated compared with 0 mmol/L LiCl group (all P<0.001), the activities of SOD [(2.42±0.11) U/mg], GSH-Px [(1.29±0.03) mU/mg] and MMP [(57.24±1.69)%] were lower than those of 0 mmol/L LiCl group [(3.11±0.09) U/mg, (1.54±0.01) mU/mg, (100.00±0)%, all P<0.001], the level of lipid peroxidation [(211.18±3.60)%] and the concentration of Fe 2+ [(26.44±0.94) μmol/L] were higher than those of 0 mmol/L LiCl group [(100.00±0)%, (7.12±0.29) μmol/L, all P<0.001], and the expression of ferroptosis regulatory proteins was significantly down-regulated compared with 0 mmol/L LiCl group (all P<0.001). Compared with 20 mmol/L LiCl group cells [lipid peroxidation: (194.46±3.16)%, (194.70±3.93)%; MMP: (78.74±0.52)%, (75.32±1.29)%], ferroptosis inhibitors Fer-1 and quercetin significantly decreased the level of lipid peroxidation [(181.71±3.80)%, P=0.004; (166.88±3.22)%, P<0.001], increased the level of MMP [(86.26±0.79)%, P=0.040; (81.09±1.32)%, P=0.001], and significantly up-regulated the expression of key enzymes in testosterone synthesis and ferroptosis regulatory proteins (all P<0.05). Conclusion:Quercetin may protect LiCl-induced Leydig cells injury and testosterone production dysfunction by inhibiting cell ferroptosis.

Objective:To analyze and compare the practical value of serum cystatin C(Scys C)and serum creatinine(SCr)in the assessment of kidney function in older adults.Methods:A retrospective, cross-sectional study was performed in 2 450 participants who were divided into a non-elderly group(<65 years)and an elderly group(≥65 years).Glomerular filtration rate(GFR), Scys C and SCr were measured by 99mTc-DTPA clearance, particle-enhanced immunoturbidimetry and an oxidase method, respectively.The χ2 test was used to compare increases in percentage of Scys C and SCr at the same GFR level.The screening value of Scys C and SCr for GFR<60 ml·min -1·1.73m -2was evaluated by the area under curve(AUC)of the receiver operating characteristic(ROC)curve.Values of 95% reference ranges were established for Scys C and SCr at different GFR levels. Results:The proportions of the general population with increased Scys C were 82.74%(556/672)and 94.74%(90/95), respectively, for GFR levels between 30~59 ml·min -1·1.73m -2and <30 ml·min -1·1.73m -2, while only 38.24%(257/672)and 75.79%(72/95)had elevated SCr levels( χ2=278.328, 13.571, both P<0.001).For the above GFR intervals, the proportions of older adults with increased Scys C were 84.81%(240/283)and 100.00%(43/43)respectively, and the proportions for non-elderly adults with increased Scys C were 81.23%(316/389)and 90.38%(47/52)( χ2=1.463, 4.364, P=0.226, 0.037), respectively.The screening value of Scys C for GFR<60 ml·min -1·1.73m -2was slightly better than SCr in terms of sensitivity, specificity and the Youden index.However, the sensitivity and specificity of Scys C in older adults were 76.4% and 75.7%, respectively, both lower than 78.7% and 84.0% in non-older adults.The variability of Scys C increased progressively with age.The reference range for Scys C was higher in older adults than in non-older adults at the same GFR level. Conclusions:When screening for GFR<60 ml·min -1·1.73m -2, the sensitivity and specificity of Scys C are slightly better than those of SCr, but are lower in older adults than in non-older adults.Scys C levels are higher and more variable in older adults.Using Scys C to assess GFR may lead to over-diagnosis of chronic kidney disease in older adults.

Objective:To explore the effect of health education based on the forgetting curve in maintenance hemodialysis patients.Methods:A total of 84 patients undergoing maintenance hemodialysis in the First Affiliated Hospital of Nanjing Medical University from March 2017 to March 2020 were enrolled and divided into in the observation group (43 cases) and in the control group (41 cases) using the random number table. The control group received conventional health education, and the observation group conducted health education based on the forgetting curve. Six months after intervention, disease cognition level, self-management ability, quality of life of the two groups were compared.Results:Six months after intervention, disease cognition level, self-management ability, quality of life scores of the observation group were higher than those of the control group, and the differences were statistically significant ( P<0.05) . Conclusions:Health education based on the forgetting curve can help maintenance hemodialysis patients enhance the awareness of diseases, promote the development of self-management ability, and improve their quality of life.

Objective:To investigate the current status of medical record coders and their resignation tendency, and put forward suggestions for the training and team building of medical record coders.Methods:From April to May 2020, a questionnaire survey was conducted among the medical record coders in 27 DRG payment pilot hospitals in Anyang, Henan. Statistical analysis was conducted on the profile of the medical case department, basic personal information, professional quality, job satisfaction and resignation intention. Kendall coefficient was used to analyze the factors related to the resignation intention.Results:There were 98 coders in the surveyed hospitals, including 43 full-time coders and 55 part-time coders. Eighty-three valid questionnaires were recovered, and the effective recovery rate was 84.7%. Among them, 29(34.9%) had education level of junior college or below, 51(61.4%) had junior title or below, 15(18.1%) had medical information or case management as their major, 29(34.9%) coders had not obtained a coding certificate, 31(37.3%) were dissatisfied with the salary system, and 14(16.9%) had a tendency to leave. The medical record coders with lower professional titles, coding certificates and lower salary satisfaction were more likely to have the intention of resignation.Conclusions:Medical record coders in Anyang were challenged with such problems as shortage of manpower, low percentage of coding certificate holders and low salary satisfaction. Title, code certificate issuance and salary satisfaction were correlated with their resignation intention. Hospital managers should strengthen the importance of medical record coding and coders, and establish a reasonable salary, title evaluation and performance incentive system, so as to improve the attractiveness of coder position.

Objective:To compare the health knowledge of newly admitted patients with recurrent hypertensive stroke around 10 years, and analyze the influencing factors of health knowledge of patients.Methods:From September 2008 to December 2009 and from September 2018 to December 2019, convenience sampling was used to select newly admitted patients with recurrent hypertensive stroke from 3 ClassⅢ Grade A hospitals in Guangzhou as the research objects, and the Stroke Knowledge Questionnaire (SKQ) was used for investigation. In the two stages around 10 years, 120 and 130 questionnaires were distributed, and 109 and 116 valid questionnaires were returned with the valid response rates of 90.83% and 89.23%, respectively.Results:The education level, per capita monthly income of the family, family history of stroke, diabetes mellitus and hyperlipidemia, and smoking of the patients before and after 10 years were statistically significant ( P<0.05) . After 10 years, SKQ total scores, dimension scores in exercise, medication, stroke risk factors, stroke auras, and stroke management of patients were higher than those of 10 years ago, and dimension scores low-salt diet and smoking were lower than 10 years ago, and the differences were statistically significant ( P<0.05) . Conclusions:The level of health knowledge of newly admitted patients with recurrent hypertensive stroke is higher than that of 10 years ago, but education on low-salt diet and smoking cessation knowledge should be further strengthened.

Objective To investigate the relationship between perineural invasion (PNI) and the clinicopathological features of cervical cancer,and to explore its influence on the prognosis of cervical cancer patients.Methods Retrospective analysis the clinical data of 76 patients with early cervical cancer who received radical hysterectomy + pelvic lymphadenectomy ± paraaortic lymphadenectomy from November 2015 to December 2019 in Xuanwu Hospital.The clinicopathological characteristics of the postoperative pathological results of cervical cancer patients were analyzed to explore the related factors of PNI and its influence on prognosis.Results Single factor analysis suggested that the incidence of PNI was related to tumor diameter,depth of stroma invasion,lymph-vascular space invasion,lymph node metastasis and clinical stage (P ＜ 0.05).Multivariate analysis suggested that the infiltration depth of stroma invasion was the risk factor of PNI (P ＜ 0.05).K-M analysis showed that there was no significant difference in disease-free survival and overall survival between PNI positive and PNI negative patients (P ＞ 0.05).Conclusions PNI is related to the depth of interstitial infiltration.PNI has no significant effect on the short-term recurrence rate and short-term survival rate of cervical cancer patients.