1.Heterologous Expression,Purification and Enzymatic Characterization of Xylitol Dehydrogenase from the Thermophilic Fungus Talaromyces emersonii
Er MENG ; Cong QU ; Ke YI ; Hui-Min LI ; Xin-Yi DUAN ; Zhe-Yuan ZHANG ; Shao-Long HE ; Yu-Tao LUO ; Lei WU ; Dong-Yi ZHANG ; Chang-Jun LIU
Chinese Journal of Biochemistry and Molecular Biology 2025;41(7):1007-1018
The xylitol dehydrogenase(XDH)is a crucial enzyme involved in the xylose utilization in pentose-catabolizing yeasts and fungi.In addition to producing xylulose,XDH can also be employed to develop a biosensor for monitoring xylitol concentration.In this study,the gene encoding the thermophilic fungus Talaromyces emersonii XDH(TeXDH)was heterologously expressed in Escherichia coli BL21(DE3)at 16 ℃ in the soluble form.Recombinant TeXDH with high purity was purified by using a Ni-NTA affinity column.Size-exclusion chromatography and SDS-PAGE analysis demonstrated that the puri-fied recombinant TeXDH exists as a native trimer with a molecular mass of approximately 116 kD,and is composed of three identical subunits,each with a molecular weight of around 39 kD.The TeXDH strictly preferred NAD+as a coenzyme to NADP+.The optimal temperature and pH of the TeXDH were 40 ℃and 10.0,respectively.After EDTA treatment,the enzyme activity of TeXDH decreased to 43.26%of the initial enzyme activity,while the divalent metal ions Mg2+or Ca2+could recover the enzyme activity of TeXDH,reaching 103.32%and 110.69%of the initial enzyme activity,respectively,making them the optimal divalent metal ion cofactors for TeXDH enzyme.However,the divalent metal ions of Mn2+,Ni2+,Cu2+,Zn2+,Co2+,and Cd2+significantly inhibited the activity of TeXDH.ICP-MS and molecular doc-king studies revealed that 1 mol/L of TeXDH bound 2 mol/L Zn2+ions and 1 mol/L Mg2+ion.Further-more,TeXDH exhibited a high specificity for xylitol,laying the foundation for the development of future xylitol biosensors.
2.Detection and clinical significance of serum thymic stromal lymphopoietin levels in patients with dermatomyositis
Yongheng CHEN ; Kaile HE ; Shile CHEN ; Xinlin SHAO ; Chenyu WU ; Ling JIN ; Weiru YUAN ; Hua CAO
Chinese Journal of Dermatology 2025;58(8):729-735
Objective:To explore the relationship between serum thymic stromal lymphopoietin (TSLP) levels and the occurrence of pruritus as well as interstitial lung diseases (ILD) in patients with dermatomyositis (DM) or clinically amyopathic dermatomyositis (CADM) .Methods:A retrospective analysis was conducted on the clinical data from 61 diagnosed DM/CADM patients at Ruijin Hospital, Shanghai Jiao Tong University School of Medicine between February 2019 and March 2022. Thirty-five age- and sex-matched healthy individuals were included as controls. Serum TSLP and anti-melanoma differentiation-associated gene 5 (MDA5) antibody levels were measured using enzyme-linked immunosorbent assay (ELISA) . A multiplexed flow cytometry assay was performed to detect the serum levels of 12 cytokines, including interferon (IFN) -α2, IFN-γ, interleukin (IL) -4, IL-5, etc. Pruritus severity in DM/CADM patients was assessed using the visual analog scale (VAS) . Relationships of serum TSLP levels with skin lesions, inflammatory markers, and the severity of ILD were analyzed using GraphPad Prism 8.0.1 and R (version 4.3.2) software.Results:Among the 61 patients with DM/CADM, 18 were males (13 with DM and 5 with CADM) and 43 were females (30 with DM and 13 with CADM) , with ages of 53.08 ± 14.97 years and disease duration of 6.27 ± 4.27 months. No significant differences were observed in sex or age between the DM/CADM group and the healthy control group (both P > 0.05) . Of the 61 patients, 15 (24.59%) were positive for anti-MDA5 antibodies, and 31 (51.67%) had ILD, including 11 with acute/subacute interstitial pneumonia (A/SIP) . The pruritus VAS score ( M[ Q1, Q3]) among the 61 patients was 3 (0, 6) points; 20 patients (32.79%) reported no pruritus, while 11 (18.32%) , 17 (27.87%) , and 13 (21.31%) experienced mild, moderate, and severe pruritus, respectively. Serum TSLP levels were significantly higher in the DM/CADM patients than in the healthy controls (91.0 [34.6, 273.0] vs. 25.0 [0.0, 83.5] pg/ml, Z = 1 502.50, P < 0.001]. The TSLP levels were significantly positively correlated with serum levels of alanine aminotransferase (ALT) , lactate dehydrogenase, C-reactive protein, erythrocyte sedimentation rate and ferritin, and significantly negatively correlated with pruritus VAS scores, eosinophil percentages, and eosinophil counts (all P < 0.05) . Serum TSLP levels were significantly higher in the anti-MDA5 antibody-positive patients than in the anti-MDA5 antibody-negative patients (273.0 [103.8, 2 576.0] pg/ml vs. 67.4 [16.9, 218.7] pg/ml, P = 0.002) , as well as in the patients with A/SIP than in those without A/SIP (390.0 [168.2, 2 869.6] vs. 67.5 [21.8, 235.9] pg/ml, P < 0.001) . ROC analysis showed that the combined detection of serum TSLP levels and anti-MDA5 antibodies had a higher predictive value for A/SIP in DM/CADM patients (AUC = 0.922) than either the detection of anti-MDA5 antibodies (AUC = 0.831) or TSLP (AUC = 0.824) alone did. Conclusions:Serum TSLP levels were significantly higher in patients with DM/CADM than in healthy controls, with particularly higher levels observed in anti-MDA5 antibody-positive patients and patients with coexisting A/SIP, and were correlated with several indicators of disease activity. The combination of serum TSLP levels and anti-MDA5 antibodies provided greater predictive value for A/SIP than either of them alone did.
3.Associations of Exposure to Typical Environmental Organic Pollutants with Cardiopulmonary Health and the Mediating Role of Oxidative Stress: A Randomized Crossover Study.
Ning GAO ; Bin WANG ; Ran ZHAO ; Han ZHANG ; Xiao Qian JIA ; Tian Xiang WU ; Meng Yuan REN ; Lu ZHAO ; Jia Zhang SHI ; Jing HUANG ; Shao Wei WU ; Guo Feng SHEN ; Bo PAN ; Ming Liang FANG
Biomedical and Environmental Sciences 2025;38(11):1388-1403
OBJECTIVE:
The study aim was to investigate the effects of exposure to multiple environmental organic pollutants on cardiopulmonary health with a focus on the potential mediating role of oxidative stress.
METHODS:
A repeated-measures randomized crossover study involving healthy college students in Beijing was conducted. Biological samples, including morning urine and venous blood, were collected to measure concentrations of 29 typical organic pollutants, including hydroxy polycyclic aromatic hydrocarbons (OH-PAHs), bisphenol A and its substitutes, phthalates and their metabolites, parabens, and five biomarkers of oxidative stress. Health assessments included blood pressure measurements and lung function indicators.
RESULTS:
Urinary concentrations of 2-hydroxyphenanthrene (2-OH-PHE) ( β = 4.35% [95% confidence interval ( CI): 0.85%, 7.97%]), 3-hydroxyphenanthrene ( β = 3.44% [95% CI: 0.19%, 6.79%]), and 4-hydroxyphenanthrene (4-OH-PHE) ( β = 5.78% [95% CI: 1.27%, 10.5%]) were significantly and positively associated with systolic blood pressure. Exposures to 1-hydroxypyrene (1-OH-PYR) ( β = 3.05% [95% CI: -4.66%, -1.41%]), 2-OH-PHE ( β = 2.68% [95% CI: -4%, -1.34%]), and 4-OH-PHE ( β = 3% [95% CI: -4.68%, -1.29%]) were negatively associated with the ratio of forced expiratory volume in the first second to forced vital capacity. These findings highlight the adverse effects of exposure to multiple pollutants on cardiopulmonary health. Biomarkers of oxidative stress, including 8-hydroxy-2'-deoxyguanosine and extracellular superoxide dismutase, mediated the effects of multiple OH-PAHs on blood pressure and lung function.
CONCLUSION
Exposure to multiple organic pollutants can adversely affect cardiopulmonary health. Oxidative stress is a key mediator of the effects of OH-PAHs on blood pressure and lung function.
Humans
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Oxidative Stress/drug effects*
;
Male
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Cross-Over Studies
;
Female
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Young Adult
;
Environmental Pollutants/toxicity*
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Environmental Exposure/adverse effects*
;
Biomarkers/blood*
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Adult
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Blood Pressure/drug effects*
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Polycyclic Aromatic Hydrocarbons/urine*
;
Beijing
4.Ionizing Radiation Alters Circadian Gene Per1 Expression Profiles and Intracellular Distribution in HT22 and BV2 Cells.
Zhi Ang SHAO ; Yuan WANG ; Pei QU ; Zhou Hang ZHENG ; Yi Xuan LI ; Wei WANG ; Qing Feng WU ; Dan XU ; Ju Fang WANG ; Nan DING
Biomedical and Environmental Sciences 2025;38(11):1451-1457
5.A comparative study of endoscopic super minimally invasive surgery and laparoscopic minimally invasive excision for gastric glomus tumors
Yaoqian YUAN ; Xin WU ; Qun SHAO ; Bo NING ; Peifa LIU ; Jiafeng WANG ; Kunming LYU ; Qianqian CHEN
Chinese Journal of Postgraduates of Medicine 2025;48(8):678-682
Objective:To compare the efficacy and safety of the gastrointestinal endoscopic super minimally invasive surgery (eSMIS) and the laparoscopic minimally invasive procedure (LS) for the treatment of gastric glomus tumors(GGT).Methods:This study retrospectively included 15 patients with GGT who underwent eSMIS at the First Medical Center of Chinese PLA General Hospital from May 2014 to May 2024. Data on demographic characteristics, surgical indexes, efficacy evaluation indexes, postoperative medical disposition, health economics indexes and postoperative laboratory indexes were collected and analyzed, the patients were followed up for survival outcomes and the postoperative quality of life, efficacy and safety of the two groups were compared.Results:The amount of bleeding in the eSMIS group was lower than that in the LS group: 1.25 (0, 3.75) ml vs. 41.56 (10.00, 50.00) ml, and the surgical cost in the eSMIS group was lower than that in the LS group: 10 792.87 (8 424.90, 12 730.30) yuan vs. 21 773.06 (19 940.60, 24 843.10) yuan, there were statistical differences ( P<0.05). The surgical efficacy, postoperative laboratory indicators and medical treatment convenience between the two groups had no statistical differences ( P>0.05). Conclusions:In the choice of surgical procedures for the treatment of GGT, eSMIS is superior to LS in terms of bleeding and surgical cost, and not inferior to LS in other indicators. This suggested that the treatment of GGT with eSMIS is feasible, safe and effective.
6.Analyzing the influencing factors of occupational burnout among disease control and prevention staffs in Sichuan Province
Chaoxue WU ; Shuang DONG ; Liang WANG ; Xunbo DU ; Lin ZHAO ; Dan SHAO ; Quanquan XIAO ; Lijun ZHOU ; Chongkun XIAO ; Heng YUAN
China Occupational Medicine 2025;52(3):288-292
Objective To assess the situation and influencing factors of occupational burnout among the staff at the Center for Disease Control and Prevention (CDC) in Sichuan Province. Methods A total of 1 038 CDC staff members in Sichuan Province were selected as the study subjects using the stratified random sampling method. Occupational burnout of the staff was assessed using the Maslach Burnout Inventory General Survey via an online questionnaire. Results The detection rate of occupational burnout was 42.3% (439/1 038). Binary logistic regression analysis result showed that, after controlling for confounding factors such as education level and alcohol consumption, CDC staffs aged at 20-<31, 31-<41, and 41-<51 years were at higher risk of occupational burnout compared with those ≥51 years (all P<0.05). CDC staffs with 5-<10 or ≥10 years of service had higher occupational burnout risk compared with those with <5 years (both P<0.05). CDC staffs with poor or fair health status, irregular diet, and poor sleep quality had higher risk of occupational burnout compared with those healthy, have regular diet, and good sleep quality (all P<0.05). The risk of occupational burnout increased with higher overtime frequency (all P<0.05). Conclusion Occupational burnout among CDC staffs in Sichuan Province is relatively high. Age, years of service, health status, diet, sleep quality, and overtime frequency are key influencing factors.
7.Heterologous Expression,Purification and Enzymatic Characterization of Xylitol Dehydrogenase from the Thermophilic Fungus Talaromyces emersonii
Er MENG ; Cong QU ; Ke YI ; Hui-Min LI ; Xin-Yi DUAN ; Zhe-Yuan ZHANG ; Shao-Long HE ; Yu-Tao LUO ; Lei WU ; Dong-Yi ZHANG ; Chang-Jun LIU
Chinese Journal of Biochemistry and Molecular Biology 2025;41(7):1007-1018
The xylitol dehydrogenase(XDH)is a crucial enzyme involved in the xylose utilization in pentose-catabolizing yeasts and fungi.In addition to producing xylulose,XDH can also be employed to develop a biosensor for monitoring xylitol concentration.In this study,the gene encoding the thermophilic fungus Talaromyces emersonii XDH(TeXDH)was heterologously expressed in Escherichia coli BL21(DE3)at 16 ℃ in the soluble form.Recombinant TeXDH with high purity was purified by using a Ni-NTA affinity column.Size-exclusion chromatography and SDS-PAGE analysis demonstrated that the puri-fied recombinant TeXDH exists as a native trimer with a molecular mass of approximately 116 kD,and is composed of three identical subunits,each with a molecular weight of around 39 kD.The TeXDH strictly preferred NAD+as a coenzyme to NADP+.The optimal temperature and pH of the TeXDH were 40 ℃and 10.0,respectively.After EDTA treatment,the enzyme activity of TeXDH decreased to 43.26%of the initial enzyme activity,while the divalent metal ions Mg2+or Ca2+could recover the enzyme activity of TeXDH,reaching 103.32%and 110.69%of the initial enzyme activity,respectively,making them the optimal divalent metal ion cofactors for TeXDH enzyme.However,the divalent metal ions of Mn2+,Ni2+,Cu2+,Zn2+,Co2+,and Cd2+significantly inhibited the activity of TeXDH.ICP-MS and molecular doc-king studies revealed that 1 mol/L of TeXDH bound 2 mol/L Zn2+ions and 1 mol/L Mg2+ion.Further-more,TeXDH exhibited a high specificity for xylitol,laying the foundation for the development of future xylitol biosensors.
8.Regulatory effect of neutrophils in microglial polarization after permanent ischemic stroke
Min-Hua HUANG ; Xin-Yan YE ; Si-Yu WU ; Shao-Tong LUO ; Zhi-Shan WU ; Yuan CHEN ; Su-Ning PING
Acta Anatomica Sinica 2025;56(2):136-142
Objective To investigate the effects of peripheral blood neutrophil infiltration on the polarization regulation of cerebral resident microglia under a permanent ischemic stroke model.Methods Fifty-eight C57BL/6 mice were divided into two groups.One group was sham group,and the other group of mice was subjected to permanent middle cerebral artery occlusion surgery.Mice were euthanized 48 hours,7 days,14 days,and 30 days after surgery for tissue collection.Western blotting was used to detect expression levels of M1 microglia markers CD 16,M2 microglia marker arginase 1(Arg1),inflammatory cytokine interleukin-1 β(IL-1β),and neutrophil marker myeloperoxidase(MPO)in brain tissue.Immunofluorescence histochemical staining was used to assess neutrophil infiltration and M2 microglial distribution around the infarct area in brain sections.In vitro,purified neutrophils were co-cultured with BV2 microglial cells.After lipopolysaccharide stimulation,the phagocytosis of neutrophils by BV2 cells was observed,and the expression levels of CD16 and Arg1 proteins in BV2 cells were detected.Results Western blotting showed that the levels of CD16(P<0.05),IL-1β(P<0.001),and MPO(P<0.05)in brain tissue increased significantly 48 hours and 7 days after surgery,then decreased,with MPO expression returning to normal levels 30 days after surgery.Immunofluorescence showed a significant increase of MPO-positive cells around the infarct area of the mouse cerebral cortex 48 hours after surgery(P<0.001),followed by a decrease(P<0.05).The number of ionized calcium binding adapter molecule 1(Iba1)and MPO double-positive cells gradually increased after surgery,and reached their peak at 14 days(P<0.05).Iba1 and Arg1 double-positive cells also increased significantly 7 days(P<0.05)and 14 days(P<0.01)after surgery.In vitro,co-culture experiments showed that after BV2 phagocytosing neutrophils,CD 16(P<0.05)significantly decreased and Arg1 significantly upregulated(P<0.05).Conclusion In a permanent ischemic stroke model,microglia transition from M1 to M2 type after phagocytosing neutrophils,and the injured brain area changes from pro-inflammatory state to anti-inflammatory state.
9.Analysis of the impact of phase Ⅱ cardiac rehabilitation on the rehabilitation needs and physical activity of patients with coronary heart disease after interventional surgery
Ying ZHANG ; Xiwei ZHANG ; Jiahui WU ; Wenxian LIU ; Yan FENG ; Nan LI ; Yong SHAO ; Lixin ZHANG ; Yuan TIAN ; Zhiying LI ; Yin LI
Chongqing Medicine 2025;54(5):1080-1084
Objective To investigate the impact of phase Ⅱ cardiac rehabilitation on the rehabilitation needs and physical activity status of patients after coronary heart disease intervention.Methods A total of 90 patients with coronary heart disease who underwent percutaneous coronary intervention(PCI)in the Coronary Heart Disease Center of the hospital from August 2023 to August 2024 were selected as the research subjects.They were subjected to a 12-week standardized phase Ⅱ cardiac rehabilitation training.General data survey forms,cardiac rehabilitation scales,and the International Physical Activity Questionnaire were used for scale surveys to understand the patients' needs before and after rehabilitation and their weekly physical activity en-ergy expenditure.The cardiopulmonary exercise test gold standard,which reflects exercise capacity through three indicators-maximum oxygen uptake(VO2 max),anaerobic threshold(AT),and metabolic equivalents(MET),were used to compare the physical activity status before and after cardiac rehabilitation.Results Compared with before the implementation,after the implementation of cardiac rehabilitation,the autonomy score in-creased(21.36±1.85 vs.16.73±3.28),the process anxiety(12.60±3.87 vs.14.27±2.12)and outcome anxiety scores(2.31±1.76 vs.4.56±3.56)decreased,the level of low-intensity physical activity decreased[(2 711.62±1 487.09)min/week vs.(3 845.97±2 083.71)min/week],the levels of moderate-intensity[(1 314.67±783.54)min/week vs.(686.22±126.79)min/week],high-intensity[(1 861.33±798.27)min/week vs.(112.00±40.77)min/week],and total physical activity increased[(5 887.62±2 843.54)min/week vs.(4 644.19±2 287.16)min/week].The levels of VO2 max[(28.11±14.28)mL·min-1·kg-1 vs.(23.82±12.34)mL·min-1·kg-1],AT[(16.06±5.41)mL·min-1·kg-1 vs.(13.53±4.56)mL·min-1·kg-1],and MET[(6.89±1.59)mL·min-1·kg-1 vs.(5.78±1.21)mL·min-1·kg-1]all in-creased,with statistically significant differences(P<0.05).Conclusion Phase Ⅱ rehabilitation after PCI can effectively improve patients' physical activity levels.
10.Explanation and interpretation of blood transfusion provisions for children with hematological diseases in the national health standard "Guideline for pediatric transfusion".
Ming-Yi ZHAO ; Rong HUANG ; Rong GUI ; Qing-Nan HE ; Ming-Yan HEI ; Xiao-Fan ZHU ; Jun LU ; Xiao-Jun XU ; Tian-Ming YUAN ; Rong ZHANG ; Xu WANG ; Jin-Ping LIU ; Jing WANG ; Zhi-Li SHAO ; Yong-Jian GUO ; Xin-Yin WU ; Jia-Rui CHEN ; Qi-Rong CHEN ; Jia GUO ; Ming-Hua YANG
Chinese Journal of Contemporary Pediatrics 2025;27(1):18-25
To guide clinical blood transfusion practices for pediatric patients, the National Health Commission has issued the health standard "Guideline for pediatric transfusion" (WS/T 795-2022). Blood transfusion is one of the most commonly used supportive treatments for children with hematological diseases. This guideline provides guidance and recommendations for blood transfusions in children with aplastic anemia, thalassemia, autoimmune hemolytic anemia, glucose-6-phosphate dehydrogenase deficiency, acute leukemia, myelodysplastic syndromes, immune thrombocytopenic purpura, and thrombotic thrombocytopenic purpura. This article presents the evidence and interpretation of the blood transfusion provisions for children with hematological diseases in the "Guideline for pediatric transfusion", aiming to assist in the understanding and implementing the blood transfusion section of this guideline.
Humans
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Child
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Hematologic Diseases/therapy*
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Blood Transfusion/standards*
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Practice Guidelines as Topic

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