1.USP25 ameliorates vascular remodeling by deubiquitinating FOXO3 and promoting autophagic degradation of FOXO3.
Yanghao CHEN ; Bozhi YE ; Diyun XU ; Wante LIN ; Zimin FANG ; Xuefeng QU ; Xue HAN ; Wu LUO ; Chen CHEN ; Weijian HUANG ; Hao ZHOU ; Gaojun WU ; Yi WANG ; Guang LIANG
Acta Pharmaceutica Sinica B 2025;15(3):1643-1658
Long-term hypertension causes excessive vascular remodeling and leads to adverse cardiovascular events. Balance of ubiquitination and deubiquitination has been linked to several chronic conditions, including pathological vascular remodeling. In this study, we discovered that the expression of ubiquitin-specific protease 25 (USP25) is significantly up-regulated in angiotensin II (Ang II)-challenged mouse aorta. Knockout of Usp25 augments Ang II-induced vascular injury such as fibrosis and endothelial to mesenchymal transition (EndMT). Mechanistically, we found that USP25 interacts directly with Forkhead box O3 (FOXO3) and removes the K63-linked ubiquitin chain on the K258 site of FOXO3. We also showed that this USP25-mediated deubiquitination of FOXO3 increases its binding to light chain 3 beta isoform and autophagosomic-lysosomal degradation of FOXO3. In addition, we further validated the biological function of USP25 by overexpressing USP25 in the mouse aorta with AAV9 vectors. Our studies identified FOXO3 as a new substrate of USP25 and showed that USP25 may be a potential therapeutic target for excessive vascular remodeling-associated diseases.
2.Preparation of monoclonal antibody against PRRSV-2 N protein and identification of antigenic epitopes
Yanli PANG ; Jianguang QIN ; Muyang LIU ; Tongwei REN ; Jiaqi LIU ; Lingshan ZHOU ; Ying CHEN ; Kang OUYANG ; Weijian HUANG ; Zuzhang WEI
Chinese Journal of Veterinary Science 2025;45(1):16-21,45
To prepare monoclonal antibody to the N protein of porcine reproductive and respiratory syndrome virus(PRRSV),BALB/c mice were immunized with the purified N protein of the PRRSV-2 strain expressed by prokaryotic expression system.Mouse splenocytes were fused with myeloma cells using hybridoma technique,hybridoma cells were identified by indirect ELISA method and indirect immunofluorescence assay(IFA),and positive hybridoma cells were screened for subclones using the limited dilution method.The results showed that a monoclonal antibody cell line 4A7 was successfully obtained,and the results of Western blot and IFA indicated that the monoclonal antibody could accurately recognize the N proteins of type 1 and type 2 PRRSV.Mean-while,the N protein gene was truncated and expressed by prokaryotic expression system,and the amino acid sequence of the B-cell antigenic epitope recognized by 4A7 was screened as 51EKPHF55 using Western blot.Comparison of epitope amino acids in the N protein gene sequences of different strains revealed that the antigenic epitope 51EKPHF55 recognized by the monoclonal antibody 4A7 has no amino acid difference in the sequences of three subtypes among the PRRSV-1 strains and nine lineages among the PRRSV-2 strains,indicating a high degree of conservation.The results of the study provide a foundation the development of PRRSV diagnostic kits and novel vaccines.
3.Evaluation index system of life support equipment of hospital based on fuzzy comprehensive evaluation model
Liangjun NI ; Weijian CHEN ; Weijie HUANG ; Xia ZHANG
China Medical Equipment 2025;22(6):113-118
Objective:To construct a fuzzy comprehensive evaluation model for life support equipment of hospital,so as to improve quality of management for equipment.Methods:Based on the operating status of life support equipment in Jiangmen Maternal and Child Health Care Hospital of Guangdong Province,a fuzzy comprehensive evaluation model was constructed,and the indicators of different levels were set.The index weights were calculated according to the Analytic Hierarchy Process(AHP),and the equipment management strategy was formulated based on the evaluated results.A total of 98 life support equipment in clinical use in our hospital from January 2023 to July 2024 were selected.In them,49 equipment were managed under the conventional management mode,and the other 49 equipment were managed under the assessment system management mode of the kind of life support equipment based on fuzzy comprehensive evaluation model(model management mode)according to the random number table method.The timeliness,management effect and management efficiency of the two management modes for equipment were compared.A self-made satisfaction questionnaire was used to investigate the satisfaction scores of equipment engineers,clinicians and managers of relevant departments in usage management for equipment in the equipment usage of the two management modes.Results:The timeliness of equipment early warning,the timeliness of maintenance personnel's arrival,and the timeliness of proposing maintenance opinions were respectively(0.78±0.14)min,(5.14±2.11)min,and(10.69±3.11)min in the model management mode,all which were shorter than those in the conventional management mode,and the differences were statistically significant(t=16.697,11.857,16.117,P<0.05).The regular maintenance rate of equipment adopting the model management mode was higher than that adopting the conventional management mode,and the failure rate was lower than that adopting the conventional management mode,and the differences were statistically significant(x2=12.274,12.261,P<0.05),respectively.The maintenance time of the model management mode was shorter than that of the conventional management mode,and the operation and maintenance cost of the model management mode was lower than that of the conventional management mode,and the differences were statistically significant(t=17.451,12.707,P<0.05).The scores of management efficiencies of resource allocation,technical support,environmental guarantee and information infrastructure of the model management mode were all higher than those of the conventional management mode,and the differences were statistically significant(t=12.268,14.908,16.128,13.920,P<0.05).In usage management for equipment,the satisfaction scores of equipment engineers,clinicians and managers of relevant departments for equipment management adopting the model management mode were higher than those adopting the conventional management mode,and the differences were statistically significant(t=13.118,16.209,12.557,P<0.05).Conclusion:The assessment system of hospital for life support equipment based on the fuzzy comprehensive evaluation model can enhance the quality of management and operation for equipment,and improve the service level of equipment,and increase the operational efficiency of equipment.
4.Preparation of monoclonal antibody against PRRSV-2 N protein and identification of antigenic epitopes
Yanli PANG ; Jianguang QIN ; Muyang LIU ; Tongwei REN ; Jiaqi LIU ; Lingshan ZHOU ; Ying CHEN ; Kang OUYANG ; Weijian HUANG ; Zuzhang WEI
Chinese Journal of Veterinary Science 2025;45(1):16-21,45
To prepare monoclonal antibody to the N protein of porcine reproductive and respiratory syndrome virus(PRRSV),BALB/c mice were immunized with the purified N protein of the PRRSV-2 strain expressed by prokaryotic expression system.Mouse splenocytes were fused with myeloma cells using hybridoma technique,hybridoma cells were identified by indirect ELISA method and indirect immunofluorescence assay(IFA),and positive hybridoma cells were screened for subclones using the limited dilution method.The results showed that a monoclonal antibody cell line 4A7 was successfully obtained,and the results of Western blot and IFA indicated that the monoclonal antibody could accurately recognize the N proteins of type 1 and type 2 PRRSV.Mean-while,the N protein gene was truncated and expressed by prokaryotic expression system,and the amino acid sequence of the B-cell antigenic epitope recognized by 4A7 was screened as 51EKPHF55 using Western blot.Comparison of epitope amino acids in the N protein gene sequences of different strains revealed that the antigenic epitope 51EKPHF55 recognized by the monoclonal antibody 4A7 has no amino acid difference in the sequences of three subtypes among the PRRSV-1 strains and nine lineages among the PRRSV-2 strains,indicating a high degree of conservation.The results of the study provide a foundation the development of PRRSV diagnostic kits and novel vaccines.
5.Evaluation index system of life support equipment of hospital based on fuzzy comprehensive evaluation model
Liangjun NI ; Weijian CHEN ; Weijie HUANG ; Xia ZHANG
China Medical Equipment 2025;22(6):113-118
Objective:To construct a fuzzy comprehensive evaluation model for life support equipment of hospital,so as to improve quality of management for equipment.Methods:Based on the operating status of life support equipment in Jiangmen Maternal and Child Health Care Hospital of Guangdong Province,a fuzzy comprehensive evaluation model was constructed,and the indicators of different levels were set.The index weights were calculated according to the Analytic Hierarchy Process(AHP),and the equipment management strategy was formulated based on the evaluated results.A total of 98 life support equipment in clinical use in our hospital from January 2023 to July 2024 were selected.In them,49 equipment were managed under the conventional management mode,and the other 49 equipment were managed under the assessment system management mode of the kind of life support equipment based on fuzzy comprehensive evaluation model(model management mode)according to the random number table method.The timeliness,management effect and management efficiency of the two management modes for equipment were compared.A self-made satisfaction questionnaire was used to investigate the satisfaction scores of equipment engineers,clinicians and managers of relevant departments in usage management for equipment in the equipment usage of the two management modes.Results:The timeliness of equipment early warning,the timeliness of maintenance personnel's arrival,and the timeliness of proposing maintenance opinions were respectively(0.78±0.14)min,(5.14±2.11)min,and(10.69±3.11)min in the model management mode,all which were shorter than those in the conventional management mode,and the differences were statistically significant(t=16.697,11.857,16.117,P<0.05).The regular maintenance rate of equipment adopting the model management mode was higher than that adopting the conventional management mode,and the failure rate was lower than that adopting the conventional management mode,and the differences were statistically significant(x2=12.274,12.261,P<0.05),respectively.The maintenance time of the model management mode was shorter than that of the conventional management mode,and the operation and maintenance cost of the model management mode was lower than that of the conventional management mode,and the differences were statistically significant(t=17.451,12.707,P<0.05).The scores of management efficiencies of resource allocation,technical support,environmental guarantee and information infrastructure of the model management mode were all higher than those of the conventional management mode,and the differences were statistically significant(t=12.268,14.908,16.128,13.920,P<0.05).In usage management for equipment,the satisfaction scores of equipment engineers,clinicians and managers of relevant departments for equipment management adopting the model management mode were higher than those adopting the conventional management mode,and the differences were statistically significant(t=13.118,16.209,12.557,P<0.05).Conclusion:The assessment system of hospital for life support equipment based on the fuzzy comprehensive evaluation model can enhance the quality of management and operation for equipment,and improve the service level of equipment,and increase the operational efficiency of equipment.
6.Source and application of seed cells in bioartificial liver support system
Xuejing ZHU ; Weijian HUANG ; Hexin YAN
Journal of Clinical Hepatology 2024;40(2):246-251
So far, there are still no specific treatment methods for severe hepatitis and liver failure, resulting in a mortality rate of over 70%, and they are the difficulties in the treatment of critical illness in China and globally. Liver transplantation is currently the most effective treatment method for end-stage liver disease, but only 1% — 2% of patients can receive the opportunity for organ transplantation. The bioartificial liver support system utilizes external mechanical, physical, and biological devices to remove various harmful substances accumulated in the patient’s body, compensate for the metabolic functions of the liver, supplement necessary substances, improve internal environment, promote the recovery of liver function, help patients get through the critical period, and save time for liver transplantation, and therefore, it is considered one of the important methods for the treatment of end-stage liver disease. Since hepatocytes are the core element of bioartificial liver, this article summarizes the sources of liver seed cells, 3D culture methods, and corresponding bioreactor culture systems and hopes to gradually solve the core issue of large-scale in vitro preparation of hepatocytes to obtain hepatocytes with adequate quantity and quality, which urgently needs to be addressed in clinical application.
7.Effect of individualized PEEP regulated by driving pressure on infant one-lung ventilation during thoracic surgery
Weijian HUANG ; Yang LI ; Haiyan WANG ; Jing LIU ; Hui LUO ; Zurong HU
The Journal of Practical Medicine 2024;40(3):360-364
Objective To explore the effect of protective ventilation strategies based on individualized positive end expiratory pressure(PEEP)regulated by driving pressure(Pd)in infants under OLV.Methods Sixty infants undergoing elective thoracoscopic surgery were randomly divided into the control group and driving pressure group,with 30 cases in each group.The two groups were compared in terms of MAP,HR,tidal volume(Vt),PEEP,Pd,airway peak pressure(Ppeak),and static lung compliance(Cs),and arterial blood gas during OLV before the artificial pneumothorax(T0),10 min(T1),30 min(T2),and 60 min(T3)after the artificial pneumothorax,and right at the end of the artificial pneumothorax(T4).Results There were no significant differences in MAP,HR,and Vt between the two groups at all the time points(P>0.05).Both groups showed the increase in Pd and Ppeak and decresase in Cs at T1,T2,and T3(all P<0.05).PaO2 and OI in both group were decreased and PaCO2 was increased at T2(all P<0.05).Compared with the control group,the driving pressure group presented lower Pd and Ppeak,higher PEEP and Cs at T1,T2,and T3(P<0.05),and higher PaO2 and OI(P<0.05)and no significant differences in PaCO2 and FiO2 at T2(P<0.05).The rate of rescue ventilation during OLV was lower in the driving pressure group(2 cases)than in the control group(9 cases)(6.9%versus 32.4%,P<0.05).There was no significant difference in the inci-dence of postoperative complications between the two groups(P<0.05).Conclusion The individualized positive end expiratory pressure(PEEP)regulated by Pd can optimize the PEEP settings upon infant individuals under OLV and improve the compliance and oxygenation of ventilated lung.
8.Study on lipidomics of CD4+T cells in mice with diabetic kidney disease
Xiaoyu ZHANG ; Haibo TAN ; Minyi HUANG ; Weijian BEI ; Yiqi YANG
Chinese Journal of Immunology 2024;40(3):471-477
Objective:To investigate the lipidomics differences of CD4+T immune cells in diabetic kidney disease(DKD)mice,and screen out the differential metabolites with biological significance.Methods:CD4(L3T4)MicroBeads immunomagnetic beads were used to isolate CD4+T immune cells from spleen of BKS.Cg-Dock7m+/+Leprdb/J mice with spontaneous DKD;the purity of CD4+T cells were identified by flow cytometry.The non-targeted lipidomics of CD4+T cells were detected by LC-MS/MS,and the differ-ential metabolites were analyzed.Results:A total of 463 metabolites were detected by LC-MS.PCA and OPLS-DA analysis showed that the metabolic components were significantly separated;twenty-four differential metabolites were screened out.KEGG and enrich-ment analysis showed that the differential metabolites involved in the disorder of glycerol phospholipid metabolism.Conclusion:Phos-pholipid metabolism of CD4+T cells is closely related to the occurrence of DKD.Phospholipid metabolism targeting DKD CD4+T cells in DKD may be a new direction of DKD treatment.
9.Construction and operation of hospital radiation protection management information system
Zhenhua ZHAO ; Fei CAI ; Jiaqian ZHANG ; Min HAN ; Weijian ZHU ; Yanan HUANG ; Qianling LI
Chinese Journal of Hospital Administration 2022;38(12):927-931
The construction and operation of the hospital-wide radiation protection management information system is of great significance for improving the efficiency and quality of radiation protection management. A hospital has built a hospital-wide radiation protection management system, which consisted of four modules: radiation staff management module, protection items management module, site and equipment management module, and self-inspection module. The system adopted B/S mode, carried the lightweight J2EE framework-SpringBoot framework, and used mybatis, which was flexible and practical, as the persistence layer of the system, to store the data completely in the SQLServer database.Java language was used as the system development language, and the front and back-end separation architecture was used. Each of the four modules implemented the management responsibility departments and responsible personnel to carry out the all-round management of radiation protection, and realized the integrated, information-based and standardized management of radiation protection management in the hospital.
10.Altered inter- and intrahemispheric functional connectivity dynamics in long-term smokers
Mengzhe ZHANG ; Huiyu HUANG ; Xinyu GAO ; Zhengui YANG ; Mengmeng WEN ; Weijian WANG ; Jingliang CHENG ; Yong ZHANG
Chinese Journal of Psychiatry 2022;55(2):98-105
Objective:To explore the neuro mechanism of tobacco addiction, this study applied the dynamic functional connectivity density (dFCD) to analyze the dynamic changes of brain functional networks at both whole-brain and hemisphere levels in long-term male smokers.Methods:Thirty-seven smokers met the criteria for substance addiction according to the Diagnostic and Statistical Manual of Mental Disorders-Four Edition (DSM-Ⅳ) and 47 healthy controls matched with age and education were recruited. Resting-state functional magnetic resonance imaging (rs-fMRI) data and clinical data were collected in both smokers and healthy controls. The whole-brain functional connectivity was decomposed into ipsilateral and contralateral functional connectivity. Sliding-window analysis was utilized to capture the intra- and interhemispheric dFCD patterns. The temporal variability of the functional connectivity dynamics was further quantified using the standard deviation (SD) of intra- and inter-hemispheric dFCD over time. The two-sample t-test was used to compare dFCD variance between the two groups. Results:Compared with healthy controls, smokers showed increased global dFCD variance in the left insula and left putamen ( t=3.99, 4.49). Increased dFCD variability was observed in the left insula and left putamen among smokers compared with healthy controls when comparing dFCD variability at both intra-( t=4.20, 4.33) and interhemispheric ( t=3.81, 4.33) level (Gaussian Random Field GRF corrected, voxel P<0.005, cluster P<0.05). Conclusion:Among long-term smokers, a time-varying characteristic of functional connectivity patterns was observed in the left insula and left putamen, which reflects the abnormal communication between these brain areas and other brain areas. The alteration of intra- and interhemispheric dynamic functional connectivity may be the underlying neuro mechanism of tobacco addiction.

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