1.Study on the angiogenic ability of exosomes derived from dental pulp stem cells modified by circRNA SIPA1L1
Jing LIU ; Chuntao LENG ; Yan WANG
The Journal of Practical Medicine 2024;40(9):1211-1217
Objective To investigate the effect of human pulp stem cells(hDPSC)-derived exosomes(Exo)modified by circRNA(circRNA)signal-induced proliferation-associated protein-like 1(SIPA1L1)on the angiogenesis of human umbilical vein endothelial cells(HUVEC).Methods hDPSC was isolated and cultured from pulp tissue,the circSIPA1L1 overexpression plasmid was transfected into hDPSC,and Exo was isolated and identified.HUVEC was divided into control group,hDPSC Exo group and circSIPA1L1-hDPSC Exo group.After culture for 48 h,the vasculoformation ability was detected by Matrigel matrix gel vasculoformation test,the expres-sion levels of vascular endothelial growth factor(VEGF),vascular endothelial growth factor receptor 2(VEGFR2)and placental growth factor(PGF)were determined by qRT-PCR and Western blot.Results The Exo was success-fully isolated from the untransfected hDPSC and the hDPSC transfected with circSIPA1L1,and compared with the HDPSC-derived Exo,the relative expression of circSIPA1L1 in the hDPSC Exo transfected with circSIPA1L1 was significantly up-regulated(P<0.05).Compared with hDPSC Exo group,the number of HUVEC tubular structures in circSIPA1L1-hDPSC Exo group was significantly increased(P<0.05),and the mRNA and protein relative expressions of VEGF,VEGFR2 and PGF were also significantly up-regulated(P<0.05).Conclusions Modification of hDPSC-derived Exo by circRNA SIPA1L1 can promote angiogenesis,and the mechanism may be related to up-regulation of VEGF,VEGFR2 and PGF expression levels.
2.Evolution and innovation of preservation fluid for donor liver
Guotao YU ; Yanfeng YIN ; Chuntao YAN ; Guangxu ZOU ; Huangyan ZHANG ; Li MA ; Zongqiang HU
Organ Transplantation 2024;15(1):131-137
Organ preservation fluid could mitigate cold ischemia injury and maintain normal function of the grafts. At present, how to reduce a series of injury caused by cold ischemia of donor liver and improve the preservation quality of grafts are the hot and challenging spots in this field. Currently, preservation fluid in clinical practice has not achieved ideal preservation effect, especially for the protection of marginal donor organs. In the context of severe donor shortage, the key solution is still to explore the optimal preservation protocol for donor liver to prevent grafts from cold ischemia injury. In this article, the mechanism of donor liver injury during cold ischemia, the classification and evolution of donor liver preservation fluid were summarized, the development direction and challenges of donor liver preservation fluid were discussed, aiming to provide novel ideas and references for the research and development of donor liver preservation fluid.
3.Delphi method investigation and research on the revision of Diagnosis of Occupational Arsenic Poisoning (GBZ 83-2013)
Juan YUAN ; Yan LAI ; Lei HUANG ; Chuntao HUANG ; Fangkun TANG ; Yimei YANG
Chinese Journal of Industrial Hygiene and Occupational Diseases 2024;42(9):679-683
Objective:To explore the expert opinions on the revision of the Diagnosis of Occupational Arsenic Poisoning (GBZ 83-2013) .Methods:In March 2023, the improved Delphi expert consultation method was adopted, in the first round of consultation, a pre-survey was conducted on 20 experts, and the questionnaire was improved according to the experts' opinions. Then, a second round of expert consultation questionnaire was formed to conduct a questionnaire survey and consultation of 50 experts engaged in occupational disease diagnosis and related work. The feedback of experts was collected and analyzed.Results:The average score for the scientificity and progressiveness of the main technical content of the original standard was 3.33, and the average score for the rationality and operability of the main technical content of the original standard was 3.25. The importance of individual indicators with specific connotations were ranged from 4.20 to 4.45, with coefficients of variation <0.25, and the experts' opinions were relatively concentrated. The experts have provided feedback indicating that the original standard had issues such as lack of continuity in diagnostic gradation, the need to integrate biomarkers with urinary and hair arsenic levels, a scarcity of objective diagnostic indicators, the removal of exposure response from the main text, and a low level of consistency in standard usage. These issues need to be revised urgently.Conclusion:The Diagnosis of Occupational Arsenic Poisoning (GBZ 83-2013) should be revised based on experts' feedback and suggestions to meet the current real demand for occupational arsenic poisoning diagnosis.
4.Delphi method investigation and research on the revision of Diagnosis of Occupational Arsenic Poisoning (GBZ 83-2013)
Juan YUAN ; Yan LAI ; Lei HUANG ; Chuntao HUANG ; Fangkun TANG ; Yimei YANG
Chinese Journal of Industrial Hygiene and Occupational Diseases 2024;42(9):679-683
Objective:To explore the expert opinions on the revision of the Diagnosis of Occupational Arsenic Poisoning (GBZ 83-2013) .Methods:In March 2023, the improved Delphi expert consultation method was adopted, in the first round of consultation, a pre-survey was conducted on 20 experts, and the questionnaire was improved according to the experts' opinions. Then, a second round of expert consultation questionnaire was formed to conduct a questionnaire survey and consultation of 50 experts engaged in occupational disease diagnosis and related work. The feedback of experts was collected and analyzed.Results:The average score for the scientificity and progressiveness of the main technical content of the original standard was 3.33, and the average score for the rationality and operability of the main technical content of the original standard was 3.25. The importance of individual indicators with specific connotations were ranged from 4.20 to 4.45, with coefficients of variation <0.25, and the experts' opinions were relatively concentrated. The experts have provided feedback indicating that the original standard had issues such as lack of continuity in diagnostic gradation, the need to integrate biomarkers with urinary and hair arsenic levels, a scarcity of objective diagnostic indicators, the removal of exposure response from the main text, and a low level of consistency in standard usage. These issues need to be revised urgently.Conclusion:The Diagnosis of Occupational Arsenic Poisoning (GBZ 83-2013) should be revised based on experts' feedback and suggestions to meet the current real demand for occupational arsenic poisoning diagnosis.
5.Effects of NLRP3 inflammasome activation on the proliferation, migration and extracellular matrix deposition of pancreatic stellate cells
Haitao GU ; Hanguang DONG ; Jiuliang YAN ; Zihao QI ; Beiyuan HU ; Chuntao WU ; Jiang LONG
Chinese Journal of Pancreatology 2023;23(2):108-113
Objective:To investigate the effects of NOD-like receptor protein 3(NLRP3) inflammasome activation on the proliferation, migration and extracellular matrix desposition of activated pancreatic stellate cells(PSCs).Methods:The rat PSCs were isolated, cultured and identified, and were divided into control group or LPS group based on the pretreatment with LPS (10 μg/ml for 24 hours) or without. The expression of NLRP3 inflammasome associated molecules in PSCs culture medium was detected by ELISA. The PSCs with NLRP3 inhibition were constructed by shRNA carrying lentivirus infection and were divided into LPS+ negative control group and LPS+ lentivirus group based on whether the cells were treated with LPS and infected by lentivirus or not. The alteration in cell proliferation and migration were detected by CCK-8 kit and transwell chamber method. The expression of extracellular matrix α-SMA and collagen in PSCs was detected by immunofluorescence staining and the expression of TGF-β mRNA was analyzed by RT-qPCR.Results:The cytoplasm of PSCs which were cultured for 24 hours was rich in bright annular lipid droplets, and the cells expressed desmin. After 7 days of culture, the cell became larger in size, the lipid droplets basically disappeared, and the cells were activated and expressed α-SMA. The expression of caspase-1, IL-1β and IL-18 in the supernatant of PSCs culture medium in LPS group were significantly higher than those in control group (1.55±0.04 vs 0.65±0.03), (2.02±0.04 vs 1.05±0.05) and (1.70±0.05 vs 0.97±0.03), respectively. After inhibiting by lentivirus infection, the expression of NLRP3 in the lentivirus group (0.25±0.04) was significantly lower than that in negative control group (0.68±0.05). In control group, LPS group, LPS+ negative control group and LPS+ lentivirus group, the A490 values was 0.61±0.02, 1.15±0.06, 0.96±0.05, and 0.56±0.01, respectively; the migrating PSCs number was (64.12±4.58), (121.67±8.02), (111.67±4.67) and (69.67±8.08)/HF, respectively; the relative expression of α-SMA was 0.78±0.05, 4.12±0.04, 3.81±0.06 and 0.88±0.05, respectively; the relative expression of collagen was 0.65±0.03, 3.43±0.02, 2.67±0.02 and 0.48±0.03, respectively; and the expression of TGF-β mRNA was 0.22±0.03, 0.89±0.01, 0.86±0.03 and 0.43±0.02, respectively. The A490 value, the migrating cells number, the expression of α-SMA, collagen and the expression of TGF-β mRNA in LPS group and LPS+ negative control group was significantly higher than those in control group and LPS+ lentivirus group, and all the differences were statistically significant (all P value <0.05). Conclusions:NLRP3 inflammasome activation may accelerate the extracellular matrix deposition and pancreatic fibrogenesis by promoting PSCs proliferation and migration ability via regulating the biological functions.
6.Application value of CT and MRI radiomics based on machine-learning method in diagnosing pancreatic cancer
Qingguo WANG ; Jiang LONG ; Wei TANG ; Tao CHEN ; Chuntao WU ; Haitao GU ; Zihao QI ; Jiuliang YAN ; Beiyuan HU ; Yan ZHENG ; Hanguang DONG
Chinese Journal of Pancreatology 2023;23(2):128-133
Objective:To investigate the application value of CT and MRI imageomics based on machine learning method in the diagnosis of pancreatic cancer.Methods:The clinical data of 62 patients with surgically resected and pathologically confirmed pancreatic cancer, who underwent enhanced CT scan, MRI plain or enhanced scan in Shanghai General Hospital between January 2014 and December 2021 were collected. According to the chronological order of surgery, 49 patients from January 2014 to December 2020 were enrolled in the training set and 13 patients from January 2021 to December 2021 were enrolled in the validation set. 3D-slicer 4.8.1 software was used to draw the region of interest in each layer of CT and MRI images for cancerous and paracancerous tissue segment. Image features were extracted by Python and the optimal feature set from the training set data was obtained by using Lasso regression model. The machine learning decision tree model was constructed. The receiver operating characteristic curve(ROC) curve was drawn, and the area under the curve (AUC) was calculated to evaluate the value of these three kinds of imageomics models in the diagnosis of pancreatic cancer.Results:The 1 767 CT features and 1 674 MRI features were obtained from enhanced CT scan, MRI plain scan and enhanced MRI scan, respectively. For the differential diagnosis model of cancerous tissue and paracancerous tissue, the enhanced CT scan data model obtained the optimal feature set involving 6 features, the MRI plain scan model obtained the optimal feature set involving 16 features, and the enhanced MRI scan model obtained the optimal feature set involving 15 features. The diagnostic model based on enhanced CT scan had an AUC of 0.98 in the training set and 1 in the verification group. The AUC of the MRI plain scan and enhanced MRI scan models in both the training set and the validation set was 1. The specificity and sensitivity of machine learning decision tree model based on the three kinds of imageomics models in the diagnosis of cancerous tissue and paracancerous tissue were 100%. For the differential diagnosis model of splenic artery wrapping, the enhanced CT scan model didn′t obtain the optimal features and had no diagnostic efficacy. The MRI plain scan model and enhanced MRI scan model obtained the optimal feature set involving 5 and 4 features, respectively. The AUC of the MRI plain scan model in the training set and the validation set were 0.862 and 0.750, respectively, with diagnostic sensitivity of 93.8% and 50.0%, and specificity of 78.6% and 100%, respectively. The AUC of the enhanced MRI scan model in the training set and the validation set were 0.950 and 0.861, respectively, with diagnostic sensitivity of 90.0% and 93.6%, and specificity of 100% and 78.6%, respectively.Conclusions:Based on the radiomics of CT enhanced, MRI plain scan and enhanced MRI scan, the machine learning diagnostic model has an accuracy of more than 90% in differentiating pancreatic cancer from paracancerous tissue. For the differentiation of splenic artery wrapping in pancreatic cancer, the diagnostic model based on enhanced MRI scan haS the best diagnostic efficiency.
7.Interpretation of the group standard for Standard strains of pathogenic microorganism-technical specifications for establishment of HIV strains
Chuntao MA ; Qiang WEI ; Yan JIANG
Chinese Journal of Experimental and Clinical Virology 2023;37(3):265-270
Human immunodeficiency virus (HIV) strains are important national biological resources, and the establishment of HIV standard strains forms material basis for HIV diagnostic testing, epidemic surveillance, disease prevention and control, reagent evaluation, drug research and vaccine development. The National Center for AIDS/STD Control and Prevention of China took the lead in development and publication of the Chinese Preventive Medicine Association (CPMA) group standard, Standard strains of pathogenic microorganism-technical specifications for establishment of HIV strains (T/CPMA 027—2023). Published on February 2, 2023, the standard, for the first time, specifies all technical requirements for establishment of HIV standard strains, covering sampling, isolation, culture, titration, identification, evaluation, preservation, archiving, quality control and biosafety management, and it also standardizes HIV isolation and characterization procedure, and relevant information description. This paper provides in-depth views on development of HIV standard strains, formulation and application of relevant standards, in order to pave solid ground for establishment of HIV standard strain banks and improvement of repository quality control, utilization and biosafety management.
8.Primary construction and application of National bank for HIV-1 standard strains
Li REN ; Lin HE ; Meiling ZHU ; Yan WANG ; Desheng KONG ; Chen WANG ; Yi FENG ; Yiming SHAO ; Hui XING ; Chuntao MA ; Liying MA
Chinese Journal of Experimental and Clinical Virology 2023;37(3):280-285
Objective:To construct standard strains representing the main epidemic clades of HIV-1 in China, amplify the virus strains, and establish a seed lot.Methods:Six isolates of HIV-1 virus were identified and analyzed in genotype and phenotype, according to " interpretation for the social organization of the Standard strains of pathogenic microorganism- technical specifications for establishment of HIV strains". The isolates were amplified and cultivated to generate the secondary generation primary seed lot and the third generation working seed lot as frozen storage in liquid nitrogen. Results:Six HIV-1 standard strains were obtained, of which 3 strains are CRF_ 07BC (NRPC2.4.9003, NRPC2.4.9005, NRPC2.4.9006), 1 strain is CRF_ 01AE (NRPC2.4.9001), 1 strain is CRF_ 08BC (NRPC2.4.9002), and 1 strain is URF (NRPC2.4.9004). Phenotypic detection showed that all six strains are CCR5 tropics and Non syncytia inducing virus. TCID 50 were all greater than 1 × 10 5/ml, and concentrations of p24 antigen were all higher than 2 ng/ml. A primary seed lot with no less than 20 vials per strain and a working seed lot with no less than 50 vials per strain were constructed. The standard virus strains were used in evaluating antiviral drugs PEG2kC34, PEG5kC34, LP-19, and neutralizing antibody LSEVh LS-F. Conclusions:Six standard strains of HIV-1 virus covering the three main epidemic subtypes of HIV-1 in China have been obtained, and a storage of HIV-1 standard strain was constructed. It meets the need of the preservation of HIV-1 standard strains in China and provides support for drug and vaccine evaluation.
9.Clinical analysis of 11 cases of sub-acute arsenic poisoning
Juan YUAN ; Yan LAI ; Chuntao HUANG ; Lei HUANG ; Fangkun TANG ; Lun HONG ; Yimei YANG
Chinese Journal of Industrial Hygiene and Occupational Diseases 2020;38(12):921-924
Objective:To investigate the clinical manifestations, treatments and prognosis of subacute arsenic poisoning.Methods:In January 2020, a retrospective analysis was carried out on 11 patients hospitalized with subacute arsenic poisoning caused by arsenic contaminated drinking water. We observed manifestations, treatments and prognosis.Results:The main clinical presentations of subacute arsenic poisoningin were gastroenteritis in early phase, some of them had other organ damage, such as skin, blood, liver, kidney, cardiovascular and so on. The later phase was mainly peripheral nervous system damage. The treatment was mainly to chelate arsenic, protect target organs and treat toxic peripheral neuropathy. Most were significantly recoveried, but the recovery of severe toxic peripheral neuropathy was tardy.Conclusion:Acute gastroenteritis is the mainly early manifestation of subacute arsenic poisoning caused by digestive tract, and toxic peripheral neuropathy in the later phase. The prognosis is good, but the recovery of severe toxic peripheral neuropathy is tardy.
10. Implementation of centralized quarantine for close contacts in a community of Shanghai during COVID-19 epidemic
Yunfeng YANG ; Ting LI ; Wenqin GU ; Yan WANG ; Chuntao YI
Chinese Journal of General Practitioners 2020;19(0):E012-E012
Since the key point of prevention and control of new coronavirus infection (COVID-19) in the community is to block the spread, the centralized quarantine for close contacts with confirmed or suspected cases is the most effective way in breaking the chain of transmission. A community centralized quarantine site was set up by Fenglin Community Health Service Center for medical observation of close contacts. The detailed plan and management of the centralized quarantine site, and the implementation experiences are introduced in this article to provide reference for medical institutions involved in the prevention and control of COVID-19 epidemic.

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