1.Mechanism of DUX-induced differentiation of mESC into extraembryonic endoderm
Lei HONG ; Chuanliang GUO ; Qin CAI ; Wanrui LI ; Yitao ZENG ; Yan XUE ; Fanyi ZENG
Journal of Shanghai Jiaotong University(Medical Science) 2024;44(11):1359-1369
Objective·To explore the effect of double homeobox(DUX)protein on the differentiation potential of mouse embryonic stem cells(mESCs)into extraembryonic endoderm(XEN)and the possible mechanism of its action.Methods·Overexpression of DUX cell lines in mESCs was achieved by using a lentiviral system.The proportion of 2-cell-like cells(2CLCs)before and after DUX overexpression was detected by flow cytometry,and the expression of 2-cell stage-specific genes,Dux,zinc finger and SCAN domain containing 4c(Zscan4c),zinc finger protein 352(Zfp352)and murine endogenous retrovirus-L polymerase(MERVL-pol),were detected by real-time quantitative reverse transcription polymerase chain reaction(RT-qPCR).RT-qPCR assay was used to detect the expression of pluripotency factors,nanog homeobox(Nanog),kruppel-like transcription factor 4(Klf4),sex determining region Y-box 2(Sox2),and octamer-binding transcription factor 4(Oct4),in pluripotent state,as well as the expression of signature genes for different germ layers in the differentiated state[endodermal:GATA binding protein 4(Gata4),GATA binding protein 6(Gata6),and sex determining region Y-box 17(Sox17);ectodermal:Nestin and tubulin beta 3 class Ⅲ(Tubb3);mesodermal:heart and neural crest derivatives expressed 1(Hand1),myogenic differentiation 1(Myod1),and kinase insert domain protein receptor(Flk1)].Public RNA sequencing(RNA-seq)data were mined to further clarify the effect of DUX on the differentiation of mESCs into extraembryonic endoderm.Functional and pathway enrichment analyses of differentially expressed genes were performed using Gene Ontology(GO),Kyoto Encyclopedia of Genes and Genomes(KEGG),and gene set enrichment analysis(GSEA)to identify the signaling pathways regulated by DUX.Additionally,an in-depth analysis of existing chromatin immunoprecipitation sequencing(ChIP-seq)data was conducted to explore the potential target genes of DUX.Results·Molecular biology experiments showed that overexpression of DUX could effectively maintain the pluripotency of mESCs,which was consistent with the analysis of public RNA-seq data.Differential gene analysis revealed that endodermal genes were specifically upregulated.After differentiation assay of mESCs,RT-qPCR assay experiments showed that mRNA expression of the XEN marker genes(Gata4,Gata6,Sox17)was significantly upregulated(P<0.001).In contrast,there was no specific change in mesodermal and ectodermal genes.GSEA enrichment analysis indicated that DUX might activate the retinoid metabolism signaling pathway,and the analysis of the ChIP-seq data further revealed the presence of a large number of known retinoic acid receptor motif in DUX-bound peaks,which could activate downstream target genes related to the development of the XEN.Conclusion·DUX has a strong correlation with the retinoic acid signaling pathway and it is predicted to activate the retinoic acid signaling pathway,which could promote the tendency of mESCs toward XEN differentiation.
2.Factor analysis of effective platelet-producing ability of fetal liver-derived cells
Mengru YU ; Guanheng YANG ; Guanghui LIU ; Yitao ZENG ; Yan XUE ; Qingwen MA ; Fanyi ZENG
Chinese Journal of Internal Medicine 2022;61(6):664-672
Objective:To study the different factors affecting platelet production post transplantation of hematopoietic stem cells (HSCs) isolated from different sources in order to explore novel options for treating platelet depletion following HSCs transplantation.Methods:HSCs and their downstream derivatives including myeloid and lymphoid cells (i.e., collective of mononuclear cells (MNCs)), were isolated from E14.5 fetal liver (FL) and bone marrow (BM) of 8-week-old mice by Ficoll separation technique. These cells were subsequently transplanted into the tibia bone marrow cavity of recipient mice post lethal myeloablative treatment in order to construct the FL-MNCs and BM-MNCs transplantation mouse model. Routine blood indices were examined in these recipient mice. The chimeric rate of donor cells in recipient peripheral blood cells were determined by flow cytometry. Different groups of cells involved in platelet reconstruction were analyzed. CD41 +megakaryocytes were sorted from fetal liver or bone marrow using magnetic beads, which were then induced to differentiate into platelets in an in vitro assay . Quantitative RT-PCR was used to detect the expression of platelet-related genes in CD41 +megakaryocytes from the two sources. Results:Both the FL-MNCs and the BM-MNCs transplantation groups resumed normal hematopoiesis at the 4th week after transplantation, and the blood cells of the recipient mice were largely replaced by the donor cells. Compared with the mice transplanted with BM-MNCs, the platelet level of mice transplanted with FL-MNCs recovered faster and were maintained at a higher level. At week 4, the PLT level of the FL-MNCs group was (1.45±0.37)×10 12/L, and of the BM-MNCs group was (1.22±0.24)×10 12/L, P<0.05. The FL-MNCs contain a higher proportion of hematopoietic stem cells (Lin -Sca-1 +c-Kit +)(7.60%±1.40%) compared to the BM-MNCs (1.10%±0.46%), P<0.01; the proportion of the megakaryocyte progenitor cells (Lin -Sca-1 -c-Kit +CD41 +CD150 +) and mature megakaryocyte cells (CD41 +CD42b +), also differ significantly between the FL-MNCs (3.05%±0.22%, 1.60%±0.06%, respectively) and the BM-MNCs (0.15%±0.02%, 0.87%±0.11%, respectively) groups, both P<0.01. In vitro functional studies showed that FL-MNCs-CD41 +megakaryocytes could produce proplatelet-like cells more quickly after induction, with proplatelet-like cells formation on day 3 and significant platelet-like particle formation on day 5, in contrast to bone marrow-derived BM-MNCs-CD41 +megakaryocytes that failed to form proplatelet-like cell on day 5. In addition, FL-MNCs-CD41 +cells expressed higher levels of platelet-related genes, Mpl (3.25-fold), Fog1 (3-fold), and Gata1 (1.5-fold) ( P<0.05). Conclusion:Compared with the BM-MNCs group, the FL-MNCs transplantation group appears to have a more efficient platelet implantation effect in the HSCs transplantation recipient in vivo , as well as a higher platelet differentiation rate in vitro. This might be related to a higher proportion of megakaryocytes and higher expression levels of genes such as Mpl, Fog1, and Gata1 that could be important for platelet formation in FL-MNCs-CD41 +cells. Further exploration of the specific functions of these genes and the characteristics of the different proportions of the donor cells will provide valuable clues for the future treatment of platelets reconstitution after HSCs transplantation clinically.
3.Health hazards and hearing loss risk assessment of workers exposed to noise in an automobile manufacturing enterprise
Tao LIU ; Jing LIU ; Cheng HAN ; Yitao LIU ; Qiang ZENG ; Qing GU
Chinese Journal of Industrial Hygiene and Occupational Diseases 2022;40(6):434-438
Objective:To investigate the current situation of occupational exposure to noise among noise workers in an automobile manufacturing enterprise in Tianjin, understand the impact of noise on workers' nervous system and hearing, and assess the risk of hearing loss among noise workers.Methods:In May 2021, 3516 workers in an automobile manufacturing enterprise were investigated by using a self-made questionnaire"Noise Workers Questionnaire" and cluster sampling method. The occupational noise hygiene survey and occupational hazards detection were carried out in their workplaces. They were divided into noise exposure group and non-noise exposure group according to whether they were exposed to noise or not. The general characteristics, hearing and nervous system symptoms of the two groups of workers were compared, and the risk of hearing loss was assessed.Results:There were 758 workers in the noise exposure group, aged (26±5) years old, with a working age of 3.0 (2.0, 6.0) years exposed to noise. 2758 workers in the non-noise exposure group, aged (25±6) years old, with a working age of 2.0 (1.0, 4.0) years. There were statistically significant differences in the distribution of workers'education level, working age and memory loss between the two groups (χ 2=37.98, 38.70, 5.20, P<0.05). The workers in the noise exposure group showed a decreasing trend of insomnia, dreaminess, sweating and fatigue with the increase of working age (χ 2trend=6.16, 7.99, P<0.05). The risk classification of binaural high-frequency hearing loss for workers in all noise positions until the age of 50 and 60 was negligible, the risk of occupational noise deafness was low for workers in stamping and welding noise positions until the age of 60. Conclusion:The occupational noise exposed to automobile manufacturing workers may cause certain harm to their nervous and auditory systems. Noise protection measures should be taken to reduce the risk of hearing loss and occupational noise deafness.
4.Health hazards and hearing loss risk assessment of workers exposed to noise in an automobile manufacturing enterprise
Tao LIU ; Jing LIU ; Cheng HAN ; Yitao LIU ; Qiang ZENG ; Qing GU
Chinese Journal of Industrial Hygiene and Occupational Diseases 2022;40(6):434-438
Objective:To investigate the current situation of occupational exposure to noise among noise workers in an automobile manufacturing enterprise in Tianjin, understand the impact of noise on workers' nervous system and hearing, and assess the risk of hearing loss among noise workers.Methods:In May 2021, 3516 workers in an automobile manufacturing enterprise were investigated by using a self-made questionnaire"Noise Workers Questionnaire" and cluster sampling method. The occupational noise hygiene survey and occupational hazards detection were carried out in their workplaces. They were divided into noise exposure group and non-noise exposure group according to whether they were exposed to noise or not. The general characteristics, hearing and nervous system symptoms of the two groups of workers were compared, and the risk of hearing loss was assessed.Results:There were 758 workers in the noise exposure group, aged (26±5) years old, with a working age of 3.0 (2.0, 6.0) years exposed to noise. 2758 workers in the non-noise exposure group, aged (25±6) years old, with a working age of 2.0 (1.0, 4.0) years. There were statistically significant differences in the distribution of workers'education level, working age and memory loss between the two groups (χ 2=37.98, 38.70, 5.20, P<0.05). The workers in the noise exposure group showed a decreasing trend of insomnia, dreaminess, sweating and fatigue with the increase of working age (χ 2trend=6.16, 7.99, P<0.05). The risk classification of binaural high-frequency hearing loss for workers in all noise positions until the age of 50 and 60 was negligible, the risk of occupational noise deafness was low for workers in stamping and welding noise positions until the age of 60. Conclusion:The occupational noise exposed to automobile manufacturing workers may cause certain harm to their nervous and auditory systems. Noise protection measures should be taken to reduce the risk of hearing loss and occupational noise deafness.
5. Analysis of the related factors of nosocomial infection in patients with non-surgical basal ganglia intracranial hemorrhage in acute phase
Hui ZHANG ; Nianlong SUN ; Siyan CHEN ; Silin ZENG ; Yitao HE
Chinese Journal of Postgraduates of Medicine 2020;43(2):112-117
Objective:
To assess the risk factors associated with nosocomial infection in patients with non-surgical basal ganglia intracranial hemorrhage (ICH) in the acute phase to provide evidence for prevention and intervention of nosocomial infections.
Methods:
Clinical data of 224 patients with non-surgical basal ganglia ICH from January 2014 to December 2018 in the Shenzhen People′s Hospital were analyzed. Patients were divided into 2 groups based on the presence or absence of infection. Clinical data between the two groups were compared including gender, age, past medical history, bleeding volume, hematoma growth rate, systolic blood pressure, diastolic blood pressure, GCS, NIHSS, WBC, RBC, FBI, PLT, CR, BUN, GLU, CRP, UA, CHOL, TG, LDL, HCY. Multivariate Logistic regression analysis and the area under the ROC curve were performed on meaningful variables (
6.Epidemic features of coronavirus disease 2019 in Hunan Province.
Yitao MAO ; Huihui ZENG ; Ying WANG ; Juxiong XIAO ; Wei YANG ; Gaofeng ZHOU ; Weihua LIAO
Journal of Central South University(Medical Sciences) 2020;45(5):576-581
OBJECTIVES:
To explore and analyze the epidemic features of coronavirus disease 2019 (COVID-19) in Hunan Province from January 21, 2020 to March 14, 2020, as well as to investigate the COVID-19 epidemics in each city of Hunan Province.
METHODS:
The epidemic data was obtained from the official website of Hunan Province's Health Commission. The data of each city of Hunan Province was analyzed separately. Spatial distribution of cumulative confirmed COVID-19 patients and the cumulative occurrence rate was drawn by ArcGIS software for each city in Hunan Province. Some regional indexes were also compared with that in the whole country.
RESULTS:
The first patient was diagnosed in January 21, sustained patient growth reached its plateau in around February 17. Up to March 14, the cumulative confirmed COVID-19 patients stopped at 1 018. The cumulative occurrence rate of COVID-19 patients was 0.48 per 0.1 million person. The number of cumulative severe patients was 150 and the number of cumulative dead patients was 4. The mortality rate (0.39%) and the cure rate (99.6%) in Hunan Province was significantly lower and higher respectively than the corresponding average rate in the whole country (0.90% and 96.2%, Hubei excluded). The first 3 cities in numbers of the confirmed patients were Changsha, Yueyang, and Shaoyang. While sorted by the cumulative occurrence rate, the first 3 cities in incidence were Changsha, Yueyang, and Zhuzhou.
CONCLUSIONS
The epidemic of COVID-19 spread out smoothly in Hunan Province. The cities in Hunan Province implement anti-disease strategies based on specific situations on their own and keep the epidemic in the range of controllable.
Betacoronavirus
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China
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epidemiology
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Cities
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epidemiology
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Coronavirus Infections
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epidemiology
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mortality
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Humans
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Pandemics
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Pneumonia, Viral
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epidemiology
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mortality
7. The preliminary study on the combined application model of serum miRNAs in the diagnosis of pneumoconiosis
Yitao LIU ; Qiang ZENG ; Jing LIU ; Ming ZHANG ; Rongming MIAO
Chinese Journal of Industrial Hygiene and Occupational Diseases 2019;37(1):26-29
Objective:
To explore the role of serum microRNA (miRNAs) levels in the detection of pneumoconiosis, and to establish a combined application model of multiple serums miRNAs for pneumoconiosis diagnosis.
Methods:
152 cases were selected in the case group and the control group respectively. The TaqMan Low Density Array method was used to screen out the candidate miRNAs for early screening of pneumoconiosis, and RT-qPCR was used to verify. According to the area under the curve (AUC) , the sensitivity and specificity of the candidate indicators were investigated. The logistic regression model was established by the two-class logistic regression model.
Results:
The expression of 7 candidate miRNAs in the serum of pneumoconiosis patients was significantly different (
8. Influencing factors on two types of occupational stress in workers of electronics manufacturing industry
Xueying YANG ; Jing LIU ; Pei LI ; Yitao LIU ; Meili LI ; Shuang LI ; Chao WANG ; Qiang ZENG
China Occupational Medicine 2018;45(01):60-65
OBJECTIVE: To investigate occupational stress and its influencing factors in workers of electronic manufacturing.METHODS: The judgment sampling method was used to select 2 251 workers as study subjects. The Simple Occupational Stress Questionnaire and the Effort-Reward Imbalance( ERI) Questionnaire were used to investigate job demand-control(JDC) and ERI occupational stress respectively. RESULTS: Among the 2 251 workers,the prevalence rates of high occupational stress in JDC and ERI models were 56. 4% and 10. 1%,respectively. The positive rate of JDC occupational stress was higher than that of ERI( P < 0. 01). The multivariate logistic analysis indicated that workers with assembly line had higher risk of JDC and ERI occupational stress than workers without assembly line( P < 0. 05). Workers with labor dispatching had higher risk of JDC and ERI occupational stress than workers with long-term contract( P < 0. 05). Workers exposed to occupational hazards had higher risk of JDC and ERI occupational stress than workers not exposed to occupational hazards( P < 0. 05). Workers with disease had higher risk of JDC and ERI occupational stress than healthy workers( P < 0. 05). CONCLUSION: JDC is the main occupational stress model in workers of electronics manufacturing factory. The main influencing factors are assembly line,labor dispatching,weekly work time,exposure to occupational hazards and illness.
9. Experimental study of miRNA200a regulating Wnt/β-catenin signaling pathway in silica-induced mouse lung epithelial cells
Xin WANG ; Qiang ZENG ; Xueying YANG ; Jing LIU ; Yitao LIU
Chinese Journal of Industrial Hygiene and Occupational Diseases 2018;36(1):47-50
Objective:
To observe the effect of overexpression of miRNA200a (miR-200a) recombinant lentivirus on the expression of Wnt/β-catenin signaling pathway in mouse lung epithelial cell line MLE-12 induced by silica (SiO2) .
Methods:
The mice were divided into SiO2 control group (SiO2) , virus control group (SiO2+Lv-NC) group and overexpressing miR-200a virus group (SiO2+Lv-miR-200a). The expression of β-catenin, MMP2, MMP9, TCF-4 and Cyclin D1 mRNA and protein were detected by realtime-PCR and western blot after incubating cells for 18 h stimulating at the final concentration of 200 μg/ml of SiO2.
Results:
The expression of miR-200a in MLE-12 cells of SiO2+Lv-miR-200 a group was significantly higher than that in SiO2 group and SiO2+Lv-NC group. The mRNA and protein expression of β-catenin, MMP2, MMP9, TCF-4 and Cyclin D1 in MLE-12 cells of SiO2+Lv-miR-200a group were significantly lower than those in SiO2 group and SiO2+Lv-NC group (
10.The expression of serum miRNAs in pneumoconiosis complicated with pulmonary tuberculosis patients
Yitao LIU ; Qiang ZENG ; Shuxin LI ; Deyi YANG ; Weiyi XING ; Linmin FENG ; Ming ZHANG
Chinese Journal of Industrial Hygiene and Occupational Diseases 2016;34(7):525-527
Objective To inquiry the changes of serum miRNAs expression in pneumoconiosis complicated with pulmonary tuberculosis patients.Methods The study subjects (all subjects are male) were divided into three groups:I stage pneumoconiosis (38 cases)、Pneumoconiosis complicated with pulmonary tuberculosis (12 cases) and healthy control (40cases).The first two groups come from Tianjin worker's sanatorium and healthy control group came from a Hospital Health Screening Center.RT-qPCR was used to detect the expression levels of miR-21,miR-200c,miR-16,miR-204,miR-206,miR-155,let-7g,miR-30b,miR-192,miR-29a in serum.Results Compared with the healthy control group,the expression levels of miR-204,miR-206,miR-21,miR-16,miR-29a,miR-155,miR-200c in other two groups are different (P<0.05);Compared with the I stage pneumoconiosis,the expression levels of miR-155 is significantly higher (P<0.05) and miR-29a is significantly lower (P<0.05).Conclusion Compared with the I stage pneumoconiosis,the expression levels of miR-155 and miR-29a are significantly different,suggesting that these two miRNAs may play a important role in the process of pneumoconiosis complicated with pulmonary tuberculosis

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