1.Prokaryotic expression of human mitochondrial transcription termination factor 3 and preparation of its polyclonal antibody
Yongqin YANG ; Chenggui ZHANG ; Meitao SUN ; Yun WANG ; Zefang YANG ; Xiaojuan ZHANG ; Wei XIONG
Journal of Medical Postgraduates 2016;29(10):1020-1025
Objective Human mitochondrial transcription termination factor 3 ( MTERF3 ) is a negative regulator of mito?chondrial gene expression and energy metabolism. This study was to construct a prokaryotic expression system for MTERF3 in Esche?richia coli ( E. coli ) and prepare its mouse?anti?human polyclonal antibody. Methods The complete open reading frame ( ORF) of human MTERF3 cDNA was amplified by RT?PCR and subcloned into prokaryotic expression vector pET28b. Then the recombinant plasmid pET28b?MTERF3 was transformed into competent E.coli BL21(DE3) and IPTG induced the expression of 6×his fusion protein. The recom?binant human MTERF3 protein was purified through Ni2+?NTA agar?ose gel column affinity chromatography and the purified recombinant protein was used as immunogen to immunize the BALB/c mice to pre?pare its specific polyclonal antibody. The titer and specificity of the antibody were analyzed by ELISA, Western blot and cellular immuno?fluorescence, respectively. Results The recombinant human MTERF3 protein was successfully expressed in E. coil and the mouse?anti?human MTERF3 polyclonal antibody with high quality was successfully prepared. ELISA showed that the titer of the antibody was 1:105 . Western blot and immunofluorescence detection revealed that the mouse?anti?human MTERF3 antibody could recognize the native MTERF3 antigen specifically. Conclusion Human MTERF3 expressed in the prokaryotic system has strong immunogenicity and the polyclonal antibody obtained from immunizing mice has high titer and specificity. The prokaryotic expression of human MTERF3 and the preparation of its antibody lay the foundation for further function research of human MTERF3.
2.Correlation between 24 h urinary protein quantitation and pregnancy outcome in patients with pre-eclampsia
Xun GONG ; Meitao YANG ; Haiyi LIU ; Fuyuan QIAO ; Ling FENG ; Dongrui DENG
Journal of Xinxiang Medical College 2017;34(9):808-812
Objective To investigate the correlation between 24 h urinary protein quantitation and pregnancy outcome in patients with pre-eclampsia.Methods A total of 332 pre-eclampsia patients were selected in Tongji Hospital from January 2014 to December 2016.The patients were divided into microalbuminuria group(24 h urinary protein quantification < 0.3 g,n =46),mild proteinuria group (0.3 g ≤ 24 h urinary protein quantification < 2.0 g,n =98),moderate proteinuria group (2.0 g ≤ 24 h urinary protein quantification < 5.0 g,n =71) and severe proteinuria group(24 h urinary protein quantification ≥ 5.0 g,n =117) according to the results of 24 h urinary protein quantification.The pregnancy outcomes were compared between the four groups.Results The 24 h urinary protein quantification and the serum creatinine,urea nitrogen,uric acid levels in the mild proteinuria group,moderate proteinuria group and severe proteinuria group were significantly higher than those in the microalbuminuria group (P < 0.05);and gestational week was significantly shorter than that in the microalbuminuria group (P < 0.05).The 24 h urinary protein quantification and serum urea nitrogen,uric acid levels in the moderate proteinuria group were significantly higher than those in the mild proteinuria group (P < 0.05);and gestational week was significantly shorter than that in the mild proteinuria group (P < 0.05);but there was no significant difference in serum creatinine level between the two groups (P > 0.05).The 24 h urinary protein quantification,serum creatinine,urea nitrogen and uric acid levels in the severe proteinuria group were significantly higher than those in the mild proteinuria group (P < 0.05);and the gestational week was significantly lower than that in the mild albuminuria group (P < 0.05).The 24 h urinary protein quantification in the severe proteinuria group was significantly higher than that in the moderate proteinuria group (P < 0.05),but there was no significant difference in the gestational week and serum creatinine,urea nitrogen,uric acid levels between the two groups (P >0.05).There was no significant difference in the rates of cesarean section and spontaneous labor between the four groups (P >0.05).The rate of induced labor in the moderate proteinuria group and the severe proteinuria group was significantly higher than that in the mild albuminuria group and the microalbuminuria group (P < 0.05).There was no significant difference in the rate of induced labor between the mild proteinuria group and the microalbuminuria group (P > 0.05).There was no significant difference in the rate of induced labor between the severe proteinuria group and the moderate proteinuria group (P > 0.05).The incidence of complications in microalbuminuria group,mild proteinuria group,moderate proteinuria group and severe proteinuria group was 30.43% (14/46),47.96% (47/98),74.65% (53/71) and 74.36% (87/117) respectively;the incidence of complications in the moderate proteinuria group and the severe proteinuria group was significantly higher than that in the microalbuminuria group and the mild albuminuria group (P < 0.05),but there was no significant difference in the incidence of complications between microalbuminuria group and mild albuminuria group (P > 0.05),there was no significant difference in the incidence of complications between the moderate proteinuria group and the severe proteinuria group (P >0.05).The incidences of premature birth and neonatal asphyxia in the mild proteinuria group were significantly higher than that in the microalbuminuria group (P < 0.05),and the body mass of the neonates was significantly lower than that in the microalbuminuria group (P <0.05),but there was no significant difference in the perinatal mortality rate and the incidences of fetal growth restriction(FGR) and poor neonatal resuscitation between the two groups (P > 0.05).The incidences of FGR,premature birth,neonatal asphyxia,poor neonatal resuscitation and the perinatal mortality in the moderate proteinuria group and severe proteinuria group were significantly higher than those in the microalbuminuria group (P < 0.05);and neonatal body mass was significantly lower than that in the mieroalbuminuria group (P < 0.05).The incidences of FGR,premature birth and poor neonatal resuscitation and perinatal mortality in the moderate proteinuria group were significantly higher than those in the mild proteinuria group (P < 0.05);and the neonatal body mass was significantly lower than that in the mild proteinuria group (P < 0.05);but there was no significant difference in the neonatal asphyxia incidence between the two groups (P > 0.05).The incidences of FGR,premature birth,neonatal asphyxia,poor neonatal resuscitation and perinatal mortality in the severe proteinuria group were significantly higher than those in the mild proteinuria group (P < 0.05);and the body mass of the newborns was significantly lower than that in the mild albuminuria group (P < 0.05).The incidence of neonatal asphyxia in the severe proteinuria group was significantly higher than that in the moderate proteinuria group (P < 0.05),but there was no significant difference in the incidences of FGR,premature birth,poor neonatal resuscitation,perinatal mortality and neonatal body mass between the two groups (P > 0.05).Conclusions The of 24 h urinary protein quantitation is closely related to the pregnancy outcome in patients with pre-eclampsia,the 24 h urinary protein quantification should be regularly detected in the patients with pre-eclampsia.When the urinary protein quantitation is more than 2.0 g,the incidences of maternal complications and poor prognosis of the perinatal infants is significantly higher,but the boundary value of the 24 h urinary protein quantitation for the diagnosis of severe pre-eclampsia still needs further large sample study.
3.Expression of human mitochondrial transcription termination factor-3 in non-small-cell lung cancer and its clinicopathological significance
Jiaji ZI ; Yongqin YANG ; Meitao SUN ; Wen MEI ; Xiaojuan ZHANG ; Wei XIONG
Journal of Medical Postgraduates 2017;30(2):160-164
Objective The purpose of this study was to investigate the expression of human mitochondrial transcription termi-nation factor-3 ( hMTERF3) in non-small cell lung cancer ( NSCLS) and to analyze its clinicopathological significance. Methods The paraffin block samples used in this study included 65 cases of NSCLC and 32 cases of normal alveolar epithelial tissues. We determined the expressions of hMTERF3 in NSCLC and normal alveolar epithelial tis-sues by immunohistochemistry, calculate the survival rate using the Kaplan-Meier method, and analyzed the risk factors affecting the prognosis of NSCLC using the Cox Proportional Hazard Model. Results In the 65 cases of NSCLC, 31 ( 47. 69%) showed positive expression of hMTERF3. The total survival time was significantly shor-ter in the patients with a high than in those with a low hMTERF3 ex-pression ([30.39±3.35] vs [57.61±7.12] mo, P<0.05). The riskfactors affecting the prognosis of NSCLC included positive expression of hMTERF3 (HR=3.302, 95% CI:1.598-6.905) and lymph node metastasis (HR=4.052, 95% CI: 1.212-12.398). Conclusion hMTERF3 is overexpressed in NSCLC. Highly expressed hMTERF3 and lymph node metastasis reduce the survival time of NSCLC patients, suggesting that hMTERF3 may be a potential bio-marker for the prognosis of NSCLC.
4.Bioinformatics analysis of the human MTERF1 gene promoter
Yongqin YANG ; Meitao SUN ; Yue LI ; Yun WANG ; Xiaojuan ZHANG ; Wei XIONG
Journal of Medical Postgraduates 2016;29(4):348-353
Objective The proteins encoded by mitochondrial transcription termination factor 1 ( MTERF1) plays important roles in regulating the mitochondrial gene expression and oxidative phosphorylation .This study was to investigate the characteristics and regulation mechanisms of the human MTERF1 gene with bioinformatics tools . Methods Using online bioinformatics software and phylogenetic foot-printing, we analyzed the promoter distribution , GpG island, transcription factors , and binding sites of the human MTERF1 gene. Results The human MTERF1 gene was located on 7q21.2, with a full length of 7845 bp, consisting of 4 exons and 3 introns.There were at least 2 promoters in the 5′region of the gene.The core promoter of the MTERF1 gene was located between 1878 and 2447 bp, which played a key role in its transcription .An 812 bp CpG island was observed in the gene promoter region .In addi-tion, 26 transcription factor binding sites were found in the conserved promoter region of human and mouse homologous MTERF1 genes. Conclusion Gene promoter-related online bioinformatics software can improve the efficiency of human MTERF1 gene promoter resear-ches and provide significant information for the prediction of gene pro-moter function.
5.Alteration of heat shock protein 20 expression in preeclamptic patients and its effect in vascular and coagulation function.
Fanfan LI ; Mengzhou HE ; Meitao YANG ; Yao FAN ; Yun CHEN ; Xi XIA ; Yin XIE ; Dongrui DENG
Frontiers of Medicine 2018;12(5):542-549
Preeclampsia (PE) is a pregnancy-specific, multi-system disorder and the leading cause of maternal and perinatal morbidity and mortality in obstetrics worldwide. Excessive vasoconstriction and dysregulated coagulation function are closely associated with PE. Heat shock protein 20 (HSP20) is ubiquitously expressed under normal physiological conditions and has important roles in vascular dilatation and suppression of platelet aggregation. However, the role of HSP20 in the pathogenesis of PE remains unclear. In this study, we collected chorionic plate resistance arteries (CPAs) and serum from 118 healthy pregnant women and 80 women with PE and detected the levels of HSP20 and its phosphorylated form. Both HSP20 and phosphorylated HSP20 were downregulated in CPAs from women with PE. Comparison of the vasodilative ability of CPAs from the two groups showed impaired relaxation responses to acetyl choline in preeclamptic vessels. In addition to the reduced HSP20 in serum from women with PE, the platelet distribution width and mean platelet volume were also decreased, and the activated partial thromboplastin time and thromboplastin time were elevated.With regard to the vital roles of HSP20 in mediating vasorelaxation and coagulation function, the decreased HSP20 might contribute to the pathogenesis of PE.
Adult
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Case-Control Studies
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Chorion
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blood supply
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Female
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HSP20 Heat-Shock Proteins
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metabolism
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Humans
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Phosphorylation
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Placenta
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blood supply
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Platelet Function Tests
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Pre-Eclampsia
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metabolism
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Pregnancy
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Vasoconstriction
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Vasodilation
6.Research hotspots and trends of tigecycline drug resistance: A study based on CiteSpace
Xinjing JIA ; Yanding WANG ; Chunyuan DUAN ; Lisha LIU ; Di WU ; Xinran GONG ; Zhiqiang LI ; Meitao YANG ; Dayang ZOU ; Yong WANG
Journal of Public Health and Preventive Medicine 2024;35(1):16-19
Objective To explore the research progress, research hotspot and development trend of tigecycline resistance based on the quantitative analysis and visualization function of CiteSpace. Methods The data were collected from 4,263 Chinese and English articles on tigecycline resistance in CNKI, Wanfang, VIP and Web of Science (WOS) databases from 2012 to 2022. CiteSpace 5.8.R3 software was used to analyze the cooperative network of authors, the cooperative network of countries and institutions, the total citation times of journals, and keywords included in the literature, to reveal the hotspots and trends of tigecycline resistance research. Results The number of articles published in English literature was higher than that in Chinese literature. China had the largest number of published documents, showing a significant international academic influence in this research field. Countries all over the world were concerned about the resistance of tigecycline, but Chinese literatures focused more on the clinical infection and prevention of tigecycline resistance, while English literatures placed special emphasis on the research about the drug resistance mechanism of tigecycline. Conclusion The research direction at home and abroad is basically the same, but the research focus has gradually shifted from the clinical treatment and monitoring of tigecycline to the molecular level of drug resistance mechanism.