Study on CYP2E1 gene polymorphism and the relationship between methylation level of its promoter region and liver injury induced by anti-tuberculosis drugs in Mongolian tuberculosis patients in Inner Mongolia / 医学研究生学报

ObjectiveThere are few studies on whether the occurrence of anti-tuberculosis drug-induced liver injury (ADIH) is associated with the polymorphism of CYP2E gene and methylation level. This study aims to CYP2E1 gene polymorphism and the relationship between the methylation level of the promoter region and ADIH in Mongolian tuberculosis (TB) patients.Methods A total of 135 Mongolian TB patients who received standardized treatment at the Tuberculosis Research Institute of Tongliao City, Inner Mongolia from November 2015 to June 2018 were selected. According to the ADIH criteria, TB patients with liver injury were selected as the ADIH group (n=45), and TB patients without liver injury were matched as the control group based on a ratio of 1∶2 (n=90). DNA extraction and polymerase chain reaction (PCR) were performed to amplify the CYP2E1 gene to determine the CYP2E1 rs2031920 genotype, and to analyze the CYP2E1 gene polymorphism and relationship between ADIH and promoter methylation level.Results There were no significant differences in the distribution of CYP2E1 rs2031920 genotype, C1 and C2 gene frequencies between the ADIH group and the control group (P>0.05). The overall methylation level in the promoter region of CYP2E1 gene in ADIH group (0.711±0.085) was significantly lower than that of the control group (0.759±0.062). Results of Logistic regression showed that the overall methylation level in the promoter region of CYP2E1 gene was the influencing factor for the occurrence of ADIH (P<0.005). For each 0.1 unit increase of methylation level, the risk of ADIH occurrence reduced by 0.388 times, and the OR (95% CI) value was 0.388 (between 0.204 and 0.739).Conclusion The overall methylation level in the promoter region of CYP2E1 gene was reduced in Mongolian ADIH patients, but the polymorphism of CYP2E1 gene was not related to the occurrence of ADIH. These results suggested that CYP2E1 methylation could be applied to the prevention and treatment of ADIH in patients with tuberculosis.

Study on CYP2E1 gene polymorphism and the relationship between methylation level of its promoter region and liver injury induced by anti-tuberculosis drugs in Mongolian tuberculosis patients in Inner Mongolia / 医学研究生学报

ObjectiveThere are few studies on whether the occurrence of anti-tuberculosis drug-induced liver injury (ADIH) is associated with the polymorphism of CYP2E gene and methylation level. This study aims to CYP2E1 gene polymorphism and the relationship between the methylation level of the promoter region and ADIH in Mongolian tuberculosis (TB) patients.Methods A total of 135 Mongolian TB patients who received standardized treatment at the Tuberculosis Research Institute of Tongliao City, Inner Mongolia from November 2015 to June 2018 were selected. According to the ADIH criteria, TB patients with liver injury were selected as the ADIH group (n=45), and TB patients without liver injury were matched as the control group based on a ratio of 1∶2 (n=90). DNA extraction and polymerase chain reaction (PCR) were performed to amplify the CYP2E1 gene to determine the CYP2E1 rs2031920 genotype, and to analyze the CYP2E1 gene polymorphism and relationship between ADIH and promoter methylation level.Results There were no significant differences in the distribution of CYP2E1 rs2031920 genotype, C1 and C2 gene frequencies between the ADIH group and the control group (P>0.05). The overall methylation level in the promoter region of CYP2E1 gene in ADIH group (0.711±0.085) was significantly lower than that of the control group (0.759±0.062). Results of Logistic regression showed that the overall methylation level in the promoter region of CYP2E1 gene was the influencing factor for the occurrence of ADIH (P<0.005). For each 0.1 unit increase of methylation level, the risk of ADIH occurrence reduced by 0.388 times, and the OR (95% CI) value was 0.388 (between 0.204 and 0.739).Conclusion The overall methylation level in the promoter region of CYP2E1 gene was reduced in Mongolian ADIH patients, but the polymorphism of CYP2E1 gene was not related to the occurrence of ADIH. These results suggested that CYP2E1 methylation could be applied to the prevention and treatment of ADIH in patients with tuberculosis.

Etiological characteristics of an imported Chikungunya fever epidemic in Fujian province in 2018 / 中华实验和临床病毒学杂志

Objective@#To study the etiological characteristics of an imported Chikungunya fever (CHIK) epidemic in Fujian province in 2018.@*Methods@#Serum samples collected at different days after the onset of the two CHIK cases were detected by real-time RT-PCR and ELISA. Structural protein E1 gene was amplified by RT-PCR and sequenced for nucleotide characteristics analysis and phylogenetic tree analysis.@*Results@#RNA of Chikungunya virus (CHIKV) was detected in the 4 serum samples collected on the first 5 days of the disease, and the earliest IgM antibodies were detected in specimens on the 5th day of the disease, however, IgG antibodies were only detected in specimen on 10th day. Compared with the S27-African prototype strain, 12 mutant points were found in the amino acids of E1 genes in this study. The E1 genes of the two CHIK cases were exactly the same, and they were closest to the evolutionary relationship with the strain isolated in the Philippines in 2014. Their genotype was Asian genotype.@*Conclusions@#This epidemic was confirmed to have been imported from the Philippines after the infection with the Asian genotype CHIKV, which suggests that Fujian province should strengthen the monitoring of persons entering from the CHIK epidemic area, so as to prevent imported cases from causing local outbreaks.

Nr2e1 Downregulation Is Involved in Excess Retinoic Acid-induced Developmental Abnormality in the Mouse Brain / 生物医学与环境科学（英文）

<p><b>OBJECTIVE</b>This study aimed to investigate the expression pattern and function of Nuclear receptor subfamily 2 group E member 1 (Nr2e1) in retinoic acid (RA)-induced brain abnormality.</p><p><b>METHODS</b>The mouse model of brain abnormality was established by administering 28 mg/kg RA, and neural stem cells (NSCs) were isolated from the mouse embryo and cultured in vitro. Nr2e1 expression was detected by whole mount in situ hybridization, RT-PCR, and Western blotting. Nr2e1 function was determined by transducing Nr2e1 shRNA into NSCs, and the effect on the sonic hedgehog (Shh) signaling pathway was assessed in the cells. In addition, the regulation of Nr2e1 expression by RA was also determined in vitro.</p><p><b>RESULTS</b>Nr2e1 expression was significantly downregulated in the brain and NSCs of RA-treated mouse embryos, and knockdown of Nr2e1 affected the proliferation of NSCs in vitro. In addition, a similar expression pattern of Nr2e1 and RA receptor (RAR) α was observed after treatment of NSCs with different concentrations of RA.</p><p><b>CONCLUSION</b>Our study demonstrated that Nr2e1 could be regulated by RA, which would aid a better understanding of the mechanism underlying RA-induced brain abnormality.</p>

The effects of targeted forkhead box E1 gene interference on epithelial-mesenchymal transition in thyroid cancer cells / 中华内分泌外科杂志

Objective To investigate the effects of targeted forkhead box E1( FOXE1) gene on epitheli-al-mesenchymal transition(EMT) of papillary thyroid cancer (PTC) cell line TPC-1, and to study its role in in-vasion and migration of TPC-1 cells.Methods The lentiviral expression vector for RNA interference of FOXE 1 gene was constructed to silence the expression of FOXE 1.Real-time polymerase chain reaction ( RT-PCR) and Western blot were used to detect the expression of such EMT markers as E-cadherin , N-cadherin and Vimentin . Transwell assay and Scratch assay were used to analyze TPC-1 cells migration and invasion .Results After RNA interference of FOXE1, the morphology of TPC-1 cells indicated a transformation of EMT .The expression of epi-thelial phenotype marker E-cadherin decreased remarkably while mesenchymal marker Vimentin was significantly up-regulated(P<0.01).Compared with the control group , the expression of Vimentin mRNA in the experimen-tal group was 2.24 times higher .The migration and invasion ability of TPC-1 cells increased significantly , and the number of cells in transwell was 2.11 times of the control's.Conclusion The silence of FOXE1 gene probably increases the invasion potential of human PTC cells through EMT .

Transgenic plants of Petunia hybrida harboring the CYP2E1 gene efficiently remove benzene and toluene pollutants and improve resistance to formaldehyde

The CYP2E1 protein belongs to the P450 enzymes family and plays an important role in the metabolism of small molecular and organic pollutants. In this study we generated CYP2E1 transgenic plants of Petunia using Agrobacterium rhizogenes K599. PCR analysis confirmed that the regenerated plants contained the CYP2E1 transgene and the rolB gene of the Ri plasmid. Southern blotting revealed the presence of multiple copies of CYP2E1 in the genome of transgenic plants. Fluorescent quantitative PCR revealed exogenous CYP2E1 gene expression in CYP2E1 transgenic plants at various levels, whereas no like expression was detected in either GUS transgenic plants or wild-types. The absorption of benzene and toluene by transgenic plants was analyzed through quantitative gas chromatography. Transgenic plants with high CYP2E1 expression showed a significant increase in absorption capacity of environmental benzene and toluene, compared to control GUS transgenic and wild type plants. Furthermore, these plants also presented obvious improved resistance to formaldehyde. This study, besides being the first to reveal that the CYP2E1 gene enhances plant resistance to formaldehyde, also furnishes a new method for reducing pollutants, such as benzene, toluene and formaldehyde, by using transgenic flowering horticultural plants.

Genetic polymorphism of alcohol-metabolizing enzyme and alcohol dependence in Polish men

Alcohol dependence poses a serious medical and sociological problem. It is influenced by multiple environmental and genetic factors, which may determine differences in alcohol metabolism. Genetic polymorphism of the enzymes involved in alcohol metabolism is highly ethnically and race dependent. The purpose of this study was to investigate the differences, if present, in the allele and genotype frequency of alcohol dehydrogenase 1B (ADH1B), ADH1C and the microsomal ethanol-oxidizing system (MEOS/CYP2E1) between alcohol-dependent individuals and controls and also to determine if these genotypes cause a difference in the age at which the patients become alcohol dependent. The allele and genotype frequencies of ADH1B, ADH1C, and CYP2E1 were determined in 204 alcohol dependent men and 172 healthy volunteers who do not drink alcohol (control group). Genotyping was performed by PCR-RFLP methods on white cell DNA. ADH1B*1 (99.3 percent) and ADH1C*1 (62.5 percent) alleles and ADH1B*1/*1 (N = 201) and ADH1C*1/*1 (N = 85) genotypes were statistically more frequent among alcohol-dependent subjects than among controls (99.3 and 62.5 percent, N = 201 and 85 vs 94.5 and 40.7 percent, N = 153 and 32, respectively). Differences in the CYP2E1 allele and genotype distribution between groups were not significant. The persons with ADH1C*1/*1 and CYP2E1*c1/*c2 genotypes became alcohol dependent at a considerably younger age than the subjects with ADH1C*1/*2, ADH1C*2/*2 and CYP2E1*c1/*c1 genotypes (28.08, 25.67 years vs 36.0, 45.05, 34.45 years, respectively). In the Polish men examined, ADH1C*1 and ADH1B*1 alleles and ADH1C*1/*1 and ADH1B*1/*1 genotypes favor alcohol dependence. The ADH1B*2 allele may protect from alcohol dependence. However, subjects with ADH1C*1/*1 and CYP2E1*c1/*c2 genotypes become alcohol dependent at a considerably younger age than the subjects with ADH1C*1/*2, ADH1C*2/*2 and CYP2E1*c1/*c1 genotypes.

Study oil the heredity and variance of E1 gene of rubella virus Matsuba strain / 中华微生物学和免疫学杂志

Objecflve To study the heredity and variance of E1 gene of rubella villls Matsuba vac-cine strain.and to evaluate the protective efficacy of the vaccine prepared by Matsuba vaccine strain on genelevel.Methods The Matsuba strain was passaged on primary rabbit kidnev cells from 14 to 23 generations.and the E1 genes of 14,16,17,18,23 passages amplified by RT-PCR were cloned into the T-A cloning vector pGEM-T and sequenced,respectively.The sequences of E1 gene of the passaged viruses were tom-pared with rubella virus Matsuba reference strain E1 gene(Accession No.D50673)and other rubella strains in GenBank.respectively.Results The Matanba passaged shared higher homology of nucleotide and amino acid with Matsuba reference strain(D50673).,The sequences of nucleotide and amino acid of RV14,RV16.RV18 were identical to D50673.The homology of nucleotide sequences of RV17,RV23 and D50673 was 99.9%and 99.7%,respectively,andthe homology of amino acidwgg 99.8% and 99.2%,respective-ly.The amino acid related to glycosylation and epitopes of the passage viruses were highly conservative.The phylogenetic tree showed Matsuba strain belonged to la genotype.The rubella virus genotypes circulated in China recent years were 1E,lF,2A and 2B,and 1E genotype was the predominant genotype.The homology of nucleotide and amino acid of Matsuba strain and the other genotype reference strains was 92.1%-99.6% and 98.1%-99.8%.respectively.Condusion The Matsuba vaccine strain possesses highly genetic stabil-ity.which indicates that the Matsuba strain and its vaccine are stable on molecular level.And the vaccine prepared by Matsuba vaccine strain can prevent the infection of various genotype rubella viruses.

Functional polymorphism of CYP2E1 gene and alcohol use disorders in a Tibetan population / 中南大学学报(医学版)

Objective To investigate the relationship between the CYPphism and alcohol use disorders, and the potential influence of the CYP2E1 * c1/* c2 polymorphismon the severity and dimensions of alcohol use disorders in Tibetan. Methods Three hundred and forty Tibetans with Alcohol Use Disorders Identification Test (AUDIT) score I>10 and another 315 matched control subjects with AUDIT score ≤5 were enrolled. The CYP2E1 * c1/* c2 polymorphism was de-termined by the standard PCR-RFLP method. Results The frequency of the CYP2E1 * c2 allele in subjects with alcohol use disorders ( 16. 2 % ) was significantly higher than that of the controls(10.8%) , with a P value of 0. 005 and OR value of 1.60 (95% CI: 1.15～2.21). There was also a significant difference in genotype frequencies between the 2 groups ( χ2= 8.75, P = 0.01).Subjects with alcohol use disorders had higher frequencies of genotypes with at least one copy of allele c2 (28.5% vs. 18.7%; χ2=8.65, P=0.003; 0R=1.73) than the control group. The associ-ation of CYP2E1 * c2 allele with alcohol use disorders was much stronger in males than in females,with a male OR value of 2.30. CYP2E1 * c2 allele was associated with increased alcohol consumption and alcohol use disorders in males. Conclusion There is the positive association among CYP2E1 * c2 allele, alcohol use disorders, and the amount of alcohol consumption in Tibetan population.

CONSTRUCTION,IDENTIFICATION AND EXPRESSION OF FRAMEWORK PLASMID pAd.HCV-CE1 OF ADENOVIRUS EXPRESSION VECTOR OF HCV-CE1 / 解放军医学杂志

The HCV core gene and E1 gene derived from the plasmid pBRTM/HCV1 3011 by using polymerase chain reaction(PCR) was inserted into the backward position of cytomegalovirus(CMV) immediate early promotor element of framework plasmid (pAd.CMV Link 1) of adenovirus expression vector, then the recombinant plasmid pAd.HCV CE1 was obtained. The inserted DNA of pAd.HCV CE1 was confirmed to be HCV core and E1 gene by endonuclease, PCR and sequencing. HCV core gene was expressed transiently with lipofectamine 2000 coated in human hepatoblastoma 7721 cells which was confirmed by immunofluorescence. The results indicate that the recombinant framework plasmid of adenovirus expression vector pAd.HCV CE1 can express HCV core and E1 gene. This should be useful to pack adenovirus expression vector which can express HCV core and E1 gene.

Expression of cyp2e1 gene in liver tissue in patients with hepatic cirrhosis and obstuctive jaundice / 第二军医大学学报

In order to compare the difference of cyp2e1 gene expression between patients with hepatic cirrhosis and obstructive jaundice,and to investigate the pharmacologic significance behind this difference,liver samples were obtained from patients undergoing hepatic surgery with hepatic cirrhosis( n =7),obstructive jaundice( n =6) and normal tissue of hepatic angioma (controls, n =6). Total hepatic RNA were extracted using the one step method, cyp2e1 cDNA probe prepared by primer randomly marked method, and difference of cyp2e1 expression was compared among patients by Northern blotting.Compared with control group, the expressions of cyp2e1 in liver tissue among patients with obstructive jaundice were evidently reduced, while expression of this gene in cirrhosis liver was unaltered. CYP2E1 isoenzyme encoded by cyp2e1 decreased among patients with obstructive jaundice as expression of cyp2e1 gene significantly reduced, liver ability for metabolism of inhalant anesthetics declined consequently, under some special conditions, hepato nephric toxicity of anesthetics might be aggravated.