1.A preliminary study on regulation of HMOX1, RPS27a, MT2A and UBB by HSPC238
Jiayu TAN ; Xiang HUANG ; Jinglin CHEN ; Yuheng ZHONG
Chinese Journal of Immunology 2016;32(4):509-512
Objective:To investigate the effect of HSPC 238 overexpression or low expression on the regulation of the target protein ( HMOX1, MT2A, UBB, RPS27a ) and the regulation pathways.Methods: To construct the interference vector and overexpression vector of HSPC238 respectively,transfected the HEPG2 cell lines by the liposome method,detect the expression of mRNA and protein of the HSPC 238 and the target proteins by the RT-PCR and Western blot , further to transfer the overexpression plasmids of the target proteins into the HEPG 2 cell lines which had been transfected with interference vector and overexpression vector of HSPC238,the experimental group cell lines were treated with proteasome inhibitor MG 132,to purify the target proteins with nickel column,do immunoblotting with HSPC238 to detect the accumulation situation of the target proteins.Results: The HMOX1,MT2A, RPS27a were downregulated obviously when the HSPC 238 was interfered exogenous;and the MT2A,UBB,RPS27a were up-regulated after the HSPC238 overexpressed.The overexpression plasmid of target proteins were transfected into the HEPG 2 cell lines which have been transfected with interference vector and overexpression vector of HSPC 238 ,compared with the control group ,the target protein band in the experimental group was significantly increased after treatment with the proteasome inhibitor MG 132.Conclusion:The HSPC238 overexpression can upregulate the expression of MT 2A,UBB and RPS27a,and interfering HSPC238 can downregulate the expression of HMOX1,MT2A and RPS27a;the HSPC238 target protein may play a regulatory role through the UPP pathway;the HSPC238 may play a regulatory role on the target proteins through the UPP pathway.
2.Preliminary screening of RPL5 interacting with HSPC238
Jinglin CHEN ; Xiang HUANG ; Jiayu TAN ; Yuheng ZHONG ; Zhidan WAN
Chinese Journal of Immunology 2015;(11):1494-1497
Objective:To construct a bait vector for HSPC238, and to screen the target proteins which interact with the HSPC238.Methods:Gene synthesis method was used to synthetic gene HSPC238, then connected with the pGBKT7 vector after digesting by the sfiIA and sfiIB,to obtain the bait plasmid pGBKT7-HSPC238,then transferred into the yeast strains AH109 with the empty plasmid pGBKT7after sequencing,to observe its self-activating effect in the nutrient deficiencies medium,and further to screen the target proteins which interact with HSPC238 from the human fetal liver cDNA library.Results:The bait vector pGBKT7-HSPC238 was successfully constructed,and it had no self-activating effect through the phenotypic screening,after the yeast two-hybrid technology with literature analysis,we preliminary screened and found that the ribosomal protein L5(RPL5) may be the one of the target proteins which interacted with HSPC238 from the human fetal liver cDNA library.Conclusion: We successfully constructed the bait plasmid vector PGBKT7-HSPC238,and after the yeast two-hybrid technology with literature analysis,we preliminary screened and found that the ribosomal protein L5( RPL5) may be the one of the target proteins which interacted with HSPC238.
3.The effects of HSPC238 on the expression of RB in hepatoma carcinoma cells
Chunlei YUAN ; Jiayu TAN ; Xiang HUANG ; Jinglin CHEN
International Journal of Laboratory Medicine 2016;37(18):2505-2506,2510
Objective To investigate the effect of up‐regulation and down‐regulation of HSPC238 on the RB gene mRNA and pRb protein .Methods PcDNA3 .1‐HSPC238 vector and PLL3 .7‐HSPC238 vector was transiently transfected into HepG2 cells re‐spectively .The level of RB mRNA was detected by real‐time PCR and pRb was detected by Western blotting .Results The level of RB mRNA and pRb in up‐regulation group both increased compared with control grops respectively .Conversely ,the level of RB mRNA and pRb in up‐regulation group both decreased compared with control grops respectively .Above results were all statistically significant(P<0 .05) .Conclusion HSPC238 could have a positive effect on the expression of the RB gene .
4.The progress in research for human pulse system modeling and pulse condition information analysis.
Journal of Biomedical Engineering 2002;19(2):329-333
This article introduces the significance, methods and applications in human pulse study, with an emphasis on the history, current situation and progress of pulse system modeling and pulse signal analysis. The problems in present study and the methods for the future investigations were analyzed.
Heart Rate
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Models, Cardiovascular
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Research
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trends
5.Expression and significance of HSPC238 in cervical intraepithelial neoplasia and cervical cancer
Yuheng ZHONG ; Xiang HUANG ; Jinglin CHEN ; Jiayu TAN ; Xianjun MI ; Xiangyu LIU
Chinese Journal of Immunology 2016;(2):223-225
Objective: To investigate the expression of HSPC238 in cervical intraepithelial neoplasia and cervical cancer.Methods:We collected 76 cases of cervical cancer,105 cases of CIN and 28 cases of normal cervical epithelial.Then we inves-tigated the expression of HSPC238 by using immunohistochemistry and compared the significant differences between them.Results:There was no significant difference in the expression of HSPC238 between the cervical cancer and normal cervical epithelial ( Z=-0.242,P>0.05).However,there was significant difference between the cervical intraepithelial neoplasia and normal cervical epithelial (χ2=19.159,P<0.01) and the expression of HSPC238 was correlated with the grades of CIN.The expression of HSPC238 decreased when the grade of CIN was increasing.( rs=-0.327,P<0.01 ).Conclusion:The low expression of HSPC238 might be correlated with the development of cervical neoplasia.
6.Diagnostic value of serum AFU for primary hepatic carcinoma:a meta-analysis
Dongxiu LI ; Jinglin CHEN ; Xiang HUANG ; Jiayu TAN ; Chunlei YUAN ; Zhian WAN
International Journal of Laboratory Medicine 2015;(1):1-4
Objective To evaluate the diagnostic value of serum α-L-fucosidase (AFU)in primary hepatic carcinoma(PHC)by using the meta analysis method.Methods The databases of Cochrane Library,PubMed,web of knowledge(Medline,BIOSIS Pre-views),Springer link and Science Direct were retrieved from January 1997 to December 2013.The domestic databases of CBMdisc (Chinese BioMedical Literature on disc),CNKI,Wangfang and VIP(VIP Database for Chinese Technical Periodicals)were retrieved from January 1984 to December 2013.Retrieval languages were limited to Chinese and English.The related literatures on the study of serum AFU diagnostic value for PHC were collected and the included studies meeting the inclusion criteria were performed the quality assessment.The meta-Disc 1 .4 software was adopted to conduct the analysis for comprehensively evaluating serum AFU value in the diagnosis of PHC.Results 20 articles were included (15 articles in Chinese and 5 articles in English).A total of 2 114 cases in the patients groups and 5 718 cases in the control groups were analyzed.The heterogeneity test was carried out on the in-cluded 20 articles,suggesting that the heterogeneity of the included studies was caused by the non-threshold effect,so the random effects model was selected for conducting the meta-analysis.And the pooled sensitivity was 0.77,the pooled specificity was 0.87, the pooled positive likelihood ratio was 5.37,the pooled negative likelihood ratio was 0.28,the pooled diagnostic odds ratio was 20. 47 and the SROC area under the curve(AUC)was 0.87.Conclusion Serum AFU has highly sensitivity and specificity for the diag-nosis of PHC and has reference value for its clinical diagnosis.
7.Construction of recombinant eukaryotic expression vector pCDNA3.1-MT2A and cellular localization of MT2A protein in different cell lines
Jiayu TAN ; Jinglin CHEN ; Xiang HUANG ; Dongxiu LI ; Chunlei YUAN ; Zhidan WAN
Chinese Journal of Immunology 2014;(11):1504-1507
Objective:To construct the recombinant eukaryotic expression vector pCDNA3.1-MT2A and to investigate the cellular localization of MT2A protein in 293T and SMCC7721cell lines.Methods: Gene synthesis method was used to synthetic gene MT2A,added a Kozak sequence and His tag sequence at the N-terminus,the amplified target gene was connected to the pcDNA3.1(+) vector which was double digested between the BamH Ⅰ and Not Ⅰ.After transformation to E.coli DH5α, the positive clones were picked for plasmid extraction then Electrophoretic and sequenced.The pCDNA3.1-MT2A plasmids which passed through electrophoretic and sequenced were transfected 293T and SMMC7721 cell lines by liposome method,and then observed their expression and localization in eukaryotic cells by laser confocal microscopy.Results: The recombinant plasmid pCDNA3.1-MT2A was confirmed by restriction analysis and DNA sequencing,the sequence of the target gene MT2A was entirely correct,eukaryotic expression vector was successfully constructed and cell lines which had transfected recombinants could see the expression of green fluorescent protein in the cytoplasm.Conclusion:Successfully constructed fusion gene of pCDNA3.1-MT2A and expressed in eukaryotic cells,we found that the MT2A was mainly localized in the cytoplasm of 293T and SMMC7721 cell lines.The findings can help us to lay the foundation for the functions of MT2A in hepatoma cells.
8.Association between ADRB2 rs1042713 gene polymorphism and susceptibility of asthma in Chinese Population:a meta-analysis
Jinglin CHEN ; Xiang HUANG ; Jiayu TAN ; Zhidan WAN ; Xuewei WU ; Dongxiu LI
Chinese Journal of Immunology 2015;(8):1037-1039,1048
Objective:To investigate the association between rs1042713 polymorphisms of ADRB2 gene and the susceptibility of asthma in Chinese Population by meta-analysis.Methods: The Pubmed database,Emabase database,Web of Knowledge database, CNKI database,Wanfang database and Weipu database were searched for all publications about the susceptibility of asthma and the rs1042713 polymorphisms of ADRB2 gene in Chinese Population.The article which met the inclusion criteria were assessed by the STA-TA12.0 software.Results:12 studies were included,with 2 193 asthmatic patients and 2 033 controls.All the included articles were satisfied to the Hardy-Weinberg equilibrium.The results of Meta-analysis was showed that the risk of asthma of the mutations G carriers ( GG +GA) on ADRB2 gene rs1042713 loci in Chinese people compared with the wild-type homozygotes( AA) was not significantly in-creased overall(OR=1.08,95% CI=0.82-1.44).However,subgroup analysis showed that the risk of G carriers of children had a relatively higher incidence(OR=1.69,95% CI 0.99-2.87),while the risk of adult-onset have a relatively lower incidence(OR=0.88,95%CI 0.68-1.15).Conclusion:The ADRB2 gene rs1042713 polymorphisms have a certain correlation with the susceptibility of asthma in Chinese children,the mutant gene G carriers may relatively increase the risk of asthma in childhood.
9.Application of capillary electrophoresis by dried filter blood paper for screening of α-thalassemia in neonates
Zhidan WAN ; Jinglin CHEN ; Xiang HUANG ; Xuewei WU ; Dongxiu LI ; Haixin YANG
International Journal of Laboratory Medicine 2016;37(6):731-732,735
Objective To investigate the application of capillary electrophoresis by dried filter blood paper for screening of α-thalassemia in neonates .Methods The hemoglobin (Hb) of 46 718 cases of neonatal dried heel blood spots were analyzed by the capillary electrophoresis and the content of HbA ,HbF ,HbA2 and abnormal Hb were detected ,the phenotype cases which was screened positive were recalled for genetic analysis .Results A total of 2 598 cases of Bart hemoglobin (Hb Bart′s) positive were detected in 46 718 cases of neonatal heel blood dried blood spots .The screening positive rate was 5 .56% (2 598/46 718) .A total of 477 cases of α-thalassemia gene carriers were confirmed by genetic analysis in the 544 cases which were recalled .The coincidence rate of Hb Bart′s screening and genetic diagnosis was 87 .68% (477/544) .By analyzing the relationship between the clinical pheno-types and the content of Hb Bart′s ,we found the Hb Bart′s content gradually increased with the severity of clinical phenotype ,and the difference was statistically significant (P= 0 .000) .Conclusion There is a good consistency between the capillary electrophore-sis of dried filter blood paper and the genetic analysis .It could be determined α-thalassemia clinical type according to the Hb Bart′s content .
10.Detection of IgG antibody affinity in suspected cases of measles and rubella in Beijing
Xiang GAO ; Jie GAO ; Ping ZHANG ; Jinglin ZHOU ; Lin ZOU ; Chong ZHANG ; Ling TONG ; Guofeng ZHANG ; Yanchun WANG ; Jianguo WANG ; Meng CHEN
Chinese Journal of Microbiology and Immunology 2020;40(6):465-469
Objective:To investigate the causes of immune failure in the population with high vaccination rate of measles and rubella vaccine in Beijing by detecting the IgG antibody affinity in suspected cases of measles and rubella.Methods:Serum samples of 276 suspected cases of measles and rubella were tested for IgM and IgG antibodies by enzyme-linked immunosorbent assay (ELISA). The affinity of IgG antibody was detected, and the relative affinity index was calculated.Results:Among the 276 suspected cases, 104 were measles and 108 were rubella. Six measles cases had vaccination history and were caused by primary immunization failure ( n=3) and secondary immunization failure ( n=3). Twelve rubella cases had vaccination history and were due to primary immunization failure ( n=4) and secondary immunization failure ( n=8). Specific high-affinity antibodies were detected in nine measles cases and seven rubella cases without vaccination history, which indicated that these cases were reinfected. In the cases without measles or rubella, other pathogenic infections including mixed infections were detected, which were mainly caused by EB virus. Conclusions:Both primary and secondary immunization failure occurred in the population with immunization history. Reinfection was found in the patients who had not received vaccination against measles or rubella. Other pathogenic infections were existed among the cases without measles or rubella. Thus, misdiagnosis was responsible for the increased proportion of measles and rubella patients with immunization history in suspected cases in recent years. Full-course vaccination was conducive to produce high-affinity antibodies against measles and rubella. A supplementary vaccination campaign should be launched to consolidate the immune barrier against measles and rubella in key population or high-risk population, aiming to block the circulation of measles virus and achieve the goal of eliminating measles.