BACKGROUND A growing number of clinical examples suggest that coronavirus disease 2019(COVID-19)appears to have an impact on the treatment of patients with liver cancer compared to the normal population,and the preval...BACKGROUND A growing number of clinical examples suggest that coronavirus disease 2019(COVID-19)appears to have an impact on the treatment of patients with liver cancer compared to the normal population,and the prevalence of COVID-19 is significantly higher in patients with liver cancer.However,this mechanism of action has not been clarified.Gene sets for COVID-19(GSE180226)and liver cancer(GSE87630)were obtained from the Gene Expression Omnibus database.After identifying the common differentially expressed genes(DEGs)of COVID-19 and liver cancer,functional enrichment analysis,protein-protein interaction network construction and scree-ning and analysis of hub genes were performed.Subsequently,the validation of the differential expression of hub genes in the disease was performed and the regulatory network of transcription factors and hub genes was constructed.RESULTS Of 518 common DEGs were obtained by screening for functional analysis.Fifteen hub genes including aurora kinase B,cyclin B2,cell division cycle 20,cell division cycle associated 8,nucleolar and spindle associated protein 1,etc.,were further identified from DEGs using the“cytoHubba”plugin.Functional enrichment analysis of hub genes showed that these hub genes are associated with P53 signalling pathway regulation,cell cycle and other functions,and they may serve as potential molecular markers for COVID-19 and liver cancer.Finally,we selected 10 of the hub genes for in vitro expression validation in liver cancer cells.CONCLUSION Our study reveals a common pathogenesis of liver cancer and COVID-19.These common pathways and key genes may provide new ideas for further mechanistic studies.展开更多
Seed weight is a component of seed yield in rapeseed(Brassica napus L.).Although quantitative trait loci(QTL)for seed weight have been reported in rapeseed,only a few causal quantitative trait genes(QTGs)have been ide...Seed weight is a component of seed yield in rapeseed(Brassica napus L.).Although quantitative trait loci(QTL)for seed weight have been reported in rapeseed,only a few causal quantitative trait genes(QTGs)have been identified,resulting in a limitation in understanding of seed weight regulation.We constructed a gene coexpression network at the early seed developmental stage using transcripts of 20,408 genes in QTL intervals and 1017 rapeseed homologs of known genes from other species.Among the 10 modules in this gene coexpression network,modules 1 and 2 were core modules and contained genes involved in source–flow–sink processes such as synthesis and transportation of fatty acid and protein,and photosynthesis.A hub gene SERINE CARBOXYPEPTIDASE-LIKE 19(SCPL19)was identified by candidate gene association analysis in rapeseed and functionally investigated using Arabidopsis T-DNA mutant and overexpression lines.Our study demonstrates the power of gene coexpression analysis to prioritize candidate genes from large candidate QTG sets and enhances the understanding of molecular mechanism for seed weight at the early developmental stage in rapeseed.展开更多
Background:To provide a reference for the clinical development of drugs to suppress severe acute respiratory syndrome coronavirus 2(SARS-CoV-2).Methods:Retrieving genes related to SARS-CoV-2 with Genecards database an...Background:To provide a reference for the clinical development of drugs to suppress severe acute respiratory syndrome coronavirus 2(SARS-CoV-2).Methods:Retrieving genes related to SARS-CoV-2 with Genecards database and then importing the obtained gene data into the database of Database for Annotation,Visualization and Integrated Discovery(DAVID)(Version 6.8)to collect relevant information on pathways and genes.Genes enriched in the first 20 most significant pathways and genes with gene occurrence frequency≥6 were respectively imported into the STRING database to construct protein-protein interaction(PPI)network diagrams,and the two network diagrams were compared.Results:In the two network graphs,RELA,MAPK1,MAPK3,PIK3CA,PIK3R1,MAPK8,JAK1,STAT1,TNF,IL6,MAPK14,and IL1B ranked higher,and the occurrence frequency of the first 20 pathways was≥10.Conclusion:The pathogenesis of SARS-CoV-2 is associated with multiple pathways such as influenza A,TNF signaling pathway,chemokine signaling pathway,toll-like receptor signaling pathway,T cell receptor signaling pathway et al.RELA,MAPK1,MAPK3,PIK3CA,PIK3R1,MAPK8,JAK1,STAT1,TNF,IL6,MAPK14 and IL1B are closely related to SARS-CoV-2 and need further study.Gene interaction network and pathway analysis of diseaseassociated genes will help us to screen the key target genes of SARS-CoV-2 and provide a reference for the clinical development of effective drugs.展开更多
Objectives Clopidogrel is a prodrug that has to be converted to an active metabolite by hepatic cytochrome P450(CYP) isoenzymes to inhibit platelet aggregation.Individualvariability of platelet inhibition by clopidogr...Objectives Clopidogrel is a prodrug that has to be converted to an active metabolite by hepatic cytochrome P450(CYP) isoenzymes to inhibit platelet aggregation.Individualvariability of platelet inhibition by clopidogrel suggests a possibility for genetic factors having a significant influence on clopidogrel responsiveness.In this study,we sought to determine the association between the single nucleotide polymorphism of CYP 2C19 681G】A and the occurrence of clopidogrel resistance(CR) in Chinese.Methods The study enrolled 614 hospitalized patients who underwentsuccessful percutaneouscoronary intervention with drug-eluting stents were received the treatmentwith dual antiplatelet regimen(aspirin plus clopidogrel).All patients received loading doses of 600 mg clopidogrel and 300 mg aspirin.20μmol/L ADP-induced platelet aggregation ratio(PAR ) was assessed 24 h after clopi- dogrel administration.The maximum residual PAR≥70%was defined as CR.Genomic DNA was extracted from whole blood samples according to standard protocols,the single nucleotide polymorphism of the CYP2C19 681G】A was genotyped by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) in all the patients.Results CR was found in 126 patients(20.5%).There was CYP2C19 681G】A polymorphism in the study population.The frequencies of the three kinds of genotypes(GG,GA,A A) in CR group and non-CR (NCR)group were 32.5%,47.6%,19.8%and 48.0%, 45.0%,7.0%,respectively.The frequency of AA genotype was significantly higher in NCR group than that in CR group (OR =3.03,95%CI:1.889~5.784,P=0.003).The A allele carriers were more likely to develop clopidogrel resistance compared with that of G allele carriers(OR=1.85,95%CI: 1.392~2.459,P=0.002).Conclusions CYP2C19 681G/A polymorphism is associated with the risk of CR,and the A allele carriers may be a possible genetic susceptibility factor for patients with CR.展开更多
Fasciclin-like arabinogalactan proteins(FLAs),a subclass of arabinogalactan proteins(AGPs),are usually involved in cell development in plants.To investigate the expression profiling as well
Temperature-dependent sex determination(TSD) is a type of environmental sex determination in which the sex of the embryos depends on the ambient temperature; however,the molecular mechanisms governing this process r...Temperature-dependent sex determination(TSD) is a type of environmental sex determination in which the sex of the embryos depends on the ambient temperature; however,the molecular mechanisms governing this process remain unknown.Aromatase,encoded by the cyp19a1 gene,which converts androgens into estrogens in animals,was considered to be the key gene for TSD.In this study,the 5'-flanking region of the cyp19a1 gene in Reeves' turtle(Mauremys reevesii) was cloned,and the promoter region was identified using the luciferase reporter assay.Then the eggs of Reeves' turtle were incubated at different temperatures(26°C: male-biased temperature; 29°C: non-sex-biased temperature and 32°C: female-biased temperature).During the thermosensitive period,the adrenal kidney gonad complexes(AKG) were sampled.DNA methylation analysis of the AKG samples showed that the promoter region of the cyp19a1 gene was significantly de-methylated in the female-biased temperature regime(P<0.01).Quantitative analysis of the cyp19a1 gene and estrogen by q PCR and Elisa assay showed that the expression level of the cyp19a1 gene and estrogen content were both upregulated significantly at the female-biased temperature(P<0.01).These results indicated that the de-methylation response of the cyp19a1 gene to incubation temperature,especially at the female-biased temperature,could lead to temperature-specific expression of aromatase and increased estrogen levels,which may further determine gonadal development in Reeves' turtle.These findings provide insights into the genetic mechanisms underlying TSD.展开更多
61 varieties of wheat collected in the gene fund of the Research Institute of Crop Husbandry were screened using SCAR-markers associated with the gene of resistance to brown leaf rust, Lr19. As a result of PCR analysi...61 varieties of wheat collected in the gene fund of the Research Institute of Crop Husbandry were screened using SCAR-markers associated with the gene of resistance to brown leaf rust, Lr19. As a result of PCR analysis using SCS123 marker the 737 bp locus was detected in 48 genotypes. The expected fragment of the 688 bp was detected in 53 genotypes using the SCS253 marker. The results obtained using both markers indicate that the Lr19 gene is present on 7D chromosomes of 45 genotypes. The existence of the Lr19 gene has not been proven only for 5 from the 61 analyzed wheat genotypes.展开更多
Extended stage small cell lung cancer (SCLC) responds to platinum/vepeside-based first-line chemotherapy but relapses early as drug-resistant tumor associated with a dismal prognosis. A pair of SCLC cell lines obtaine...Extended stage small cell lung cancer (SCLC) responds to platinum/vepeside-based first-line chemotherapy but relapses early as drug-resistant tumor associated with a dismal prognosis. A pair of SCLC cell lines obtained from a single patient at different time points during treatment allows for the investigation of the changes in gene expression prior to (GLC14) and following cycles of chemotherapy and irradiation (GLC19). GLC19 cells were reported to reveal an increased doubling time and exhibit increased chemoresistance to doxorubicin, etoposide, melphalan and vinblastine. Upregulated transcripts in GLC19, as assessed by microarray analysis, comprised proteins involved in regulation of cellular growth (NGFRAP1/BEX3), adhesion, glutathione metabolism and, in particular, WNT/Notch pathways and the putative cancer stem cell phenotype (CD44, ALDH1A1, and AKR1C1/13). Metallothioneins, tubulins TUBA3/4 and tumor protein p53 inducible protein 11 (TP53IP11) were downregulated in this cell line compared to GLC14. Except increased expression of glutathione transferases no classical markers of chemoresistance were found, pointing to a role of altered growth control/differentiation and reduced accessibility of this SCLC tumor cells growing as multicellular spheroids. In conclusion, treatment of this single SCLC with cyclophosphamide, doxorubicin and etoposide (CDE) followed by radiotherapy ultimately resulted in an enrichment of tumor cells displaying the typical signature of tumor-initiating or cancer stem cells (CIC/CSC).展开更多
Objective The aim of this study was to explore the clinical significance of the expression of proteins human bone marrow endothelial cell markers(HBME-1), Galectin-3, and cytokeratin19(CK19), as well as the status of ...Objective The aim of this study was to explore the clinical significance of the expression of proteins human bone marrow endothelial cell markers(HBME-1), Galectin-3, and cytokeratin19(CK19), as well as the status of v-raf murine sarcoma viral oncogene homolog B1(BRAF) mutation in papillary thyroid carcinoma(PTC). Methods Immunohistochemical staining was performed in 82 specimens each of PTC and papillary benign lesions to detect the expression of HBME-1, Galectin-3, and CK19. Polymerase chain reaction(PCR) and gene sequencing were performed on 60 specimens each of PTC and papillary benign lesions to detect the status of BRAF mutation. Results The positive expression ratios of HBME-1, Galectin-3, and CK19 in PTC were 98.8%, 97.6% and 100% respectively, which were significantly higher than the expressions in papillary benign lesions(P < 0.05). No significant relationship was observed between the expression of these makers and the clinicopathological features of PTC. The sensitivity of co-expression of HBME-1 and CK19 or HBME-1 and Galectin-3 as diagnostic criteria of PTC was 99.9%, with a specificity of 95.4%. BRAF mutation was detected in 40 of 60 PTC(66.7%) specimens. There was a statistical difference in BRAF mutations between PTC and papillary benign lesions(P < 0.05); there were no associations between BRAF mutation and the clinicopathological features of PTC. Conclusion Combined immunohistochemical staining of HBME-1, Galectin-3, and CK19 can further improve the sensitivity and specificity of differential diagnosis of PTC. BRAF mutation is a significant genetic event, which may have diagnostic value for PTC.展开更多
Objective: To investigate the changes of cyclin L2(CCNL2) gene mRNA and protein during the differentiation of P19 cells to cardiac myocytes, and to explore the relationship between CCNL2 gene and the differentiatio...Objective: To investigate the changes of cyclin L2(CCNL2) gene mRNA and protein during the differentiation of P19 cells to cardiac myocytes, and to explore the relationship between CCNL2 gene and the differentiation of cardiac myocytes. Methods: P19 cells were cultured with 0.9% DMSO for 18 days. Western blots of cardiac troponin I (cTnI) were used to identify cell differentiation. Total RNA was extracted from P19 cells during the process of differentiation at various time points:pre-differentiation(Day 0), and Day 1 to Day 18. The expression levels of CCNL2 gene mRNA and protein were evaluated by RT-PCR and Western blot, respectively. Results: After being induced to differentiate by DMSO for 4 days in suspension, spontaneously and rhythmically beating ceils were seen at 8 day, which were cTnI-positive. In P19 cells, both the expression level of CCNL2 gene mRNA and protein were gradually down-regulated. Conclusion: Both the expression of CCNL2 gene and protein were down-regulated during the process of the differentiation of P19 cells into cardiac myocytes, suggesting a possible role for this cyclin in their differentiation.展开更多
This study is aimed at establishing a sensitive approach to detect disseminated tumor cells in peripheral blood and evaluate its clinical significance. A total of 198 blood samples including 168 from colorectal carcin...This study is aimed at establishing a sensitive approach to detect disseminated tumor cells in peripheral blood and evaluate its clinical significance. A total of 198 blood samples including 168 from colorectal carcinoma (CRC) patients and 30 from healthy volunteers were examined by quantitative real-time reverse transcription-polymerase chain reaction (RT-PCR) to evaluate the expression of carcinoembryonic antigen (CEA), cytokeratin 20 (CK20) and cytokeratin 19 (CK19) mRNA. CEA mRNA was detected in 35.8% of patients and 3.3% of controls, CK20 mRNA in 28.3% of patients and 6.7% of controls, and CK19 mRNA in 41.9% of patients and 3.3% of controls. CEA and CK20 mRNA positive ratio increased with the advancing Dukes stages, but there was no significant difference in positive ratio between any two stages (P>0.05). Also, relatively high positive ratio of CEA, CK20 and CK19 mRNA expression was observed in some CRC patients with earlier Dukes stages. A higher positive ratio was obtained when two or three detection markers were combined compared to a single marker. Our study indicates that quanti-tative real-time RT-PCR detection for CEA, CK20 and CK19 mRNA in peripheral blood is a valuable tool for monitoring early stage dissemination of CRC cells in blood circulation.展开更多
PCR enzyme linked immune sorbent assay (ELISA) was developed for detection of RT PCR amplified cytokeratin 19 (CK19) mRNA and carcinoembryonic antigen (CEA) mRNA. The non radioactive hybridization was performed in ...PCR enzyme linked immune sorbent assay (ELISA) was developed for detection of RT PCR amplified cytokeratin 19 (CK19) mRNA and carcinoembryonic antigen (CEA) mRNA. The non radioactive hybridization was performed in a streptavidin coated microwell with digoxigenin labeled PCR products and with biotin labeled capture probe. PCR ELISA was proved to be expedient, simple, sensitive and safe for identification of CK19 , CEA RT PCR products. These results were proven by sequencing.展开更多
Objective: The aim of this study was to investigate the expressions of CD147 and CK19 in hepatocellular car- cinoma (HCC) and their clinical significance. Methods: The expressions of CD147 and CK19 were determined by ...Objective: The aim of this study was to investigate the expressions of CD147 and CK19 in hepatocellular car- cinoma (HCC) and their clinical significance. Methods: The expressions of CD147 and CK19 were determined by tissue microarray and immunohistochemistry (IHC) in 272 cases of HCC and 81 cases of adjacent tumorous tissue. Results: The positive expression of CD147 in HCC and adjacent tumorous tissue was 73.53% (200/272) and 13.58% (11/81) with signifi- cant difference (P < 0.05). The positive expression of CK19 in HCC and adjacent tumorous tissue was 14.34% (39/272) and 0 (0/81) with significant difference (P < 0.05). The positive expression of CD147 were closely correlated to the histological grade, clinical stage, tumor-free survival, diameter of tumor and embolus of cancer in aqueduct or portal vein; but not to the patients' sex, age, liver cirrhosis, AFP level, infection of HBV, lymph node metastasis, number of tumor, invasion liver involucrum and the micro-satellites (P > 0.05). The expression of CK19 in HCC were closely correlate to the tumor-free survival, histological grade, diameter of tumor, liver cirrhosis, micro-satellites, lymph node metastasis and clinical stage; but not to patients' sex, age, number of tumor, invasion liver involucrum, AFP level, infection of HBV and embolus of cancer in aqueduct or portal vein (P > 0.05). Among the patients of positive expression of CD147, the median replacing time and overall survival were 13 and 24 months, lower than 48 and 60 months in the patients of negative expression (P < 0.05). Among the patients of positive expression of CK19, the median replacing time and overall survival were 7 and 13 months, lower than 31 and 42 months in the patients of negative expression (P < 0.05). The expression of CD147 had no correlation with the expression of CK19 (r = 0.061, P = 0.317). Conclusion: The positive of CD147 and CK19 closely correlate with the clinical prognosis of HCC, it may indicate poor prognosis of HCC.展开更多
Objective: Using nested reverse transcription-polymerase chain reaction(Nested RT-PCR) to test the mRNA level in peripheral blood CK19 and MUC-1 in colorectal cancer patients and it's clinical significance, to dis...Objective: Using nested reverse transcription-polymerase chain reaction(Nested RT-PCR) to test the mRNA level in peripheral blood CK19 and MUC-1 in colorectal cancer patients and it's clinical significance, to discuss the feasibility of colorectal carcinoma micro-metastasis detection of molecular markers. Methods: The expression level was detected by nested RT-PCR in 20 healthy people, 20 patients with colorectal adenoma and 90 cases of patients with colorectal cancer disease peripheral blood CK19 mRNA and MUC-1 mRNA. Results: The positive expression rate of CK19 mRNA and MUC-1 mRNA were: 58.89%(53/90) and 52.22%(47/90). No CK19 mRNA healthy people 20 cases in the control group in the peripheral blood, the expression of MUC-1 mRNA in 12 cases, the expression rate of 60%(12/20). In 20 cases of colorectal adenoma diseases have the expression of CK19 mRNA in 1 cases, the expression rate of 5%(1/20), the expression of MUC-1 mRNA in 10 cases, the expression rate of 50%. Patients with colorectal cancer CK19 mRNA, MUC-1 mRNA expression rate was significantly correlated with tumor staging, the degree of differentiation of the tumor cells and tumor metastasis(P < 0.05). Conclusion: Marker CK19 mRNA as the detection of micro-metastasis in peripheral blood of patients with colorectal cancer has good sensitivity and specificity, but CK19 mRNA, MUC-1 mRNA can be used to judge the prognosis of patients with colorectal cancer index.展开更多
文摘BACKGROUND A growing number of clinical examples suggest that coronavirus disease 2019(COVID-19)appears to have an impact on the treatment of patients with liver cancer compared to the normal population,and the prevalence of COVID-19 is significantly higher in patients with liver cancer.However,this mechanism of action has not been clarified.Gene sets for COVID-19(GSE180226)and liver cancer(GSE87630)were obtained from the Gene Expression Omnibus database.After identifying the common differentially expressed genes(DEGs)of COVID-19 and liver cancer,functional enrichment analysis,protein-protein interaction network construction and scree-ning and analysis of hub genes were performed.Subsequently,the validation of the differential expression of hub genes in the disease was performed and the regulatory network of transcription factors and hub genes was constructed.RESULTS Of 518 common DEGs were obtained by screening for functional analysis.Fifteen hub genes including aurora kinase B,cyclin B2,cell division cycle 20,cell division cycle associated 8,nucleolar and spindle associated protein 1,etc.,were further identified from DEGs using the“cytoHubba”plugin.Functional enrichment analysis of hub genes showed that these hub genes are associated with P53 signalling pathway regulation,cell cycle and other functions,and they may serve as potential molecular markers for COVID-19 and liver cancer.Finally,we selected 10 of the hub genes for in vitro expression validation in liver cancer cells.CONCLUSION Our study reveals a common pathogenesis of liver cancer and COVID-19.These common pathways and key genes may provide new ideas for further mechanistic studies.
基金provided by the National Natural Science Foundation of China(32201776)the Natural Science Foundation of Chongqing(cstc2019jcyj-bsh X0055,cstc2019jcyj-zdxm X0012)。
文摘Seed weight is a component of seed yield in rapeseed(Brassica napus L.).Although quantitative trait loci(QTL)for seed weight have been reported in rapeseed,only a few causal quantitative trait genes(QTGs)have been identified,resulting in a limitation in understanding of seed weight regulation.We constructed a gene coexpression network at the early seed developmental stage using transcripts of 20,408 genes in QTL intervals and 1017 rapeseed homologs of known genes from other species.Among the 10 modules in this gene coexpression network,modules 1 and 2 were core modules and contained genes involved in source–flow–sink processes such as synthesis and transportation of fatty acid and protein,and photosynthesis.A hub gene SERINE CARBOXYPEPTIDASE-LIKE 19(SCPL19)was identified by candidate gene association analysis in rapeseed and functionally investigated using Arabidopsis T-DNA mutant and overexpression lines.Our study demonstrates the power of gene coexpression analysis to prioritize candidate genes from large candidate QTG sets and enhances the understanding of molecular mechanism for seed weight at the early developmental stage in rapeseed.
文摘Background:To provide a reference for the clinical development of drugs to suppress severe acute respiratory syndrome coronavirus 2(SARS-CoV-2).Methods:Retrieving genes related to SARS-CoV-2 with Genecards database and then importing the obtained gene data into the database of Database for Annotation,Visualization and Integrated Discovery(DAVID)(Version 6.8)to collect relevant information on pathways and genes.Genes enriched in the first 20 most significant pathways and genes with gene occurrence frequency≥6 were respectively imported into the STRING database to construct protein-protein interaction(PPI)network diagrams,and the two network diagrams were compared.Results:In the two network graphs,RELA,MAPK1,MAPK3,PIK3CA,PIK3R1,MAPK8,JAK1,STAT1,TNF,IL6,MAPK14,and IL1B ranked higher,and the occurrence frequency of the first 20 pathways was≥10.Conclusion:The pathogenesis of SARS-CoV-2 is associated with multiple pathways such as influenza A,TNF signaling pathway,chemokine signaling pathway,toll-like receptor signaling pathway,T cell receptor signaling pathway et al.RELA,MAPK1,MAPK3,PIK3CA,PIK3R1,MAPK8,JAK1,STAT1,TNF,IL6,MAPK14 and IL1B are closely related to SARS-CoV-2 and need further study.Gene interaction network and pathway analysis of diseaseassociated genes will help us to screen the key target genes of SARS-CoV-2 and provide a reference for the clinical development of effective drugs.
文摘Objectives Clopidogrel is a prodrug that has to be converted to an active metabolite by hepatic cytochrome P450(CYP) isoenzymes to inhibit platelet aggregation.Individualvariability of platelet inhibition by clopidogrel suggests a possibility for genetic factors having a significant influence on clopidogrel responsiveness.In this study,we sought to determine the association between the single nucleotide polymorphism of CYP 2C19 681G】A and the occurrence of clopidogrel resistance(CR) in Chinese.Methods The study enrolled 614 hospitalized patients who underwentsuccessful percutaneouscoronary intervention with drug-eluting stents were received the treatmentwith dual antiplatelet regimen(aspirin plus clopidogrel).All patients received loading doses of 600 mg clopidogrel and 300 mg aspirin.20μmol/L ADP-induced platelet aggregation ratio(PAR ) was assessed 24 h after clopi- dogrel administration.The maximum residual PAR≥70%was defined as CR.Genomic DNA was extracted from whole blood samples according to standard protocols,the single nucleotide polymorphism of the CYP2C19 681G】A was genotyped by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) in all the patients.Results CR was found in 126 patients(20.5%).There was CYP2C19 681G】A polymorphism in the study population.The frequencies of the three kinds of genotypes(GG,GA,A A) in CR group and non-CR (NCR)group were 32.5%,47.6%,19.8%and 48.0%, 45.0%,7.0%,respectively.The frequency of AA genotype was significantly higher in NCR group than that in CR group (OR =3.03,95%CI:1.889~5.784,P=0.003).The A allele carriers were more likely to develop clopidogrel resistance compared with that of G allele carriers(OR=1.85,95%CI: 1.392~2.459,P=0.002).Conclusions CYP2C19 681G/A polymorphism is associated with the risk of CR,and the A allele carriers may be a possible genetic susceptibility factor for patients with CR.
文摘Fasciclin-like arabinogalactan proteins(FLAs),a subclass of arabinogalactan proteins(AGPs),are usually involved in cell development in plants.To investigate the expression profiling as well
基金supported financially by the National Natural Science Foundation of China(Nos.31401053 and 31471966)Guangdong Provincial Natural Science Foundation of China(No.2015A030313903)+1 种基金GDAS Special Project of Science and Technology Development(2017GDASCX-0107)the Funds for Environment Construction and Capacity Building of GDAS’Research Platform(2016GDASPT-0107)
文摘Temperature-dependent sex determination(TSD) is a type of environmental sex determination in which the sex of the embryos depends on the ambient temperature; however,the molecular mechanisms governing this process remain unknown.Aromatase,encoded by the cyp19a1 gene,which converts androgens into estrogens in animals,was considered to be the key gene for TSD.In this study,the 5'-flanking region of the cyp19a1 gene in Reeves' turtle(Mauremys reevesii) was cloned,and the promoter region was identified using the luciferase reporter assay.Then the eggs of Reeves' turtle were incubated at different temperatures(26°C: male-biased temperature; 29°C: non-sex-biased temperature and 32°C: female-biased temperature).During the thermosensitive period,the adrenal kidney gonad complexes(AKG) were sampled.DNA methylation analysis of the AKG samples showed that the promoter region of the cyp19a1 gene was significantly de-methylated in the female-biased temperature regime(P&lt;0.01).Quantitative analysis of the cyp19a1 gene and estrogen by q PCR and Elisa assay showed that the expression level of the cyp19a1 gene and estrogen content were both upregulated significantly at the female-biased temperature(P&lt;0.01).These results indicated that the de-methylation response of the cyp19a1 gene to incubation temperature,especially at the female-biased temperature,could lead to temperature-specific expression of aromatase and increased estrogen levels,which may further determine gonadal development in Reeves' turtle.These findings provide insights into the genetic mechanisms underlying TSD.
文摘61 varieties of wheat collected in the gene fund of the Research Institute of Crop Husbandry were screened using SCAR-markers associated with the gene of resistance to brown leaf rust, Lr19. As a result of PCR analysis using SCS123 marker the 737 bp locus was detected in 48 genotypes. The expected fragment of the 688 bp was detected in 53 genotypes using the SCS253 marker. The results obtained using both markers indicate that the Lr19 gene is present on 7D chromosomes of 45 genotypes. The existence of the Lr19 gene has not been proven only for 5 from the 61 analyzed wheat genotypes.
文摘Extended stage small cell lung cancer (SCLC) responds to platinum/vepeside-based first-line chemotherapy but relapses early as drug-resistant tumor associated with a dismal prognosis. A pair of SCLC cell lines obtained from a single patient at different time points during treatment allows for the investigation of the changes in gene expression prior to (GLC14) and following cycles of chemotherapy and irradiation (GLC19). GLC19 cells were reported to reveal an increased doubling time and exhibit increased chemoresistance to doxorubicin, etoposide, melphalan and vinblastine. Upregulated transcripts in GLC19, as assessed by microarray analysis, comprised proteins involved in regulation of cellular growth (NGFRAP1/BEX3), adhesion, glutathione metabolism and, in particular, WNT/Notch pathways and the putative cancer stem cell phenotype (CD44, ALDH1A1, and AKR1C1/13). Metallothioneins, tubulins TUBA3/4 and tumor protein p53 inducible protein 11 (TP53IP11) were downregulated in this cell line compared to GLC14. Except increased expression of glutathione transferases no classical markers of chemoresistance were found, pointing to a role of altered growth control/differentiation and reduced accessibility of this SCLC tumor cells growing as multicellular spheroids. In conclusion, treatment of this single SCLC with cyclophosphamide, doxorubicin and etoposide (CDE) followed by radiotherapy ultimately resulted in an enrichment of tumor cells displaying the typical signature of tumor-initiating or cancer stem cells (CIC/CSC).
文摘Objective The aim of this study was to explore the clinical significance of the expression of proteins human bone marrow endothelial cell markers(HBME-1), Galectin-3, and cytokeratin19(CK19), as well as the status of v-raf murine sarcoma viral oncogene homolog B1(BRAF) mutation in papillary thyroid carcinoma(PTC). Methods Immunohistochemical staining was performed in 82 specimens each of PTC and papillary benign lesions to detect the expression of HBME-1, Galectin-3, and CK19. Polymerase chain reaction(PCR) and gene sequencing were performed on 60 specimens each of PTC and papillary benign lesions to detect the status of BRAF mutation. Results The positive expression ratios of HBME-1, Galectin-3, and CK19 in PTC were 98.8%, 97.6% and 100% respectively, which were significantly higher than the expressions in papillary benign lesions(P < 0.05). No significant relationship was observed between the expression of these makers and the clinicopathological features of PTC. The sensitivity of co-expression of HBME-1 and CK19 or HBME-1 and Galectin-3 as diagnostic criteria of PTC was 99.9%, with a specificity of 95.4%. BRAF mutation was detected in 40 of 60 PTC(66.7%) specimens. There was a statistical difference in BRAF mutations between PTC and papillary benign lesions(P < 0.05); there were no associations between BRAF mutation and the clinicopathological features of PTC. Conclusion Combined immunohistochemical staining of HBME-1, Galectin-3, and CK19 can further improve the sensitivity and specificity of differential diagnosis of PTC. BRAF mutation is a significant genetic event, which may have diagnostic value for PTC.
文摘Objective: To investigate the changes of cyclin L2(CCNL2) gene mRNA and protein during the differentiation of P19 cells to cardiac myocytes, and to explore the relationship between CCNL2 gene and the differentiation of cardiac myocytes. Methods: P19 cells were cultured with 0.9% DMSO for 18 days. Western blots of cardiac troponin I (cTnI) were used to identify cell differentiation. Total RNA was extracted from P19 cells during the process of differentiation at various time points:pre-differentiation(Day 0), and Day 1 to Day 18. The expression levels of CCNL2 gene mRNA and protein were evaluated by RT-PCR and Western blot, respectively. Results: After being induced to differentiate by DMSO for 4 days in suspension, spontaneously and rhythmically beating ceils were seen at 8 day, which were cTnI-positive. In P19 cells, both the expression level of CCNL2 gene mRNA and protein were gradually down-regulated. Conclusion: Both the expression of CCNL2 gene and protein were down-regulated during the process of the differentiation of P19 cells into cardiac myocytes, suggesting a possible role for this cyclin in their differentiation.
基金Project (No. 021103004) supported by the Science and TechnologyDevelopment Program of Zhejiang Province, China
文摘This study is aimed at establishing a sensitive approach to detect disseminated tumor cells in peripheral blood and evaluate its clinical significance. A total of 198 blood samples including 168 from colorectal carcinoma (CRC) patients and 30 from healthy volunteers were examined by quantitative real-time reverse transcription-polymerase chain reaction (RT-PCR) to evaluate the expression of carcinoembryonic antigen (CEA), cytokeratin 20 (CK20) and cytokeratin 19 (CK19) mRNA. CEA mRNA was detected in 35.8% of patients and 3.3% of controls, CK20 mRNA in 28.3% of patients and 6.7% of controls, and CK19 mRNA in 41.9% of patients and 3.3% of controls. CEA and CK20 mRNA positive ratio increased with the advancing Dukes stages, but there was no significant difference in positive ratio between any two stages (P>0.05). Also, relatively high positive ratio of CEA, CK20 and CK19 mRNA expression was observed in some CRC patients with earlier Dukes stages. A higher positive ratio was obtained when two or three detection markers were combined compared to a single marker. Our study indicates that quanti-tative real-time RT-PCR detection for CEA, CK20 and CK19 mRNA in peripheral blood is a valuable tool for monitoring early stage dissemination of CRC cells in blood circulation.
文摘PCR enzyme linked immune sorbent assay (ELISA) was developed for detection of RT PCR amplified cytokeratin 19 (CK19) mRNA and carcinoembryonic antigen (CEA) mRNA. The non radioactive hybridization was performed in a streptavidin coated microwell with digoxigenin labeled PCR products and with biotin labeled capture probe. PCR ELISA was proved to be expedient, simple, sensitive and safe for identification of CK19 , CEA RT PCR products. These results were proven by sequencing.
文摘Objective: The aim of this study was to investigate the expressions of CD147 and CK19 in hepatocellular car- cinoma (HCC) and their clinical significance. Methods: The expressions of CD147 and CK19 were determined by tissue microarray and immunohistochemistry (IHC) in 272 cases of HCC and 81 cases of adjacent tumorous tissue. Results: The positive expression of CD147 in HCC and adjacent tumorous tissue was 73.53% (200/272) and 13.58% (11/81) with signifi- cant difference (P < 0.05). The positive expression of CK19 in HCC and adjacent tumorous tissue was 14.34% (39/272) and 0 (0/81) with significant difference (P < 0.05). The positive expression of CD147 were closely correlated to the histological grade, clinical stage, tumor-free survival, diameter of tumor and embolus of cancer in aqueduct or portal vein; but not to the patients' sex, age, liver cirrhosis, AFP level, infection of HBV, lymph node metastasis, number of tumor, invasion liver involucrum and the micro-satellites (P > 0.05). The expression of CK19 in HCC were closely correlate to the tumor-free survival, histological grade, diameter of tumor, liver cirrhosis, micro-satellites, lymph node metastasis and clinical stage; but not to patients' sex, age, number of tumor, invasion liver involucrum, AFP level, infection of HBV and embolus of cancer in aqueduct or portal vein (P > 0.05). Among the patients of positive expression of CD147, the median replacing time and overall survival were 13 and 24 months, lower than 48 and 60 months in the patients of negative expression (P < 0.05). Among the patients of positive expression of CK19, the median replacing time and overall survival were 7 and 13 months, lower than 31 and 42 months in the patients of negative expression (P < 0.05). The expression of CD147 had no correlation with the expression of CK19 (r = 0.061, P = 0.317). Conclusion: The positive of CD147 and CK19 closely correlate with the clinical prognosis of HCC, it may indicate poor prognosis of HCC.
文摘Objective: Using nested reverse transcription-polymerase chain reaction(Nested RT-PCR) to test the mRNA level in peripheral blood CK19 and MUC-1 in colorectal cancer patients and it's clinical significance, to discuss the feasibility of colorectal carcinoma micro-metastasis detection of molecular markers. Methods: The expression level was detected by nested RT-PCR in 20 healthy people, 20 patients with colorectal adenoma and 90 cases of patients with colorectal cancer disease peripheral blood CK19 mRNA and MUC-1 mRNA. Results: The positive expression rate of CK19 mRNA and MUC-1 mRNA were: 58.89%(53/90) and 52.22%(47/90). No CK19 mRNA healthy people 20 cases in the control group in the peripheral blood, the expression of MUC-1 mRNA in 12 cases, the expression rate of 60%(12/20). In 20 cases of colorectal adenoma diseases have the expression of CK19 mRNA in 1 cases, the expression rate of 5%(1/20), the expression of MUC-1 mRNA in 10 cases, the expression rate of 50%. Patients with colorectal cancer CK19 mRNA, MUC-1 mRNA expression rate was significantly correlated with tumor staging, the degree of differentiation of the tumor cells and tumor metastasis(P < 0.05). Conclusion: Marker CK19 mRNA as the detection of micro-metastasis in peripheral blood of patients with colorectal cancer has good sensitivity and specificity, but CK19 mRNA, MUC-1 mRNA can be used to judge the prognosis of patients with colorectal cancer index.