A new method for the preparation of oligonucleotide microarray for gene expression detection was found. The key techniques and standards of quality controlling for preparation of oligonucleotide microarray was explore...A new method for the preparation of oligonucleotide microarray for gene expression detection was found. The key techniques and standards of quality controlling for preparation of oligonucleotide microarray was explored using gene of human 23kD highly protein and Luciferase and mouse cytokine-associated genes. By the using of a software system MProbe, oligonucleotide probes were designed and BLAST. All the probes have a very high specificity, i.e. except target sequence, the similarity between the probe and non-target sequences is less than 70% and the hairpin structure are not exist in all probes.All the probes have the same length 40. GC contents in all probes are in a narrow scope (from 45% to 55%). All the probes are modified with amino at 5′ or 3′ terminal. The satisfied images with good sensativity and very high specificity were obtained by the using of the methods above and also using of positive and negative controls and some internal controls(house keeping gene) to quantitate and balance expression of genes. High specificity, good sensativity and stablity have been verified by three continuous experiments using the oligonucleotide microarray to study gene expression profile of normal mouse breast grand tissue .The oligonucleotide microarray for expression detection prepared using our method have high specificity, good sensativity and stablity et al . It may be a more advanced method for analysis of gene expression profile.展开更多
Sex determination and sex differentiation are important phenomena in fish, but the mechanisms of sex determination in Takifugu rubripes are poorly understood. In our study, the expression patterns of genes for DMRTs ...Sex determination and sex differentiation are important phenomena in fish, but the mechanisms of sex determination in Takifugu rubripes are poorly understood. In our study, the expression patterns of genes for DMRTs （DMRT1, DMRT2 and DMRT3）,sox9a and sox9b in T. rubripes tissues were verified with the Reverse Transcription （RT）-PCR detection. It is showed that DMRT1 expressions in testis and ovaries were much lower, and no expressions were fotmd in muscle, blood and tailfin. However, expressions for DMRT2 and DMRT3 were not found in the tissues stated above. Transcripts of sox9a were detected in muscle, fin, ovary and testis, but not in blood, whereas sox9b expression was only detected in ovary. The expression patterns of DMRTs, sox9a and sox9b in T. rubripes gonads suggest that these genes may not be sex-specific.展开更多
OBJECTIVE To quantitatively explore the expression of Syndecan-1 and heparanase-1 in esophageal cancer tissue as well as their relationship with the clinicopathological factors, in order to evaluate their roles in tum...OBJECTIVE To quantitatively explore the expression of Syndecan-1 and heparanase-1 in esophageal cancer tissue as well as their relationship with the clinicopathological factors, in order to evaluate their roles in tumor invasion and metastasis.METHODS Real-time fluorescence quantitative PCR （Q-PCR） was used to analyze the expression levels of Syndecan-1 and heparanase-1 genes￡？participants included 67 cases with esophageal cancers and 32 healthy volunteers.RESULTS The expression of Heparanase-1 gene in esophageal cancers was higher than that in normal esophageal tissue （P 〈 0.001）, and the expression of Syndecan-1 gene in the normal esophageal tissue was higher compared with esophageal cancers （P 〈 0.001）. The positive rates of Syndecan-1 and Heparanase-1 gene in esophageal cancer were 13.4% （9/67） and 85.1% （57/67）.The expression of Syndecan-1 and Heparanase-1 genes was signifi cantly related to di. erentiation, depth of infi ltration, lymph node metastasis, vessel metastasis, and TNM stages of disease （P 〈 0.05）. In an attempt to measure the association between the 2 agents, this study found that the expression of Syndecan-1 mRNA had a significantly negative correlation with the expression of Heparanase-1 mRNA by using Spearman rank correlation test （OR = -0.572, P 〈 0.001）.CONCLUSION Syndecan-1 and Heparanase-1 play important roles in the invasion and metastasis of esophageal cancer. The reduction of Syndecan-1 and/or the increase of Heparanase-1 may influence the invasion and metastasis of malignant tumors.Thus the combination assay of Syndecan-1 and Heparanase-1 may contribute to the diagnosis and treatment of malignant tumors.展开更多
OBJECTIVE Tob is a member of Tob/BTG antiproliferative family. To date, Tob expression in human carcinoma using clinical specimens has not been studied in depth except for lung carcinoma and thyroid carcinoma. This st...OBJECTIVE Tob is a member of Tob/BTG antiproliferative family. To date, Tob expression in human carcinoma using clinical specimens has not been studied in depth except for lung carcinoma and thyroid carcinoma. This study is the first to investigate the expression levels of Tob gene in human colorectal cancer tissues, and their corresponding para-cancerous tissues. The correlation of expression of the Tob gene with clinicopathological characteristics of colorectal cancer was also analyzed. METHODS Quantitative real time RT-PCR was used to detect the expression of Tob mRNA in 31 colorectal cancers. RESULTS Compared with normal tissues, up-regulation of Tob mRNA was observed in 31 colorectal cancer tissues （P = 0.020）. The expression level of Tob at Dukes C ＋ D phase was higher than Dukes A ＋ B phase, and the difference was significant （P 〈 0.05）. However, in this study, it was found that the expression of Tob mRNA was not related with age, gender, and pathological type of colorectal cancer. CONCLUSION The up-regulation of Tob may be closely associated with tumorigenesis of colorectal carcinoma.展开更多
Using PCR-denaturing gradient gel electrophoresis （DGGE） targeting the RNA polymerase beta subunit （rpoB） gene, a simultaneous detection method for Vibrio species was established, rpoB gene-based PCR-DGGE was carr...Using PCR-denaturing gradient gel electrophoresis （DGGE） targeting the RNA polymerase beta subunit （rpoB） gene, a simultaneous detection method for Vibrio species was established, rpoB gene-based PCR-DGGE was carried out with eight Vibrio reference strains （each from different species）, mixed sample （including these Vibrio reference strains）, two non Vibrio strains, four environmental Vibrio strains, and three unidentified environmental strains. For comparison, 16S rRNA gene-based PCR-DGGE of the eight Vibrio reference strains was performed with universal primers. In addition, three unidentified strains were identified by 16S rRNA and gyrB gene sequencing and API20E system in order to confirm the accuracy of rpoB gene-based PCR-DGGE detection. Results revealed that rpoB-based PCR-DGGE could well discriminate eight Vibrio reference strains and could not discriminate different strains within the same species. The bands derived from two non Vibrio strains could not match with any bands in reference marker. Meanwhile, 16S rRNA gene-based DGGE failed to distinguish these reference strains. Further-more, four out of eight Vibrio species exhibited heterogenous bands in 16S rRNA gene-based DGGE. Sequencing and API 20E identification of unidentified strains coincided with the detection by rpoB gene-based PCR-DGGE. The results demonstrated that rpoB-based PCR-DGGE provided a rapid and efficient method for simultaneous detection of muhiple Vibrio species, which can avoid the limitations inherent in 16S rRNA gene-based PCR-DGGE.展开更多
When gene expression profile is used for gene detection,the probe on the chip can emit fluorescence with different wavelengths.Under the action of confocal laser scanner,a clear gene change image can be obtained,by wh...When gene expression profile is used for gene detection,the probe on the chip can emit fluorescence with different wavelengths.Under the action of confocal laser scanner,a clear gene change image can be obtained,by which the gene changes of the sample to be tested can be observed directly.First,the knee osteoarthritis(KOA)models of mice are established by the method of collateral ligament and meniscus resection(MLI-OA).Then,Bushen Huoxue formula is given by gavage,and ribonucleic acid(RNA)is routinely extracted and purified.Finally,the gene expression changes of KOA tissues of mice are detected by Agilent SurePrint G3 Mouse GE V2.0 gene expression profile.The results show that Bushen Huoxue formula has significant regulation effect on gene expression of KOA tissue.Among the genes with significant up-regulation effect of Bushen Huoxue formula,there are 56 genes of traditional Chinese medicine(TCM)groups up-regulated more than twice compared with model groups.Among the genes with significant down-regulation effect,there are 119 genes of TCM groups down-regulated more than twice compared with model groups.The experimental results indicate that Bushen Huoxue formula may promote the metabolism of arthritic factors and delay cartilage degeneration to treat KOA by regulating genes that are currently unknown in the pathological process of KOA.展开更多
Objective:The aim of this study was to investigate the expression of BRAF of prostate cancer,and to explore its relations with clinic-pathological factors.Methods:Immunohistochemistry was used to detect the expression...Objective:The aim of this study was to investigate the expression of BRAF of prostate cancer,and to explore its relations with clinic-pathological factors.Methods:Immunohistochemistry was used to detect the expression of BRAF in 74 cases of prostate cancer,and 51 cases of benign prostate hyperplasia(BPH).Results:The positive rate of BRAF had significant difference between patients with prostate cancer and BPH(P < 0.05).The expression of BRAF was correlated with grade and stage of prostate cancer(P < 0.05),but not to age of onset(P > 0.05).Conclusion:The expression of BRAF may play a role in the tumorigenesis and progression of colorectal cancer.BRAF could be a factor to diagnose the typing of prostate cancers and predict the prognosis of prostate cancers.展开更多
Objective： To study the effect and mechanism of aspirin alone or combined with cisplatin （DDP） on human cervical carcinoma HeLa cells. Methods： HeLa cells were treated by different concentrations of aspirin, DDP a...Objective： To study the effect and mechanism of aspirin alone or combined with cisplatin （DDP） on human cervical carcinoma HeLa cells. Methods： HeLa cells were treated by different concentrations of aspirin, DDP alone or both. The inhibitory effect on cell growth was analyzed by MTT and colony-forming assay. Cell apoptosis was measured with flow cytometry. The mRNA levels ofBcl-2, Bax and NF-κB（P65） were studied by RT-PCR. Results： MTT assay showed that aspirin inhibited HeLa cell proliferation in a time-and dose-dependant maoner. Aspirin decreased clone numbers in colony formation assay. Aspirin also induced apoptosis of HeLa cells in a dose- and time-dependent manner as detected by flow cytometry. The inhibition effects on proliferation, colony formation and apoptosis were significantly enhanced when cells were treated with both aspirin and DDP. RT-PCR demonstrated that aspirin decreased the transcription of Bcl-2 and NF-κB, and increased expression of Bax gene. Conclusion： Aspirin can induce apoptosis in HeLa cells. Combination of aspirin and DDP displays a synergistic effect. The possible mechanism might be that aspirin downregulates the mRNA levels of Bcl-2 and NF-κB gene and upregulates the expression of Bax.展开更多
文摘A new method for the preparation of oligonucleotide microarray for gene expression detection was found. The key techniques and standards of quality controlling for preparation of oligonucleotide microarray was explored using gene of human 23kD highly protein and Luciferase and mouse cytokine-associated genes. By the using of a software system MProbe, oligonucleotide probes were designed and BLAST. All the probes have a very high specificity, i.e. except target sequence, the similarity between the probe and non-target sequences is less than 70% and the hairpin structure are not exist in all probes.All the probes have the same length 40. GC contents in all probes are in a narrow scope (from 45% to 55%). All the probes are modified with amino at 5′ or 3′ terminal. The satisfied images with good sensativity and very high specificity were obtained by the using of the methods above and also using of positive and negative controls and some internal controls(house keeping gene) to quantitate and balance expression of genes. High specificity, good sensativity and stablity have been verified by three continuous experiments using the oligonucleotide microarray to study gene expression profile of normal mouse breast grand tissue .The oligonucleotide microarray for expression detection prepared using our method have high specificity, good sensativity and stablity et al . It may be a more advanced method for analysis of gene expression profile.
文摘Sex determination and sex differentiation are important phenomena in fish, but the mechanisms of sex determination in Takifugu rubripes are poorly understood. In our study, the expression patterns of genes for DMRTs （DMRT1, DMRT2 and DMRT3）,sox9a and sox9b in T. rubripes tissues were verified with the Reverse Transcription （RT）-PCR detection. It is showed that DMRT1 expressions in testis and ovaries were much lower, and no expressions were fotmd in muscle, blood and tailfin. However, expressions for DMRT2 and DMRT3 were not found in the tissues stated above. Transcripts of sox9a were detected in muscle, fin, ovary and testis, but not in blood, whereas sox9b expression was only detected in ovary. The expression patterns of DMRTs, sox9a and sox9b in T. rubripes gonads suggest that these genes may not be sex-specific.
文摘OBJECTIVE To quantitatively explore the expression of Syndecan-1 and heparanase-1 in esophageal cancer tissue as well as their relationship with the clinicopathological factors, in order to evaluate their roles in tumor invasion and metastasis.METHODS Real-time fluorescence quantitative PCR （Q-PCR） was used to analyze the expression levels of Syndecan-1 and heparanase-1 genes￡？participants included 67 cases with esophageal cancers and 32 healthy volunteers.RESULTS The expression of Heparanase-1 gene in esophageal cancers was higher than that in normal esophageal tissue （P 〈 0.001）, and the expression of Syndecan-1 gene in the normal esophageal tissue was higher compared with esophageal cancers （P 〈 0.001）. The positive rates of Syndecan-1 and Heparanase-1 gene in esophageal cancer were 13.4% （9/67） and 85.1% （57/67）.The expression of Syndecan-1 and Heparanase-1 genes was signifi cantly related to di. erentiation, depth of infi ltration, lymph node metastasis, vessel metastasis, and TNM stages of disease （P 〈 0.05）. In an attempt to measure the association between the 2 agents, this study found that the expression of Syndecan-1 mRNA had a significantly negative correlation with the expression of Heparanase-1 mRNA by using Spearman rank correlation test （OR = -0.572, P 〈 0.001）.CONCLUSION Syndecan-1 and Heparanase-1 play important roles in the invasion and metastasis of esophageal cancer. The reduction of Syndecan-1 and/or the increase of Heparanase-1 may influence the invasion and metastasis of malignant tumors.Thus the combination assay of Syndecan-1 and Heparanase-1 may contribute to the diagnosis and treatment of malignant tumors.
文摘OBJECTIVE Tob is a member of Tob/BTG antiproliferative family. To date, Tob expression in human carcinoma using clinical specimens has not been studied in depth except for lung carcinoma and thyroid carcinoma. This study is the first to investigate the expression levels of Tob gene in human colorectal cancer tissues, and their corresponding para-cancerous tissues. The correlation of expression of the Tob gene with clinicopathological characteristics of colorectal cancer was also analyzed. METHODS Quantitative real time RT-PCR was used to detect the expression of Tob mRNA in 31 colorectal cancers. RESULTS Compared with normal tissues, up-regulation of Tob mRNA was observed in 31 colorectal cancer tissues （P = 0.020）. The expression level of Tob at Dukes C ＋ D phase was higher than Dukes A ＋ B phase, and the difference was significant （P 〈 0.05）. However, in this study, it was found that the expression of Tob mRNA was not related with age, gender, and pathological type of colorectal cancer. CONCLUSION The up-regulation of Tob may be closely associated with tumorigenesis of colorectal carcinoma.
基金the National Basic Research Programme of China(No.2006CB101803)the National Natural Science Foundation of China(No.30700016)
文摘Using PCR-denaturing gradient gel electrophoresis （DGGE） targeting the RNA polymerase beta subunit （rpoB） gene, a simultaneous detection method for Vibrio species was established, rpoB gene-based PCR-DGGE was carried out with eight Vibrio reference strains （each from different species）, mixed sample （including these Vibrio reference strains）, two non Vibrio strains, four environmental Vibrio strains, and three unidentified environmental strains. For comparison, 16S rRNA gene-based PCR-DGGE of the eight Vibrio reference strains was performed with universal primers. In addition, three unidentified strains were identified by 16S rRNA and gyrB gene sequencing and API20E system in order to confirm the accuracy of rpoB gene-based PCR-DGGE detection. Results revealed that rpoB-based PCR-DGGE could well discriminate eight Vibrio reference strains and could not discriminate different strains within the same species. The bands derived from two non Vibrio strains could not match with any bands in reference marker. Meanwhile, 16S rRNA gene-based DGGE failed to distinguish these reference strains. Further-more, four out of eight Vibrio species exhibited heterogenous bands in 16S rRNA gene-based DGGE. Sequencing and API 20E identification of unidentified strains coincided with the detection by rpoB gene-based PCR-DGGE. The results demonstrated that rpoB-based PCR-DGGE provided a rapid and efficient method for simultaneous detection of muhiple Vibrio species, which can avoid the limitations inherent in 16S rRNA gene-based PCR-DGGE.
基金National Natural Science Foundation of China(No.81673782)Shanghai Putuo District Health System“315”Project Talent Training Program(No.14Q-RC-11)。
文摘When gene expression profile is used for gene detection,the probe on the chip can emit fluorescence with different wavelengths.Under the action of confocal laser scanner,a clear gene change image can be obtained,by which the gene changes of the sample to be tested can be observed directly.First,the knee osteoarthritis(KOA)models of mice are established by the method of collateral ligament and meniscus resection(MLI-OA).Then,Bushen Huoxue formula is given by gavage,and ribonucleic acid(RNA)is routinely extracted and purified.Finally,the gene expression changes of KOA tissues of mice are detected by Agilent SurePrint G3 Mouse GE V2.0 gene expression profile.The results show that Bushen Huoxue formula has significant regulation effect on gene expression of KOA tissue.Among the genes with significant up-regulation effect of Bushen Huoxue formula,there are 56 genes of traditional Chinese medicine(TCM)groups up-regulated more than twice compared with model groups.Among the genes with significant down-regulation effect,there are 119 genes of TCM groups down-regulated more than twice compared with model groups.The experimental results indicate that Bushen Huoxue formula may promote the metabolism of arthritic factors and delay cartilage degeneration to treat KOA by regulating genes that are currently unknown in the pathological process of KOA.
文摘Objective:The aim of this study was to investigate the expression of BRAF of prostate cancer,and to explore its relations with clinic-pathological factors.Methods:Immunohistochemistry was used to detect the expression of BRAF in 74 cases of prostate cancer,and 51 cases of benign prostate hyperplasia(BPH).Results:The positive rate of BRAF had significant difference between patients with prostate cancer and BPH(P < 0.05).The expression of BRAF was correlated with grade and stage of prostate cancer(P < 0.05),but not to age of onset(P > 0.05).Conclusion:The expression of BRAF may play a role in the tumorigenesis and progression of colorectal cancer.BRAF could be a factor to diagnose the typing of prostate cancers and predict the prognosis of prostate cancers.
基金Supported by the Scientific and Technological Project in Shaanxi Province (2005K09-G6-2)
文摘Objective： To study the effect and mechanism of aspirin alone or combined with cisplatin （DDP） on human cervical carcinoma HeLa cells. Methods： HeLa cells were treated by different concentrations of aspirin, DDP alone or both. The inhibitory effect on cell growth was analyzed by MTT and colony-forming assay. Cell apoptosis was measured with flow cytometry. The mRNA levels ofBcl-2, Bax and NF-κB（P65） were studied by RT-PCR. Results： MTT assay showed that aspirin inhibited HeLa cell proliferation in a time-and dose-dependant maoner. Aspirin decreased clone numbers in colony formation assay. Aspirin also induced apoptosis of HeLa cells in a dose- and time-dependent manner as detected by flow cytometry. The inhibition effects on proliferation, colony formation and apoptosis were significantly enhanced when cells were treated with both aspirin and DDP. RT-PCR demonstrated that aspirin decreased the transcription of Bcl-2 and NF-κB, and increased expression of Bax gene. Conclusion： Aspirin can induce apoptosis in HeLa cells. Combination of aspirin and DDP displays a synergistic effect. The possible mechanism might be that aspirin downregulates the mRNA levels of Bcl-2 and NF-κB gene and upregulates the expression of Bax.
