DNA chip technology employs light- directed in situ oligonucleotide synthesis and/or DNA microarray printing device to produce arrays of large number of probes in the tiny surface of silicon substrates, which makes it...DNA chip technology employs light- directed in situ oligonucleotide synthesis and/or DNA microarray printing device to produce arrays of large number of probes in the tiny surface of silicon substrates, which makes it possible that the gene detection be conducted efficiently with high speed and sensitivity. The DNA chip may take important part in genome research, gene diagnoses and so on.展开更多
Objective: To identify the differential gene expression profiles between the normal and aspermia human testes utilizing cDNA microarray. Methods: cDNA probes were prepared by labeling mRNA of aspermia testes tissues w...Objective: To identify the differential gene expression profiles between the normal and aspermia human testes utilizing cDNA microarray. Methods: cDNA probes were prepared by labeling mRNA of aspermia testes tissues with Cy5-dUTP and mRNA of normal testes tissues with Cy3-dUTP respectively through reverse transcription. The mixed cDNA probes were then hybridized with 4096 cDNA arrays (4096 unique human cDNA sequences), and the fluorescent signals were scanned by ScanArray 3000 scanner (General Scanning, Inc.). The values of Cy5-dUTP and Cy3-dUTP on each spot were analyzed and calculated by ImaGene 3.0 software (BioDiscovery, Inc.). Differentially expressed genes were screened according to the criterion that the absolute value of natural logarithm of the ratio of Cy5-dUTP to Cy3-dUTP was greater-than 2.0 or less-than 0.5. A randomly chosen gene RAP1A was studied by in situ hybridization to evaluate the accuracy of the results. Results: 623 differential expressed genes related to aspermia were found. There were 303 up-expressed genes and 320 down-expressed genes. A distinct up-expressed gene RAP1A was confirmed by in situ hybridization. Conclusions: Screening the differential gene expression profiles between the normal and aspermia human testis by cDNA microarray can be used in the study of aspermia-related genes and the further research due to its properties, RAP1A may play some roles in the development and progression of aspermia.展开更多
Objective: To evaluate p63 expression at mRNA transcripts and protein levels in lung squamous cell cancer (SCC), adenocarcinoma, large cell lung cancer (LCLC) and small cell lung cancer (SCLC) and their matched metast...Objective: To evaluate p63 expression at mRNA transcripts and protein levels in lung squamous cell cancer (SCC), adenocarcinoma, large cell lung cancer (LCLC) and small cell lung cancer (SCLC) and their matched metastatic tumors. The association between p63 expression and p63 locus at chromosomal 3q27 q29 was also investigated. Methods: p63 mRNA expression levels in a large series of lung cancers including SCC, adenocarcinoma, LCLC, SCLC and their matched metastatic tumors were analyzed by cDNA microarray technology. A tissue microarray from 150 primary lung cancer specimens was constructed and used for immunohistochemical detection of p63 protein expression. Chromosomal imbalances at the p63 locus in 70 primary lung cancers samples were studied by comparative genomic hybridization (CGH) technology. Results: mRNA levels were 10 fold in SCC compared to LCLC, SCLC, and adenocarcinoma. Interestingly, the mRNA expression of p63 in metastatic carcinomas was significantly higher than that in their matched primary tumors ( P <0 001). Immunohistochemistry demonstrated that p63 expression was 94.64% in SCC but only 1 79% in lung adenocarcinoma and 2 of 4 LCLC were positive staining. All the results in of SCLC were negative. There was a statistically significant difference for p63 positivity between pT1 tumors and those of higher stage ( P =0 035). The CGH results indicated that p63 locus at chromosomal 3q27 q29 was overrepresented in SCC. p63 immunopositivity correlated significantly with pronounced gains of the p63 locus at chromosomal 3q27 q29 (P=0.0001), indicating that strong expression of p63 in lung SCC correlated with increased gene amplification. Conclusion: p63 might play an important role not only in squamous differentiation of lung cancer but also in tumor development and progression.展开更多
目的:探究厦门地区结核病患者对利福平与异烟肼的耐药情况及其耐药基因突变位点,为临床耐药结核病患者的抗结核治疗提供参考。方法:选取2019年1月—2020年12月厦门大学附属第一医院收治的1029例结核病患者作为研究对象,采集患者的各种...目的:探究厦门地区结核病患者对利福平与异烟肼的耐药情况及其耐药基因突变位点,为临床耐药结核病患者的抗结核治疗提供参考。方法:选取2019年1月—2020年12月厦门大学附属第一医院收治的1029例结核病患者作为研究对象,采集患者的各种临床标本并通过DNA微阵列芯片技术检测结核分枝杆菌(Mycobacterium tuberculosis,MTB)对利福平与异烟肼的耐药情况及其耐药基因突变位点的分布情况。结果:1029例结核病患者分离出的MTB对利福平和/或异烟肼的总耐药率为18.56%(191/1029);男性患者对利福平和/或异烟肼的总耐药率略高于女性(19.24%vs 16.60%,P>0.05);在各年龄段中,40~<60岁患者对利福平和/或异烟肼的总耐药率最高(19.80%),而<20岁患者对利福平和/或异烟肼的总耐药率最低(12.82%),但各年龄段结核病患者对利福平和/或异烟肼的总耐药率经比较其差异无统计学意义(χ^(2)=1.553,P>0.05);共有127例患者的利福平耐药相关基因rpo B发生突变,突变率为12.34%,位点531(C→T)突变频率最高(为52.76%),共有141例患者的异烟肼耐药相关基因发生突变,其中kat G基因突变100例,突变率为9.72%,位点315(G→C)突变频率最高(为68.09%),inh A基因启动子突变40例,突变率为3.89%,kat G基因和inh A基因启动子同时突变1例;1029例结核病患者中耐多药结核病(multidrug-resistant tuberculosis,MDR-TB)的发生率为7.48%,基因rpo B 531(C→T)-kat G 315(G→C)的突变率最高(为45.45%),有1例患者的MTB的rpo B、kat G和inh A基因均发生了突变。结论:厦门地区MTB耐多药和利福平耐药的形势严峻,rpo B 531、kat G 315和rpo B 531-kat G 315是其利福平耐药、异烟肼耐药和耐多药的主要突变位点。展开更多
Polyurethane based on polypropylene glycol(PPG) and TDI using 3,3\|dichloride\|4,4\|methylenedianiline(MOCA) as the crosslinker is presented for the first time to fabricate molecular stamps(PU stamps) for the synt...Polyurethane based on polypropylene glycol(PPG) and TDI using 3,3\|dichloride\|4,4\|methylenedianiline(MOCA) as the crosslinker is presented for the first time to fabricate molecular stamps(PU stamps) for the synthesis of DNA microarray with contact procedure. The predictability of the process is achieved by utilizing commercially available starting materials. SEM analysis of the morphology of PU stamps and photoresist motherboard showed that PU elastomer could replicate subtly the motherboards patterns with a high fidelity. It was proved from the contact angle measurement that PU stamps surface has a good affinity with acetonitrile, which guarantee the well\|distribution of DNA monomers on patterned stamps. Laser confocal fluorescence microscopy images of oligonucleotide arrays confirm that polyurethane is an excellent material for molecular stamps.展开更多
基金This work was supported by the China National Science Foundation (No.39880032)
文摘DNA chip technology employs light- directed in situ oligonucleotide synthesis and/or DNA microarray printing device to produce arrays of large number of probes in the tiny surface of silicon substrates, which makes it possible that the gene detection be conducted efficiently with high speed and sensitivity. The DNA chip may take important part in genome research, gene diagnoses and so on.
文摘Objective: To identify the differential gene expression profiles between the normal and aspermia human testes utilizing cDNA microarray. Methods: cDNA probes were prepared by labeling mRNA of aspermia testes tissues with Cy5-dUTP and mRNA of normal testes tissues with Cy3-dUTP respectively through reverse transcription. The mixed cDNA probes were then hybridized with 4096 cDNA arrays (4096 unique human cDNA sequences), and the fluorescent signals were scanned by ScanArray 3000 scanner (General Scanning, Inc.). The values of Cy5-dUTP and Cy3-dUTP on each spot were analyzed and calculated by ImaGene 3.0 software (BioDiscovery, Inc.). Differentially expressed genes were screened according to the criterion that the absolute value of natural logarithm of the ratio of Cy5-dUTP to Cy3-dUTP was greater-than 2.0 or less-than 0.5. A randomly chosen gene RAP1A was studied by in situ hybridization to evaluate the accuracy of the results. Results: 623 differential expressed genes related to aspermia were found. There were 303 up-expressed genes and 320 down-expressed genes. A distinct up-expressed gene RAP1A was confirmed by in situ hybridization. Conclusions: Screening the differential gene expression profiles between the normal and aspermia human testis by cDNA microarray can be used in the study of aspermia-related genes and the further research due to its properties, RAP1A may play some roles in the development and progression of aspermia.
文摘Objective: To evaluate p63 expression at mRNA transcripts and protein levels in lung squamous cell cancer (SCC), adenocarcinoma, large cell lung cancer (LCLC) and small cell lung cancer (SCLC) and their matched metastatic tumors. The association between p63 expression and p63 locus at chromosomal 3q27 q29 was also investigated. Methods: p63 mRNA expression levels in a large series of lung cancers including SCC, adenocarcinoma, LCLC, SCLC and their matched metastatic tumors were analyzed by cDNA microarray technology. A tissue microarray from 150 primary lung cancer specimens was constructed and used for immunohistochemical detection of p63 protein expression. Chromosomal imbalances at the p63 locus in 70 primary lung cancers samples were studied by comparative genomic hybridization (CGH) technology. Results: mRNA levels were 10 fold in SCC compared to LCLC, SCLC, and adenocarcinoma. Interestingly, the mRNA expression of p63 in metastatic carcinomas was significantly higher than that in their matched primary tumors ( P <0 001). Immunohistochemistry demonstrated that p63 expression was 94.64% in SCC but only 1 79% in lung adenocarcinoma and 2 of 4 LCLC were positive staining. All the results in of SCLC were negative. There was a statistically significant difference for p63 positivity between pT1 tumors and those of higher stage ( P =0 035). The CGH results indicated that p63 locus at chromosomal 3q27 q29 was overrepresented in SCC. p63 immunopositivity correlated significantly with pronounced gains of the p63 locus at chromosomal 3q27 q29 (P=0.0001), indicating that strong expression of p63 in lung SCC correlated with increased gene amplification. Conclusion: p63 might play an important role not only in squamous differentiation of lung cancer but also in tumor development and progression.
文摘目的:探究厦门地区结核病患者对利福平与异烟肼的耐药情况及其耐药基因突变位点,为临床耐药结核病患者的抗结核治疗提供参考。方法:选取2019年1月—2020年12月厦门大学附属第一医院收治的1029例结核病患者作为研究对象,采集患者的各种临床标本并通过DNA微阵列芯片技术检测结核分枝杆菌(Mycobacterium tuberculosis,MTB)对利福平与异烟肼的耐药情况及其耐药基因突变位点的分布情况。结果:1029例结核病患者分离出的MTB对利福平和/或异烟肼的总耐药率为18.56%(191/1029);男性患者对利福平和/或异烟肼的总耐药率略高于女性(19.24%vs 16.60%,P>0.05);在各年龄段中,40~<60岁患者对利福平和/或异烟肼的总耐药率最高(19.80%),而<20岁患者对利福平和/或异烟肼的总耐药率最低(12.82%),但各年龄段结核病患者对利福平和/或异烟肼的总耐药率经比较其差异无统计学意义(χ^(2)=1.553,P>0.05);共有127例患者的利福平耐药相关基因rpo B发生突变,突变率为12.34%,位点531(C→T)突变频率最高(为52.76%),共有141例患者的异烟肼耐药相关基因发生突变,其中kat G基因突变100例,突变率为9.72%,位点315(G→C)突变频率最高(为68.09%),inh A基因启动子突变40例,突变率为3.89%,kat G基因和inh A基因启动子同时突变1例;1029例结核病患者中耐多药结核病(multidrug-resistant tuberculosis,MDR-TB)的发生率为7.48%,基因rpo B 531(C→T)-kat G 315(G→C)的突变率最高(为45.45%),有1例患者的MTB的rpo B、kat G和inh A基因均发生了突变。结论:厦门地区MTB耐多药和利福平耐药的形势严峻,rpo B 531、kat G 315和rpo B 531-kat G 315是其利福平耐药、异烟肼耐药和耐多药的主要突变位点。
文摘Polyurethane based on polypropylene glycol(PPG) and TDI using 3,3\|dichloride\|4,4\|methylenedianiline(MOCA) as the crosslinker is presented for the first time to fabricate molecular stamps(PU stamps) for the synthesis of DNA microarray with contact procedure. The predictability of the process is achieved by utilizing commercially available starting materials. SEM analysis of the morphology of PU stamps and photoresist motherboard showed that PU elastomer could replicate subtly the motherboards patterns with a high fidelity. It was proved from the contact angle measurement that PU stamps surface has a good affinity with acetonitrile, which guarantee the well\|distribution of DNA monomers on patterned stamps. Laser confocal fluorescence microscopy images of oligonucleotide arrays confirm that polyurethane is an excellent material for molecular stamps.
