The effects of BTHP on Ca 2+ independent action potential and the two components of delayed rectifier potassium currents were studied in guinea pig single ventricular myocytes by using whole cell patch clamp tec...The effects of BTHP on Ca 2+ independent action potential and the two components of delayed rectifier potassium currents were studied in guinea pig single ventricular myocytes by using whole cell patch clamp technique. BTHP 30 μmol·L -1 significantly prolonged APD 90 from 143±16 ms to 184±21 ms ( P 【0.01, n=5) without affecting either the RP or APA, and the APD prolonging effects of BTHP were independent of extracellular Ca 2+ . BTHP inhibited both I kr (IC 50 =7 9 μmol·L -1 ) and I ks (IC 50 =22 4 μmol·L -1 ) in a concentration dependent fashion. The results demon strated that BTHP had no obvious selectivity for I kr and I ks .展开更多
Rapid,high-throughput and reliable methods are urgently required to accurately detect and monitor harmful algae,which are responsible for algal blooms,such as red and green tides. In this study,we successfully develop...Rapid,high-throughput and reliable methods are urgently required to accurately detect and monitor harmful algae,which are responsible for algal blooms,such as red and green tides. In this study,we successfully developed a multiplex PCR-based DNA microarray method capable of detecting nine harmful algal species simultaneously,namely A lexandrium tamarense,Gyrodinium instriatum,Heterosigma akashiwo,Karenia mikimotoi,Prorocentrum donghaiense,Prorocentrum minimum,Ulva compressa,Ulva ohnoi and Ulva prolifera. This method achieved a limit of detection(LOD) of 0.5 ng of genomic DNA(orders of magnitude of the deci-nanogram range) in the tested algae cultures. Altogether,230 field samples from ship ballast waters and seaport waters were used to evaluate the DNA microarray. The clinical sensitivity and specificity of the DNA microarray assay in detecting field samples were 96.4% and 90.9%,respectively,relative to conventional morphological methods. This indicated that this high-throughput,automatic,and specific method is well suited for the detection of algae in water samples.展开更多
Objective: The aim of the study was to observe the expressions of genes related to genome stability and DNA repair in the members of nasopharyngeal carcinoma (NPC) clustedng families. Methods: In the Zhongshan Cit...Objective: The aim of the study was to observe the expressions of genes related to genome stability and DNA repair in the members of nasopharyngeal carcinoma (NPC) clustedng families. Methods: In the Zhongshan City where there is highly incidence rate of NPC, we chose the members of the NPC clustering families as objects, and the patients of nasopharyngitis and NPC as the control group. We isolated the RNA from the nasopharyngeal tissue, and synthesized its cRNA, the genome stability and DNA repair genes chip technique, chemiluminescent detection and real-time fluorescence quantita- tive technique were used to examine the genome stability and DNA repair genes in the nasopharyngeal tissue. Results: More genome stability and DNA repair genes were up-regulated in the members of the NPC clustering families than the NPC patients, and the range of up-regulated was high, with the over up-regulated 100 times genes including TEP1, MSH4, PMS2LI. Fewer genome stability and DNA repair genes were down-regulated in the members of the NPC clustering families than the NPC patients, the ubiquitin genes almost were down-regulated, the results also could be confirmed by real-time fluorescence quantitative PCR. Conclusion: There are specially expression character of genome stability and DNA repair genes in the members of NPC clustering families.展开更多
AIM:To assess human epidermal growth factor receptor-2 (HER2)-status in gastric cancer and matched lymph node metastases by immunohistochemistry (IHC) and chromogenic in situ hybridization (CISH).METHODS:120 cases of ...AIM:To assess human epidermal growth factor receptor-2 (HER2)-status in gastric cancer and matched lymph node metastases by immunohistochemistry (IHC) and chromogenic in situ hybridization (CISH).METHODS:120 cases of primary gastric carcinomas and 45 matched lymph node metastases from patients with full clinicopathological features were mounted onto multiple-punch and single-punch tissue microarrays,respectively,and examined for HER2 overexpression and gene amplification by IHC and CISH.RESULTS:Twenty-four tumors (20%) expressed HER2 immunohistochemically.An IHC score of ≥ 2+ was observed in 20 tumors (16.6%).HER2 amplification was detected by CISH in 19 tumors (15.8%) and in their matched lymph node metastases.A high concordancerate was found between HER2 positivity (as detected by IHC) and HER2 gene amplification (as detected by CISH),since 19 of the 20 IHC positive cases were amplified (95%).All amplified cases had 2+ or 3+ IHC results.Amplification was associated with intestinal phenotype (P < 0.05).No association with grading,staging or survival was found.CONCLUSION:In gastric cancer,HER2 amplification is the main mechanism for HER2 protein overexpression and is preserved in lymph node metastases.展开更多
OBJECTIVE This study aimed to explore the expression and significance of transforming growth factor β1(TGF-β1),extracellular signal-regulated kinases 1/2 (ERK1/2), and K-ras in colorectal cancer (CRC) using ti...OBJECTIVE This study aimed to explore the expression and significance of transforming growth factor β1(TGF-β1),extracellular signal-regulated kinases 1/2 (ERK1/2), and K-ras in colorectal cancer (CRC) using tissue microarray technology.METHODS The expressions of TGF-β1, ERK1/2, and K-ras in colon cancer cells taken from the specimens of 92 CRC patients (stage Ⅰ: 16 cases, stage Ⅱ: 28 cases, stage Ⅲ: 24 cases, and stage Ⅳ:24 cases) were analyzed using tissue microarray technology and immunohistochemistry, and compared with those of 20 normal colon tissue samples.RESULTS High immunoreactive scores (IRS) of TGF-β1,p-ERK1/2, and K-ras protein in CRC were obtained, which were 66.3% (61/92), 59.8% (55/92), and 48.9% (45/92), respectively, and those in normal epithelial cells of colon were 10% (2/20), 20% (4/20), and 30% (6/20), respectively (P 〈 0.05). The expressions of TGF-β1 and ERK1/2 in CRC at stage Ⅰwere 37.5% and 31.3%,respectively, and those in CRC at stage Ⅳ were 83.3% and79.3%, respectively, with statistically significant differences. No significant relationship was found between K-ras expression and tumor stages (P〉0.05).CONCLUSION High level expressions of TGF-β1 and ERK1/2 are closely related to the clinical stages of colon cancer and crosstalk may exist between the 2 signal pathways.展开更多
Microarray technologies are widely used all over the world for the gene expression analysis in various tissues, but still this technology has some limitations. The problem of eliminated reproducibility of the results,...Microarray technologies are widely used all over the world for the gene expression analysis in various tissues, but still this technology has some limitations. The problem of eliminated reproducibility of the results, obtained in different laboratories using different platforms, is very relevant nowadays. For revelation of problems ofmicroarrays, the comparative analysis of hepatic and renal gene expression profiles (GEP) was carried out by using swine protein annotated oligonucleotide microarrays (SPAM). Revealed differences in GEP between kidney and liver of pigs were correlated with functional and histological distinctions of these organs. It was shown that sources of errors in the comparative analysis of organ-specific GEP could be connected to the cross hybridization of one probe to transcripts (cDNA of mRNA) of different genes and to individual variability in gene expression between animals, related with the changeability of influences of exo- and endogenous regulation factors.展开更多
Based on rpoB gene micro array as target gene, we are going to use gene chip technology to test 24 mycobacterium standard specimens, 8 non-mycobacterium specimens and 86 mycobacterium clinical isolated specimens. As a...Based on rpoB gene micro array as target gene, we are going to use gene chip technology to test 24 mycobacterium standard specimens, 8 non-mycobacterium specimens and 86 mycobacterium clinical isolated specimens. As a result, after mycobacterium and non-mycobacterium standard specimens were duplicated by PCR, mycobacterium standard specimens reproduced 360bp DNA fragments; on the other hand, non-mycobacterium specimens did not reproduce any fragments except for hemolytic streptococcus and corynebacterium pseudodiphtheriticum which had the same results as mycobacterium standard specimens. Sensitive test is able to detect lpg tuberculosis mycobacterium DNA. The probe test showed that, among 21 oligonucleotide probes, probe-M. fortuitum and M. marinum were cross-hybrid; the other probes were specific. We used the new method to identify 126 mycobacterium clinical isolated specimens. The test results of this new method matched with conventional method. In conclusion, compared to the traditional method, the use of rpob gene chip technology to identify mycobacterium species will be faster, more accurate and higher value in application.展开更多
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
Gene expression profiling using cDNA or high-density oligonucleotide microarray contributes signifi cantly to our understanding on the transcriptome of a given biological condition. Using this technology, huge number...Gene expression profiling using cDNA or high-density oligonucleotide microarray contributes signifi cantly to our understanding on the transcriptome of a given biological condition. Using this technology, huge number of differentially-expressed genes of interest have been identified in a broad range of circumstances. Making sense biologically on these genes using the recently-improved functional annotation and data integration has leveraged our understanding in diseases and their biological mechanisms. However, understanding the codes encrypt- ed in the cis-aeting regulatory regions and gaining insights into the circuitry of functional regulatory networks on the genomic scale will require additional empirical data sets that are capable of revealing the cohorts or regulons of the transcription and the dynamic progression of molecular events responsible for certain biological function.展开更多
OBJECTIVE:To explore electro-acupuncture's(EA's)effect on gene expression in heart of rats with stress-induced pre-hypertension and try to reveal its biological mechanism based on gene chip technology.METHODS:...OBJECTIVE:To explore electro-acupuncture's(EA's)effect on gene expression in heart of rats with stress-induced pre-hypertension and try to reveal its biological mechanism based on gene chip technology.METHODS:Twenty-seven Wistar male rats were randomly divided into 3 groups.The stress-induced hypertensive rat model was prepared by electric foot-shocks combined with generated noise.Molding cycle lasted for 14 days and EA intervene was applied on rats in model + EA group during model preparation.Rat Gene 2.0 Sense Target Array technology was used for the determination of gene expression profiles and the screened key genes were verified by real-time quantitative polymerase chain reaction(RT-PCR) method.RESULTS:Compared with blank control group,390 genes were changed in model group;compared with model control group,330 genes were changed in model + EA group.Significance analysis of gene function showed that the differentially expressed genes are those involved in biological process,molecular function and cellular components.RT-PCR result of the screened key genes is consistent with that of gene chip test.CONCLUTION:EA could significantly lower blood pressure of stress-induced pre-hypertension rats and affect its gene expression profile in heart.Genes that related to the contraction of vascular smooth muscle may be involved in EA's anti-hypertensive mechanism.展开更多
Microarray technology has been proved to be greatly helpful for biomedical and biological diagnosis. And the evaluation of its biological applications lies in the detection sensitivity, which requires high intensity a...Microarray technology has been proved to be greatly helpful for biomedical and biological diagnosis. And the evaluation of its biological applications lies in the detection sensitivity, which requires high intensity and stability of the signal. Recently, several nanomaterials, especially semiconductor nanomaterials, due to their excellent fluorescence properties, have been widely used to construct microarrays for biosensors. Here, we presented an approach for constructing CdSe/ZnS quantum dot (QD) microarray in microfluidic channels on a glass slide by photolithography. The conditions for immobilizing stable and uniform QD microarray on the glass slide were optimized. Several types of QD microarrays with different emission wavelengths and modified groups were constructed using silanization and lithography technology. Based on the fluorescence quenching effect of Cu2+ on QDs, the microfluidic chip with QD microarray was applied for the determination of Cu2+. 1 nmol/L Cu2+ could be detected by this method.展开更多
OBJECTIVE:To investigate the molecular effect of Socheongryong Tang(SCRT,Xiaoqinglong Tang in Chinese) on whole genome level in asthma mouse model by microarray technology.METHODS:Asthma was induced by intranasal inst...OBJECTIVE:To investigate the molecular effect of Socheongryong Tang(SCRT,Xiaoqinglong Tang in Chinese) on whole genome level in asthma mouse model by microarray technology.METHODS:Asthma was induced by intranasal instillation of ovalbumin in mouse.After administration of SCRT on asthma-induced mouse,the expression of genes in lung tissue was measured using whole genome microarray.The functional implication of differentially expressed genes was performed using ontological analysis and the similarity of promoter structure of genes was also analyzed.RESULTS:Treatment of SCRT restored expression level of many up- or down-regulated genes in asthma model,and this recovery rate means SCRT could regulate a set of genes having specific TFBS binding sites.CONCLUSION:In this study,we identified a set of genes subjected to similar regulation by SCRT in asthma model in mice.展开更多
文摘The effects of BTHP on Ca 2+ independent action potential and the two components of delayed rectifier potassium currents were studied in guinea pig single ventricular myocytes by using whole cell patch clamp technique. BTHP 30 μmol·L -1 significantly prolonged APD 90 from 143±16 ms to 184±21 ms ( P 【0.01, n=5) without affecting either the RP or APA, and the APD prolonging effects of BTHP were independent of extracellular Ca 2+ . BTHP inhibited both I kr (IC 50 =7 9 μmol·L -1 ) and I ks (IC 50 =22 4 μmol·L -1 ) in a concentration dependent fashion. The results demon strated that BTHP had no obvious selectivity for I kr and I ks .
基金Supported by the National High Technology Research and Development Program of China(863 Program)(No.2012AA092001)the Science and Technology Project of Zhejiang Province(No.2013C03045-1)+2 种基金the Zhejiang Marine Biotechnology Innovation Team(No.2010R50029-12)the Natural Science Foundation of Ningbo City of China(No.2013A610168)the KC Wong Magna Fund in Ningbo University
文摘Rapid,high-throughput and reliable methods are urgently required to accurately detect and monitor harmful algae,which are responsible for algal blooms,such as red and green tides. In this study,we successfully developed a multiplex PCR-based DNA microarray method capable of detecting nine harmful algal species simultaneously,namely A lexandrium tamarense,Gyrodinium instriatum,Heterosigma akashiwo,Karenia mikimotoi,Prorocentrum donghaiense,Prorocentrum minimum,Ulva compressa,Ulva ohnoi and Ulva prolifera. This method achieved a limit of detection(LOD) of 0.5 ng of genomic DNA(orders of magnitude of the deci-nanogram range) in the tested algae cultures. Altogether,230 field samples from ship ballast waters and seaport waters were used to evaluate the DNA microarray. The clinical sensitivity and specificity of the DNA microarray assay in detecting field samples were 96.4% and 90.9%,respectively,relative to conventional morphological methods. This indicated that this high-throughput,automatic,and specific method is well suited for the detection of algae in water samples.
基金Supported by a grant from the National Natural Science Foundation of China (No.30672713)
文摘Objective: The aim of the study was to observe the expressions of genes related to genome stability and DNA repair in the members of nasopharyngeal carcinoma (NPC) clustedng families. Methods: In the Zhongshan City where there is highly incidence rate of NPC, we chose the members of the NPC clustering families as objects, and the patients of nasopharyngitis and NPC as the control group. We isolated the RNA from the nasopharyngeal tissue, and synthesized its cRNA, the genome stability and DNA repair genes chip technique, chemiluminescent detection and real-time fluorescence quantita- tive technique were used to examine the genome stability and DNA repair genes in the nasopharyngeal tissue. Results: More genome stability and DNA repair genes were up-regulated in the members of the NPC clustering families than the NPC patients, and the range of up-regulated was high, with the over up-regulated 100 times genes including TEP1, MSH4, PMS2LI. Fewer genome stability and DNA repair genes were down-regulated in the members of the NPC clustering families than the NPC patients, the ubiquitin genes almost were down-regulated, the results also could be confirmed by real-time fluorescence quantitative PCR. Conclusion: There are specially expression character of genome stability and DNA repair genes in the members of NPC clustering families.
文摘AIM:To assess human epidermal growth factor receptor-2 (HER2)-status in gastric cancer and matched lymph node metastases by immunohistochemistry (IHC) and chromogenic in situ hybridization (CISH).METHODS:120 cases of primary gastric carcinomas and 45 matched lymph node metastases from patients with full clinicopathological features were mounted onto multiple-punch and single-punch tissue microarrays,respectively,and examined for HER2 overexpression and gene amplification by IHC and CISH.RESULTS:Twenty-four tumors (20%) expressed HER2 immunohistochemically.An IHC score of ≥ 2+ was observed in 20 tumors (16.6%).HER2 amplification was detected by CISH in 19 tumors (15.8%) and in their matched lymph node metastases.A high concordancerate was found between HER2 positivity (as detected by IHC) and HER2 gene amplification (as detected by CISH),since 19 of the 20 IHC positive cases were amplified (95%).All amplified cases had 2+ or 3+ IHC results.Amplification was associated with intestinal phenotype (P < 0.05).No association with grading,staging or survival was found.CONCLUSION:In gastric cancer,HER2 amplification is the main mechanism for HER2 protein overexpression and is preserved in lymph node metastases.
基金This work was supported by grants from Soochow University Students Innovation Foundation (No. 5731512810), Chinese National Natural Science Foundation (No.81072186), and Jiangsu Provincial Higher Institution Natural Science Foundation (No.10KJB320018).
文摘OBJECTIVE This study aimed to explore the expression and significance of transforming growth factor β1(TGF-β1),extracellular signal-regulated kinases 1/2 (ERK1/2), and K-ras in colorectal cancer (CRC) using tissue microarray technology.METHODS The expressions of TGF-β1, ERK1/2, and K-ras in colon cancer cells taken from the specimens of 92 CRC patients (stage Ⅰ: 16 cases, stage Ⅱ: 28 cases, stage Ⅲ: 24 cases, and stage Ⅳ:24 cases) were analyzed using tissue microarray technology and immunohistochemistry, and compared with those of 20 normal colon tissue samples.RESULTS High immunoreactive scores (IRS) of TGF-β1,p-ERK1/2, and K-ras protein in CRC were obtained, which were 66.3% (61/92), 59.8% (55/92), and 48.9% (45/92), respectively, and those in normal epithelial cells of colon were 10% (2/20), 20% (4/20), and 30% (6/20), respectively (P 〈 0.05). The expressions of TGF-β1 and ERK1/2 in CRC at stage Ⅰwere 37.5% and 31.3%,respectively, and those in CRC at stage Ⅳ were 83.3% and79.3%, respectively, with statistically significant differences. No significant relationship was found between K-ras expression and tumor stages (P〉0.05).CONCLUSION High level expressions of TGF-β1 and ERK1/2 are closely related to the clinical stages of colon cancer and crosstalk may exist between the 2 signal pathways.
文摘Microarray technologies are widely used all over the world for the gene expression analysis in various tissues, but still this technology has some limitations. The problem of eliminated reproducibility of the results, obtained in different laboratories using different platforms, is very relevant nowadays. For revelation of problems ofmicroarrays, the comparative analysis of hepatic and renal gene expression profiles (GEP) was carried out by using swine protein annotated oligonucleotide microarrays (SPAM). Revealed differences in GEP between kidney and liver of pigs were correlated with functional and histological distinctions of these organs. It was shown that sources of errors in the comparative analysis of organ-specific GEP could be connected to the cross hybridization of one probe to transcripts (cDNA of mRNA) of different genes and to individual variability in gene expression between animals, related with the changeability of influences of exo- and endogenous regulation factors.
文摘Based on rpoB gene micro array as target gene, we are going to use gene chip technology to test 24 mycobacterium standard specimens, 8 non-mycobacterium specimens and 86 mycobacterium clinical isolated specimens. As a result, after mycobacterium and non-mycobacterium standard specimens were duplicated by PCR, mycobacterium standard specimens reproduced 360bp DNA fragments; on the other hand, non-mycobacterium specimens did not reproduce any fragments except for hemolytic streptococcus and corynebacterium pseudodiphtheriticum which had the same results as mycobacterium standard specimens. Sensitive test is able to detect lpg tuberculosis mycobacterium DNA. The probe test showed that, among 21 oligonucleotide probes, probe-M. fortuitum and M. marinum were cross-hybrid; the other probes were specific. We used the new method to identify 126 mycobacterium clinical isolated specimens. The test results of this new method matched with conventional method. In conclusion, compared to the traditional method, the use of rpob gene chip technology to identify mycobacterium species will be faster, more accurate and higher value in application.
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Gene expression profiling using cDNA or high-density oligonucleotide microarray contributes signifi cantly to our understanding on the transcriptome of a given biological condition. Using this technology, huge number of differentially-expressed genes of interest have been identified in a broad range of circumstances. Making sense biologically on these genes using the recently-improved functional annotation and data integration has leveraged our understanding in diseases and their biological mechanisms. However, understanding the codes encrypt- ed in the cis-aeting regulatory regions and gaining insights into the circuitry of functional regulatory networks on the genomic scale will require additional empirical data sets that are capable of revealing the cohorts or regulons of the transcription and the dynamic progression of molecular events responsible for certain biological function.
基金the National Natural Science Foundation of China(Based on the Systems Biology of Acupuncture on Irritable Early Hypertension Control Mechanism and Intervention Study Targets Research,No.81072861)
文摘OBJECTIVE:To explore electro-acupuncture's(EA's)effect on gene expression in heart of rats with stress-induced pre-hypertension and try to reveal its biological mechanism based on gene chip technology.METHODS:Twenty-seven Wistar male rats were randomly divided into 3 groups.The stress-induced hypertensive rat model was prepared by electric foot-shocks combined with generated noise.Molding cycle lasted for 14 days and EA intervene was applied on rats in model + EA group during model preparation.Rat Gene 2.0 Sense Target Array technology was used for the determination of gene expression profiles and the screened key genes were verified by real-time quantitative polymerase chain reaction(RT-PCR) method.RESULTS:Compared with blank control group,390 genes were changed in model group;compared with model control group,330 genes were changed in model + EA group.Significance analysis of gene function showed that the differentially expressed genes are those involved in biological process,molecular function and cellular components.RT-PCR result of the screened key genes is consistent with that of gene chip test.CONCLUTION:EA could significantly lower blood pressure of stress-induced pre-hypertension rats and affect its gene expression profile in heart.Genes that related to the contraction of vascular smooth muscle may be involved in EA's anti-hypertensive mechanism.
基金supported by the National Basic Research Program of China (2011CB933600)the Science Fund for Creative Research Groups (20921062)+1 种基金the National Natural Science Foundation of China (21175100)the Program for New Century Excellent Talents in University (NCET-10-0656)
文摘Microarray technology has been proved to be greatly helpful for biomedical and biological diagnosis. And the evaluation of its biological applications lies in the detection sensitivity, which requires high intensity and stability of the signal. Recently, several nanomaterials, especially semiconductor nanomaterials, due to their excellent fluorescence properties, have been widely used to construct microarrays for biosensors. Here, we presented an approach for constructing CdSe/ZnS quantum dot (QD) microarray in microfluidic channels on a glass slide by photolithography. The conditions for immobilizing stable and uniform QD microarray on the glass slide were optimized. Several types of QD microarrays with different emission wavelengths and modified groups were constructed using silanization and lithography technology. Based on the fluorescence quenching effect of Cu2+ on QDs, the microfluidic chip with QD microarray was applied for the determination of Cu2+. 1 nmol/L Cu2+ could be detected by this method.
文摘OBJECTIVE:To investigate the molecular effect of Socheongryong Tang(SCRT,Xiaoqinglong Tang in Chinese) on whole genome level in asthma mouse model by microarray technology.METHODS:Asthma was induced by intranasal instillation of ovalbumin in mouse.After administration of SCRT on asthma-induced mouse,the expression of genes in lung tissue was measured using whole genome microarray.The functional implication of differentially expressed genes was performed using ontological analysis and the similarity of promoter structure of genes was also analyzed.RESULTS:Treatment of SCRT restored expression level of many up- or down-regulated genes in asthma model,and this recovery rate means SCRT could regulate a set of genes having specific TFBS binding sites.CONCLUSION:In this study,we identified a set of genes subjected to similar regulation by SCRT in asthma model in mice.