BACKGROUND: The good therapeutic effects of large dose of dexamethasone on severe acute pancreatitis (SAP) patients have been proved. This study was designed to investigate the influence of dexamethasone on apoptosis ...BACKGROUND: The good therapeutic effects of large dose of dexamethasone on severe acute pancreatitis (SAP) patients have been proved. This study was designed to investigate the influence of dexamethasone on apoptosis of acinar cells in the pancreas of rats with SAP and the protein expression of the apoptosis-regulating genes Bax and Bcl-2. METHODS: Ninety Sprague-Dawley rats with SAP were randomly divided into a model group and a dexamethasone treated group (45 rats in each group), and another 45 rats formed the sham operation group. Survival rates were calculated and gross pathological changes in the pancreas of each group were observed under a light microscope 3, 6 and 12 hours after operation. Tissue microarray technology was applied to prepare pancreatic tissue sections. The changes in Bax and Bcl-2 protein expression levels of pancreatic tissues from each group were assessed by immunohistochemical staining, and TUNEL staining was used to evaluate changes in apoptosis index. RESULTS: The model and treated groups did not differ in mortality at each time point. The pathological score for the pancreas in the treated group was significantly lower than that in the model group at 3 and 6 hours. The positive rates of Bax protein expression in the head and tail of the pancreas in the treated group at all time points were all markedly higher than those of the model group. The positive rate of Bcl-2 protein expression in the head of the pancreas in the treated group was significantly higher than that of the model group at 3 hours. TUNEL staining showed that the pancreas head and tail apoptosis indices of the treated group were markedly higher than those of the model group after 6 hours. CONCLUSIONS: Apoptosis may be a protective response. to pancreatic cell injury. The mechanism of action of dexamethasone in treating SAP may be related to the apoptosis of acinar cells in the pancreas induced by apoptosis-regulating genes such as Bax and Bcl-2. The advantages of tissue microarrays in pathological examination of the pancreas include saving of time and energy, efficiency and highly representative.展开更多
The genome sequence information in combination with DNA microarrays promises to revolutionize the way of cellu-lar and molecular biological research by allowing complex mixtures of RNA and DNA to interrogated in a par...The genome sequence information in combination with DNA microarrays promises to revolutionize the way of cellu-lar and molecular biological research by allowing complex mixtures of RNA and DNA to interrogated in a parallel and quantita-tive fashion. DNA microarrays can be used to measure levels of gene expression for tens of thousands of gene simultane-ously and take advantage of all available sequence information for experimental design and data interpretation in pursuit of biological understanding. Recent progress in experimental genomics allows DNA microarrays not simply to provide a cata-logue of all the genes and information about their function, but to understand how the components work together to comprise functioning cells and organisms. This brief review gives a survey of DNA microarrays technology and its applications in ge-nome and gene function analysis, gene expression studies, biological signal and defense system, cell cycle regulation, mechanism of transcriptional regulation, proteomics, and the functionality of food component.展开更多
AIM: To study the pathogenetic processes and the role of gene expression by microarray analyses in expediting our understanding of the molecular pathophysiology of pancreatic adenocarcinoma, and to identify the novel ...AIM: To study the pathogenetic processes and the role of gene expression by microarray analyses in expediting our understanding of the molecular pathophysiology of pancreatic adenocarcinoma, and to identify the novel cancer-associated genes.METHODS: Nine histologically defined pancreatic head adenocarcinoma specimens associated with clinical data were studied. Total RNA and mRNA were isolated and labeled by reverse transcription reaction with Cy5 and Cy3 for cDNA probe. The cDNA microarrays that represent a set of 4 096 human genes were hybridized with labeled cDNA probe and screened for molecular profiling analyses.RESULTS: Using this methodology, 184 genes were screened out for differences in gene expression level after nine couples of hybridizations. Of the 184 genes,87 were upregulated and 97 downregulated, including 11 novel human genes. In pancreatic adenocarcinoma tissue, several invasion and metastasis related genes showed their high expression levels, suggesting that poor prognosis of pancreatic adenocarcinoma might have a solid molecular biological basis.CONCLUSION: The application of cDNA microarray technique for analysis of gene expression patterns is a powerful strategy to identify novel cancer-associated genes, and to rapidly explore their role in clinical pancreatic adenocarcinoma. Microarray profiles provide us new insights into the carcinogenesis and invasive process of pancreatic adenocarcinoma. Our results suggest that a highly organized and structured process of tumor invasion exists in the pancreas.展开更多
Objective： The aim of this study was to investigate the MRP-1/CD9mRNA expression in lung cancer and normal lung tissues and the relationship between its expression and pathologic grades, clinical stages, metastasis a...Objective： The aim of this study was to investigate the MRP-1/CD9mRNA expression in lung cancer and normal lung tissues and the relationship between its expression and pathologic grades, clinical stages, metastasis and prognosis. Methods： To observe MRP-1/C9mRNA expression, tissue microarray （TMA） containing 54 lung cancers and 10 normal lung tissues was prepared and Fluorescence in situ hybridization was used. Results： The positive rate of MRP-1/CD9 expression was 48.1% in lung cancer, lower than that of normal lung tissues. The statistical difference was significant （P〈0.05）. Its protein expression had no relationship with the patients＇ ages, sex and the macroscopic type of tumor, but had relationships with the histological type, clinical stage, differentiated degree and metastasis. The expression in non-small cell lung cancer （NSCLC） was higher than that in small cell lung cancer （SCLC）; in well-moderately differentiated group was higher than that in poorly differentiated group; Earlier period group （I＋II） was higher than in later period group （Ⅲ＋Ⅳ）; and in group without lymphoid metastasis was higher than in patients with lymphoid metastasis. Conclusion： The progression of the lung cancer maybe related with the descended MRP-1/Cd9 expression, which may be useful in evaluating the prognosis of cancer patients.展开更多
Objective:The aim of the study was to screen the differentially expressed genes of Peutz-Jeghers syndrome (PJS) and colorectal carcinoma (CRC).Methods:This study used cDNA microarray to comparatively analyze the gene ...Objective:The aim of the study was to screen the differentially expressed genes of Peutz-Jeghers syndrome (PJS) and colorectal carcinoma (CRC).Methods:This study used cDNA microarray to comparatively analyze the gene expression profiles of 4 cases of PJS combined with colorectal adenocarcinoma vs.normal mucosae.The cDNA microarray contained 8064 human genes,and then using RT-PCR to test three genes of all.Results:The experimental data showed that fourteen genes were differentially expressed,which were up-regulated in PJS.Fifty-one genes expressions were altered in CRCs,of which 32 were up-regulated,as compared to the normal mucosae.In addition,5 genes were similarly altered in both PJS and CRCs.RT-PCR analyses confirmed the cDNA microarray data for three of those genes:LATS2,APC and MADH4.Conclusion:LCN2,USP4,GRO3,HYAL1 and APC-these differentially expressed genes likely represent biomarkers for early detection of CRC and may be potential therapeutic targets.展开更多
Advanced diagnostic methods and algorithms for immune disorders provide qualitative and quantitative multiplex measurement for pre-clinical prognostic and clinical diagnostic biomarkers specific for diseases.Choice of...Advanced diagnostic methods and algorithms for immune disorders provide qualitative and quantitative multiplex measurement for pre-clinical prognostic and clinical diagnostic biomarkers specific for diseases.Choice of therapy is confirmed by modulating diagnostic efficacy of companion, theranotic drug concentrations.Assay methods identify, monitor and manage autoimmune diseases, or risk thereof, in subjects who have,or who are related to individuals with autoimmune disease. These same diagnostic protocols also integratequalitative and quantitative assay test protocol designs for responder patient assessment, risk analysis and management of disease when integrating multiplex planar microarray diagnostic tests, patient theranostic companion diagnostic methods and test panels for simultaneous assessment and management of dysimmune and inflammatory disorders, autoimmunity, allergy and cancer. Proprietary assay methods are provided to identify, monitor and manage dysimmune conditions, or risk thereof, in subjects with pathological alterations in the immune system, or who are related to individuals with these conditions. The protocols can be used for confirmatory testing of subjects who exhibit symptoms of dysimmunity, as well as subjects who are apparently healthy and do not exhibit symptoms of altered immune function. The protocols also provide for methods of determining whether a subject has, is at risk for, or is a candidate for disease therapy, guided by companion diagnosis and immunosuppressive therapy, as well as therapeutic drug monitoring and theranostic testing of disease biomarkers in response to immunoabsorption therapy. The multiplex test panels provide the components that are integral for performing the methods to recognized clinical standards.展开更多
In cancer genomes, there are frequent copy number aberration (CNA) events, some of which are believed to be tumori-genic. While copy numbers can be detected by a number of technologies, e.g., SNP arrays, their relatio...In cancer genomes, there are frequent copy number aberration (CNA) events, some of which are believed to be tumori-genic. While copy numbers can be detected by a number of technologies, e.g., SNP arrays, their relations with gene expressions are not well clarified. Here, we describe an approach to visualize the global relations between copy numbers and gene expressions using expression microarrays. We mapped the gene expression signals detected by microar-ray probesets onto a reference human genome, the RefSeq, based on their annotated physical positions, resulting in a landscape that we called expressogram. To study the expressograms under various conditions and their relations with cytogenetic events, such as CNAs, we obtained three classes of array samples, namely samples of a cancer (e.g., liver cancer), normal samples in the same tissue, and normal samples of other tissues. We developed a Bayesian based algorithm to estimate a background signal from the latter two sources for the cancer samples. By subtracting the estimated background from the raw signals of the cancer samples, and subjecting the differences to a kernel-based smoothing scheme, we produced an expressogram that shows strong consistency with the copy numbers. This indicates that copy numbers are on average positively correlated with and have strong impacts on gene expressions. To further explore the applicability of these findings, we submit the expressograms to the significant CNA detection algorithm GISTIC. The results strongly indicate that expressogram can also be used to infer copy number events and significant regions of CNA affected dysregulation.展开更多
It has been previously reported that small mother against decapentaplegic 3 （Smad3） gene knockout （Smad3^ex8/ex8） mice displays phenotypes similar to human osteoarthritis, as characterized by abnormal hypertrophic...It has been previously reported that small mother against decapentaplegic 3 （Smad3） gene knockout （Smad3^ex8/ex8） mice displays phenotypes similar to human osteoarthritis, as characterized by abnormal hypertrophic differentiation of articular chondrocytes. To further clarify the crucial target genes that mediate transformation growth factor-β （TGF-β）/Smad3 signals on articular chondrocytes differentiation and investigate the underlying molecular mechanism of osteoarthritis, microarrays were used to perform comparative transcriptional profiling in the articular cartilage between Smad3^ex8/ex8and wild-type mice on day five after birth. The gene profding results showed that the activity of bone morphogenetic protein （BMP） and TGF-β/cell division cycle 42 （Cdc42） signaling pathways were enhanced in Smad3^ex8/ex8 chondrocytes. Moreover, there was altered gene expression in growth hormone/insulin-like growth factor 1 （Igfl） axis and fibroblast growth factor （Fgf） signaling pathway. Notably, protein synthesis related genes and electron transport chain related genes were upregulated in Smad3^ex8/ex8 chondrocytes, implying that accelerated protein synthesis and enhanced cellular respiration might contribute to hypertrophic differentiation of articular chondrocytes and the pathogenesis of osteoarthritis.展开更多
AIM:To compare the performance of three commercially available anti-human epidermalgrowth factor receptor 2(HER2)antibodies in whole-tissue sections and tissue microarrays(TMAs)of a series of gastric tumors.METHODS:We...AIM:To compare the performance of three commercially available anti-human epidermalgrowth factor receptor 2(HER2)antibodies in whole-tissue sections and tissue microarrays(TMAs)of a series of gastric tumors.METHODS:We present a comparative analysis of three anti-HER2 antibodies(HercepTest,4B5 and SP3)using TMA and whole-tissue sections prepared from the same paraffin blocks of 199 gastric adenocarcinomas operated upon between January 2004 and December2008 at a Brazilian cancer hospital.The data on the patients’age,sex,the anatomical location of the tumor and the Lauren’s histological classification were collected from clinical and pathological records.The immunohistochemical(IHC)results were examined by two pathologists and the cases were classified as positive(3+),equivocal(2+)and negative(0 or 1+),according to the criteria of the IHC scoring system of gastric cancer.TMAs and whole-tissue sections were evaluated separately and independently.All cases yielding discordant IHC results and/or scored as 2+were subjected to dual-color in situ hybridization in order to determine the final HER2 status.Besides determining the sensitivity and predictive value for HER2-positive status,we measured the accuracy of each antibody by calculating the area under the receiver operating characteristic(ROC)curve.The agreement between the results obtained using the TMAs and those obtained using the whole-tissue sections was assessed by means of Kappa coefficient.RESULTS:Intratumoral heterogeneity of HER2 expression was observed with all antibodies.HER2-positive expression(3+)in the whole-tissue sections was observed in 23 cases(11.6%)using the 4B5 antibody,in 18 cases(9.1%)using the SP3 antibody and in 10 cases(5.1%)using the HercepTest antibody.In the TMAs,11 positive cases(5.6%)were identified using SP3 antibody,9(4.6%)using the 4B5 antibody and 6(3%)using the HercepTest antibody.The sensitivity using whole-tissue sections and TMA,respectively,was 95.2%and 42.9%with 4B5,90.5%and 66.7%with SP3 and 47.6%and42.9%with HercepTest.The accuracy,calculated from the area under the ROC curve,using whole-tissue sections and TMA,respectively,was 0.91 and 0.79 by 4B5,0.86 and 0.80 by SP3 and 0.73 and 0.71 by HercepTest.The concordance of the results obtained using wholetissue sections and TMA was 97.4%(Kappa 0.75)using HercepTest,85.6%(Kappa 0.56)using SP3 and 84.1%(Kappa 0.38)using 4B5.CONCLUSION:The use of the 4B5 antibody on wholetissue sections was the most accurate IHC method for evaluating HER2 expression in gastric adenocarcinoma.展开更多
Conidial fungi or molds and mildews are widely used in modern biotechnology as producers of antibiotics and other secondary metabolites,industrially important enzymes,chemicals and food.They are also important pathoge...Conidial fungi or molds and mildews are widely used in modern biotechnology as producers of antibiotics and other secondary metabolites,industrially important enzymes,chemicals and food.They are also important pathogens of animals including humans and agricultural crops.These various applications and extremely versatile natural phenotypes have led to the constantly growing list of complete genomes which are now available.Functional genomics and proteomics widely exploit the genomic information to study the cell-wide impact of altered genes on the phenotype of an organism and its function.This allows for global analysis of the information flow from DNA to RNA to protein,but it is usually not sufficient for the description of the global phenotype of an organism.More recently,Phenotype MicroArray (PM) technology has been introduced as a tool to characterize the metabolism of a (wild) fungal strain or a mutant.In this article,we review the background of PM applications for fungi and the methodic requirements to obtain reliable results.We also report examples of the versatility of this tool.展开更多
The rapid detection of pathogenic bacteria is vital for the prevention of outbreaks of infectious diseases, including infections by the common foodborne bacteria E.coli and Salmonella Carbohydrate microarrays have bee...The rapid detection of pathogenic bacteria is vital for the prevention of outbreaks of infectious diseases, including infections by the common foodborne bacteria E.coli and Salmonella Carbohydrate microarrays have been developed as a powerful method to investigate carbohydrate-protein interaction with only very small amounts of glycans, which show great potential for detect the carbohydrate mediated interaction with pathogens. Here, different mannose-coated microarrays were constructed and tested with E.coli(K-12 and BL-21) and Salmonella enterica strains(ATCC9184 and ATCC31685) exhibiting different mannose binding affinities. The optimized carbohydrate microarray was then applied to test the binding of 12 Salmonella enterica and 9 E.coli isolates from local patients for the first time and showed strong binding with certain serovars or subtypes. The results showed that microarray probed with the single mannose structure is not enough for the detection of bacteria with various serovars or subtypes, which contain a high degree of allelic variation in adhesin. We suggest that a complex carbohydrate microarray containing different glycan conformation may be needed for detection of different bacteria isolates.展开更多
Objective: A single mechanistic pathway cannot explain the genesis of drug resistance in cancer. Drug resistance in cancer is a major obstacle to successful chemotherapy. KB cells provide a useful starting point for s...Objective: A single mechanistic pathway cannot explain the genesis of drug resistance in cancer. Drug resistance in cancer is a major obstacle to successful chemotherapy. KB cells provide a useful starting point for selection of the multidrug resistant (MDR) cell lines. Methods: We used cDNA microarrays containing 12,720 sequences of known genes, expressed sequence tags and unknown clones to monitor gene expression profiles in MDR KB cells. Results: Preliminary data analysis showed that 18 genes were up-regulated and 18 genes were down-regulated by comparison of expression patterns between KB 3-1 and MDR KB-V1 cells. Furthermore, the highly over-expressed CGA, CLU genes in MDR KB-V1 cell were verified with conventional Northern blot analysis. These genes contain information predictive of drug resistance of cancer cells. Conclusion: Our study demonstrates that genome-wide gene expression profiling by using cDNA microarray technique is a valuable approach in obtaining molecular mechanism of drug resistance in cancer cells.展开更多
Objective： Plasminogen activator inhibitor-1 （PAI-1）, one crucial component of the plasminogen activator system, is a major player in the pathogenesis of many vascular diseases as well as in cancer. High levels of ...Objective： Plasminogen activator inhibitor-1 （PAI-1）, one crucial component of the plasminogen activator system, is a major player in the pathogenesis of many vascular diseases as well as in cancer. High levels of PAI-1 in breast cancer tissue are associated with poor prognosis. The aim of this study is to evaluate rigorously the potential of serum PAI-1 concentration functioning as a general screening test in diagnostic or prognostic assays. Methods： A protein-microarray-based sandwich fluorescence immunoassay （FIA） was developed to detect PAI-1 in serum. Several conditions of this microarray-based FIA were optimized to establish an efficacious method. Serum specimens of 84 healthy women and 285 women with breast cancer were analyzed using the optimized FIA microarray. Results： The median serum PAI-1 level of breast cancer patients was higher than that of healthy women （109.7 ng/ml vs. 63.4 ng/ml）. Analysis of covariance revealed that PAI-1 levels of the two groups were significantly different （P0.001） when controlling for an age effect on PAI-1 levels. However, PAI-1 values in TNM stage I？IV patients respectively were not significantly different from each other. Conclusion： This microarray-based sandwich FIA holds potential for quantitative analysis of tumor markers such as PAI-1.展开更多
Objective： To explore the construction of metastatic spinal cancer （MSC） tissue microarrays and validate its value in immunohistochemical study of MSC. Methods： Paraffin-embedded specimens from 71 MSC cases and 6 ...Objective： To explore the construction of metastatic spinal cancer （MSC） tissue microarrays and validate its value in immunohistochemical study of MSC. Methods： Paraffin-embedded specimens from 71 MSC cases and 6 primary tumor cases were selected as donor blocks and prepared into MSC tissue microarrays by tissue array arrangement, the steps of which included location, punching, sampling, sample seeding, and re-diagnosis by hematoxylin-eosin （HE） as well as MMP-9 and MMP-14 immunohistoehemical staining. Results： The MSC tissue microarrays thus constructed were intact and craekless, containing 154 complete and well arranged microarray points. None of the sectioned tissue microarrays was lost, and the results of HE staining was consistent with the primary pathologic diagnoses. Immunohistochemical staining was also good without non-specific or marginal effect. Conclusion： The MSC tissue microarrays have a high value in the immunohistochemical study of MSC.展开更多
Objective: We investigated the expression levels of KiSS-1 and KAI-1 in gallbladder carcinoma and their relationship with clinicopathologic parameters and metastasis potential. Methods: Pathological specimens from 59 ...Objective: We investigated the expression levels of KiSS-1 and KAI-1 in gallbladder carcinoma and their relationship with clinicopathologic parameters and metastasis potential. Methods: Pathological specimens from 59 gallbladder carcinoma tissues (including 23 invasion tissues), matched 7 para-tumor and 6 normal gallbladder tissues were examined for the expression of KiSS-1 and KAI-1 protein by tissue microarray technique and immunohistochemistry (EnVision). Results: The positive rate of KiSS-1 and KAI-1 was down-regulated (P < 0.05) in tumor tissues, compared with non-tumor tissues. The positive rate of KiSS-1 and KAI-1 were down-regulated (P < 0.01) in invasion tissues, compared with para-tumor tissues. Down-regulating expression levels of KiSS-1 and KAI-1 were significantly correlated with some malignant behavior of gallbladder carcinoma. The positive rate of KiSS-1 and KAI-1 were all negatively correlated with the Nevin staging of gallbladder carcinoma (P < 0.01). Moreover, a strong positive correlation was found between the positive expression of KiSS-1 and KAI-1 in gallbladder carcinoma (P < 0.001). It was indicated that KiSS-1 and KAI-1 were all associated with high metastasis potential of gallbladder carcinoma. Conclusion: KiSS-1 and/or KAI-1 may be important markers of reflecting invasive and metastatic potential and prognosis in gallbladder carcinoma, and may be the target genes to inhibit metastasis of gallbladder carcinoma. Detection of the expression of KiSS-1 and/or KAI-1 may have important clinical value for finding early-stage and evaluating the prognosis of gallbladder carcinoma.展开更多
The application of microarray-based techniques for the diagnosis of genomic rearrangements has been steadily growing in popularity since its introduction in 2004.Given the many advantages of these techniques over conv...The application of microarray-based techniques for the diagnosis of genomic rearrangements has been steadily growing in popularity since its introduction in 2004.Given the many advantages of these techniques over conventional cytogenetics,there is increasing pressure towards their application in prenatal diagnosis.However,there remain several important issues that must be addressed.For example,microarray-based techniques(comparative genomic hybridization-based arrays and single nucleotide polymorphism-based arrays) allow detection of even very small genomic imbalances that can determine pathological clinical conditions.In addition,there are other copy number variations which represent normal variation,with no detectable effects on phenotype.Given the still incomplete knowledge of the changes in our genome and the associated phenotypes,microarray-based diagnosis is likely to find variants of uncertain and unknown clinical significance.The interpretation of these variants is now a major challenge for the medical geneticist,who often find it difficult to establish precise correlations between genotype and phenotype.There is sufficient available evidence to justify the use of microarray-based diagnostics for a select number of specific conditions,but there is also an inevitable trend towards ever wider application.It is very important that this drift does not progress in an unchecked and uncontrolled manner under the thrust of commercial interests.Therefore,we recommend that scientific societies be vigilant and take an advisory role in the adopting of these technologies as new scientific knowledge becomes available.展开更多
Quantum dots color conversion(QDCC)is considered as a facial and versatile way to achieve full-color organic light emitting diode(OLED)and micro-LED display due to the wide color gamut performance and easy integration...Quantum dots color conversion(QDCC)is considered as a facial and versatile way to achieve full-color organic light emitting diode(OLED)and micro-LED display due to the wide color gamut performance and easy integration.However,the aggregation of QDs and coffee-ring effects after solvent evaporation lowers the light conversion efficiency and emission uniformity in QDs microarrays,raising blue-light leakage or optical crosstalk.Here,we report the fabrication of perovskite quantum dots(PQDs)microarrays by combining the inkjet printing and in situ fabrication of PQDs during the photopolymerization of precursor ink.The resulting PQDs microarrays exhibit three-dimensional(3D)morphology with hemisphere shape as well as strong photoluminescence,which is desirable for QDCC applications.We demonstrate the dominant role of ultraviolet(UV)curable precursors and surface functionalized substrate in controlling the shape of microarrays,where significantly increased contact angle(100°)and large height to diameter ratio(0.42)can be achieved.We further demonstrate the potential use of the in situ direct print photopolymerization method for fabricating large-area multicolor patterned pixel microarrays with a wide color gamut and high resolution.The fabrication of 3D PQDs microarrays opens up new opportunities in a variety of applications including photonics integration,micro-LED,and near-field display.展开更多
Mouse-Immunoglobulin G(mouse-IgG) with different concentrations in a range from 1000 to 0.0128 μg/mL and a specific hybridization with goat anti-mouse IgG were detected successfully by using an oblique-incidence refl...Mouse-Immunoglobulin G(mouse-IgG) with different concentrations in a range from 1000 to 0.0128 μg/mL and a specific hybridization with goat anti-mouse IgG were detected successfully by using an oblique-incidence reflectivity difference(OI-RD) method.Two detection signals,consisting of an imaginary part(Im{Δp-Δs}) and a real part(Re{Δp-Δs}) of OI-RD,were obtained simultaneously.The detection results of hybridization by OI-RD were in accord with that of traditional fluorescent scans.In particular,we label-freely detected the washed mouse-IgG microarray with a series of concentrations and acquired a linear correlation between OI-RD intensities and the protein concentrations in logarithmic coordinates.The detection sensitivity of OI-RD can reach 14 fg.These experimental results suggest that the OI-RD method has potential applications in proteomics and clinical diagnosis.展开更多
Background Oligonucleotide microarrays are increasingly being used to identify gene expression profiles that associated with complex genetic diseases. Peripheral lymphocytes communicate with cells and extracellular ma...Background Oligonucleotide microarrays are increasingly being used to identify gene expression profiles that associated with complex genetic diseases. Peripheral lymphocytes communicate with cells and extracellular matrixes in almost all tissues and organs in human body, suggesting that the gene expression profiles in peripheral lymphocytes may reflect the presence of disease in the body. This study aimed to identify molecular biomarkers for cervical cancer in peripheral blood lymphocytes by using oligonucleotide microarrays. Methods Total RNA was extracted from peripheral blood lymphocytes of 24 early stage cervical cancer patients and 18 healthy controls. We used 22K Human Genome microarrays to profile peripheral blood lymphocytes from 4 early stage cervical cancer patients and compared their gene expression profiles with those from 3 healthy controls. Differentially expressed genes would be identified if they had adjusted P values of less than 0.05 and a groupwise average fold change greater than 1.5 or less than 0.67. Then the selected 5 genes were validated in the remaining 20 early stage cervical cancer patients and the 15 healthy controls by using real-time reverse-transcription polymerase chain reaction （RT-PCR）. Results Genes identified by the gene selection program expressed differently between the blood samples of the early stage cervical cancer patients and those of the healthy controls. To validate the gene expression data, 5 genes were analyzed by real-time RT-PCR. In three of the 5 identified genes, tenasin-c （TNC）, nuceolin （NCL）, and enolase 2 （EN02） showed a significant up-regulation in the blood samples of the early stage cervical cancer patients versus that of the healthy controls. Conclusions The up-regulation of TNC, NCL, and EN02 in peripheral blood may be used to identify novel blood biomarkers for detecting cervical cancer in a clinically accessible surrogate tissue, and thus to provide a possibility to develop a noninvasive and predictive diagnosis for the disease.展开更多
文摘BACKGROUND: The good therapeutic effects of large dose of dexamethasone on severe acute pancreatitis (SAP) patients have been proved. This study was designed to investigate the influence of dexamethasone on apoptosis of acinar cells in the pancreas of rats with SAP and the protein expression of the apoptosis-regulating genes Bax and Bcl-2. METHODS: Ninety Sprague-Dawley rats with SAP were randomly divided into a model group and a dexamethasone treated group (45 rats in each group), and another 45 rats formed the sham operation group. Survival rates were calculated and gross pathological changes in the pancreas of each group were observed under a light microscope 3, 6 and 12 hours after operation. Tissue microarray technology was applied to prepare pancreatic tissue sections. The changes in Bax and Bcl-2 protein expression levels of pancreatic tissues from each group were assessed by immunohistochemical staining, and TUNEL staining was used to evaluate changes in apoptosis index. RESULTS: The model and treated groups did not differ in mortality at each time point. The pathological score for the pancreas in the treated group was significantly lower than that in the model group at 3 and 6 hours. The positive rates of Bax protein expression in the head and tail of the pancreas in the treated group at all time points were all markedly higher than those of the model group. The positive rate of Bcl-2 protein expression in the head of the pancreas in the treated group was significantly higher than that of the model group at 3 hours. TUNEL staining showed that the pancreas head and tail apoptosis indices of the treated group were markedly higher than those of the model group after 6 hours. CONCLUSIONS: Apoptosis may be a protective response. to pancreatic cell injury. The mechanism of action of dexamethasone in treating SAP may be related to the apoptosis of acinar cells in the pancreas induced by apoptosis-regulating genes such as Bax and Bcl-2. The advantages of tissue microarrays in pathological examination of the pancreas include saving of time and energy, efficiency and highly representative.
文摘The genome sequence information in combination with DNA microarrays promises to revolutionize the way of cellu-lar and molecular biological research by allowing complex mixtures of RNA and DNA to interrogated in a parallel and quantita-tive fashion. DNA microarrays can be used to measure levels of gene expression for tens of thousands of gene simultane-ously and take advantage of all available sequence information for experimental design and data interpretation in pursuit of biological understanding. Recent progress in experimental genomics allows DNA microarrays not simply to provide a cata-logue of all the genes and information about their function, but to understand how the components work together to comprise functioning cells and organisms. This brief review gives a survey of DNA microarrays technology and its applications in ge-nome and gene function analysis, gene expression studies, biological signal and defense system, cell cycle regulation, mechanism of transcriptional regulation, proteomics, and the functionality of food component.
基金Supported by the National Natural Science Foundation of China,No. 30000160
文摘AIM: To study the pathogenetic processes and the role of gene expression by microarray analyses in expediting our understanding of the molecular pathophysiology of pancreatic adenocarcinoma, and to identify the novel cancer-associated genes.METHODS: Nine histologically defined pancreatic head adenocarcinoma specimens associated with clinical data were studied. Total RNA and mRNA were isolated and labeled by reverse transcription reaction with Cy5 and Cy3 for cDNA probe. The cDNA microarrays that represent a set of 4 096 human genes were hybridized with labeled cDNA probe and screened for molecular profiling analyses.RESULTS: Using this methodology, 184 genes were screened out for differences in gene expression level after nine couples of hybridizations. Of the 184 genes,87 were upregulated and 97 downregulated, including 11 novel human genes. In pancreatic adenocarcinoma tissue, several invasion and metastasis related genes showed their high expression levels, suggesting that poor prognosis of pancreatic adenocarcinoma might have a solid molecular biological basis.CONCLUSION: The application of cDNA microarray technique for analysis of gene expression patterns is a powerful strategy to identify novel cancer-associated genes, and to rapidly explore their role in clinical pancreatic adenocarcinoma. Microarray profiles provide us new insights into the carcinogenesis and invasive process of pancreatic adenocarcinoma. Our results suggest that a highly organized and structured process of tumor invasion exists in the pancreas.
基金This work was supported by a grant from Tianjin Science and Technology Committee (No. 033804211)
文摘Objective： The aim of this study was to investigate the MRP-1/CD9mRNA expression in lung cancer and normal lung tissues and the relationship between its expression and pathologic grades, clinical stages, metastasis and prognosis. Methods： To observe MRP-1/C9mRNA expression, tissue microarray （TMA） containing 54 lung cancers and 10 normal lung tissues was prepared and Fluorescence in situ hybridization was used. Results： The positive rate of MRP-1/CD9 expression was 48.1% in lung cancer, lower than that of normal lung tissues. The statistical difference was significant （P〈0.05）. Its protein expression had no relationship with the patients＇ ages, sex and the macroscopic type of tumor, but had relationships with the histological type, clinical stage, differentiated degree and metastasis. The expression in non-small cell lung cancer （NSCLC） was higher than that in small cell lung cancer （SCLC）; in well-moderately differentiated group was higher than that in poorly differentiated group; Earlier period group （I＋II） was higher than in later period group （Ⅲ＋Ⅳ）; and in group without lymphoid metastasis was higher than in patients with lymphoid metastasis. Conclusion： The progression of the lung cancer maybe related with the descended MRP-1/Cd9 expression, which may be useful in evaluating the prognosis of cancer patients.
基金Supported by the grants from Funded Project based by the Health Innovation of the Xiamen Municipal Science and Technology Programme (No.3502Z20084031)Medical Technology Innovation Project of Nanjing Military Region (No.09MA066)
文摘Objective:The aim of the study was to screen the differentially expressed genes of Peutz-Jeghers syndrome (PJS) and colorectal carcinoma (CRC).Methods:This study used cDNA microarray to comparatively analyze the gene expression profiles of 4 cases of PJS combined with colorectal adenocarcinoma vs.normal mucosae.The cDNA microarray contained 8064 human genes,and then using RT-PCR to test three genes of all.Results:The experimental data showed that fourteen genes were differentially expressed,which were up-regulated in PJS.Fifty-one genes expressions were altered in CRCs,of which 32 were up-regulated,as compared to the normal mucosae.In addition,5 genes were similarly altered in both PJS and CRCs.RT-PCR analyses confirmed the cDNA microarray data for three of those genes:LATS2,APC and MADH4.Conclusion:LCN2,USP4,GRO3,HYAL1 and APC-these differentially expressed genes likely represent biomarkers for early detection of CRC and may be potential therapeutic targets.
文摘Advanced diagnostic methods and algorithms for immune disorders provide qualitative and quantitative multiplex measurement for pre-clinical prognostic and clinical diagnostic biomarkers specific for diseases.Choice of therapy is confirmed by modulating diagnostic efficacy of companion, theranotic drug concentrations.Assay methods identify, monitor and manage autoimmune diseases, or risk thereof, in subjects who have,or who are related to individuals with autoimmune disease. These same diagnostic protocols also integratequalitative and quantitative assay test protocol designs for responder patient assessment, risk analysis and management of disease when integrating multiplex planar microarray diagnostic tests, patient theranostic companion diagnostic methods and test panels for simultaneous assessment and management of dysimmune and inflammatory disorders, autoimmunity, allergy and cancer. Proprietary assay methods are provided to identify, monitor and manage dysimmune conditions, or risk thereof, in subjects with pathological alterations in the immune system, or who are related to individuals with these conditions. The protocols can be used for confirmatory testing of subjects who exhibit symptoms of dysimmunity, as well as subjects who are apparently healthy and do not exhibit symptoms of altered immune function. The protocols also provide for methods of determining whether a subject has, is at risk for, or is a candidate for disease therapy, guided by companion diagnosis and immunosuppressive therapy, as well as therapeutic drug monitoring and theranostic testing of disease biomarkers in response to immunoabsorption therapy. The multiplex test panels provide the components that are integral for performing the methods to recognized clinical standards.
文摘In cancer genomes, there are frequent copy number aberration (CNA) events, some of which are believed to be tumori-genic. While copy numbers can be detected by a number of technologies, e.g., SNP arrays, their relations with gene expressions are not well clarified. Here, we describe an approach to visualize the global relations between copy numbers and gene expressions using expression microarrays. We mapped the gene expression signals detected by microar-ray probesets onto a reference human genome, the RefSeq, based on their annotated physical positions, resulting in a landscape that we called expressogram. To study the expressograms under various conditions and their relations with cytogenetic events, such as CNAs, we obtained three classes of array samples, namely samples of a cancer (e.g., liver cancer), normal samples in the same tissue, and normal samples of other tissues. We developed a Bayesian based algorithm to estimate a background signal from the latter two sources for the cancer samples. By subtracting the estimated background from the raw signals of the cancer samples, and subjecting the differences to a kernel-based smoothing scheme, we produced an expressogram that shows strong consistency with the copy numbers. This indicates that copy numbers are on average positively correlated with and have strong impacts on gene expressions. To further explore the applicability of these findings, we submit the expressograms to the significant CNA detection algorithm GISTIC. The results strongly indicate that expressogram can also be used to infer copy number events and significant regions of CNA affected dysregulation.
基金This work was supported by the National Key Program on Basic Research of China (No. 2006BAI23B01-3)National Natural Scie- nce Foundation of China (No. 30430350, 30500)National High-Tech Research and Development Program (No. 2006AA 02Z168, Z000 6303041231).
文摘It has been previously reported that small mother against decapentaplegic 3 （Smad3） gene knockout （Smad3^ex8/ex8） mice displays phenotypes similar to human osteoarthritis, as characterized by abnormal hypertrophic differentiation of articular chondrocytes. To further clarify the crucial target genes that mediate transformation growth factor-β （TGF-β）/Smad3 signals on articular chondrocytes differentiation and investigate the underlying molecular mechanism of osteoarthritis, microarrays were used to perform comparative transcriptional profiling in the articular cartilage between Smad3^ex8/ex8and wild-type mice on day five after birth. The gene profding results showed that the activity of bone morphogenetic protein （BMP） and TGF-β/cell division cycle 42 （Cdc42） signaling pathways were enhanced in Smad3^ex8/ex8 chondrocytes. Moreover, there was altered gene expression in growth hormone/insulin-like growth factor 1 （Igfl） axis and fibroblast growth factor （Fgf） signaling pathway. Notably, protein synthesis related genes and electron transport chain related genes were upregulated in Smad3^ex8/ex8 chondrocytes, implying that accelerated protein synthesis and enhanced cellular respiration might contribute to hypertrophic differentiation of articular chondrocytes and the pathogenesis of osteoarthritis.
文摘AIM:To compare the performance of three commercially available anti-human epidermalgrowth factor receptor 2(HER2)antibodies in whole-tissue sections and tissue microarrays(TMAs)of a series of gastric tumors.METHODS:We present a comparative analysis of three anti-HER2 antibodies(HercepTest,4B5 and SP3)using TMA and whole-tissue sections prepared from the same paraffin blocks of 199 gastric adenocarcinomas operated upon between January 2004 and December2008 at a Brazilian cancer hospital.The data on the patients’age,sex,the anatomical location of the tumor and the Lauren’s histological classification were collected from clinical and pathological records.The immunohistochemical(IHC)results were examined by two pathologists and the cases were classified as positive(3+),equivocal(2+)and negative(0 or 1+),according to the criteria of the IHC scoring system of gastric cancer.TMAs and whole-tissue sections were evaluated separately and independently.All cases yielding discordant IHC results and/or scored as 2+were subjected to dual-color in situ hybridization in order to determine the final HER2 status.Besides determining the sensitivity and predictive value for HER2-positive status,we measured the accuracy of each antibody by calculating the area under the receiver operating characteristic(ROC)curve.The agreement between the results obtained using the TMAs and those obtained using the whole-tissue sections was assessed by means of Kappa coefficient.RESULTS:Intratumoral heterogeneity of HER2 expression was observed with all antibodies.HER2-positive expression(3+)in the whole-tissue sections was observed in 23 cases(11.6%)using the 4B5 antibody,in 18 cases(9.1%)using the SP3 antibody and in 10 cases(5.1%)using the HercepTest antibody.In the TMAs,11 positive cases(5.6%)were identified using SP3 antibody,9(4.6%)using the 4B5 antibody and 6(3%)using the HercepTest antibody.The sensitivity using whole-tissue sections and TMA,respectively,was 95.2%and 42.9%with 4B5,90.5%and 66.7%with SP3 and 47.6%and42.9%with HercepTest.The accuracy,calculated from the area under the ROC curve,using whole-tissue sections and TMA,respectively,was 0.91 and 0.79 by 4B5,0.86 and 0.80 by SP3 and 0.73 and 0.71 by HercepTest.The concordance of the results obtained using wholetissue sections and TMA was 97.4%(Kappa 0.75)using HercepTest,85.6%(Kappa 0.56)using SP3 and 84.1%(Kappa 0.38)using 4B5.CONCLUSION:The use of the 4B5 antibody on wholetissue sections was the most accurate IHC method for evaluating HER2 expression in gastric adenocarcinoma.
基金Project (No.FWF P-P17859-B06) supported by the Austrian Science Foundation
文摘Conidial fungi or molds and mildews are widely used in modern biotechnology as producers of antibiotics and other secondary metabolites,industrially important enzymes,chemicals and food.They are also important pathogens of animals including humans and agricultural crops.These various applications and extremely versatile natural phenotypes have led to the constantly growing list of complete genomes which are now available.Functional genomics and proteomics widely exploit the genomic information to study the cell-wide impact of altered genes on the phenotype of an organism and its function.This allows for global analysis of the information flow from DNA to RNA to protein,but it is usually not sufficient for the description of the global phenotype of an organism.More recently,Phenotype MicroArray (PM) technology has been introduced as a tool to characterize the metabolism of a (wild) fungal strain or a mutant.In this article,we review the background of PM applications for fungi and the methodic requirements to obtain reliable results.We also report examples of the versatility of this tool.
基金National Natural Science Foundation of China(Nos.31700706,21877052)Natural Science Foundation of Jiangsu Province(No.BK20180030)+5 种基金the Fundamental Research Funds for the Central Universities(No.JUSRP51712B)the Public Health Research Center at Jiangnan University(No.JUPH201502)Max Planck Society International Partner Group ProgramHigh-end Foreign Experts Recruitment Programthe Thousand Talents Plan(Young Professionals)the China Scholarship Council.
文摘The rapid detection of pathogenic bacteria is vital for the prevention of outbreaks of infectious diseases, including infections by the common foodborne bacteria E.coli and Salmonella Carbohydrate microarrays have been developed as a powerful method to investigate carbohydrate-protein interaction with only very small amounts of glycans, which show great potential for detect the carbohydrate mediated interaction with pathogens. Here, different mannose-coated microarrays were constructed and tested with E.coli(K-12 and BL-21) and Salmonella enterica strains(ATCC9184 and ATCC31685) exhibiting different mannose binding affinities. The optimized carbohydrate microarray was then applied to test the binding of 12 Salmonella enterica and 9 E.coli isolates from local patients for the first time and showed strong binding with certain serovars or subtypes. The results showed that microarray probed with the single mannose structure is not enough for the detection of bacteria with various serovars or subtypes, which contain a high degree of allelic variation in adhesin. We suggest that a complex carbohydrate microarray containing different glycan conformation may be needed for detection of different bacteria isolates.
基金This work is supported by the City University of Hong Kong through a Strategic Research Grant (CityU Project No. 7001113).
文摘Objective: A single mechanistic pathway cannot explain the genesis of drug resistance in cancer. Drug resistance in cancer is a major obstacle to successful chemotherapy. KB cells provide a useful starting point for selection of the multidrug resistant (MDR) cell lines. Methods: We used cDNA microarrays containing 12,720 sequences of known genes, expressed sequence tags and unknown clones to monitor gene expression profiles in MDR KB cells. Results: Preliminary data analysis showed that 18 genes were up-regulated and 18 genes were down-regulated by comparison of expression patterns between KB 3-1 and MDR KB-V1 cells. Furthermore, the highly over-expressed CGA, CLU genes in MDR KB-V1 cell were verified with conventional Northern blot analysis. These genes contain information predictive of drug resistance of cancer cells. Conclusion: Our study demonstrates that genome-wide gene expression profiling by using cDNA microarray technique is a valuable approach in obtaining molecular mechanism of drug resistance in cancer cells.
基金supported by the National "863" High Technology Project Found of China (No.2006AA02A402)
文摘Objective： Plasminogen activator inhibitor-1 （PAI-1）, one crucial component of the plasminogen activator system, is a major player in the pathogenesis of many vascular diseases as well as in cancer. High levels of PAI-1 in breast cancer tissue are associated with poor prognosis. The aim of this study is to evaluate rigorously the potential of serum PAI-1 concentration functioning as a general screening test in diagnostic or prognostic assays. Methods： A protein-microarray-based sandwich fluorescence immunoassay （FIA） was developed to detect PAI-1 in serum. Several conditions of this microarray-based FIA were optimized to establish an efficacious method. Serum specimens of 84 healthy women and 285 women with breast cancer were analyzed using the optimized FIA microarray. Results： The median serum PAI-1 level of breast cancer patients was higher than that of healthy women （109.7 ng/ml vs. 63.4 ng/ml）. Analysis of covariance revealed that PAI-1 levels of the two groups were significantly different （P0.001） when controlling for an age effect on PAI-1 levels. However, PAI-1 values in TNM stage I？IV patients respectively were not significantly different from each other. Conclusion： This microarray-based sandwich FIA holds potential for quantitative analysis of tumor markers such as PAI-1.
基金supported by the research funds of Changzheng Hospital,Shanghai,Chinathe research funds of health bureau in Shanghai (320.2745)medical and health science research program of People’s Liberation Army in the eleventh 5-year plan (06MB218)
文摘Objective： To explore the construction of metastatic spinal cancer （MSC） tissue microarrays and validate its value in immunohistochemical study of MSC. Methods： Paraffin-embedded specimens from 71 MSC cases and 6 primary tumor cases were selected as donor blocks and prepared into MSC tissue microarrays by tissue array arrangement, the steps of which included location, punching, sampling, sample seeding, and re-diagnosis by hematoxylin-eosin （HE） as well as MMP-9 and MMP-14 immunohistoehemical staining. Results： The MSC tissue microarrays thus constructed were intact and craekless, containing 154 complete and well arranged microarray points. None of the sectioned tissue microarrays was lost, and the results of HE staining was consistent with the primary pathologic diagnoses. Immunohistochemical staining was also good without non-specific or marginal effect. Conclusion： The MSC tissue microarrays have a high value in the immunohistochemical study of MSC.
基金Supported by a grant from the Technology Program of Social Development Foundation of Kunshan (No. KS0703)
文摘Objective: We investigated the expression levels of KiSS-1 and KAI-1 in gallbladder carcinoma and their relationship with clinicopathologic parameters and metastasis potential. Methods: Pathological specimens from 59 gallbladder carcinoma tissues (including 23 invasion tissues), matched 7 para-tumor and 6 normal gallbladder tissues were examined for the expression of KiSS-1 and KAI-1 protein by tissue microarray technique and immunohistochemistry (EnVision). Results: The positive rate of KiSS-1 and KAI-1 was down-regulated (P < 0.05) in tumor tissues, compared with non-tumor tissues. The positive rate of KiSS-1 and KAI-1 were down-regulated (P < 0.01) in invasion tissues, compared with para-tumor tissues. Down-regulating expression levels of KiSS-1 and KAI-1 were significantly correlated with some malignant behavior of gallbladder carcinoma. The positive rate of KiSS-1 and KAI-1 were all negatively correlated with the Nevin staging of gallbladder carcinoma (P < 0.01). Moreover, a strong positive correlation was found between the positive expression of KiSS-1 and KAI-1 in gallbladder carcinoma (P < 0.001). It was indicated that KiSS-1 and KAI-1 were all associated with high metastasis potential of gallbladder carcinoma. Conclusion: KiSS-1 and/or KAI-1 may be important markers of reflecting invasive and metastatic potential and prognosis in gallbladder carcinoma, and may be the target genes to inhibit metastasis of gallbladder carcinoma. Detection of the expression of KiSS-1 and/or KAI-1 may have important clinical value for finding early-stage and evaluating the prognosis of gallbladder carcinoma.
文摘The application of microarray-based techniques for the diagnosis of genomic rearrangements has been steadily growing in popularity since its introduction in 2004.Given the many advantages of these techniques over conventional cytogenetics,there is increasing pressure towards their application in prenatal diagnosis.However,there remain several important issues that must be addressed.For example,microarray-based techniques(comparative genomic hybridization-based arrays and single nucleotide polymorphism-based arrays) allow detection of even very small genomic imbalances that can determine pathological clinical conditions.In addition,there are other copy number variations which represent normal variation,with no detectable effects on phenotype.Given the still incomplete knowledge of the changes in our genome and the associated phenotypes,microarray-based diagnosis is likely to find variants of uncertain and unknown clinical significance.The interpretation of these variants is now a major challenge for the medical geneticist,who often find it difficult to establish precise correlations between genotype and phenotype.There is sufficient available evidence to justify the use of microarray-based diagnostics for a select number of specific conditions,but there is also an inevitable trend towards ever wider application.It is very important that this drift does not progress in an unchecked and uncontrolled manner under the thrust of commercial interests.Therefore,we recommend that scientific societies be vigilant and take an advisory role in the adopting of these technologies as new scientific knowledge becomes available.
基金the National Key Research and Development Program of China(No.2020YFB2009303)the National Natural Science Foundation of China(Nos.62105025 and 61935001)Beijing Institute of Technology Research Fund Program for Young Scholars(No.3040011182113)。
文摘Quantum dots color conversion(QDCC)is considered as a facial and versatile way to achieve full-color organic light emitting diode(OLED)and micro-LED display due to the wide color gamut performance and easy integration.However,the aggregation of QDs and coffee-ring effects after solvent evaporation lowers the light conversion efficiency and emission uniformity in QDs microarrays,raising blue-light leakage or optical crosstalk.Here,we report the fabrication of perovskite quantum dots(PQDs)microarrays by combining the inkjet printing and in situ fabrication of PQDs during the photopolymerization of precursor ink.The resulting PQDs microarrays exhibit three-dimensional(3D)morphology with hemisphere shape as well as strong photoluminescence,which is desirable for QDCC applications.We demonstrate the dominant role of ultraviolet(UV)curable precursors and surface functionalized substrate in controlling the shape of microarrays,where significantly increased contact angle(100°)and large height to diameter ratio(0.42)can be achieved.We further demonstrate the potential use of the in situ direct print photopolymerization method for fabricating large-area multicolor patterned pixel microarrays with a wide color gamut and high resolution.The fabrication of 3D PQDs microarrays opens up new opportunities in a variety of applications including photonics integration,micro-LED,and near-field display.
基金supported by the National Basic Research Program of China (Grant No. 2007CB935700)
文摘Mouse-Immunoglobulin G(mouse-IgG) with different concentrations in a range from 1000 to 0.0128 μg/mL and a specific hybridization with goat anti-mouse IgG were detected successfully by using an oblique-incidence reflectivity difference(OI-RD) method.Two detection signals,consisting of an imaginary part(Im{Δp-Δs}) and a real part(Re{Δp-Δs}) of OI-RD,were obtained simultaneously.The detection results of hybridization by OI-RD were in accord with that of traditional fluorescent scans.In particular,we label-freely detected the washed mouse-IgG microarray with a series of concentrations and acquired a linear correlation between OI-RD intensities and the protein concentrations in logarithmic coordinates.The detection sensitivity of OI-RD can reach 14 fg.These experimental results suggest that the OI-RD method has potential applications in proteomics and clinical diagnosis.
文摘Background Oligonucleotide microarrays are increasingly being used to identify gene expression profiles that associated with complex genetic diseases. Peripheral lymphocytes communicate with cells and extracellular matrixes in almost all tissues and organs in human body, suggesting that the gene expression profiles in peripheral lymphocytes may reflect the presence of disease in the body. This study aimed to identify molecular biomarkers for cervical cancer in peripheral blood lymphocytes by using oligonucleotide microarrays. Methods Total RNA was extracted from peripheral blood lymphocytes of 24 early stage cervical cancer patients and 18 healthy controls. We used 22K Human Genome microarrays to profile peripheral blood lymphocytes from 4 early stage cervical cancer patients and compared their gene expression profiles with those from 3 healthy controls. Differentially expressed genes would be identified if they had adjusted P values of less than 0.05 and a groupwise average fold change greater than 1.5 or less than 0.67. Then the selected 5 genes were validated in the remaining 20 early stage cervical cancer patients and the 15 healthy controls by using real-time reverse-transcription polymerase chain reaction （RT-PCR）. Results Genes identified by the gene selection program expressed differently between the blood samples of the early stage cervical cancer patients and those of the healthy controls. To validate the gene expression data, 5 genes were analyzed by real-time RT-PCR. In three of the 5 identified genes, tenasin-c （TNC）, nuceolin （NCL）, and enolase 2 （EN02） showed a significant up-regulation in the blood samples of the early stage cervical cancer patients versus that of the healthy controls. Conclusions The up-regulation of TNC, NCL, and EN02 in peripheral blood may be used to identify novel blood biomarkers for detecting cervical cancer in a clinically accessible surrogate tissue, and thus to provide a possibility to develop a noninvasive and predictive diagnosis for the disease.
