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Objective:The aim of the study was to investigate the correlation between blood pressure response to cold pressor test (CPT) and follow-up blood pressure after 8 years in subjects, and to evaluate the predictive value of CPT for long-term blood pressure levels.Methods:A total of 365 individuals from eight natural villages were enrolled by stratified cluster sampling from Mei County, Shaanxi Province in 2004. Baseline characteristics of subjects were collected and CPTs were conducted. Subjects were followed up in 2009 and 2012, respectively. According to the maximal change of systolic response (SR), the area under the curve (AUC) of systolic blood pressure change (AUC-SBP), the maximal change of diastolic response (DR) and the AUC of diastolic blood pressure change (AUC-DBP) in CPT, the individuals were divided into four quartile groups by above parameters, respectively: group Ⅰ ( P25), group Ⅱ ( P50), group Ⅲ ( P75) and group Ⅳ ( P100). The correlation between blood pressure response to CPT and the follow-up blood pressure was analyzed. Results:(1) There were no significant differences in baseline blood pressure levels and prevalence of hypertension among four quartile groups no matter it was grouped on SR, DR, AUC-SBP or AUC-DBP. (2) The prevalence of hypertension in each group from lowest ( P25) to highest ( P100) in 2012 was 25.64%, 30.67%, 38.03%, 55.74% on SR grouping ( P<0.01), and 27.5%, 29.17%, 38.46%, 57.35% on AUC-SBP grouping ( P<0.05), respectively. (3) There were no significant differences in the prevalence of hypertension among four groups in 2012 ( P>0.05) either on DR or on AUC-DBP grouping. (4) The random effects model analysis showed that the correlation coefficient between SR, AUC-SBP and long-term systolic blood pressure increase were 1.91 ( P<0.05) and 1.44 ( P<0.05), respectively, and the correlation coefficient between DR, AUC-DBP and long-term diastolic blood pressure increase were 0.82 ( P<0.05) and 0.78 ( P>0.05), respectively. Age, male, body mass index, and fasting blood glucose were independent risk factors for long-term blood pressure elevation, and age, body mass index and fasting blood glucose positively correlated with changes in long-term blood pressure (all P<0.05). Conclusion:Individual systolic blood pressure response to CPT can be used as a predictor of long-term hypertension.
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Objective Based on the age of big data, the treatment mechanisms of Qishen-Yiqi(QSYQ) for myocardial infarction was focused. Methods All the data stemmed from Gene Expression Omnibus (GEO). For ensuring the efficacy genetic molecular, differentially expressed genes were discobered by Qlucore Omics Explorer (QOE). And then gene set enrichment analysis was performed by Database for Annotation, Visualization and Integrated Discovery online Gene Annotation system(DAVID) for showing the genes functions during bioprocess. In the end, the predicted genes and proteins interactions networks were demonstrated by Gene/protein interaction database (STRING). Results The main biological process involved up regulating the expression of some specil genes such as NFIL3, ARNTL, DBP, FGD4 and nuclear receptor genes like NR1D1, NR1D2 while using QSYQ treatment. In such case, BHLHE40/41 was regulated. Conclusion Traditional Chinese medicine, QSYQ, could influence the expression of genes of cytothesis, inflammatory and enzymatic activity as its effect of the molecular mechanism, in order to treat and cure the myocarditis infarction.
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<p><b>OBJECTIVE</b>To investigate the expression of HERC4 in human breast cancer tissues and its relationship with the clinicopathological features.</p><p><b>METHODS</b>RT-qPCR was used to detect mRNA expression of HERC4, and Western blotting and immunohistochemistry were employed to detect protein expression of HERC4 in 67 breast cancer tissues and adjacent breast tissues.</p><p><b>RESULTS</b>The results of RT-qPCR showed a significantly higher mRNA expression of HERC4 in breast cancer tissues than in the adjacent breast tissues (P<0.05). Western blotting demonstrated HERC4 over-expression in breast cancer tissues compared with the adjacent breast tissues (P<0.05). Immunohistochemistry revealed HERC4 expression located predominantly in the cell cytoplasm. Positive HERC4 expression was detected in 61.2% of the breast cancer tissues as compared to 19.4% in the adjacent breast tissues, and its expression level was closely correlated with TNM stage and the histological grade (P<0.05).</p><p><b>CONCLUSION</b>HERC4 is correlated with the tumorigenesis and progression of breast cancer and may serve as a potential biomarker for early diagnosis and also as a potential therapeutic target in breast cancer.</p>
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Female , Humans , Blotting, Western , Breast , Metabolism , Breast Neoplasms , Metabolism , Disease Progression , Gene Expression Regulation, Neoplastic , Immunohistochemistry , Real-Time Polymerase Chain Reaction , Ubiquitin-Protein Ligases , MetabolismABSTRACT
10.3969/j.issn.1007-3969.2013.05.004
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<p><b>BACKGROUND AND OBJECTIVE</b>Lung adenocarcinoma (AC) is the most common type of lung cancer, however, its mechanism of oncongenesis is still unknown. The aim of this study is to screen candidate genes of lung adenocarcinoma using bioinformatics strategy and elucidate the mechanism of lung adenocarcinoma.</p><p><b>METHODS</b>Two published microarray data (GSE7670 and GSE10072) was obtained from Gene Expression Omnibus (GEO). Significance analysis of microarrays was performed with the software dchip, and differential expression genes from dchip analysis were defined as "test gene set". Genes correlated with lung adenocarcinoma, obtained by data mining tools genecard and Fable were regarded as "train gene set". Finally, candidate genes of lung adenocarcinoma were screened by the tool "Toppgene".</p><p><b>RESULTS</b>Three hundred and forty-four differential genes were defined as "test gene set", and 277 genes correlated with lung adenocarcinoma were regarded as "train gene set". Thirty-six candidate genes were screened out by Toppgene, among them, 21 genes had nearly no report in cancer. In the following QRT-PCR experiment, CD36, PMAIP1 and FABP4 were down-regulated expression in A549, which coincided with the gene chip.</p><p><b>CONCLUSION</b>It is demonstrated that Toppgene is useful in identification of the candidate genes of lung adenocacinoma, which provides the proof for the discovery of the specific disease genes.</p>
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Humans , Adenocarcinoma , Genetics , Computational Biology , Data Mining , Lung Neoplasms , Genetics , Oligonucleotide Array Sequence AnalysisABSTRACT
Fluorescent Linear Labelling technique is an effective, single-tube and linear amplification method. The sample cDNA was synthesized from total RNA, and was labelled antisense cRNA from double-stranded cDNA with T7RNA polymerase; it can be used for hybridization to oligonucleotide biochip. Fluorescent Linear Labelling Method can result in 50- to 100-fold higher degrees of amplification, and has been shown to retain information on transcript abundance, thus making it an efficient, robust technique for fluorescent labelling on biochip.
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Humans , Fluorescent Dyes , Chemistry , Oligonucleotide Array Sequence Analysis , Methods , Staining and Labeling , MethodsABSTRACT
To obtain non-pathogenic rabies virus glycoprotein(RV-G),we expressed RV-G in Saccaromyces Cerevisiae(S.cerevisiae).In our study,tat-G fusion gene was cloned into the expression vector pYes2.0,which allows expression of a foreign gene in the yeast cells under the control of GAl1 promoter.Transforma-tion was performed by using lithium-treated yeast cells and several Ura+-tranformants were isolated.Ac-cording to the relative mobility in SDS-PAGE,we know probably two forms(designated as yGI and yGⅡ) of RV-G analogues produced in S.cerevisiae,their molecular weights were estimated as 66 kD and 56 kD,respectively.On the other hand,there was a specific band about 56 kD shown in western blot result.Com-bining precursors’ achievements,we will draw a conclusion that trans-membrane domain(TD) and cyto-plasmic domain have a negative regulation on RV-G antigen immunogenicity in S.cerevisiae.
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Based on signal to noise ratio and probabilistic neural network method associated with experimental data,all analysis model in gastric carcinoma is presented.According to the available information,the samples of gastric carcinoma can be tested and ana.Lyzed.The signal to noise ratio is first calculated．Secondly,records in the database are chosen as a training set to build a probabilistie neural network model and the feature subset is selected according to accuracy.Finally,test set is to test accuracy of model．The model is implemented using MATLAB,and it can be generalized and applied to similar disease auxiliary diagnosis region.
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Objective To study the double restriction fluorescent labeling (DRFL) method for fluorescent labeling of trace DNA samples and its effect in enhancing the pathogen detection sensitivity of microarray assays. Method SARS-CoV RNA samples were reversely transcribed and then further amplified with the restriction display (RD)-PCR and fluorescently labeled by conventional restriction labeling directly with Cy-universal primer and the novel double labeling with Cy-universal primer and CydNTP. The labeled samples were applied to the microarray with the viral probes, processed and analyzed. Results Compared with the conventional method, DRFL labeling resulted in 3. 5835 times higher fluorescent intensity of all the SARS probes on average, even though increased fluorescent intensities for different probes varied considerably. Conclusion Signal to noise ratio can be enhanced by the DRFL method which improves the sensitivity of microarray technology in trace pathogen detections.
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Objective In the treatment of acute myeloid leukemia (AML-M2a), the first CCR (continuous complete remission) has been one of the most critical indicators to the prognosis of the patients. Using microarray approaches, gene expression profiles have been studied in patients with different CCR, in order to find out the genes relevant to the progresses of the AML. Methods Bone marrow mononuclear cells were collected and used as different experimental groups respectively. Group A composed of three AML patients with CCR<6 months, while group B composed of three AML patients with CCR>12 months. mRNAs were purified and labeled with Cy3 and Cy5 respectively, which were used to hybridize against the Agilent human 1B 60mer oligonucleotide microarrays. Results In the 20173 genes tested, 21 genes were found expressed differentially between these two groups. Of these differentially expressed genes, 10 genes were up-regulated while 11 genes were down-regulated in group A. Conclusion Through microarray studies, 21 genes including APP were found to be differentially expressed in AML patients whom were treated with standard chemotherapy. Theses genes can be early indicators for the diagnosis as well as prognosis of the refractory AML.
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BACKGROUND: Chronic myeloid leukemia (CML) is characterized by the clonal expansion of hematopoietic stem cells. Without effective treat ment, individuals in the indolent, chronic phase (CP) of CML will undergo blast crisis (BC), the prognosis for which is poor. Therefore, it is important to clarify the mechanism underlying CML from a whole-genome perspec tive. OBJECTIVE: To investigate the gene expression profile of bone marrow mononuclear cells from CML with Applied Biosystems Expression Array System.DESIGN: Observation and controlled analysis.SETTING: Institute of Gene Engineering, Southern Medical University PARTICIPANTS: Samples of two cases of bone marrow (a chronic myeloid leukemia patient and a healthy person).METHODS: This experiment was conducted at the Institute of Gene Engineering, Southern Medical University from October 2004 to September 2005.The total RNAs were extracted and purified from bone marrow mononuclear cells derived from a CML patient and a healthy person. mRNAs were purified using an oligo (dT)-cellulose mRNA purification kits and labeled using reverse transcription, in vitro transcription (RT-IVT), then hybridized with microarray. Gene expression differentiation of the bone marrow mononuclear cells were examined by ABI 1700 Chemiluminescent Microarray Analyzer. Reproducibility of microarray results was assessed by comparing data sets obtained from the same sample and analyzed by two different arrays.MAIN OUTCOME MEASURES: ①Assessment of quality of total RNA and labled cRNA. ②Reproducibility of microarray. ③ Hybridization of array.④Results of semi-quantitative reverse transcription-polymerase chain reaction RESULTS: ①Using statistical data analysis tools, we identified 6 706 genes that were up- or down-regulated in CML patient compared with the healthy person. In these genes, we found that 17 genes were up-regulated while 51 genes were down-regulated among 68 genes closely related to CML. ②most differentially expressed genes in C/EBPalpha mediated path way and CD40L signaling pathway had reduced expression. ③Good repro ducibility of microarray was confirmed by analysis of correlation and detection concordance in technical replicates. The correlation coefficient of the detectable probe in technical replicates was 0.991 for the CML patient and 0.988 for the healthy person. ④The results of semi-quantitative RT-PCR experiments supported the reliability of our microarray analysis.CONCLUSION: By comparing expression patterns of CML with those of the healthy person, we identified a large number of genes that, were up- or down-regulated in CML patients. These data should provide useful information for finding candidate genes whose products might serve as molecular targets for treatment of CML patients.
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Objective To report a new method of fluorescent labeling technique in microarray studies: universal primer U2 labeling( UPL). The efficiency was compared of the UPL with that of random primer, restriction display labeling method and the reverse transcription coupled random primer spiking labeling method(RT-PSL). Methods Influenza viral RNA was labeled with both UPL and the conventional random primer labeling method as well as two other more laborious labeling methods( RD-direct and RD-incorporate), and hybridized with influenza virus oligonucleotide microarrays. The signals extracted from the microarrays were analyzed using SPSS 10.0 software. Results The fluorescent intensity, signal-to-noise ration(SNR), true positive ratio(TPR) of probes and labeling reproducibility of UPL were demonstrated to be higher than those of the Random primer approaches.Conclusion These results established that UPL is a valid new labeling protocol, which may have wide applications in the research and development of the microarray technology.
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Objectives:To preparing the probes of cDNA microarray in detection of the Hepatitis D virus. Methods:The specific primers of PCR were designed accordion to the conserved region of HDV. Results: Sequences were aligned , and the results showed that the products of PCR amplification were the specific gene fragments of HDV. Conclusions:Using PCR amplification products to prepare gene chip probe was a quickly, simple effective method.
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<p><b>OBJECTIVE</b>To study the role of neuronal nitric oxide synthase (nNOS) in aged rats' hippocampal delayed neuronal death (DND) following brain ischemia.</p><p><b>METHODS</b>Models of incomplete brain ischemia were induced by clipping common carotid artery. A total of 46 aged SD rats were divided into 8 groups: normal control group (Group A, n=5), sham-operation group (Group B, n=5), reperfusion 1, 6, 12, 24, 48, and 96 hours groups after brain ischemia for 30 minutes (Group C, D, E, F, G, and H, n=6/group). The expression of nNOS was examined by immunohistochemistry and neuronal ultrastructural changes were observed by the transmission electron microscopy (TEM) at different time points after reperfusion.</p><p><b>RESULTS</b>Immunohistochemistry showed that nNOS expression in the hippocampal neurons was high in Group E, low expression in Group D, moderate expression in Group F and G. There was nearly no expression of nNOS in Group A, B, C, and H. Ultrastructure of hippocampal neurons was damaged more severely in reperfusion over 24 hours groups.</p><p><b>CONCLUSIONS</b>Nitric oxide (NO) may be one of the important factors in inducing DND after ischemia/reperfusion.</p>
Subject(s)
Animals , Female , Male , Rats , Apoptosis , Brain Ischemia , Hippocampus , Pathology , Immunohistochemistry , Microscopy, Electron , Neurons , Nitric Oxide Synthase , Metabolism , Rats, Sprague-Dawley , Reperfusion InjuryABSTRACT
<p><b>OBJECTIVE</b>To screen for the inhibitor of vascular endothelial growth factor (VEGF) 165 from random peptide library.</p><p><b>METHODS</b>Positive phage clones were rescued after two rounds of panning and competitive elution. Its affinity activity to KDR was monitored through ELISA, immunohistochemical method, Chicken CAM assay and MTT.</p><p><b>RESULTS</b>Five specific binding positive target molecule phage clones were obtained which were able to bind to cells whose surface had high KDR, among which, clone 3 and 13 could effectively block the vascularization of the chorioallantoic membrane of chick embryo, but they were not inhibitive on the proliferation of high KDR expression cells.</p><p><b>CONCLUSION</b>The peptides, being the inhibitors of VEGF, may be useful in the treatment of cancers.</p>
Subject(s)
Animals , Humans , Binding Sites , Endothelial Growth Factors , Metabolism , Enzyme-Linked Immunosorbent Assay , Intercellular Signaling Peptides and Proteins , Metabolism , Lymphokines , Metabolism , Peptide Library , Peptides , Pharmacology , Vascular Endothelial Growth Factor A , Vascular Endothelial Growth FactorsABSTRACT
[Objective] To observe the effect of Jiedu Lifei Oral Liquid (JLOL, mainly composed of Flos Lonicerae,Fructus Forsythiae, Radix Scutellariae, Bulbus Frityllariae, etc. ) on the immune function in mice with viral pniumonia.[Methods] Sixty mice were randomized into 5 groups: normal group (A), model group (B), virazole group (C), small-dose JLOL group (D) and large-dose JLOL group (E). Mouse models of viral pheumonia were induced by infection ofinfluenza virus FM1. Effects of JLOL on serum levels of interleukin-1 (IL-1), interleukin-2 (IL-2), tumor necrosisfactor ? (TNF-?) and interferon ? (IFN-?) were observed. [Results] Compared with normal group, serum levels of IL-1 and (IFN-? were decreased and TNF-? increased (P 0.05 ) in model group.After treatment wth JLOL, serum levels of IL-2 and IFN-? were increased , TNF-? decreased (P 0.05). [ Conclusion] JLOL can increase the immune function by increasing the serumlevels of IL2 and IFN-? and can reduce the immune impairment by inhibiting the over production of TNF-?.
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Objective To investigate the applieation of restriction display (RD) technique in the preparation of HCV probes of clinical genotyping microarray. Methods Restriction enzyme Sau3A Ⅰwas chosen to digest the full-length HCV cDNAs of three distinct subtypes, i.e.1a, 1b and 2a. The resultant restrictive fragments were then ligated with universal adapters. PCR primers were designed to match the universal adapters but with one "nesting" base overhanging at the 3′- end. The PCR reactions were performed by ten pairs of different primer combinations. The differential genes were separated through polyacrylamide gel electrophoresis and silver staining. The second-round PCR was performed using the isolated bands as PCR templates. The purified PCR products were then cloned into T-vectors. The recombinant plasmids were extracted from positive recombinant clones and the target gene fragments were sequenced. Results The target HCV gene fragments ranging from 200 to 900bp were isolated and sequenced, which were correlated precisely with the RFLP (Restriction Fragment Length Polymorphism) prediction. A total of 66 different fragments were obtained, averaging about 22 for each subtypes. These fragments could be further used as probes in HCV microarray preparations. Conclusion RD technique is of great value in obtaining a large number of equal sized gene probes, which provide a swift protocol in generating DNA probes for the preparation of microarrays.
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Objective To investigate the feasibility of preparing HCV-1 diagnostic microarray probes using the technique of cDNA fragments library construction. Methods The full-length cDNAs of HCV of subtypes 1a and 1b were digested with restriction endonuclease Sau3A I, and the resulted fragments were cloned into the pMD18-T vector. Positive clones were isolated and identified by sequencing. Results A total of 57 different fragments were obtained, and sequence analysis showed that all the fragments ranging from 200 to 1000bp were specific gene fragments of HCV genotype 1, which can be efficiently used as probes in microarray prepapration. Conclusion The method of preparing microarray probes by construction of cDNA fragments library was effective, quick and simple.
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AIM: To investigate the effect of canstatin on cultured rabbit vascular smooth muscle cells(VSMC). METHODS: By means of cationic liposome mediated method, canstatin RNA was transferred into cultured VSMC. The proliferation quantity of VSMC were determined by the cell counting method and thymidine(-TdR) incorporation. RESULTS: Canstatin RNA could be effectively transferred into cultured primary rabbit aortic smooth muscle cells by the cationic liposome-Dosper and could markedly inhibit VSMC proliferation. CONCLUSION: Transfection of canstatin RNA could inhibit the growth of VSMC in vitro.
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Objective: To isolate small molecular polypeptides which bind to KDR specifically using C7 and 12 peptide libraries. Methods: KDR/IgGFc was coated directly on plates, C7 and 12 peptide libraries are then applied, followed by vigorous washings in washing buffers to remove most non-specifically bound phage and to select specific phage particals by VEGF suspension and elution in acid buffers. The positive phage clones were detected by ELISA, cell-ELISA and competitive binding ELISA. Results: After three washes, 12 positive phage clones were selected and sequenced. Conclusion: A conserved peptide motif was not found in these sequences. The peptide binding to KDR specifically may be helpful for lead drug to improve selectivity and decrease the side effect of anti cancer drug in clinical treatment.