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Next generation sequencing(NGS)has made great progress, applied from science technology to clinical diagnosis in the past several years.With a simple, fast, high-throughput, and high-resolution feature, it has been believed to be one of the most potential technology in clinical tests, especially in individual tumor diagnosis and treatment, noninvasive prenatal screening, genetic disorders screening field.This paper will briefly review the application of next generation sequencing in the field of infection disease.
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Objective To use the flow cytometry to detect the platelet vasodilator-stimulated phosphoprotein(VASP)phospho-rylation level and to evaluate the clopidogrel curative effect after PCI.Methods 17 cases in the control group without any drug in-tervention and 26 cases of acute coronary syndrome(ACS)after PCI operation with clopidogrel were selected.Platelet VASP phos-phorylation levels at being selecting and on 7 d after anti-platelet therapy were detected by the flow cytometry and the platelet reac-tivity index (PRI)was calculated.Results The PRI after anti-platelet therapy in the ACS group was decreased significantly,the difference between before treatment and after treatment had statistical significance (P <0.05 ).Conclusion The platelet VASP phosphorylation level detected by the flow cytometry can specifically evaluate the effect of clopidogrel.
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Genetic factors are important causes of male infertility and account for about 30%infertility cases.Therefore, it is necessary to carry out molecular and genetic detections in the diagnosis and treatment of male infertility.Here the most commonly used techniques for molecular and genetic diagnosis of male infertility in recent years are discussed , including chromosomal abnormities analysis , Y chromosome microdeletions detection , gene mutation screening and sperm quality and function examination.
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Objective To explore earlier diagnostic value of ischemia modified albumin(IMA)in acute coronary syndrome(ACS) patients.Methods We detected serum IMA and CK-MB in 117 patients with ACS and 100 healthy controls to analyze the diagnos-tic value of IMA in ACS.Results When the critical value of 73.25 U/mL,serum IMA testing determine the ACS of the sensitivity, specific,positive predictive value and negative predictive value of 89.6%,86.0%,84.93% and 86.0% respectively.IMA levels in serum in patients with AMI group was obviously higher than that of healthy control group(P <0.05).Sensitivity could be improved to 91.5% when detected IMA and CK-MB together.And the specific was 71.5%.Conclusion Serum IMA can be used as sensitive index of early diagnosis and risk stratification of ACS.Detecting IMA and CK-MB can improve the diagnostic sensitivity of ACS.
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Objective To investigate the value of the flow cytometry(FCM)for detecting platelet associated antibody in the di-agnosis of immune thrombocytopenia(ITP).Methods Platelet associated antibody,expression rate and the fluorescence intensity of platelet-associated immunoglobulin(PAIg)were measured in 51 patients with ITP(23 cases of newly diagnosed ITP and 28 cases of persistent ITP),21 patients with non-ITP and 60 healthy individuals.The correlation between the detection results with the platelet and megakaryocyte counts was performed;the expression rates of PAIg before and after treatment were compared.Results The fluorescence positive percentage and mean fluorescence intensity of IgG,Ig A and IgM in the patients group were significantly high-er than those in the control group with statistical difference(P 0.05);the difference between the autoimmune disease group and the healthy control group had statistical significance(P <0.01 );in the persistent ITP group,PAIg in the complete response group had statistical difference between the groups before treament.PAIg in the response group also had statistical difference between the groups before treament (P <0.05).Conclusion FCM for detecting PAIg can be used in the diagnosis,differential diagnosis and the disease condition monitoring of ITP.
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To identify the prevalence and the distribution of ticks infected with spotted fever group rickettsiae (SFGR) in Xunke Area of Heilongjiang Province ,China ,partial outer membrane protein A gene (ompA) and citrate synthase gene (gltA) specific fragments were assessed using the PCR method .The positive products were sequenced .Result showed that the pres-ence of SFGR was 14 of 60 in detection Dermacentor silvarum cases ,while the overall positive rate was 23 .33% .Its nucleotide sequence of ompA showed 99 .3% and 99 .18% similarity with nucleotide sequence of Rickettsia sp .JL-02 and Rickettsia rao-ultii respectively .And the evolutionary positions of SFGR species were similar with Rickettsiamontana and Rickettsiamassili-ae .It's concluded that the nature focus of tick-borne spotted fever did exist in the area of Xunke Area of Heilongjiang Province , China .
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Objective To verify the analytical performance of the Roche cobas 8000 automatic biochemical analyzer.Methods The intra-batch and inter-batch precisions,accuracy,linearity and reference interval were validated according to the Clinical and La-boratory Standards Institute (CLSI)documents (EP5-A2,EP15-A2,EP6-A;C28-A2 ).Results The intra-batch precision was 0.5%-2.3% and the inter-batch precision was 0.8%-3.11%,which were all less than 1/4 Tea(CLIA'88).The detection results of various items had better correlation with the target values(r2 >0.99);the linear range of various detected items were in line with the linear range provided by the specification;the quotative biological reference range conformed to the group of this laboratory services.Conclusion The various performance indexes of the Roche cobas 8000 automatic biochemical analyzer conform to the qual-ity requirements and this analyzer can be used in clinical biochemical detection work..
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microRNAs ( miRNAs) are a class of endogenous noncoding small molecular RNAs , which regulate target gene expression at the post-transcriptional level and play vital roles in cell development , proliferation, differentiation, tumorigenesis and other biological behavior.Changes in miRNA expression profiles have been observed in gastric cancer in recent studies.Further studies demonstrated that miRNAs were closely related to the carcinogenesis , development and metastasis of gastric cancer , which suggested miRNAs have broad prospective application in diagnosis and therapy of gastric carcinoma as biomarkers and therapeutic targets.
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<p><b>OBJECTIVE</b>To study the expression and mechanism of WNK4 gene regulated by aldosterone.</p><p><b>METHODS</b>Wistar rats were treated with aldosterone and potassium water. Serum aldosterone and ion as well as urine ion were measured. The expression of WNK4 gene in kidney tissues was detected by real-time PCR. Kidney-derived HEK293 cells were cultured, transfected with pGL3-WNK4, and then stimulated by aldosterone. After 24 h of transfection, luciferase activities of the plasmid were detected.</p><p><b>RESULTS</b>Compared with those of the controls, serum aldosterone and urine K(+) of experimental rats were significantly elevated, whilst urine Na(+) was significantly decreased. And urine Cl(-) was significantly increased only in the group of high K(+). Serum K(+), Na(+) and Cl(-) showed no significant difference. Expression of WNK4 gene in kidney tissues was significantly decreased. The luciferase activity of pGL3-WNK4-484 plasmid has decreased after stimulated with aldosterone, while the activity of pGL3-WNK4-275 showed no change.</p><p><b>CONCLUSION</b>Aldosterone can down-regulate the expression of WNK4 through binding with regulatory element in the upstream of the gene.</p>
Subject(s)
Animals , Humans , Male , Rats , Aldosterone , Blood , Pharmacology , Cell Line , Chlorides , Blood , Gene Expression Regulation , Kidney , Metabolism , Potassium , Blood , Promoter Regions, Genetic , Protein Serine-Threonine Kinases , Genetics , Metabolism , Sodium , Blood , Transcriptional ActivationABSTRACT
<p><b>OBJECTIVE</b>To identify and characterize laryngeal cancer related novel genes located on chromosome 6q25.</p><p><b>METHODS</b>Electric hybridization was performed in human genome database using EST (expression sequence tag) as probe. Novel genes were deduced by software from positive DNA clones and their cDNAs were amplified by RT-PCR using primers designed according to the sequence of the putative genes.</p><p><b>RESULTS</b>A novel gene was cloned successfully. The full length of this gene was about 21 kb. It contained two exons and produced a 1006 bp transcript coding a protein with 235 amino acid residues. It's 5'flanking sequence contained two binding sites of oncoprotein c-Myc, thus it was named MTLC (c-Myc target from laryngeal cancer cells). Homologous assay showed that MTLC exhibited little overall homology to known human proteins but it exhibited good overall homology to mouse MT-MC1 protein with an identity of 78%. The primary structure of MTLC protein contained a nuclear location signal motif, but it did not have other conserved domains. The results of subcellular location experiment showed that MTLC expressed in nuclei of human hepatocellular carcinoma cell line Bel7402 cells, while a wide distribution of MTLC in various tissues was demonstrated by Northern blotting.</p><p><b>CONCLUSION</b>MTLC may play an important role as a target gene of c-Myc and as a transcription factor in keeping the normal physiological process of cells.</p>