45 results about "Paraoxonase" patented technology

The invention provides methods of screening a mammalian subject to determine if the subject is at risk to develop, or is suffering from, cardiovascular disease. The methods comprise detecting an amount of at least one biomarker in a biological sample, or HDL subfraction thereof, from the subject, and comparing the detected amount of the biomarker to a predetermined value, where a difference between the detected amount and the predetermined value is indicative of the presence or risk of cardiovascular disease in the subject. In some embodiments, the biomarker comprises at least one of ApoC-IV, Paraoxonase 1, C3, C4, ApoA-IV, ApoE, ApoL1, C4B1, Histone H2A, ApoC-II, ApoM, Vitronectin, Haptoglobin-related protein, and Clusterin, or combinations thereof.

The invention provides methods and compositions for treating or preventing ischemic reperfusion injury. The methods of the instant invention comprise the administration of compositions comprising apolipoproteins, lecithin cholesterol acyltransferase or paraoxonase and lipid complexes thereof to treat, reduce or prevent ischemic reperfusion injury.

The present invention relates to antisense oligonucleotides that modulate the expression of and / or function of Paraoxonase 1 (PON1), in particular, by targeting natural antisense polynucleotides of Paraoxonase 1 (PON1). The invention also relates to the identification of these antisense oligonucleotides and their use in treating diseases and disorders associated with the expression of PON1.

The inventors have proposed a novel panel of human plasma protein biomarkers for diagnosing hepatic fibrosis and cirrhosis. Presently there is no reliable non-invasive way of assessing liver fibrosis. A 2D-PAGE based proteomics study was used to identify potential fibrosis biomarkers. Plasma from patients with hepatic cirrhosis induced by infection with the hepatitis C virus (HCV) were analysed. Several proteins associated with liver scarring and potentially also related to viral infection were identified. These proteins include 14-3-3 protein zeta / delta, adiponectin, afamin, alpha-1-antitrypsin, alpha-2-HS-glycoprotein, apolipoprotein C-III, apolipoprotein E, C4b-binding protein beta chain, intact / cleaved complement C3dg, corticosteroid-binding globulin, fibrinogen gamma chain, beta haptoglobin at pH 5.46-5.49, haptoglobin-related protein, hemopexin, immunoglobulin J chain, leucine-rich alpha-2-glycoprotein, lipid transfer inhibitor protein, retinol-binding protein 4, serum paraoxonase / arylesterase 1, sex hormone-binding globulin and zinc-alpha-2-glycoprotein. These biomarkers can be used in conjunction with polypeptides in WO / 2008 / 031051. The concentrations of these novel biomarkers can be determined using an immunoassay where the concentrations would reflect the extent of fibrosis. A fibrosis scoring scale for each of the novel biomarkers is proposed. The additive result from the scores of all the novel biomarkers would give a more reliable indication of the degree of fibrosis rather than examining individual biomarkers.

Compositions and methods relating to ApoA-1 fusion polypeptides are disclosed. The fusion polypeptides include a first polypeptide segment corresponding to an ApoA-1 polypeptide or ApoA-1 mimetic, and may also include a dimerizing domain such as, e.g., an Fc region, which is typically linked carboxyl-terminal to the first polypeptide segment via a flexible linker. In some embodiments, the fusion polypeptide further includes a second polypeptide segment located carboxyl-terminal to the first polypeptide segment and which confers a second biological activity (e.g., an RNase, paraoxonase, platelet-activating factor acetylhydrolase, cholesterol ester transfer protein, lecithin-cholesterol acyltransferase, polypeptide that specifically binds to proprotein convertase subtilisin / kexin type 9, or polypeptide that specifically binds to amyloid beta). Also disclosed are dimeric proteins comprising first and second ApoA-1 fusion polypeptides as disclosed herein. The fusion polypeptides and dimeric proteins are useful in methods for therapy.

The invention belongs to biotechnology pharmacy technology field. The recombination plasmid Pci-Hpon3 expressed in zooblast is constructed by used human mintacol enzyme 3 gene (Hpon3), Hpon3 is expressed efficiently and secreted into blood in small murine skeletal muscles organizing in electric transferring method to improve PON3 in blood serum and enforce the ability of small mouse antioxidation pressure so that it can control liver damage efficiently. The activity of Hpon3 transgene expression small mouse blood serum hydrolytic decomposition melilotol is improved by 1.4 times and can maintain 24 days. After small mouse acute or subacute liver damage mould is built by injecting carbon tetrachloride into abdominal cavities of transgene group and normal small mouse, the blood serum ALT and AST level of transgene small mouse are lower greatly, malondialdehyde (MDA) level of liver homogenate is lower, reducing glutathione (GSH) and total antioxidant ability (T-AOC) are higher than contrasting group, and transgene group small mouse liver cell damage extent indicated in nosology slicer result is lower than CC1(4) control group's indicate that recombination human PON3 plasmid transgene expression can improve efficiently blood serum PON3 activity and reduce liver oxygenizing pressure so that it can be used for preventing and treating liver damage.

Disclosed are compounds of the formula (I): wherein R3, R4, R5, R9, and R10 are selected from the group consisting of H and groups or atoms other than H, and R6 and R8 are halo or hydrogen; X1, X2, and X3 are independently O or S; provided that R9 and R10 are not simultaneously H, when all of X1, X2, and X3 are O; and of the formula (II) wherein R11-R14 are selected from the group consisting of H and groups or atoms other than H; X4-X9 are independently O or S; n and m are 0 or 1 but m and n cannot be 0 simultaneously; R15-R24 can be H or any substituent so long as the compound of formula II upon hydrolysis provides a fluorescent compound. These compounds are useful as substrates with high specificity for organophosphatase particularly human paraoxonase and bacterial organophosphorus hydrolase. Also disclosed is a method for detecting and / or measuring the paraoxonase activity in a fluid comprising contacting the fluid with a fluorescent substrate and measuring the fluorescence of the fluorescent product formed.

The invention provides methods of screening a mammalian subject to determine if the subject is at risk to develop, or is suffering from, cardiovascular disease. The methods comprise detecting an amount of at least one biomarker in a biological sample, or HDL subfraction thereof, from the subject, and comparing the detected amount of the biomarker to a predetermined value, where a difference between the detected amount and the predetermined value is indicative of the presence or risk of cardiovascular disease in the subject. In some embodiments, the biomarker comprises at least one of ApoC-IV, Paraoxonase 1, C3, C4, ApoA-IV, ApoE, ApoL1, C4B1, Histone H2A, ApoC-II, ApoM, Vitronectin, Haptoglobin-related protein, and Clusterin, or combinations thereof.

The invention belongs to the technical field of clinical diagnosis of cardiovascular diseases, and particularly relates to a PON-1 (paraoxonase-1) active detecting reagent kit and a detecting method thereof which are suitable for quantitative assay of the activity of PON-1 in body fluid such as human serum in vitro through a biochemical analyzer and used as the assisting diagnosis. The reagent kit comprises a buffer solution and a substrate solution, wherein the buffer solution contains 0.15mol / L glycine, 2mol / L NaOH (sodium hydroxide), 2.5*10<-3>mol / L CaCl2 (calcium chloride) and a water solvent; and the substrate solution is 0.15mol / L to 0.42mol / L diethyl p-nitrophenyl phosphate, and the solvent is dimethyl sulfoxide. The PON-1 active detecting reagent kit has the advantages that the detecting method of the reagent kit is fast and simple, the early cardiovascular disease can be effectively diagnosed by assaying the activity of the PON-1 in the serum; and the synthesis method of the dimethyl sulfoxide is fast, simple and safe compared with the prior art, and the yield is high.

A method of determining a level of biologically active PON enzyme is provided. The method comprising determining lactonase activity of the PON enzyme, the lactonase activity being indicative of the level of biologically active PON enzyme. Also provided are novel compounds which may be used for measuring a lactonase activity of an enzyme.

The invention discloses a method for detecting enzymatic activity of PON-1 in a blood sample and application thereof. The method comprises the following steps: (a) taking paraoxon as a substrate, and detecting the activity of paraoxonase-1, PON-1 in a blood serum or blood plasma sample; (b) detecting the apolipoprotein AI (apo AI) level in the sample by immunity transmission turbidity; (c) calculating the ratio of the activity of the PON-1 to the apo AI level; and (d) comparing the calculated ratio with at least one known index indicating existence or inexistence of coronary heart disease. The method specifically and sensitively evaluates the existence of fatalness of the coronary heart disease by reflecting the relative activity of the PON-1 in HDL particles. The detection of the enzymatic activity is applied in preparing medicaments for treating or preventing cardiovascular and cerebrovascular diseases such as the coronary heart disease, miocardial infarction and the like. The PON-1 is a characteristic antioxidant enzyme having main antioxidant function in the HDL, and the activity thereof can reflect the antioxidant function of the HDL, so activity detection of the PON-1 is taken as a method and a tool for evaluating the fatalness of the coronary heart disease.

The invention discloses a recombined Q-typed hPON1Q in the biological pharmaceutical technological domain, which is characterized by the following: transmitting expressive plasmid to do ectopic secretory expression through skeletal muscle excited by electricity; improving serum PON1 level effectively; preventing liver from damaging as new gene drug; maintaining the activity of phosphoroso enzyme over 16d; making the genome-switching mouse serum ALT and AST level lower than positive contrast group (P<0.01).

Disclosed are compounds of the formula (I): wherein R3, R4, R5, R9, and R10 are selected from the group consisting of H and groups or atoms other than H, and R6 and R8 are halo or hydrogen; X1, X2, and X3 are independently O or S; provided that R9 and R10 are not simultaneously H, when all of X1, X2, and X3 are O; and of the formula (II) wherein R11-R14 are selected from the group consisting of H and groups or atoms other than H; X4-X9 are independently O or S; n and m are 0 or 1 but m and n cannot be 0 simultaneously; R15-R24 can be H or any substituent so long as the compound of formula II upon hydrolysis provides a fluorescent compound. These compounds are useful as substrates with high specificity for organophosphatase particularly human paraoxonase and bacterial organophosphorus hydrolase. Also disclosed is a method for detecting and / or measuring the paraoxonase activity in a fluid comprising contacting the fluid with a fluorescent substrate and measuring the fluorescence of the fluorescent product formed.

The invention relates to a diethyl nitrophenyl phosphoesteraase diagnosis / determination reagent kit which utilizes an enzyme colorimetry method and an enzyme-linked method technology, simultaneously the invention also relates to a method principle for determining the diethyl nitrophenyl phosphoesteraase activity concentration, the composition and the components of a reagent, belonging to the technical field of the medicine examination and determination. The main components of the reagent kit of the invention comprise buffer solution, coenzyme, aryl dialkyl phosphoric acid, aryl alcohol dehydrogenase, and stabilizer. A sample is mixed with the reagent according to a certain volume to lead to perform a series of enzymatic reaction, a reacting substance is arranged under an ultraviolet / visible light analyzer to examine the rising degree / speed of absorbance at the position of a 340 nm dominant wavelength, thereby detecting and calculating the diethyl nitrophenyl phosphoesteraase concentration value. Through the adoption of the invention, the required determined results can be obtained completely through the ultraviolet / visible light analyzer.

The inventors have proposed a novel panel of human plasma protein biomarkers for diagnosing hepatic fibrosis and cirrhosis. Presently there is no reliable non-invasive way of assessing liver fibrosis. A 2D-PAGE based proteomics study was used to identify potential fibrosis biomarkers. Plasma from patients with hepatic cirrhosis induced by infection with the hepatitis C virus (HCV) were analysed. Several proteins associated with liver scarring and potentially also related to viral infection were identified. These proteins include 14-3-3 protein zeta / delta, adiponectin, afamin, alpha-1-antitrypsin, alpha-2-HS-glycoprotein, apolipoprotein C-M, apolipoprotein E, C4b-binding protein beta chain, intact / cleaved complement C3dg, corticosteroid-binding globulin, fibrinogen gamma chain, beta haptoglobin at pH 5.46-5.49, haptoglobin-related protein, hemopexin, immunoglobulin J chain, leucine-rich alpha-2-glycoprotein, lipid transfer inhibitor protein, retinol-binding protein 4, serum paraoxonase / arylesterase 1, sex hormone-binding globulin and zinc-alpha-2-glycoprotein.

The present disclosure provides probes for detecting a polymorphism in the PON1 gene.

The invention discloses a PON-1 (paraoxonase-1) activity detection kit and a detection method for the same. The kit comprises two reagents, namely, a buffer solution and a substrate solution, wherein the substrate solution contains 0.15-0.42 mol / L 2-chloro-p-nitrophenyl diethyl phosphate and a solvent, namely, dimethyl sulfoxide; and the buffer solution contains 0.15 mol / L glycine, 2 mol / L NaOH, 0.0025 mol / L CaCl2 and an aqueous solvent, and has a pH value of 10.0-11.0. The detection method adopting the kit is rapid, simple and convenient, and early cardiovascular diseases can be effectively by measuring the activity of PON-1 in serum; and compared with the prior art, a synthesis method for 2-chloro-p-nitrophenyl diethyl phosphate is rapid, simple and convenient, safe, and high in yield.

The present invention pertains to a new method for the diagnosis, prognosis, stratification and / or monitoring of a therapy, of cancer, preferably colorectal cancer (CRC), in a subject. The method is based on the determination of the level of a panel of least one, preferably 3, 4 and most preferably at least 5, protein biomarker selected from the group consisting of the protein biomarkers Amphiregulin (AREG), Carcinoembryonic antigen (CEA), Insulin like growth factor binding protein 2 (IGFBP2), Keratin, type I cytoskeletal 19 (KRT19), Mannan binding lectin serine protease 1 (MASP1), Osteopontin (OPN), Serum paraoxonase lactonase 3 (PON3) and Transferrin receptor protein 1 (TR), in the biological sample obtained from the subject. The new biomarker panel of the invention allows diagnosing and even stratifying various cancer diseases. Furthermore, provided are diagnostic kits for performing the non-invasive methods of the invention. Since the biomarker panel of the invention provides a statistically robust method independent of the protein detection technology used, and considering that the biomarker panel of the invention is detected in plasma samples of the subjects, the invention provides an early detection screening examination that may be applied to a larger population.

The invention relates to a human paraoxonase (PON) 2 gene 148 site G / A polymorphic detection kit. The detection kit comprises a forward primer and a reverse primer in a sequence near the amplified human PON2 gene polymorphic rs12026 site and a restriction enzyme, wherein the forward primer is shown in SEQ ID NO:12, the reverse primer is shown in SEQ ID NO:13, and the restriction enzyme is BsmAI. The invention has the advantages of simple operation process, low cost and wide operation range.

This invention is directed to a DNA sequence comprising a nucleotide sequence encoding a variant paraoxonase protein and to said variant paraoxonase protein as well as a method and a kit for detecting a risk of cancer, coronary or cerebrovascular disease, hypertension, type 2 diabetes, dementia, joint arthrosis, cataract, or sensitivity to organophosphorus compounds in a subject, the method comprising isolating genomic DNA from said subject, determining the allelic pattern for the codon 102 of the paraoxonase encoding PON1 gene in the genomic DNA, identification of Ile101Val mutation indicating said risk being increased and for targeting paraoxonase activity modulating therapies. Further this invention relates to transgenic animals comprising a human DNA molecule encoding said variant paraoxonase and to a method of phenotype-targeted gene sequencing.

Human serum paraoxonase enzyme and DNA (RNA) encoding such serum paraoxonase enzymes are disclosed. Also provided is the procedure for producing such polypeptides by recombinant techniques. Uses of such polypeptides include their use as an antidote for organophosphate poisoning and to prevent neuronal cell death.

Human serum paraoxonase enzyme and DNA (RNA) encoding such serum paraoxonase enzymes are disclosed. Also provided is the procedure for producing such polypeptides by recombinant techniques. Uses of such polypeptides include their use as an antidote for organophosphate poisoning and to prevent neuronal cell death.

The invention discloses a method for detecting atherosclerosis and / or coronary heart disease susceptibility, which includes detecting whether variation exists in individual paraoxonase 3 (PON3), transcript and / or albumen in comparison with normal. The existence of variation shows that the possibility that an individual contracts atherosclerosis and / or coronary heart disease is higher than that of a normal person. The invention also discloses a corresponding detection kit.

The present invention relates to a method of identifying a subject predisposed to lacunar stroke. The method includes the step of identifying in the subject the presence of a thymine to cytosine mutation at position -107 in both alleles of the paraoxonase 1 locus.

The invention discloses a method for detecting atherosclerosis and / or coronary heart disease susceptibility, which includes detecting whether variation exists in individual paraoxonase 3 (PON3), transcript and / or albumen in comparison with normal. The existence of variation shows that the possibility that an individual contracts atherosclerosis and / or coronary heart disease is higher than that of a normal person. The invention also discloses a corresponding detection kit.

The invention relates to a neural tube defect prenatal noninvasive diagnosis molecular marker which comprises 6 miRNA (miR-144, miR-142-3p, miR-720, miR-1275, miR-575 and miR-765) and 12 proteins (pro-protein convertase subtilisin / kexin 9, ceruloplasmin, leukaemia inhibitory factor, 14-3-3, paraoxonase, plasma alpha-globulin inhibiting factor H4, serum S protein, glycosyl-phosphatidylinositol specific phospholipase D1, fetuin A, amyloid protein P, plasma alpha-globulin inhibiting factor H2 and platelet factor 4). The invention further provides application of the molecular marker.

The invention relates to a human paraoxonase (PON) 2 gene 148 site G / A polymorphic detection kit. The detection kit comprises a forward primer and a reverse primer in a sequence near the amplified human PON2 gene polymorphic rs12026 site and a restriction enzyme, wherein the forward primer is shown in SEQ ID NO:12, the reverse primer is shown in SEQ ID NO:13, and the restriction enzyme is BsmAI. The invention has the advantages of simple operation process, low cost and wide operation range.

The invention discloses a structure of a human recombinant methionine sulfoxide reductase A (MsrA) protein, and a construction method of the structure. The structure contains a section of compositionstructure of human EpK peptides; and the structure of the MsrA is 213 amino acid chains without signal peptides which are connected to form a complete human EpK-MsrA fusion protein polypeptide chain through encoded amino acids of a BamHI restriction enzyme identification sequence. The prepared recombinant EpK-MsrA fusion protein is highly expressed in the liver in a mode of infecting scavenger receptor B-group I-type gene knockout (SR-BI- / -) mice with slow viruses and can be secreted into blood; and the secreted EpK-MsrA fusion protein can be characteristically bound to high-density lipoprotein (HDL), thus the content of apolipoprotein (AI) and the content of paraoxonase 1 of HDL functional protein components are increased, the activity of the AI and the activity of the paraoxonase 1 of HDL functional protein components are improved, inflammatory components in plasma and the HDL are reduced, and the effect of resisting an atherosclerotic cardiovascular disease is achieved.

The invention relates to protein transduction peptide-paraoxonase 1 fusion protein and a preparation method and application thereof, in particular to a mammalian cell of protein transduction peptide-paraoxonase 1 (P11-PON1) fusion protein and a bombyx mori expressing method. The fusion protein has activity that the mucosa is absorbed into the body and can penetrate through the blood brain barrier. The fusion protein has a treatment effect on organic phosphorus pesticide poisoning and tumors, atherosclerosis, the coronary disease, diabetes mellitus type 2 and other diseases.

The invention relates to a neural tube defect prenatal noninvasive diagnosis molecular marker which comprises 6 miRNA (miR-144, miR-142-3p, miR-720, miR-1275, miR-575 and miR-765) and 12 proteins (pro-protein convertase subtilisin / kexin 9, ceruloplasmin, leukaemia inhibitory factor, 14-3-3, paraoxonase, plasma alpha-globulin inhibiting factor H4, serum S protein, glycosyl-phosphatidylinositol specific phospholipase D1, fetuin A, amyloid protein P, plasma alpha-globulin inhibiting factor H2 and platelet factor 4). The invention further provides application of the molecular marker.