CN103336127A - Indicator and diagnostic agent of early-stage hepatopathy, hepatic fibrosis and hepatic cirrhosis - Google Patents
Indicator and diagnostic agent of early-stage hepatopathy, hepatic fibrosis and hepatic cirrhosis Download PDFInfo
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Abstract
The invention relates to a use of a gene or a protein kallistatin (KAL) in preparation of an indicator and a diagnostic agent of early-stage hepatopathy, hepatic fibrosis and hepatic cirrhosis. The KAL is a tissue kininase-binding protein. An experiment proves that the KAL can be used as a novel, sensitive, fast and accurate indicator and a diagnostic agent of early-stage hepatopathy/hepatic fibrosis and can be clinically used.
Description
Technical field:
The invention belongs to medical science and detect and biological technical field, relate to the application in preparation early stage hepatopathy, liver fibrosis and cirrhosis indicator and diagnosticum of a kind of albumen or gene.
Technical background:
Liver fibrosis is that liver is tackled the self-regeneration reaction that the different causes of disease (inflammation, alcohol, parasite, chemical toxicant, immunity, Nutrition and Metabolism obstacle, blood circulation disorder, chronic viral infection, bile duct obstruction etc.) damage stimulates.The liver cell secretion various kinds of cell factor of damage, follow cell factor and some chemical mediator of sinusoidal endothelial cell, secretion of platelet to activate HSCs jointly, make it be converted into liver myofibroblast (myofibroblasts, MFB), MFB is by paracrine and autocrine effect, and producing a large amount of is extracellular matrix (the extra cellular matrix of Main Ingredients and Appearance with collagen, ECM), the transition deposition of ECM, synthetic unbalance with degraded, cause generation, the development of liver fibrosis.Liver fibrosis is the common pathologic process of multiple chronic liver disease, and the sustainable development of liver fiber can change cirrhosis even liver cancer into.In a single day studies have shown that liver fibrosis is reversible, and develop into cirrhosis, just enter irreversible state, even deterioration is liver cancer.
Liver fibrosis, cirrhosis are the total pathological examinations of chronic injury hepatopathy, and the reconstruction that the lobuli hepatis structure all can take place destroys.Liver fibrosis especially cirrhosis pathology be difficult in early days be diagnosed, occur up to a series of serious symptoms, as hepatic failure, liver ascites, portal hypertension, hepatic encephalopathy, varication even liver cancer.In case develop complications, illustrate that liver fibrosis or cirrhosis have arrived the mistake compensatory phase or more late, the state of an illness has been difficult to control.For viral cirrhosis of compensatory phase, develop into decompensated liver cirrhosis, liver cancer even dead probability every year and be respectively 4%, 3%, 3%.So, find a kind of effective diagnosis technology or index, to solve the hepatopathy high incidence, high mortality is most important.Used diagnostic method and index mainly contains following several at present:
Liver biopsy:
Because the pathological characters of liver fiber or cirrhosis, liver biopsy are to describe liver histological, " goldstandard " of disease activity degree and liver fibrosis, or the unique sources of information such as steatosis and iron content state always.Liver fibrosis classification and many with semiquantitative Ishak and METAVIR system by stages.Hepatic tissue biopsy is at present all playing an important role aspect medical diagnosis on disease, evaluate its prognosis and the treatment evaluation, but also has many drawbacks: at first, biopsy is taken a sample and is brought very big misery to patient, and patient is unwilling to accept this inspection unless it is absolutely necessary; Secondly, liver fiber or cirrhosis are dynamic process, especially a liver fibrosis, reversible dynamic process especially, stimulate eliminate after, liver self-regeneration, restore funcitons.Sampling only represents the pathological condition of a time point, and the order of severity of system postulation fiberization between each stage is linear by stages increases, and is difficult to accurately dynamically reflect the true appearance of liver new city metabolism.
Secondly, there is sampling error in pathological biopsy, liver is organ maximum in the human body viscera, two leaves about having, adult's liver weight is equivalent to 2% of body weight, length and width, thick 25.8cm, 15.2cm, the 5.8cm of being about respectively, diseased region distributes not necessarily uniformly during pathology damage, and a little tissue of biopsy does not represent the pathology situation of whole liver.At last, also there is the difference between the observer in biopsy, and the evaluation of liver fibrosis remains the evaluation of observer's subjectivity, and its error is up to 24%.Adopt different points-scoring system (Ishak or METAVIRR points-scoring system), be difficult to estimate the result of different researchs.
Imaging technique:
The indirect diagnosing hepatic pathology situation of imaging technique Noninvasives such as ultrasonic, CT and MRI by the portal hypertension sign, has good susceptibility and specificity in detecting in the terminal stage of a disease.If any reporting, Ultrasonic Diagnosis sensitivity is 87%, and biopsy just has only 62%; Yet they are to slight or moderate fiberization susceptibility is not enough, and disturbing factor is a lot of in the testing process.In recent years, ultrasonic instantaneous elasticity imaging technique is subjected to extensive attention, thereby it utilizes ultrasonic measurement pulse wave-echo to measure liver hardness.The chronic hbv-infection patient adopts the instantaneous elasticity imaging, amasss below experimenter's performance curve when prediction META-VIR classification F2～F4 and cirrhosis and is respectively 0.79 and 0.97.The AUROC value brings up to 0.88 when instantaneous elasticity imaging binding fiber test prediction F2～F4 fiberization.This technique reproducible is good, and variation within laboratory is low between observed person's group.But also defectiveness has only 25～65mm as its signal penetration power, and obesity or patients with ascites imaging are undesirable, can't accurately distinguish fatty liver and liver fibrosis, can not follow the tracks of the variation for the treatment of liver fibrosis; Its sensitivity and degree of accuracy have much room for improvement, and appraisement system remains further perfect.
Blood serum designated object:
Liver fibrosis is a dynamic process, majority parameters all is efficient at the development of liver fibrosis and treatment, and can't assess liver fibrosis stage of particular point in time, multiple serologic marker thing joint-detection is expected to assess more accurately the degree of liver fibrosis.
The index of using always clinically has glutamic-oxalacetic transaminease, glutamic-pyruvic transaminase, prealbumin, total protein, albumin, globulin, cholinesterase, total bilirubin, bilirubin direct, indirect bilirubin, third glutamyl transpeptidase, alkaline phosphatase, TBA etc. at present, every index is from different aspect reflection liver functions, the liver diseases that the different causes of disease cause, detecting index also can be different, as the cholestasis hepatopathy, total bilirubin, bilirubin direct, indirect bilirubin, TBA etc. can better reflect disease severity.The rising of serum GGT also is common in non-viral hepatitis, as fatty liver, AML, liver cancer and disease of biliary tract etc.
Other indexs also have hyaluronic acid, III procollagen type, type, Laminin ELISA (LN), α
2-macroglobulin, III Collagen Type VI N-end peptide, aPoA 1, haptoglobin, human cartilage glycoprotein-39, collagen enzyme and inhibitor thereof, CD138, chemotactic factor (CF), cell factor etc., these indexs are less using clinically, and seeing in the experimental study has report.When the liver damage degree is more serious, hyaluronic acid contents is just more high in the serum.There are some researches show that hyaluronic acid is 8 times of normal liver during cirrhosis, liver I, III, IV, V and VI Collagen Type VI are respectively 8,4,14,8 and 10 times of normal liver during cirrhosis.α
2-macroglobulin is a kind of high molecular weight protein that is synthesized by hepatic parenchymal cells and HSCs, and content is quite abundant in human serum, and serum levels increases and increases along with degree of hepatic fibrosis.The haptoglobin level also can reduce along with increasing the weight of of degree of hepatic fibrosis.In the chronic hbv-infection patients serum, human cartilage glycoprotein-39 is greater than 284.8ng/mL, and the predictive hepatocirrhosis susceptibility reaches 80%, specificity reaches 71%, and negative predictive value is 78%.
Along with liver fibrosis mechanism further is familiar with, cytokine network becomes the emphasis of research, more research wishes to find some cell factors can be as the index of diagnosing liver fibrosis, wherein, Connective Tissue Growth Factor (CTGF) is as the TGF-β downstream effects factor, and Recent study is also more.Discover that CTGF raises obviously in chronic liver disease patients with liver fibrosis serum, but CTGF is lower in the patients serum who develops into cirrhosis.No matter be hepatic tissue biopsy or imaging technique and serum index, all there is corresponding drawback.Especially can react the blood serum designated object of liver function from many aspects, also be the direction that the diagnosing hepatism worker makes great efforts to explore.The serum index detect comparatively speaking simple to operate, expense is cheap, good reproducibility, and can monitor the state of an illness in real time.
China is hepatopathy big country, and high incidence and high mortality are the maximum characteristics of hepatopathy, is directly connected to national health and happy, and the research at hepatopathy both at home and abroad increases severely year by year, has also obtained gratifying achievement in research.In research pathogenesis and medicine strategy, we should return the source, the diagnosis of hepatopathy and evaluation and test.Clinical varied with diagnostic techniques, evaluation index experimental study, but all there is limitation, liver fibrosis is the common pathologic process of most of chronic liver disease, be difficult for finding but exist, the characteristics that their early stage is difficult for discovering have only when severe complications occurs and just can make a definite diagnosis, but at this time treatment is just very difficult, hepatopathy is again the disease of the common regulation and control of many paths, so an index sensitivity, the comprehensive evaluation liver function is diagnostic work research focus.In sum, the diagnosing hepatism technology of using clinically at present has imaging techniques such as hepatic tissue biopsy, ultrasonic, CT and MRI, and blood serum designated object.Yet these diagnostic methods all exist certain limitation and defective, seek a sensitivity, diagnostic method or index have become the problem that this area needs to be resolved hurrily fast and accurately.
Kallistatin(KAL), organize kininase in conjunction with albumen, mainly by the liver synthesis secretion, this just enlightens its evaluation metrics that can be used as liver function, and KAL has anti-oxidant, anti-inflammatory, anti-angiogenic generation, diastolic blood pressure, antineoplastic many activated proteins, concentrated expression liver function from many aspects concurrently.This laboratory discovered to what hepatopathy was prevented and treated that KAL was a kind of multifunctional protein that has various active concurrently through more than 10 years, was expected to become one " rising star " in diagnosis for liver disease, the treatment.Studies show that liver is the main synthesis secretion organ of KAL, behind the hepar damnification, the KAL expression descends.Discover that further have corresponding relation between hepatopathy and the serum KAL level, namely serum KAL level can be used as the investigation index of diagnosing hepatism, treatment, prognosis.The research achievement, not seeing as yet so far has relevant report.
Summary of the invention:
The invention provides Kallistatin(KAL) as a kind of novel, sensitive, application of hepatopathy/liver fibrosis indicator and diagnosticum in early days fast and accurately.
KAL of the present invention is protein Preparation thing or complete genetic fragment.
Diagnosticum of the present invention and indicator can contain protein fragments or the genetic fragment of KAL as required, and wherein said KAL is as the object that detects, its shared number percent in diagnosticum or indicator can be 0.01 ?100%.
Diagnosticum of the present invention or indicator can be any can for detection of formulation, these formulations comprise: water, solid phase, spraying phase.
The water form can be water-based or oily suspensions, solution, emulsion, syrup or elixir, perhaps can be a kind of water before use or other suitable composite dry products of carrier, and this liquid preparation can contain conventional adjuvant.
For described KAL is provided the application as a kind of early stage hepatopathy/liver fibrosis indicator and diagnosticum, the present invention has carried out following relevant experimental study:
1) the clinical blood serum sample KAL of patient with liver cirrhosis content analysis;
2) patients serum KAL changes before and after compensatory phase, decompensated liver cirrhosis patient and the treatment;
3) by the statistical analysis of serum KAL content and liver function index correlativity, studied KAL as a kind of novel, sensitive, application of hepatopathy/liver fibrosis indicator and diagnosticum in early days fast and accurately.
Result of study shows: patient with liver cirrhosis serum KAL level is starkly lower than normal group; Compensatory phase, decompensated liver cirrhosis patients serum KAL content, with increasing the weight of of the state of an illness, it expresses further downward modulation; The results of statistical analysis of serum KAL content and liver function index correlativity shows, serum KAL level can be used as diagnosing hepatism, by stages, the index estimated of clinical treatment method, for hepatopathy research and the anti-hepatic fibrosis medicines that is developed as high targeting, many target spots thereof provide a kind of sensitivity, efficiently, indicator and diagnosticum accurately.
Description of drawings:
The corresponding relation of the serum content of Fig. 1: KAL and hepatopathy state
Wherein: the CON-normal group; The LC-liver cirrhosis group; The viral liver cirrhosis group of VLC-; ALC-alcoholic cirrhosis group; P＜0.01
Fig. 2: patients serum KAL changes before and after compensatory phase, decompensated liver cirrhosis patient and the treatment
Wherein: compensatory group of 1-; 2-loses compensatory group; Before the 3-treatment; After the 4-treatment; P＜0.05
Fig. 3: serum KAL and liver function index linear regression analysis
Fig. 4: the diagnosis usefulness (ROC curve and area under curve) of the cirrhosis of experimenter's operating characteristic analytic curve serum analysis KAL
Fig. 5: the usefulness of experimenter's operating characteristic analytic curve diagnosis alcohol and viral cirrhosis
Embodiment:
Following examples are only understood the present invention better for help those skilled in the art, but do not limit the present invention in any way.
The clinical blood serum sample KAL of " embodiment 1 " patient with liver cirrhosis content analysis
The 88 routine hepatopathy human serum samples that go to a doctor in 180 hospitals of the Chinese People's Liberation Army " entire PLA's hepatopathy centre of prevention and cure " have been collected in this test of KAL, detect wherein KAL content, wherein 33 examples are viral cirrhosis, and 10 examples are alcoholic cirrhosis, 45 routine etiology unknown cirrhosis.30 healthy human serums of random collecting detect analysis as normal group equally in addition.Research hepar damnification pathology is to the influence of serum KAL content, the variation of its content and paathogenic factor, the relation of disease severity, inquire into it as the usefulness of diagnosing hepatism, anti-fibrosis treatment evaluation index, for the clinical research of hepatopathy and KAL is developed as the hepatopathy early diagnosis and the liver fibrosis indicator provides reliable experimental basis.
Test material and reagent: patient's blood sample (Quanzhou, Fujian 180 hospitals), 96 orifice plates (Corning), KAL Elisa kit (R﹠amp; D), bovine serum albumin(BSA) component five (Beijing Ding Guochang), TMB colour developing liquid (Beijing Zhuan Meng) Tween
20, dipotassium hydrogen phosphate, potassium dihydrogen phosphate, sodium chloride, potassium chloride etc. be analyze pure, traditional Chinese medicines.
The testing liquid preparation:
(1) PBS damping fluid:
Fully after the dissolving, transfer pH to 7.20-7.40, tri-distilled water is settled to 1L; 0.22 μ m membrane filtration, normal temperature is preserved;
(2) washing lotion: in 1L PBS solution, add 0.5mL Tween
20, mixing is transferred pH to 7.20-7.40,0.22 μ m membrane filtration, and normal temperature is preserved;
(3) sample diluting liquid (RD): in 100mL PBS solution, add 1g BSA, stir and evenly mix, transfer pH to 7.20-7.40,0.22 μ m filter membrane suction filtration, 4 ℃ of preservations;
(4) stop buffer: i.e. 1mol/L H
2SO
4, the 5.49mL concentrated sulphuric acid is slowly injected the beaker that fills 80mL distilled water, constantly stir with glass bar simultaneously, the distilled water constant volume is to 100mL after cooling, room temperature preservation;
Enzyme linked immunosorbent assay (ELISA):
(1) the capture antibody bag is by 96 orifice plates: press 1:180 dilution proportion capture antibody (mouse-anti people) to final concentration 2.0 μ g/mL with PBS, add 96 orifice plates with 100 μ L/ holes rapidly, seal film and seal, incubated at room is spent the night;
(2) wash plate: wash plate 3 times with washing the plate machine with 300 μ L Xi Ye ∕ holes, afterwards 96 orifice plates are inverted on the clean filter paper and firmly pat, no liquid is residual to the hole;
(3) sealing: sample diluting liquid is added in 96 orifice plates with 300 μ L ∕ holes, seal film and seal, room temperature is placed 1h at least;
(4) wash plate: with washing plate machine washing plate 4 times, other operations are with (2) step;
(5) application of sample: blood serum sample is diluted to suitable concentration with sample diluting liquid; Standard items are used 2 times of gradient dilutions of sample diluting liquid equally, to 62.5,125,250,500,1000,2000,4000, a 8000pg/mL8 concentration.Diluting good sample and standard items with application of sample, establish the blank hole, namely replace the same 100 μ L/ hole liquid feedings of sample with sample diluting liquid, other handle same sample well.Seal film and seal, room temperature is placed 2h;
(6) wash plate: with the operation of (2) step;
(7) press the 1:200 dilution proportion with RD and detect antibody to final concentration 200 μ g/mL, 100 μ L/ holes add 96 orifice plates, seal film and seal, and room temperature is placed 2h;
(8) wash plate: with the operation of (2) step;
(9) add horseradish peroxidase-labeled Streptavidin (Streptavidin-HRP) with 100 μ L/ holes, lucifuge, room temperature is placed 20min;
(10) wash plate: with the operation of (2) step;
(11) colour developing: adding tetramethyl benzidine with 100 μ L/ holes is TMB, lucifuge, and room temperature is placed 20min;
(12) add stop buffer with 50 μ L/ holes, jiggle and fully mix at the bottom of making it be paved with the hole;
(13) measure detection light absorption value OD under wavelength, the 540nm reference wavelength with the good microplate reader of prior preheating at 450nm immediately.
Test findings:
At first, cirrhosis is not considered the cause of disease, and ELISA measures KAL content in the serum; Secondly, be divided into alcoholic cirrhosis group and viral liver cirrhosis group according to paathogenic factor, whether the research cause of disease is influential to KAL content.Secondly, according to the order of severity of disease, cirrhosis can be divided into the compensatory phase and lose the compensatory phase, and research object is divided into two groups accordingly, and whether the content of analyzing KAL also have the difference of conspicuousness with the disease severity difference.On the other hand, follow-up study 21 routine patient with liver cirrhosis (not considering the cause of disease), detection was through one month or treatment for more time, the situation of change of patients serum KAL content, the result shows that patient with liver cirrhosis serum KAL level is starkly lower than normal group (16.33 ± 8.93 μ g/mL vs.29.43 ± 7.80 μ g/mL; P＜0.001), no matter be the viral cirrhosis that the alcoholic cirrhosis that causes of excessive drinking or chronic viral infection (hepatitis type B virus or hepatitis C virus) cause, all the expression of KAL is obviously reduced (20.80 ± 10.51 μ g/mL and 15.17 ± 4.06 μ g/mL vs.29.43 ± 7.80 μ g/mL; P＜0.001), but do not have a significant difference before alcoholic cirrhosis group and the viral liver cirrhosis group.The age of research object and sex can not influence the amount (table 1 and Fig. 1) of serum KAL.
The blood content of table 1.1KAL and the corresponding relation of hepatopathy state
Patients serum KAL changes before and after " embodiment 2 " compensatory phase, decompensated liver cirrhosis patient and the treatment
Test material and method are with implementing sharp 1.According to the clinical patient data that provides, sample is divided into compensatory cirrhosis and decompensated liver cirrhosis, data are carried out significance of difference analysis between two groups.In addition, 1 month treatment of the anti-cirrhosis medicine of 21 routine patients undergo detects the variation of serum KAL, analyzes compensatory phase and decompensated liver cirrhosis patients serum KAL content, discovery increases the weight of with the state of an illness, and (19.98 ± 10.14 μ g/mLvs.12.25 ± 6.22 μ g/mL are further reduced in the expression of KAL; P＜0.05).Find out that thus serum KAL level can reflect the order of severity of hepatopathy, the content of the more serious KAL of disease is more low.
Follow is analyzed patient and is accepted serum KAL content in the anti-cirrhosis drug therapy process, finds the treatment through after a while, its content (12.52 ± 6.71 μ g/mL vs.19.29 ± 10.60 μ g/mL that risen; P=0.018) (Fig. 2).The content that KAL is described can be used as one and estimates treatment effectively and the disease investigation index of back situation more.
" embodiment 3 " KAL is as a kind of novel, sensitive, application of hepatopathy/liver fibrosis indicator and diagnosticum in early days fast and accurately.
Test material and method are with implementing sharp 1.
1. experimenter's liver function index of correlation detects:
The experimenter is derived from hepatopath or the routine physical examination person of Quanzhou 180 hospital admissions.The every liver function index that is defined as the hepatopath is carried out statistical study, show that each index all has in various degree unusual, meets the hepatopathy feature; Healthy normal group is determined no hepatitis history.From hepatopathy distribution crowd, ill major part is the elderly, but the trend of rejuvenation is also arranged, and is the male sex more than 85%, and particularly for alcoholic cirrhosis and liver cancer, this may indulge in excessive drinking relevant with the male sex.Cirrhosis kind and by stages by at least two experts' of 180 hospitals the audit of making a definite diagnosis.Gather experimenter's liver function index of correlation in the table 2 and detected essential information.
Table 2 research object essential information statistical form
2. the statistical analysis of serum KAL content and liver function index correlativity:
All The data SPSS16.0 statistics softwares are handled in the experiment, and metering numerical value is with mean ± standard deviation (mean ± SD) expression.Conspicuousness between multi-group data is relatively used t-test, and P＜0.05(*) is significantly, P＜0.01(**) for extremely remarkable; With correlativity (Fig. 3) between Pearson's correlation test (table 3), Spearman rank correlation check (table 4) and linear regression analysis three kinds of methods analyst serum KAL and other liver function indexs; Experimenter's performance curve is estimated its usefulness that diagnoses the illness, sensitivity and specificity.The content of inquiring into serum KAL is investigated the feasibility of index as liver function.
Pearson's correlation test of table 3 patient with liver cirrhosis serum KAL content and serum liver function indexes
The Spearman rank correlation check of table 4 patient with liver cirrhosis serum KAL and serum liver function indexes
No matter be Pearson's correlation test or Spearman rank correlation check, all drawn consistent result: namely for patient with liver cirrhosis, serum KAL content and preceding albumen, total protein, albumin, Bai Dan Bai ∕ globulin and cholinesterase present significant positive correlation, present remarkable negative correlation with total bilirubin.
3. the diagnosis usefulness of the cirrhosis of experimenter's operating characteristic analytic curve serum analysis KAL
Experimenter's performance curve (receiver operator characteristic curve, the ROC curve), be called recipient's operating characteristic curve again, be according to a series of two different mode classifications (cut off value or decision threshold), be ordinate with True Positive Rate (sensitivity), false positive rate (1-specificity) is the curve that horizontal ordinate is drawn.When the ROC curve is easy to find any boundary value of a certain index to the recognition capability of disease, judge the diagnostic value of this index by calculating ROC area under a curve (AUC), ROC area under a curve value is between 1.0 and 0.5, and AUC illustrates that more close to 1 diagnosis effect is more good.And can calculate excellent diagnostics reference value (cut ?off values) by sensitivity and specificity.
The diagnosis effectiveness analysis of the cirrhosis of table 5KAL
Can find out very intuitively that from Fig. 4 serum kallistatin can diagnose cirrhosis exactly, the ROC area under curve reaches 0.840, is higher accuracy.The excellent diagnostics critical value is 21.64 μ g ∕ mL, is lower than this value, can think that this experimenter suffers from cirrhosis probably.Can calculate simultaneously, when this critical value, sensitivity (Se), specificity (Sp), the positive diagnosing rate (PPV) of serum KAL content diagnosis cirrhosis are respectively 84.4%, 75.6%, 91%, 62.3% (table 5) with negative diagnosis.
The usefulness of 4 experimenter's operating characteristic analytic curves diagnosis alcohol and viral cirrhosis
Above-mentioned analysis result shows that the ROC curve can be diagnosed cirrhosis more exactly, and next further whether serum analysis KAL content usefulness when the cirrhosis that the different causes of disease of diagnosis cause is identical.From interpretation, the accuracy rate of KAL serum content diagnosis alcoholic cirrhosis is than the height of viral cirrhosis.Area under curve is respectively 0.724,0.957(Fig. 5, table 6).
Table 6KAL is to the diagnosis effectiveness analysis of diagnosis alcohol and viral cirrhosis
This experimental analysis the content of 98 routine hepatopathy human serum KAL, with the expection the same, patient KAL serum-concentration is starkly lower than healthy people, illustrates in the hepar damnification pathological process, and the synthetic and secretion of KAL is had a strong impact on, and the more serious this influence of disease is more big, but through treatment, its level will rise, thus prompting, serum KAL content not only can be used as the hepatopathy evaluation metrics, also can be used as the treatment evaluation index.With at present clinically used liver function serum index carry out Correlation analysis showed, KAL level and preceding albumen (PA), total protein (STP), albumin (ALB), total bilirubin (TBIL), cholinesterase (CE) significant correlation illustrate that more it detects the feasibility of index as hepatopathy.
Not having correlativity with other indexs, may be because different indexs is subjected to due to the difference of paathogenic factor.Synthetic by liver cell as preceding albumen (PA), measure its concentration in blood plasma for understanding protein malnutrition, dyshepatia, having higher susceptibility than albumin and transferrins, especially for liver disease.Cirrhosis, liver meronecrosis is lighter, and preceding albumen changes little, and prognosis is better, and when the state of an illness was improved, preceding albumen also raise rapidly.Total protein (STP) is made up of albumin and globulin, hepatosis, and then the liver synthetic protein reduces, and is mainly obvious with the decline of albumin (ALB); Total bilirubin (TBIL) is the two summation of bilirubin direct and indirect bilirubin, and whether whether be mainly used to diagnosis has liver diseases or biliary tract to take place unusually.All can cause its rising when infringements such as liver is inflamed, necrosis, poisoning, disease of biliary tract and hemolytic disease also can cause change.Bilirubin direct raises and is common in primary Biliary Cirrhosis, Primary, obstruction of biliary tract etc.Indirect bilirubin raises and is common in hemolytic disease, icterus neonatorum or blood transfusion mistake etc.The bilirubin direct of hepatitis and patient with liver cirrhosis and indirect bilirubin can raise, and KAL is significantly relevant with TBIL, illustrate that its evaluation value in the cholestasis liver disease may be higher.Cholinesterase (CE) is made up of true choline esterase (AchE) and pseudocholinesterase (pchE) two parts, and the two is energy catalysis acetylcholine hydrolyzation all, generates choline and acetic acid..CE is to derive from the PchE of liver in the serum.PchE is that CE can reflect liver albumen complex functionality, and is relevant with liver cirrhosis patient liver material injury, can reflect the order of severity of hepatocellular damage by the synthetic hydrolysis acid of liver cell.When liver damage, the liver cell complex functionality descends, and change of serum C E vigor descends, and is the enzyme of unique decline behind the hepatic disease, can react the liver reserve capacity; Along with the increasing the weight of of fibrosis, ALT changes irregular when cirrhosis, and CE descends gradually, has certain correlativity with liver fibrosis, and it is more meaningful than measuring the ALT vigor to measure the CE vigor.The correlativity of serum KAL content and CE is up to 0.694(p＜0.001) illustrate that KAL also can synthetic, the reserve function of fine reflection liver.
The analysis of experimenter's performance curve shows, serum KAL accurately diagnoses alcohol, viral cirrhosis, and the diagnosis usefulness to the former is higher, the sensitivity of diagnosis of liver disease, specificity are all fine, and the ROC area under curve is suitable with the ultrasonic instantaneous elasticity imaging technique that now grows up clinically especially.
The present invention is based on the test result that 98 routine hepatopathy people and 30 routine healthy human serum kallistatin content are carried out, the level of proof serum KAL can be used as diagnosing hepatism, by stages, the index estimated of clinical treatment method, be expected to for hepatopathy research and be developed as the anti-hepatic fibrosis of high targeting, many target spots and the cirrhosis medicine provide a kind of sensitivity, efficiently, indicator and diagnosticum accurately.
Claims (5)
1. organize kininase in conjunction with albumen Kallistatin(KAL for one kind) in the application of preparation in early stage hepatopathy, liver fibrosis and cirrhosis indicator and the diagnosticum.
2. the described application of claim 1, indicator wherein and diagnosticum can contain protein fragments or the genetic fragment of KAL as required.
3. the described application of claim 1, wherein said KAL is as detected object, in diagnosticum or indicator shared number percent can be 0.01 ?100%.
4. the described application of claim 1, its diagnosticum and indicator can be any can for detection of formulation, these formulations comprise: water, solid phase, spraying phase.
5. claim 1 or 4 described application, its water form can be water-based or oily suspensions, solution, emulsion, syrup or elixir, or a kind of water before use or other suitable composite dry products of carrier, this liquid preparation can contain conventional adjuvant.
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Cited By (3)
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| CN109765378A (en) * | 2016-08-31 | 2019-05-17 | 鲁凤民 | A kind of new cirrhosis or hepatic fibrosis markers |
| CN111551736A (en) * | 2020-04-10 | 2020-08-18 | 广东省第二人民医院（广东省卫生应急医院） | Ovarian hyperstimulation syndrome diagnostic marker Kallistatin and application thereof |
| CN111551736B (en) * | 2020-04-10 | 2021-03-16 | 广东省第二人民医院（广东省卫生应急医院） | Ovarian hyperstimulation syndrome diagnostic marker Kallistatin and application thereof |
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