CN103432572B - A kind of preparation method of rhesus monkey diabetic nephropathy model - Google Patents
A kind of preparation method of rhesus monkey diabetic nephropathy model Download PDFInfo
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Abstract
The invention discloses a kind of preparation methods of rhesus monkey diabetic nephropathy model, include the following steps：（1）The streptozotocin that dosage is 80~100mg/kg is applied to rhesus macaque；（2）After blood sugar concentration rises to 11.1mmol/l, administration of insulin, makes rhesus macaque blood sugar concentration for 10 20mmol/L before the meal.The present invention by the way of glycemic control, successfully induces rhesus monkey diabetic nephropathy model on the basis of diabetes rhesus macaque.
Description
Technical field
The present invention relates to the preparation methods of rhesus monkey diabetic nephropathy model.
Background technology
Diabetes（Diabetes Mellitus, DM）Incidence be in rapid growth trend.It is reported that China it is existing there are about
92000000 DM and 1.48 hundred million DM patients' early period, become second onset diabetes big country after the U.S..With the state of an illness by
Gradually development aggravates, and it is adjoint serious that there are about the diabetes B patients of the type 1 diabetes patient of 30-40% and Yue 20% in these patients
Kidney function damage --- diabetic nephropathy（Diabetic Nephropathy, DN）.
DN is one of chronic complicating diseases of diabetic's most serious and the most important cause of death of diabetic.
Once significant clinical symptoms occurs in DN patient, selectable drug is less at this time, and the therapeutic effect of drug is poor, the state of an illness
It is in often to carry out sexual development until end stage Renal failure, huge financial burden is brought to patient home and society.Therefore it furthers investigate
The pathogenesis and method of early diagnosis of DN is explored delay or the effective measures of reverse disease process on this basis, be current
The hot and difficult issue problem of medical research.
Animal model is the basis for studying mankind's major disease, is established and the approximate DN animal models of mankind's height, it will
The effect got twice the result with half the effort is obtained studying its pathogenesis, early diagnosis and therapeutic intervention etc..Non-human primate is made
For the close relative of the mankind, about 96% or more is homologous in terms of gene, the same people of morphosis, physiological function and biochemical metabolism
Class is closely similar, establishes a kind of non-human primate DN models to studying occurrence and development, early diagnosis and the new drug development of DN
Deng with important scientific meaning and economic value.
Invention content
The purpose of the present invention is to provide the preparation methods of rhesus monkey diabetic nephropathy model.
The preparation method of rhesus monkey diabetic nephropathy model of the present invention, includes the following steps：
（1）The streptozotocin that dosage is 80~100mg/kg is applied to rhesus macaque；
（2）After blood sugar concentration rises to 11.1mmol/l, administration of insulin, makes rhesus macaque blood sugar concentration for 10- before the meal
20mmol/L。
The preparation method of rhesus monkey diabetic nephropathy model of the present invention, includes the following steps：
（1）The streptozotocin that dosage is 80~100mg/kg is applied to rhesus macaque；
（2）After blood sugar concentration is increased to 11.1mmol/l, high lipid food is fed, daily feeding 2 times, scale of feeding 0.3-
0.4kg/ times only, and in administration of insulin before the meal, it is 10-20mmol/L to make rhesus macaque blood sugar concentration, and the feed includes such as
The raw material of lower weight proportion：78 parts of standard monkey feed, 15 parts of animal fat, sugared 5 parts, 2 parts of cholesterol.
Standard monkey feed refers to《Chinese name republic standard GB/T 14924.8-2001》Defined monkey cooperation is raised
Material.
Animal fat refers to the fat from animal, such as:Butter, sheep oil, lard.
Step（1）In, the method for application is intravenous injection.
Step（2）In, the method for application is to be subcutaneously injected.
Step（2）In, the animal fat is lard；The sugar is sucrose.
The high lipid food of rhesus monkey diabetic nephropathy model of the present invention, it includes the raw material of following weight proportion：Standard monkey
78 parts of feed, 15 parts of animal fat, sugared 5 parts, 2 parts of cholesterol.
The animal fat is lard；The sugar is sucrose.
The purposes of animal model prepared by preceding method of the present invention in the drug of screening treatment rhesus macaque diabetic nephropathy.
The method that the present invention screens the drug for the treatment of rhesus monkey diabetic nephropathy model, it includes the following steps：
A, according to preceding method, rhesus monkey diabetic nephropathy model is established；
B, drug candidate is applied to animal model；
C, the drug of rhesus macaque diabetic nephropathy is potentially treated with animal model evaluation.
Modeling method of the present invention can induce rhesus macaque the clinic of the diabetic nephropathies such as kidney injury, Microalbuminuria occur
Symptom, medication is simple, and repeatability is strong.
Obviously, the above according to the present invention is not being departed from according to the ordinary technical knowledge and customary means of this field
Under the premise of the above-mentioned basic fundamental thought of the present invention, the modification, replacement or change of other diversified forms can also be made.
The specific implementation mode of form by the following examples remakes further specifically the above of the present invention
It is bright.But the range that this should not be interpreted as to the above-mentioned theme of the present invention is only limitted to example below.It is all to be based on the above of the present invention
The technology realized all belongs to the scope of the present invention.
Description of the drawings
Fig. 1 glycemic control situations；
Fig. 2 renal function testing results；
Fig. 3 fibrosis gene alteration situations；
Fig. 4 renal hemodynamics parameter and blood perfusion coefficient；
The blood distribution and frequency spectrum of Fig. 5 B-ultrasonic renal inspections；
Fig. 6 renal pathologies change；
Fig. 7 nephridial tissue immunofluorescences.
Specific implementation mode
The preparation of 1 rhesus macaque diabetogenous nephrosis symptom model of the present invention of embodiment
1 experiment material and instrument
1.1 test sample essential informations
IL-1 β antibody：abcam
TNF-a antibody：abcam
1 antibody of TGF-β:Bioworld
CTGF antibody：abcam
Collagen IV antibody:abcam
Fibronection antibody:Bioworld
MCP-1 antibody:ebioscience
CD68 (KP1) antibody:abcam
ENOS antibody:abcam
Total RNA from animal tissues extracts kit：Tiangeng
iScriptTMCDNA synthetic agent box：BIO-RAD
iQTM Green Supermix：BIO-RAD
Key instrument, instrument needed for 1.2 experiments
Blood-sugar detecting instrument：The full vigor type blood-sugar detecting instruments of Luo Kang, the full vigor type blood sugar test papers of Luo Kang
Fully automatic blood Biochemical Analyzer（Switzerland Roche COBAS Integra400Plus, number MEB-02-01）
BIO-RAD CFX96 fluorescence quantitative PCR instruments
BIO-RAD S200PCR instrument
LEICA DM4000B fluorescence microscopes
Olympus BX51 light microscopes
Puncture needle punctures rifle：BARD
Color ultrasound;Iu22philips
Fundus fluorescein angiography：Topcon TRC.50DX（Japan opens up general Kanggong department production）
1.3 experimental animal
1.3.1 experimental system
Rhesus macaque 14,3.5~5.5kg of weight intends breeding study base by Sichuan Chengdu safety animal（Production licence
Number：SCXK (river) 2004-013）It introduces.Enter the room the last week carry out tuberculin, parasite, salmonella, Salmonella of congratulating inspection
It looks into.
1.4 animal feeding management
1.4.1 quarantine/environment adapts to
Environment is first adapted to before the rhesus macaque that enters the room experiment and is quarantined one week, selects healthy buck as animal subject, inspection
Epidemic disease content：It is whether consistent with the quality index required when ordering；Body temperature, food ration, blood biochemistry, electrolyte；Animal general state；
Whether the weight of animals reaches the range of test requirements document weight.
1.4.2 animal feeding
Rhesus macaque is raised in National Chengdu Center For Safety Evaluation of Drugs's conventional animal room（Experimental animal room uses licensing
Number：SYXK（River）2003-030）, the stainless steel mouse cage sub-cage rearing of rhesus macaque every, rearing conditions meet national standard GB14925-
2001,21 ± 5 DEG C of room temperature（Temperature difference per day≤4 °C）, 55 ± 15%, 12 hours light and shade alternatings of relative humidity, rate of ventilation 8~10
Secondary/hour.
1.4.3 animal identification method
Rhesus macaque is marked jointly by label on chest card and cage
1.5 experiment packet
Grouping includes normal group（3）, experimental group A（After STZ Glycemia Declines, pass through 15 minutes before the meal daily subcutaneous notes
Insulin is penetrated, blood glucose is controlled<10mmol/l, 3）, experimental group B（After STZ Glycemia Declines, pass through 15 minutes skins before the meal daily
Lower insulin injection, control blood glucose is in 10-20mmol/l, 5）, experimental group C（After STZ Glycemia Declines, by daily before the meal
15 minutes subcutaneous insulin injections, control blood glucose feed high lipid food in 10-20mmol/l, 3）.
Feed formula：
Experimental group A, B：Monkey standard monkey feed --- commercialized monkey pellet（Purchased from Sichuan Academy of Medical Sciences animal
Research institute, formula with《Chinese name republic standard GB/T 14924.8-2001》Defined monkey mixed feed is identical）.
Experimental group C：High lipid food：15% refined lard（It is commercially available）, 5% sucrose（It is commercially available）, 2% cholesterol（It is commercially available）, 78% monkey
Standard monkey feed.
Feed mode：Feeding 2 times daily, scale of feeding are 0.3-0.4kg/ times.
2 experimental methods
2.1 animal general status are observed
Observe animal appearance sign（Hair, eye including animal and mucous membrane）, behavioral activity, mental status, glandular secretion,
Situations such as breathing, diet, excreta, injection site, detection the weight of animals, blood pressure and food ration.
2.2 modeling method
Experimental group A, B：
Animal different fasting 12h or more before administration, after venoclysis physiological saline 100ml dilatations, then is injected prepared
STZ(80mg/kg)；
Conventional monkey forage feed is given, daily feeding 2 times, scale of feeding is 0.3-0.4kg/ times；
Detect animal fasting blood-glucose（FBG）, continuous 2 days of FBG passes through 15 minutes skins before the meal daily higher than after 11.1mmol/l
Lower insulin injection controls blood sugar concentration.
Experimental group C：
Animal different fasting 12h or more before administration, after venoclysis physiological saline 100ml dilatations, then is injected prepared
STZ(80mg/kg)；
Conventional monkey forage feed is given, daily feeding 2 times, scale of feeding is 0.3-0.4kg/ times；
Detect animal fasting blood-glucose（FBG）, continuous 2 days of FBG passes through 15 minutes skins before the meal daily higher than after 11.1mmol/l
Lower insulin injection controls blood sugar concentration, while giving high lipid food nursing, feeding 2 times daily, scale of feeding 0.3-0.4kg/
Secondary.
2.3 hematological examination
Testing index：Red blood cell count(RBC)（RBC, capacitance method）, hemoglobin（HGB, HiCN method）, red cell volume（HCT,
Power Capacity point-score）, mean corpuscular volume（MCV is calculated）, mean corpuscular hemoglobin（MCH is calculated）, mean corpuscular
Hemoglobin concentration（MCHC is calculated）, reticulocyte count（RET, fluorescence colour）, white blood cell count(WBC)（WBC, laser method）
And classification（Neutrophil leucocyte NEU, lymphocyte LYM, monocyte MONO, eosinophil EOS, basophilic granulocyte
BASO, laser method）, platelet count（PLT, capacitance method）, granulophilocyte（RET, Liu Shi+fluorescence colour）, factor
Time（PT, Chromogenic assay）, Activated partial thromboplastin time（APTT, Chromogenic assay）.Aspartate amino transferase
（AST, IFCC P-5 '-P methods）, alanine aminotransferase（ALT, IFCC P-5 '-P methods）, gamma glutamyltransferase（GGT,
L- gamma-glutamyls -3- carboxyls-paranitroanilinum method）, cretinephosphokinase（CK, HK-G6PD method）, alkaline phosphatase（ALP, 4-
NPP methods）, lactic dehydrogenase（LDH, IFCC method）, urea nitrogen（Urea, Urease-GLDH-Kinetic method）, total protein（TP,
Biuret methods）, albumin（ALB, BCG method）, blood glucose（GLU, GLK-G6PDH method）, total bilirubin（TBIL, Diazo method）, creatinine
（Crea, JAFF method）, total cholesterol（CHOL, CHOD-P method）, triglycerides（TG, GPO-PAP method）, Na ion concentration（Na+,
Ion selective electrode method）, potassium concentration（K+, ion selective electrode method）, chlorine ion concentration（Cl-, ion selective electrode method）
Deng.
Detection time：STZ Isosorbide-5-Nitraes are given, week, each survey was primary, surveyed within every 1~2 month later primary.
Method：It takes a blood sample and detects from rhesus macaque back leg great saphenous vein.Hematological examination blood is used with EDTA anti-freezings, clotting time
Blood is with sodium citrate anti-freezing.
2.4 fasting blood-glucose（Fasting blood glucose, FBG）And postprandial blood sugar（Postprandial blood
Glucose, PBG）Detection
Detection time：FBG is detected 2 times a week（At 9 points in the morning, after empty stomach 12h）And PBG（2h after feed）.
Detection method：Finger or toe peripheral blood are taken, is detected with Roche blood glucose meter.
2.5 intravenous glucose tolerances are tested（IVGTT）
Vein gives 50% glucose injection（0.5g/kg weight）, 0,1,3,5,10,30,60 and 120 after injecting
Minute measures blood glucose level, while checking the insulin level after glucose injection in 0,1,3,5,30 minute blood sample（RIA or
ELISA）
The horizontal detection of 2.6 empty stomach glycosylated hemoglobins, serum insulin and C peptides (C-peptide, C-P)
Detection time：Virus monitory was taken every 4 weeks.
Detection method：Radioimmunology (RIA) kit, Enzyme-Linked Immunospot（ELISA）
2.7 insulin therapy
It is monitored on an empty stomach with postprandial blood sugar, type, the dosage form of diabetes monkey subcutaneous insulin injections is adjusted according to blood glucose level
And dosage.Blood Sugar Monitoring observes therapeutic effect, optimum scheme comparison within 24 hours.Regular promoting circulation of blood routine and biochemical analysis.
2.8 funduscopy
Detection time：Before administration and after administration at the end of the observation period.
Assay method：Rhesus macaque is anaesthetized through ketalar（Intramuscular injection, 15mg/kg）, U.S. Dolly mydriasis（Eye drip, 1
Drop/eye）, checked with binocular indirect ophthalmoscope, all imaging evaluations carry out under general anesthesia state（Animal Anesthesia method
With reference to rhesus macaque surgery anesthesia SOP）.With the shape, size, color and luster of eyeground sem observation optic papilla, whether edge is clear；It is yellow
Whether there is or not oedema, bleeding, exudation and pigment disorders in spot portion；Whether there is or not oedema, exudation, bleeding, stripping and new vessels etc. for retina.
2.9 kidney B ultrasound and renal angiography
Color ultrasound observes kidney form and blood perfusion analysis hemodynamic index and blood perfusion coefficient.Contrast agent
SonoVue (Bracco SpA, Milan, Italy) injects 1ml by great saphenous vein（5mg/ml）, then with 2ml physiological saline
It rinses.Do left kidney again later, midfeather is not less than 10 minutes, is subject to flash explosions halfway.
The lower renal biopsy with gun-biopsy of the super guiding of 2.10B
Prone position, drape after preserved skin disinfection, point of puncture is gone to select on left kidney after rhesus macaque ketamine IM and safe and sound anesthesia
Pole or lower pole, avoid blood vessel and renal plevis.BARD kidney puncture needle Needle aspirations, are respectively put into 10% paraformaldehyde and RNA
The EP pipes of latter preserve.All fixing organizations carry out HE, Masson, PAS normal dyeing, immunofluorescence dyeing（CTGF、
ENOS, IL-1 β, IV Collagen Type VIs, fibronectin FN, TGF-β, MCP-1, TNF-α, CD68）In microscopically observation pathological tissue
Learn variation.RNA latter preserve after tissue extraction RNA reverse transcriptions with fluorescence quantitative PCR detection smad2, smad3, smad4,
Smad7, the expression of actin, NF- κ b gene.
2.11 liquid-phase chip detects
Take the blood plasma of the different courses of disease, detecting factor（IL-1、IL-6、MCP-1、TNF-α、IL-17、IL-18）
2.12 urine detection
Rhesus macaque is put into metabolic cage, collects urina sanguinis and twenty-four-hour urine, detects routine urinalysis, urinary albumin and creatinine.
3 experimental results
3.1 glycemic control situations
As shown in Figure 1, the blood glucose of each experimental group is held in prespecified range.
3.2 renal functions detect
As shown in Fig. 2, uroscopy shows that experimental group B and C started Microalbuminuria occurred at 36th month（MAU）, and
And as course of disease passage has increase trend.Blood biochemistry shows that 42 months ABC, tri- groups of serum creatinines have been compared with urea nitrogen with normal group
Notable difference.
The blood distribution and frequency spectrum of 3.3 B-ultrasonic renal inspections
As shown in figure 5, early stage Renal vascular bed resistance increases, to make blood flow in kidney change, with aggravation, move
The vessel wall elasticity of arteries and veins hardening lowers, and luminal stenosis, peripheral vascular resistance increases, and renal blood perfusion is reduced, and makes renal hemodynamic in low
Stream, high-impedance state, wherein clearly, C groups are the most apparent for the pathological change of B groups and C groups.
3.4 renal hemodynamics parameters and blood perfusion coefficient
As shown in figure 4, compared with the control group, tri- groups of hemodynamic parameter drag indexs of ABC（RI）Increase, diastasis
Minimum flow rate（EDV）It reduces, area under blood perfusion coefficient curve（AUC）Reduce, the difference maximum of C groups and control group, B groups its
Secondary, A groups difference is minimum.
3.5 renal pathologies change
As shown in fig. 6, for A groups without substantially changeing, BC group inflammatory cells invade profit, proliferation of mesangial cells, base compared with normal group
Counterdie thickens, the deposition of collagen and glycogen, and C groups are more notable.
3.6 nephridial tissue immunofluorescences
As shown in fig. 7, nephridial tissue immunofluorescence show experimental group BC with normal group and experiment A groups compared with CTGF,
ENOS, IL-1, IV Collagen Type VI, fibronectin (FN), TGF-β expression increase, and more notable in experimental group C, test simultaneously
Also there is MCP1 and TNF-α expression in group C.
3.7 fibrosis gene alterations
As shown in figure 3, Real-time PCR show experimental group B and C started to occur at 24th month Smad2 and
Smad3mRNA is expressed, and experimental group C is more notable, and Smad7 and NF- κ b mRNA expression is occurred at 42nd month and increased.
3.8 eye examination
Except one（05539）There is eyes cataract to fail to check outer, remaining double external eyes is without exception, conjunctiva without congestion and edema,
Corneal transparency, anterior chamber is without exception, and pupil moderate dissipates big（It has dripped and has used mydriatic）.Crystal is transparent, and vitreum is without muddiness.Eyeground：
Eyes leopard line shape eyeground, optic papilla color is light red, C/D=0.3-0.7, A:V=1:2, it is seen that sieve pore and blood vessel fluctuation.It has no out
Blood, exudation, microaneurysm and oedema, central fovea of macula are reflective clear.
To sum up, experimental group A does not occur the related symptoms of diabetic nephropathy, and the allusion quotation of diabetic nephropathy occur in experimental group B and C
Type clinical symptoms：
1, experimental group B and C started Microalbuminuria occurred at 36th month（MAU）And as course of disease passage has increase
Trend；
2, nephridial tissue HE, Masson, PAS dyeing display：Experimental group A is without lesion, and experimental group B and C kidney are from the 24th
The moon starts the deposition that inflammatory cell invades profit, proliferation of mesangial cells, basement membrane thickened, collagen and glycogen occur, and it was found that experiment
Group C occurred local glomerulosclerosis at 42nd month.Same time nephridial tissue immunofluorescence show experimental group BC in normal group and
It tests A groups compared to CTGF, eNOS, IL-1, IV Collagen Type VI, fibronectin (FN), TGF-β expression to increase, and in experimental group C
More significantly, while also there is MCP1 and TNF-α expression in experimental group C；
3, Real-time PCR are shown：Experimental group B and C started Smad2 and Smad3mRNA expression occurred at 24th month,
Experimental group C is more notable, and Smad7 and NF- κ b mRNA expression occurs at 42nd month and increase；
4, renal angiography is shown, the blood perfusion coefficient AUC and hydromechanics parameter drag index RI of experimental group B and C
It dramatically increases, diastasis minimum flow rate EDV is reduced（P<0.05）, the vessel wall elasticity attenuating of artery sclerosis, luminal stenosis, outside
All vascular resistences are increased, and renal blood perfusion is reduced, and make renal hemodynamic in low stream, high-impedance state；
The present invention is 10-20mmol/L by controlling the blood glucose range of diabetes rhesus macaque for a long time so that diabetes
There is nephridial tissue inflammatory cell and invades the deposition of profit, proliferation of mesangial cells, basement membrane thickened, collagen and glycogen, kidney damage in rhesus macaque
The typical clinical symptom of the diabetic nephropathies such as wound, Microalbuminuria illustrates rhesus macaque diabetic nephropathy modeling success of the present invention.
The present invention is 10-20mmol/L by the blood glucose range for controlling diabetes rhesus macaque for a long time, in conjunction with long-term height
The mode of fat intake so that the various diabetic nephropathy classical symptoms of diabetes rhesus macaque are than individually controlling the mode of blood glucose more
It is notable, illustrates that the mode of the two Combined Treatment is more excellent.
Embodiment 2 treats the drug of diabetic nephropathy with model discrimination of the present invention
A, the rhesus monkey diabetic nephropathy model established according to 1 method of embodiment；
B, drug candidate is applied to animal model；
C, observation drug candidate evaluates potential treatment diabetogenous nephrosis to the influence situation of the various indexs of metabolic syndrome
The drug of sick disease.
To sum up, for modeling method of the present invention by way of controlling blood glucose for a long time, there is diabetic nephropathy in induction rhesus macaque
Clinical symptoms, in addition, long-term blood glucose control can also induce rhesus macaque diabetic nephropathy occur with the united mode of intake high in fat
Clinical symptoms, and symptom is more notable, illustrates that two ways of the present invention can effectively establish rhesus macaque diabetic nephropathy
Model.
Claims (7)
1. a kind of preparation method of rhesus monkey diabetic nephropathy model, it is characterised in that：Include the following steps：
（1）The streptozotocin that dosage is 80mg/Kg is applied to rhesus macaque；
（2）Continuous 2 days of fasting plasma glucose concentration is higher than after 11.1mmol/l, administration of insulin before the meal, makes the rhesus macaque blood sugar concentration be
10-20mmol/L。
2. a kind of preparation method of rhesus monkey diabetic nephropathy model, it is characterised in that：Include the following steps：
（1）The streptozotocin that dosage is 80mg/Kg is applied to rhesus macaque；
（2）Higher than high lipid food after 11.1mmol/l, is fed, feeding 2 times, scale of feeding are fasting plasma glucose concentration daily within continuous 2 days
0.3-0.4kg/ times only, and in administration of insulin before the meal, it is 10-20mmol/L, the feeding high in fat to make rhesus macaque blood sugar concentration
Material includes the raw material of following weight proportion：78 parts of monkey mixed feed, 15 parts of animal fat, 5 parts of sugar, 2 parts of cholesterol.
3. preparation method according to claim 1 or 2, it is characterised in that：Step（1）In, the method for application is quiet
Arteries and veins is injected.
4. preparation method according to claim 1 or 2, it is characterised in that：Step（2）In, the method for application is skin
Lower injection.
5. preparation method according to claim 2, it is characterised in that：Step（2）In, the animal fat is lard；Institute
It is sucrose to state sugar.
6. animal model prepared by claim 1 ~ 4 any one the method is in the medicine of screening treatment rhesus macaque diabetic nephropathy
Purposes in object.
7. a kind of method of the drug of screening treatment rhesus monkey diabetic nephropathy model, it is characterised in that：It includes the following steps：
A, according to claim 1 ~ 4 any one the method, rhesus monkey diabetic nephropathy model is established；
B, drug candidate is applied to animal model；
C, the drug of rhesus macaque diabetic nephropathy is potentially treated with animal model evaluation.
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| PCT/CN2014/080414 WO2014202024A1 (en) | 2013-06-21 | 2014-06-20 | Method for preparation of rhesus monkey diabetic nephropathy model |
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| US9171343B1 (en) | 2012-09-11 | 2015-10-27 | Aseko, Inc. | Means and method for improved glycemic control for diabetic patients |
| US9897565B1 (en) | 2012-09-11 | 2018-02-20 | Aseko, Inc. | System and method for optimizing insulin dosages for diabetic subjects |
| CN103432572B (en) * | 2013-06-21 | 2018-09-18 | 四川大学华西医院 | A kind of preparation method of rhesus monkey diabetic nephropathy model |
| US9486580B2 (en) | 2014-01-31 | 2016-11-08 | Aseko, Inc. | Insulin management |
| US9233204B2 (en) | 2014-01-31 | 2016-01-12 | Aseko, Inc. | Insulin management |
| EP3050023B1 (en) | 2014-10-27 | 2021-08-25 | Aseko, Inc. | Subcutaneous outpatient management |
| US11081226B2 (en) | 2014-10-27 | 2021-08-03 | Aseko, Inc. | Method and controller for administering recommended insulin dosages to a patient |
| CA2993275C (en) | 2015-08-20 | 2022-06-21 | Aseko, Inc. | Diabetes management therapy advisor |
| CN105726568B (en) * | 2016-04-11 | 2019-01-29 | 陕西科技大学 | A kind of construction method of diabetic nephropathy animal model |
| CN107469069A (en) * | 2017-08-18 | 2017-12-15 | 四川普莱美生物科技集团有限公司 | Modeling method, preparation, the preparation method of preparation, rhesus macaque pulmonary fibrosis model and its application of rhesus macaque pulmonary fibrosis model |
| CN115176758B (en) * | 2022-07-18 | 2024-03-26 | 四川省医学科学院.四川省人民医院实验动物研究所 | Method for establishing non-human primate model of type 1 diabetes |
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