CN108853477A - P glycoprotein is influencing 832/13 cell insulin secretion application of INS-1 - Google Patents
P glycoprotein is influencing 832/13 cell insulin secretion application of INS-1 Download PDFInfo
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K38/00—Medicinal preparations containing peptides
- A61K38/16—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- A61K38/17—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
- A61K38/1703—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from vertebrates
- A61K38/1709—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from vertebrates from mammals
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- Gastroenterology & Hepatology (AREA)
- Chemical & Material Sciences (AREA)
- Engineering & Computer Science (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Marine Sciences & Fisheries (AREA)
- Zoology (AREA)
- Pharmacology & Pharmacy (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Epidemiology (AREA)
- Animal Behavior & Ethology (AREA)
- General Health & Medical Sciences (AREA)
- Public Health (AREA)
- Veterinary Medicine (AREA)
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
Abstract
A kind of application the invention discloses P glycoprotein as protective factors in influence 832/13 cell insulin secretion of INS-1.Research illustrates P glycoprotein to the mechanism of action in terms of 832/13 cell insulin secretion of rat insulin oncocyte INS-1.In vitro test discovery, P glycoprotein can further activate PKA signal transduction pathway, increase PKA, CREB activity by improving intracellular cAMP amount.Simultaneously, this experimental verification P glycoprotein and Lipid Rafts GAP-associated protein GAP, the presence interaction of caveolin (Caveolinl), L-type calcium channel albumen 1.2 (Cav1.2), potassium-channel proteins (Kir6.2), adenyl cyclase, to influence the secretion of insulin.The present invention is research pancreas islet protective factors, improves pancreatic islets transplantation Clinical Outcome and provides fundamental basis.
Description
Technical field
The present invention relates to a kind of bioengineering fields, and in particular to one kind can influence 832/13 cell insulin of INS-1
The P glycoprotein of secretion.
Background technique
Diabetes (diabetes mellitus, DM) are one kind because of insulin resistance and (or) hypoinsulinism institute
The caused metabolic disorder disease characterized by chronic hyperglycemia seriously endangers human life and quality of life.Its pathology is raw
It is of science extremely complex, it is popular in the world, and its disease incidence also will continue to increase.It is 2014,18 years old global or more
The disease incidence of T2DM be about 9%.In the U.S., the influence of contingency is excluded, diabetes are the fifth-largest originals of women die
Cause is the fourth-largest reason of deaths in men.And the disease incidence of diabetes mellitus in China is 9.7% (9.24 hundred million adult), and forerunner is sugared
The disease incidence of urine disease is 15.5% (1,400,000,000 adult).The increase of diabetes morbidity and rapid economic development, average life span are prolonged
Long and lifestyle change is related.Poor blood glucose control will increase the relevant capilary of diabetes and macrovascular complications wind
Danger.Diabetes and complication reduce patients ' life quality, and life time shortens, and case fatality rate increases, should prevent and treat early.Treating diabetes
It is to improve glycemic control, to reduce the generation of complication.
Much research shows that diabetes can change the pharmacokinetics effect of many drugs in vivo, and this variation
It is usually closely related with the change of outlet transporter.Studies have found that the ATP of one of outlet transporter family Major Members is combined
Box (ATP-binding cassette) superfamily proteins and sugar, fat, the balance of cholesterol are closely related.ATP combination box is super
Family (ATP-binding cassette super family, ABC), mainly includes 49 kinsfolks in the mankind, these
Kinsfolk has been largely divided into 7 subtribes ABCA, ABCB, ABCC, ABCD, ABCE, ABCF and ABCG according to the homology of sequence.
Sulfonylureas receptor (sulfonyureal receptor, SUR) is the important member of ABCC subtribe, mainly includes SUR1 (ABCC8)
And SUR2 (ABCC9).SUR1 passes through to ion channel-ATP sensitive potassium-channel (K important on islet cellsATP)
Regulation plays an important role in insulin secretion.Another important member of ABC, ABCA1, report related to glycolipid metabolism
Gradually increase in recent years in road.Have been reported that the imbalance control of the first phase secretion during showing GSIS is related to Lipotoxicity, then
Person and ABCA1 have relationship closely.Prove that ABCA1 passes through regulation in the experiment of ABCA1 knock-out mice according to another report
The balance of internal cholesterol and improve islet beta cell function, maintain good insulin secretion and sugar resistance to.ABCG1 and ABCA1
It is similar, adjustment effect is generated to insulin secretion by regulation cholesterol.
P- glycoprotein (ABCB1B) be a molecular weight is about 170Kda as a member in abc transport superfamily protein
Macromolecular transmembrane protein, regulated and controled in the mankind by abcb1b single-gene, it is biradical by abcb1a and abcb1b in rodent
Because of regulation, metabolite is respectively MDR1 and MDR3.P- glycoprotein height is expressed in malignant tumor tissue, and and chemotherapeutics
Drug resistance is closely related, hence obtains one's name as multidrug resistance (MDR) albumen.It has the substrate of extensive structure type, as drug
Efflux pump, the energy generated by hydrolysising ATP enzyme, a large amount of hydrophobic drugs of outlet, such as anticancer drug, ciclosporin A, calcium channel resistance
Stagnant dose, berberine etc..P- glycoprotein is not only present in tumour cell, is also distributed widely in normal tissue, as brain, intestines, lung,
Liver, kidney, skin etc. especially have excretion and secretory endothelial cell surface, show that the albumen and cell secretion swash
Plain substance and prevent toxin invasion it is related.
All the time, tumor cell drug resistance is that clinic has one of important problem to be solved, and P- glycoprotein is to adjust
One of drug resistant main molecules target spot is controlled, therefore research hotspot has been focused into the drug efflux of P- glycoprotein mediation for a long time
In terms of pumping function.P- glycoprotein and the report of glycaemic homeostasis correlation were gradually increased both at home and abroad in recent years.Studies have reported that
Hyperglycemia state caused by diabetes has an impact for the P P-glycoprotein expression in blood-brain barrier, enteron aisle etc., and from China
Research report illustrates the expression quantity of P glycoprotein in the tissue such as brain cortex of diabetic rats, intestinal mucosa, hippocampus, kidney, liver
There is variation.Although more and more about P glycoprotein and the research of glycaemic homeostasis correlation both at home and abroad in recent years, its is right
The influence of insulin secretion is not yet clear.
Summary of the invention
Goal of the invention:To solve this problem, a kind of P- glycoprotein of Primary Study of the present invention is thin to INS-1 832/13
The influence of born of the same parents' insulin secretion and its potential mechanism provide a kind of new thinking for the prevention and treatment of diabetes.
Technical solution
Specifically, 832/13 cell of the rat insulin oncocyte INS-1 (abbreviation that the present invention passes through in vitro culture
832/13 cell);With contain various concentration (5.0 × 106, 1.0 × 107, 5.0 × 107, 1.0 × 108Pfu/mL overexpression)
The adenovirus (Ad-Abcb1b) of P- glycoprotein infection cell 4 hours under serum-free condition;Then normal incubation medium is changed into again
It is incubated for 20h;Using 3- (4,5-dimethyl-2-thiazolyl) -2,5-diphenyl-2-H-tetrazoli μM
Bromide (MTT) method detects cell activity;Cell total rna is then extracted, detects cell abcb1b using real-time quantitative PCR
The variation of mrna expression amount;832/13 epicyte protein is extracted, detects P- glycoprotein (P- using Western blot technology
Gp), insulin secretion GAP-associated protein GAP includes caveolin-1 (caveolin-1, Cav-1), 1.2 (L- of L-type calcium channel albumen
Type calcium channel protein, Cav1.2), potassium-channel proteins 6.2 (Kir6.2), chloride channel protein
3 (clcn-3), adenyl cyclase (adenylate cyclase, AC), apoptosis-related protein (Bcl-2 and Caspase-3) with
And the variation of phosphorylation PKA and CREB expression quantity;With the changes of function of Rhodamine 123 fluorescence experiments measurement P-gp;It utilizes
ELISA method measures intracellular cAMP amount;Finally high sugar stimulation pancreas is carried out with the cell after above-mentioned adenovirus infection 24 hours
Island element secretion test (glucose stimulated insulin secretion, GSIS) measures insulin using radioimmunoassay
Intracellular level and cell exocrine amount;Using co-immunoprecipitation method detection P-gp, Caveolin-1, Kir6.2, Clcn-3,
Interaction between Cav1.2, AC albumen.Then Laser Scanning Confocal is recycled to measure intracellular calcium signal
Beneficial effect
Present invention illustrates P glycoprotein can by activate cAMP-PKA-CREB-Ca2+ signal transduction pathway, increase PKA,
CREB activity, increases intracellular cAMP amount, making insulin secretion GAP-associated protein GAP includes caveolin (Caveolin1), L-type
The expression quantity increasing of calcium channel albumen 1.2 (Cav1.2), potassium-channel proteins (Kir6.2), adenyl cyclase (AC)
Add, the secretion of insulin is then influenced by the phase interaction between albumen.The present invention is research pancreas islet protective factors, improves pancreas islet
Transplanting Clinical Outcome is provided fundamental basis.
Detailed description of the invention:
Fluorescent effect figure after the different modes adenovirus infected cells of Fig. 1 fluorescence microscope identification.Fig. 1-1 show from a left side to
The right side is followed successively by respectively with (5 × 10 containing various concentration6, 10 × 106, 50 × 106, 100 × 106Pfu/mL) Ad-Abcb1b
After 832/13 cell 4h of adenovirus infection INS-1, changes normal incubation medium into and be incubated for the infectious effect observed after 20h again.Fig. 1-
2 show that adenovirus concentration is 50 × 106Under the conditions of pfu/mL, be from left to right followed successively by it is non-fluorescence under cell growth state, then
Infect the different time (12h, for 24 hours, 48h) infectious effect that microscopically observation arrives afterwards.
Fig. 2 is the histogram of cell activity after the P- glycoprotein of MTT identification is overexpressed.Control group is without containing Ad-
Adenovirus (the concentration 100 × 10 of Abcb1b6Pfu/mL), experimental group, which is used, contains various concentration (5 × 106, 10 × 106, 50 ×
106, 100 × 106Pfu/mL) the 832/13 cell 4h of adenovirus infection of Ad-Abcb1b, then changes normal incubation medium into and is incubated for again
20h.The experimental results showed that the P glycoprotein of selected concentration is to 832/13 cell of INS-1, there is no apparent cytotoxic effects.
Fig. 3 is P P-glycoprotein expression in cell after the distinct methods adenovirus infected cells of Western blot technical appraisement
The western blot figure of variation is measured, real-time round pcr identifies the variable quantity histogram of P glycoprotein gene level.3-1 shows not
With concentration 832/13 cell of adenovirus infection for 24 hours after P P-glycoprotein expression situation, negative control group is without containing Ad-Abcb1b
Adenovirus (concentration be 100 × 106Pfu/mL), experimental group is to contain various concentration Ad-Abcb1b adenovirus infection 832/13
Normal incubation medium is changed into after cell 4h is incubated for 20h again.Fig. 3-2 shows P P-glycoprotein expression feelings after adenovirus infected cells different time
Condition, negative control group and experimental group adenovirus concentration are 50 × 106pfu/mL.Fig. 3-3 show adenovirus concentration be 50 ×
106Pfu/mL, infection time is the expression of P-gp when for 24 hours, as a result, it has been found that not only P-gp gene and albumen water on cell
Flat expression increases, and the expression of P-gp gene and protein level increases (Fig. 3-4) in pancreas islet.Fig. 3-5 shows that adenovirus is dense
Degree is 50 × 106Pfu/mL, infection time be for 24 hours when cytoplasm and cell membrane on P-gp expression quantity variation, with control group phase
Than expression of the overexpression group P-gp on cell membrane obviously increases.
Fig. 4 is the expression quantity variation of Western Blot technical appraisement apoptosis-related protein Caspase3 and Bcl2.
Cell death related protein shown in Western Blot result, compared with negative control group cell, after P glycoprotein overexpression
The expression of Caspase3 and Bcl2, there is no significant changes
Fig. 5 is the immunofluorescence of Rhodamine 123 fluorescence experiments identification P-gp outlet changes of function.Adenovirus infected cells
Afterwards, by the outer row function of Rhodamine 123 (Rh-123) accumulation experiment detection P-gp, whether there is or not variations.P-gp can obviously drop after being overexpressed
The accumulation of the intracellular Rhodamine 123 of low INS-1 832/13 shows that the outlet pumping function of P-gp significantly increases.
Fig. 6 is that insulin synthesis expression quantity ins1, ins2 and pdx1 are expressed after the P glycoprotein of qPCR technical appraisement is overexpressed
Change histogram.Fig. 6-1 shows that compared with the control group overexpression group ins1 gene expression amount does not have significant change;Fig. 6-2 show with it is right
It is compared according to group, overexpression group ins2 gene expression amount does not have significant change;Fig. 6-3 shows compared with the control group, overexpression group pdx1
There is no significantly changing for gene expression amount.
Fig. 7 is that row low sugar and high sugar stimulate insulin secretion and tests carefully after ELISA method identification adenovirus infection 24 hours
Insulin content intracellular and extracellular amount of insulin secretion.Fig. 7-1 show low sugar be incubated for when, experimental group compared with the control group, base
Plinth insulin secretion does not have significant change;After Fig. 7-2 shows correction albumen, the content of control group and experimental group insulin inside cells is simultaneously
There is no difference.Fig. 7-3 shows the increased multiple of cell insulin after high sugar stimulation, as the result is shown compared with negative control group, experiment
The increased multiple of group is significantly higher, illustrates that amount of insulin secretion obviously increases.
Fig. 8 be Western blot technical appraisement P-gp, Cav-1, Cav1.2, Kir6.2, clcn-3, AC, Bcl-2 and
The western blot figure of Caspase-3, PKA and CREB expression quantity variation.As shown in Fig. 8-1, compared with the control group, P glycoprotein mistake
Cav1.2 the and Cav-1 expression quantity of expression group dramatically increases, as the result is shown the variation of Cav1.2 and Cav-1 protein expression and P sugar
Albumen variation is consistent.Fig. 8-2 display, compared with negative control group, after P glycoprotein is overexpressed, intracellular cAMP amount obviously increases
Add, AC protein expression increases, and PKA and CREB activity increases, and Kir6.2 protein expression does not change (Fig. 8-3).Furthermore pass through
ELISA kit detection discovery,.
Fig. 9 be co-immunoprecipitation method identification P-gp, Caveolin-1, Kir6.2, Clcn-3, Cav1.2, AC albumen between
The histogram of interaction.The results show that P-gp and Cav-1, Cav-1 and Kir6.2, have interaction between Cav-1 and AC
Relationship (Fig. 9-1, Fig. 9-2), but do not have temporarily between P-gp and Cav1.2, Kir6.2, AC and between Cav-1 and Cav1.2
It is found to have interaction relationship (data are not shown).
Figure 10 is that Laser Scanning Confocal measures above-mentioned intracellular calcium signal figure.As a result, it has been found that compared with negative control group,
When low sugar is incubated for, P glycoprotein overexpression group intracellular calcium signal does not change, after high sugar stimulation, P glycoprotein overexpression group cell
Interior Ca2+ oscillations increase.
Specific embodiment
Embodiment 1:The building of P glycoprotein overexpression 832/13 cell of insulinoma cell INS-1.
The full culture medium of cell and cell culture is got out often to be placed in cell room gnotobasis with equipment.Remove cell culture
Base, sterile PBS are rinsed cell 2 times.5mL cell culture medium is added, blows and beats cell with 3mL suction pipe, until the bright i.e. bottom of bottle of bottle wall
Cell has been blown down.Suspension moves in centrifuge tube, and appropriate culture medium is added, and is gently blown and beaten with electric gun and mixes suspension.It will
Cell suspension moves in centrifuge tube, appropriate culture medium is added, then mix after blowing and beating 10min with 10ml liquid-transfering gun, then will suspend
Liquid tiling is to 6 orifice plates, after growing 12-24h, when cell is placed in logarithmic growth phase, changes serum free medium into, then dense with difference
(the 5 × 10 of degree6, 10 × 106, 50 × 106, 100 × 106Pfu/mL the adenovirus infected cells 4 for) being overexpressed P- glycoprotein are small
Shi Hou changes normal incubation medium into, then after being incubated for 20h, respectively with Western blot method and real-time PCR method detection P sugar
The variation of the albumen and gene level of albumen.It is again 50 × 10 with adenovirus concentration6Then pfu/mL infects the different time
(12h, for 24 hours, 48h) detects the variation of the albumen and gene level of P glycoprotein afterwards.
Embodiment 2:When mtt assay detects cell activity with the suitable adenovirus concentration of determination and adenovirus optimal infection
Between.
The cell suspension of 100 μ L, adjustment cell density to 5 is added at cell suspending liquid in cell in each hole of 96 orifice plates
×103/ hole, often plus after 3-5 column, " 8 " word method mixes cell suspension, while corresponding control wells are arranged, i.e., contains only respectively
Cell culture fluid, MTT, DMSO group, the edge hole of 96 orifice plates are filled with sterile PBS to exclude the influence of edge effect.To for 24 hours
The adenovirus of various concentration is added afterwards, if 5 gradients, 6 multiple holes.After adenovirus infection, microscopically observation is cellular
State.The MTT solution (i.e. 5mg/ml) of 10 μ L 0.5% is added in every hole, continues to terminate MTT incubation after being incubated for 4h under the conditions of being protected from light, and
150 μ LDMSO are added in every hole, and shaking table low-speed oscillation 10min is placed under the conditions of being protected from light, with the suction in each hole at microplate reader detection 490nm
Shading value.Statistical procedures are carried out according to cell viability=experimental group A value/control group A value.The result shows that the P sugar of selected concentration
There is no apparent cytotoxic effects to 832/13 cell of INS-1 for albumen.Then further according to MTT's as a result, determining suitably
Dosage, 12h upon administration, for 24 hours, 48h collect cell, extract total protein of cell, and Western blot detects the table of P-gp
Up to amount, band finally is parsed using gray analysis
After embodiment 3P-gp is overexpressed, the outlet pumping function of P-gp is significantly increased
12 orifice plate culture cells are after 24 hours, respectively with 50 × 106The Ad-Control and Ad- of pfu/mL concentration
Abcb1b adenovirus incubated cell after infection, is washed cell 2 times with the serum-free 1640 culture medium of preheating.Every hole is added
Serum-free 1640 culture medium 1mL containing 1 μ l rhodamine mother liquor (1 μ g/ μ l), incubator are protected from light 3h.After incubation, with no blood
Clear 1640 washing cell 2 times, extract cell protein.Every hole adds the strong RIPA lysate of 220 μ L to carry out cracking 30min, 12000r/
Clear liquid is taken to be added in dedicated 96 orifice plate of fluorescence intensity that white is protected from light after min centrifugation 10min, 100 μ L supernatants are added in every hole
Liquid;Then the standard items that rhodamine is prepared are added and pass through VictorIII under the conditions of λ ex=485nm, λ em=535nm
Multiplate reader instrument surveys fluorescence intensity level.P-gp can obviously reduce intracellular sieve of INS-1 832/13 after being overexpressed
The accumulation of pellet bright 123, shows that the outlet pumping function of P-gp significantly increases.
Embodiment 3:After P glycoprotein is overexpressed, cell RNA is extracted, using related genes such as qPCR method detection abcb1b
The variation of expression.
1) primer
2) corresponding EP pipe is marked, following components (20 μ l of total volume) is then sequentially added:
Embodiment 4:High sugar stimulates lower insulin secretion to obviously increase after P glycoprotein overexpressing cell and islet cells.
It takes 12 orifice plates to carry out bed board, when cell density is to 70%-80%, changes serum free medium into, respectively with being free of
There are the negative control virus of abcb1b and the adenovirus (concentration 5.0 × 10 containing abcb1b7Pfu/mL) infection cell 4 hours
Afterwards, it changes normal incubation medium into, collects supernatant respectively after low sugar (2.5mM) is incubated for and height sugared (16.7mM) stimulates after being incubated for 20h
Liquid extracts cell protein with sour Ethanol Method, protein content is then detected, with Mercodia Rat Insulin ELISA kit
Test sample insulin content, and corrected with corresponding protein concentration.When rat Islet cells transfect, first select form it is consistent,
Size is close, circular fresh pancreas islet, and 50 are divided into one group, are placed in 6 orifice plates culture for 24 hours.Second day, pancreas islet is moved into 96 holes
Plate changes serum free medium into, and transfection method and GSIS experimental method are the same as above-mentioned cell.
Embodiment 5:Abcb1b increases the expression quantity of P-gp, Cav-1, AC, Cav1.2 albumen, increases CREB activity.
After P glycoprotein is overexpressed, epicyte protein or nucleoprotein are extracted, detects P sugar egg using Western blot technology
The variation of white, Cav-1, Cav1.2, Kir6.2, AC, Bcl-2 and Caspase-3 and phosphorylation PKA and CREB expression quantity.
Embodiment 6:Abcb1b intracellular P-gp and Cav-1, Cav-1 and Kir6.2 after being overexpressed, Cav-1 and AC is with phase
Interaction relationship
After being overexpressed using co-immunoprecipitation detection P-gp, Cav-1, Cav-1 and Kir6.2, Cav-1 and AC interact
Relationship, with P-gp precipitate C av-1, the same protein extracting method of cell lysing methods takes the cell cracking supernatant of 100 μ g, with egg
White G agarose magnetic bead takes supernatant after carrying out preincubate 2h, then respectively again with the C219 antibody or IgG antibody and Protein G of 5 μ g
Agarose magnetic bead is incubated for 2h.Finally cell supernatant and immunomagnetic beads antibody-containing are incubated overnight, then pass through Western
Blot method uses Cav-1 antibody as primary antibody.
Embodiment 7:Abcb1b increases intracellular calcium concentration and increases
Using the variation of the intracellular calcium signal of confocal experiments detection.After infection cell, the cleaning of low sugar solution is thin
Then born of the same parents 2-3 times are that 2 μM of Fura-2 fluorescence probe is added in culture plate the concentration that 2.5mmol low sugar is prepared, detection with
The fluorescence intensity and two intensity rates of the 510nm of 340nm and 380nm excitation.Low sugar solution is discarded when 200s, with low sugar solution
The probe solution that high sugared (16.7mmol) is prepared is added in 300s or so for cleaning raffinate 2-3 time, then detect with 340nm and
The fluorescence intensity and two intensity rates of the 510nm of 380 nm excitation.Intracellular calcium signal becomes after Figure 10 shows low sugar and high sugar is added
Change, (Ca2+ oscillations change maximum) intracellular calcium signal changes when Figure 10 shows 400s.
Claims (6)
- The application of 1.P glycoprotein influence 8721/13 cell insulin secretion of insulinoma cell INS-1.
- 2. according to claim 1, P glycoprotein is overexpressed the protective factors that can be used as pancreatic islets transplantation.
- 3. according to claim 1, P glycoprotein, which is overexpressed, improves intracellular cAMP amount.
- 4. according to claim 1, P glycoprotein, which is overexpressed, improves the high lower insulin secretion of sugar stimulation.
- 5. according to claim 1, P glycoprotein overexpression can stablize Lipid Rafts structure.
- 6. according to claim 1, P glycoprotein, which is overexpressed, influences calcium channel albumen expression.
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CN111440771A (en) * | 2020-03-17 | 2020-07-24 | 中山大学附属第三医院 | C L CN2 homozygous mutation-containing white matter encephalopathy patient specific induced pluripotent stem cell line |
CN111575310A (en) * | 2020-05-14 | 2020-08-25 | 江南大学 | Recombinant saccharomyces cerevisiae expressing caveolin and application thereof |
-
2017
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Title |
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汤云昭等: "大鼠胰岛β细胞中mini-P糖蛋白的表达及其功能研究", 《中国糖尿病杂志》 * |
Cited By (3)
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CN111440771A (en) * | 2020-03-17 | 2020-07-24 | 中山大学附属第三医院 | C L CN2 homozygous mutation-containing white matter encephalopathy patient specific induced pluripotent stem cell line |
CN111575310A (en) * | 2020-05-14 | 2020-08-25 | 江南大学 | Recombinant saccharomyces cerevisiae expressing caveolin and application thereof |
CN111575310B (en) * | 2020-05-14 | 2022-12-13 | 江南大学 | Recombinant saccharomyces cerevisiae expressing caveolin and application thereof |
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