CN107375344A - Lactobacillus plantarum ZDY2013 new opplication - Google Patents
Lactobacillus plantarum ZDY2013 new opplication Download PDFInfo
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- CN107375344A CN107375344A CN201710544737.3A CN201710544737A CN107375344A CN 107375344 A CN107375344 A CN 107375344A CN 201710544737 A CN201710544737 A CN 201710544737A CN 107375344 A CN107375344 A CN 107375344A
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- lactobacillus plantarum
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- 240000006024 Lactobacillus plantarum Species 0.000 title claims abstract description 37
- 235000013965 Lactobacillus plantarum Nutrition 0.000 title claims abstract description 37
- 229940072205 lactobacillus plantarum Drugs 0.000 title claims abstract description 37
- 206010009887 colitis Diseases 0.000 claims abstract description 16
- 239000002068 microbial inoculum Substances 0.000 claims description 17
- 206010009900 Colitis ulcerative Diseases 0.000 claims description 15
- 201000006704 Ulcerative Colitis Diseases 0.000 claims description 15
- 235000013305 food Nutrition 0.000 claims description 13
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- TVZRAEYQIKYCPH-UHFFFAOYSA-N 3-(trimethylsilyl)propane-1-sulfonic acid Chemical compound C[Si](C)(C)CCCS(O)(=O)=O TVZRAEYQIKYCPH-UHFFFAOYSA-N 0.000 abstract description 3
- 208000024891 symptom Diseases 0.000 abstract description 2
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- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 2
- 238000011725 BALB/c mouse Methods 0.000 description 2
- WSFSSNUMVMOOMR-UHFFFAOYSA-N Formaldehyde Chemical compound O=C WSFSSNUMVMOOMR-UHFFFAOYSA-N 0.000 description 2
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- 208000022559 Inflammatory bowel disease Diseases 0.000 description 2
- CSNNHWWHGAXBCP-UHFFFAOYSA-L Magnesium sulfate Chemical compound [Mg+2].[O-][S+2]([O-])([O-])[O-] CSNNHWWHGAXBCP-UHFFFAOYSA-L 0.000 description 2
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- KRKNYBCHXYNGOX-UHFFFAOYSA-N citric acid Natural products OC(=O)CC(O)(C(O)=O)CC(O)=O KRKNYBCHXYNGOX-UHFFFAOYSA-N 0.000 description 2
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- WUBBRNOQWQTFEX-UHFFFAOYSA-N 4-aminosalicylic acid Chemical compound NC1=CC=C(C(O)=O)C(O)=C1 WUBBRNOQWQTFEX-UHFFFAOYSA-N 0.000 description 1
- 208000011231 Crohn disease Diseases 0.000 description 1
- 206010012735 Diarrhoea Diseases 0.000 description 1
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 description 1
- 241001465754 Metazoa Species 0.000 description 1
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- 108010080698 Peptones Proteins 0.000 description 1
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- VMHLLURERBWHNL-UHFFFAOYSA-M Sodium acetate Chemical compound [Na+].CC([O-])=O VMHLLURERBWHNL-UHFFFAOYSA-M 0.000 description 1
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- 239000002253 acid Substances 0.000 description 1
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- 230000001154 acute effect Effects 0.000 description 1
- 229960004909 aminosalicylic acid Drugs 0.000 description 1
- 229940040526 anhydrous sodium acetate Drugs 0.000 description 1
- 235000015278 beef Nutrition 0.000 description 1
- 239000003833 bile salt Substances 0.000 description 1
- 239000008280 blood Substances 0.000 description 1
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- -1 citric acid hydrogen diamine Chemical class 0.000 description 1
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- ZPWVASYFFYYZEW-UHFFFAOYSA-L dipotassium hydrogen phosphate Chemical compound [K+].[K+].OP([O-])([O-])=O ZPWVASYFFYYZEW-UHFFFAOYSA-L 0.000 description 1
- 239000012153 distilled water Substances 0.000 description 1
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- 239000003862 glucocorticoid Substances 0.000 description 1
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- 210000002175 goblet cell Anatomy 0.000 description 1
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- 230000015784 hyperosmotic salinity response Effects 0.000 description 1
- 210000003405 ileum Anatomy 0.000 description 1
- 239000002955 immunomodulating agent Substances 0.000 description 1
- 229940121354 immunomodulator Drugs 0.000 description 1
- 230000002584 immunomodulator Effects 0.000 description 1
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- 238000001764 infiltration Methods 0.000 description 1
- 230000004968 inflammatory condition Effects 0.000 description 1
- 239000002054 inoculum Substances 0.000 description 1
- 230000000968 intestinal effect Effects 0.000 description 1
- 230000008944 intestinal immunity Effects 0.000 description 1
- 208000002551 irritable bowel syndrome Diseases 0.000 description 1
- 230000003902 lesion Effects 0.000 description 1
- 229910052943 magnesium sulfate Inorganic materials 0.000 description 1
- 235000019341 magnesium sulphate Nutrition 0.000 description 1
- 230000003211 malignant effect Effects 0.000 description 1
- 229940099596 manganese sulfate Drugs 0.000 description 1
- 235000007079 manganese sulphate Nutrition 0.000 description 1
- 239000011702 manganese sulphate Substances 0.000 description 1
- SQQMAOCOWKFBNP-UHFFFAOYSA-L manganese(II) sulfate Chemical compound [Mn+2].[O-]S([O-])(=O)=O SQQMAOCOWKFBNP-UHFFFAOYSA-L 0.000 description 1
- 244000005700 microbiome Species 0.000 description 1
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- 235000010482 polyoxyethylene sorbitan monooleate Nutrition 0.000 description 1
- 229920000053 polysorbate 80 Polymers 0.000 description 1
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- 208000016261 weight loss Diseases 0.000 description 1
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Classifications
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
- A61K35/66—Microorganisms or materials therefrom
- A61K35/74—Bacteria
- A61K35/741—Probiotics
- A61K35/744—Lactic acid bacteria, e.g. enterococci, pediococci, lactococci, streptococci or leuconostocs
- A61K35/747—Lactobacilli, e.g. L. acidophilus or L. brevis
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
- A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
- A23L33/135—Bacteria or derivatives thereof, e.g. probiotics
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2400/00—Lactic or propionic acid bacteria
- A23V2400/11—Lactobacillus
- A23V2400/169—Plantarum
Abstract
The invention discloses Lactobacillus plantarum ZDY2013 new opplication, the Lactobacillus plantarum ZDY2013 has the mouse gram lain disease for being capable of TNBS inductions, improves the apparent symptom of inflammation, adjusts oxidative stress, regulate and control the expression of inflammatory factor, reach the colitis in mice of effectively preventing and treating DSS inductions.
Description
Technical field
The invention belongs to microorganism field, is specifically related to a kind of Lactobacillus plantarum ZDY2013 new opplication.
Background technology
Colitis (inflammatory bowel disease, IBD) is divided into ulcerative colitis (ulecrative
Colitis, UC) and gram lain disease (Crohn ' s disease, CD) two types.With morbidity and mortality in recent years not
Disconnected to rise, colitis has turned into a kind of malignant disease of serious threat human life and health.Modern times are classified as by the World Health Organization
One of refractory treatment illness.Its common clinical manifestation is Body weight loss, diarrhoea, bloody stool, has the tendency of canceration, repeatability is strong.Research
Prove, the medicine currently used for treating colitis mainly has aminosalicylic acid, glucocorticoid and immunomodulator etc., medicine valency
Height, and side effect is uncontrollable.
Lactobacillus plantarum can adjust gut flora balance, suppress the life of harmful bacteria as enteron aisle Gram-positive probiotics
It is long, occupy critical role in intestine immunity effect.
The Lactobacillus plantarum ZDY2013 that applicant of the present invention is separated to before was stored in Wuhan on April 28th, 2014
Wuhan University of city China typical culture collection center, preserving number are CCTCC M2014170.And invent the Lactobacillus plantarum
ZDY2013 poles acid and bile salt tolerance, higher survival rate can be kept in gastro-intestinal Fluid, and the growth of food-borne pathogens can be suppressed.
Moreover it is possible to because gut flora diversity can be adjusted, reaching improves host gastrointestinal tract health.But in view of enterogastric diseases
Complexity and diversity, the Lactobacillus plantarum ZDY2013 also knows nothing in the treatment function of specific enterogastric diseases.
The content of the invention
An object of the present invention is to provide Lactobacillus plantarum ZDY2013 new opplication.
To realize object above, the invention provides descend technical scheme：
Applications of the Lactobacillus plantarum ZDY2013 in the medicine or health products or food for preparing preventing and treating ulcerative colitis,
The preserving number of the Lactobacillus plantarum ZDY2013 is CCTCC M2014170.
It is a further object of the present invention to provide a kind of medicine for treating ulcerative colitis or health products or food.
Realize that the technical scheme of above-mentioned purpose is as follows.
A kind of medicine for treating ulcerative colitis or health products or food, its active component include Lactobacillus plantarum
ZDY2013, the Lactobacillus plantarum ZDY2013 preserving number are CCTCC M2014170.
Preferably, the food is the survivable food of a variety of lactobacillus such as beverage, Yoghourt, jelly, as long as plant breast bar
In the survivable food of bacterium can, with specific reference to being actually needed.
Preferably, the lactobacillus plantarum microbial inoculum dosage of the treatment ulcerative colitis used in the present invention is 1 × 1010 cfu/
Kg, calculate use as the case may be, i.e., ZDY2013 containing Lactobacillus plantarum reaches in microbial inoculum used in every kg animals, human body
1010cfu。
The invention discloses a kind of probiotics microbial inoculum for being used to treat ulcerative colitis, by Lactobacillus plantarum ZDY2013
It is prepared, exists in the form of a kind of fresh viable bacteria thalline in stationary phase.
The Lactobacillus plantarum ZDY2013 that the present invention uses was stored in Wuhan City's Chinese Typical Representative culture on April 28th, 2014
Thing collection, preserving number are CCTCC M2014170.
The present invention further discloses the preparation method of the lactobacillus plantarum microbial inoculum for the treatment of colitis, comprise the following steps：
1) 37 ± 1 DEG C of 24 ± 2h of Anaerobic culturel of Lactobacillus plantarum ZDY2013MRS fluid nutrient mediums, reach stationary phase
Thalline, the preserving number of the Lactobacillus plantarum ZDY2013 is CCTCC M2014170；
2) thalline in step 1) is collected by centrifugation, is washed 2-4 times with sterile 1 × PBS；
3) with sterile 1 × PBS set-up procedures 2) gained cell concentration is to 1-2 × 109cfu/mL；
4) medicine or food or health products made of proper auxiliary materials are added into above-mentioned gained thalline.
The present inventor is studied by substantial amounts of the Lactobacillus plantarum ZDY2013 obtained before, described in discovery
Lactobacillus plantarum ZDY2013 has the mouse gram lain disease for being capable of TNBS inductions, improves the apparent symptom of inflammation, regulation oxidation should
Swash, regulate and control the expression of inflammatory factor, reach the colitis in mice of effectively preventing and treating DSS inductions.
Brief description of the drawings
Fig. 1 separate groups of mice changes of weight.
Fig. 2 separate groups of mice colon HE stained slices.
The different group ulcerative colitis mouse inflammatory Cytokines Expressions of Fig. 3.
The different group gram lain disease mouse inflammatory Cytokines Expressions of Fig. 4.
Embodiment
For the ease of understanding the present invention, the present invention will be described more fully below.The present invention can be with many not
With form realize, however it is not limited to embodiment described herein.On the contrary, the purpose for providing these embodiments is made to this
The understanding of disclosure of the invention content more thorough and comprehensive.
The experimental method of unreceipted actual conditions in the following example, generally according to normal condition, such as Sambrook etc.
People, molecular cloning：Laboratory manual (New York:Cold Spring Harbor Laboratory Press, 1989) institute in
The condition stated, or according to the condition proposed by manufacturer.Used various conventional chemical reagent, are commercially available in embodiment
Product.
Unless otherwise defined, the technical field of all technologies used in the present invention and scientific terminology with belonging to the present invention
The implication that is generally understood that of technical staff it is identical.The term used in the description of the present invention is intended merely to describe specific reality
The purpose of example is applied, is not used in the limitation present invention.Term "and/or" used in the present invention includes the listed of one or more correlations
The arbitrary and all combination of project.
Embodiment 1
The activation of thalline：
10 μ L, which are drawn, after the Lactobacillus plantarum ZDY2013 vortexs being stored in glycerine are mixed is coated on MRS solid cultures
In base, 37 DEG C of Anaerobic culturel 24h, picking single bacterium colony is inoculated in 4mL liquid MRS culture mediums, 37 DEG C of Anaerobic culturel 24h, is pressed
1% inoculum concentration is inoculated in 10mL MRS fluid nutrient mediums, 37 DEG C of Anaerobic culturel 24h, reaches stationary phase.
MRS culture mediums are prepared：Peptone 5g/L, beef extract 5g/L, tryptone 10g/L, dusty yeast 5g/L, glucose
20g/L, anhydrous sodium acetate 5g/L, citric acid hydrogen diamine 2g/L, dipotassium hydrogen phosphate 2g/L, magnesium sulfate 0.58g/L, manganese sulfate
0.25g/L, 1ml/L, pH6.3 ± 0.1,121 DEG C of Tween-80, sterilize within 15 minutes
Embodiment 2
The preparation of lactobacillus plantarum microbial inoculum：
The Lactobacillus plantarum ZDY2013 in stationary phase activated is centrifuged, new fresh thalli is collected, with sterile 1 × PBS
Washing 3 times, cell concentration is adjusted to 10 with sterile 1 × PBS9Cfu/mL, microbial inoculum needed for experiment is made, it is standby.
Embodiment 3
The structure of ulcerative colitis mouse model and experimental design：
DSS distilled water solutions are prepared, is freely drunk 7 days after BALB/c mouse adapts to environment, builds ulcerative colitis
Model, judge whether modeling succeeds by Apparent character.When mouse weight drops to 10%, start gavage Lactobacillus plantarum
ZDY2013 microbial inoculums, by 10mL/Kg, it is administered once daily, successive administration 5 days.To colitis naive mice gavage PBS, treatment
After end, pluck eyeball and take blood and put to death mouse, take terminal ileum and total colectomy.Physiological saline rinses rapidly.Sample in
10% formalin solution soaks, and does HE dyeing.Remainder colon preserves to RNA and protects liquid, and -80 DEG C save backup.
Experimental result：
(1) present invention is obviously improved mouse weight
After gavage Lactobacillus plantarum ZDY2013 microbial inoculums are treated 5 days, treatment group mouse has compared with colitis group mouse weight
Obvious to improve, as shown in figure 1, after treatment end, probiotics microbial inoculum treatment group mouse can recover to original body mass substantially.Explanation
Body weight loss situation caused by the present invention can effectively alleviate colitis, improve the body weight of mouse.
(2) present invention significantly improves mouse pathology
Pathology section is made in separate groups of mice colon, colitis group mouse Colon tissue enters with natural control group
Row contrast, figure it is seen that colitis group colon goblet cell major injury, partial loss, inflammatory cell infiltration, viscous
Film lower floor is more complete in continuously distributed, lamina propria generation lesion, of the present invention group of (lactobacillus plantarum microbial inoculum treatment group) colon morphology
It is whole, inflammatory cell partial wetting.Illustrate that microbial inoculum of the present invention can effectively alleviate colon's inflammatory conditions of colitis mice, have
There is the effect of repairing intestinal mucosal injury.
(3) microbial inoculum of the present invention significantly adjusts the expression of ulcerative colitis inflammatory factor
The colon for being stored in -80 DEG C is subjected to RNA extractions, after reverse transcription operation, detects inflammatory factor TNF-α,
IL-10 expression.
As seen from Figure 3, of the present invention group (Lactobacillus plantarum group) mouse is compared with ulcerative colitis group mouse inflammatory factor
Expression is significantly adjusted, and has lowered proinflammatory inflammation factor TNF-α expression, and has raised suppression inflammatory factor IL-10's
Expression.Illustrate that microbial inoculum of the present invention can be by adjusting Cytokine Expression Level, so as to reach protection body anti-inflammatory
Purpose.
Embodiment 4
The structure of gram lain disease model mice and experimental design：
TNBS ethanol solution is prepared, the BALB/c mouse of use fasting 24h, No. 9 straight gastric perfusion needles after environment is adapted to are entered
Row bowel lavage, establish acute gram of lain disease model.When mouse weight drops to 10%, start gavage Lactobacillus plantarum ZDY2013
Microbial inoculum, by 10mL/Kg, it is administered once daily, successive administration 8 days.After treatment end, colonic segment is chosen, preserves to RNA and protects
Liquid, -80 DEG C freeze it is standby.
Experimental result：The inflammatory Cytokines Expression of significantly regulation gram lain disease colitis of the invention
By the colon for being stored in -80 DEG C by liquid nitrogen grinding, RNA extractions are carried out using RNA extracts kits, are incited somebody to action
RNA reverse transcriptions pass through the expression of RT-qPCR methods detection inflammatory factor TNF-α into cDNA.As seen from Figure 4, this hair
Bright group of mouse is significantly adjusted compared with a gram lain disease colitis group mouse inflammatory Cytokines Expression level, has lowered proinflammatory inflammation factor
TNF-α expression.Illustrate that microbial inoculum of the present invention can be by adjusting Cytokine Expression Level, so as to reach protection body anti-inflammatory
The purpose of disease.
The specific embodiment of the present invention is these are only, protection scope of the present invention is not limited with this；Do not violating this hair
Any replacement made on the basis of bright design belongs to protection scope of the present invention with improving.
Claims (5)
1. applications of the Lactobacillus plantarum ZDY2013 in the medicine or health products or food for preparing preventing and treating ulcerative colitis, institute
The preserving number for stating Lactobacillus plantarum ZDY2013 is CCTCC M2014170.
2. a kind of medicine for treating ulcerative colitis or health products or food, it is characterized in that, its active component includes plant
Lactobacillus ZDY2013, the Lactobacillus plantarum ZDY2013 preserving number are CCTCC M2014170.
3. the medicine or health products or food for the treatment of ulcerative colitis according to claim 2, it is characterized in that, the food
Product are beverage, Yoghourt, jelly.
4. the medicine or health products or food for the treatment of ulcerative colitis according to claim 2, it is characterized in that, the plant
Thing lactobacillus microbial inoculum dosage is 1 × 1010cfu/kg。
5. a kind of preparation method for the lactobacillus plantarum microbial inoculum for treating colitis, it is characterized in that, comprise the following steps：
1) 37 ± 1 DEG C of 24 ± 2h of Anaerobic culturel of Lactobacillus plantarum ZDY2013MRS fluid nutrient mediums, the bacterium of stationary phase must be reached
Body, the preserving number of the Lactobacillus plantarum ZDY2013 is CCTCC M2014170；
2) thalline in step 1) is collected by centrifugation, is washed 2-4 times with sterile 1 × PBS；
3) with sterile 1 × PBS set-up procedures 2) gained cell concentration is to 1-2 × 109cfu/mL；
4) medicine or food or health products made of proper auxiliary materials are added into above-mentioned gained thalline.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201710544737.3A CN107375344A (en) | 2017-07-06 | 2017-07-06 | Lactobacillus plantarum ZDY2013 new opplication |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201710544737.3A CN107375344A (en) | 2017-07-06 | 2017-07-06 | Lactobacillus plantarum ZDY2013 new opplication |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN107375344A true CN107375344A (en) | 2017-11-24 |
Family
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Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201710544737.3A Pending CN107375344A (en) | 2017-07-06 | 2017-07-06 | Lactobacillus plantarum ZDY2013 new opplication |
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Cited By (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108504597A (en) * | 2018-03-30 | 2018-09-07 | 南昌大学 | A kind of allergic reaction lactobacillus and its application for alleviating beta lactoglobulin induction |
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108504597A (en) * | 2018-03-30 | 2018-09-07 | 南昌大学 | A kind of allergic reaction lactobacillus and its application for alleviating beta lactoglobulin induction |
| CN108504597B (en) * | 2018-03-30 | 2021-03-26 | 南昌大学 | Lactobacillus for relieving beta-lactoglobulin-induced anaphylactic reaction and application thereof |
| WO2020084051A1 (en) * | 2018-10-24 | 2020-04-30 | Novozymes A/S | Probiotic supplement for metabolic health comprising lactobacillus |
| US11963986B2 (en) | 2018-10-24 | 2024-04-23 | Novozymes A/S | Probiotic supplement for metabolic health |
| CN110964665A (en) * | 2019-12-20 | 2020-04-07 | 哈尔滨美华生物技术股份有限公司 | Probiotic composition for post-gastrectomy treatment and preparation method thereof |
| CN110964665B (en) * | 2019-12-20 | 2020-09-25 | 哈尔滨美华生物技术股份有限公司 | Probiotic composition for post-gastrectomy treatment and preparation method thereof |
| CN111529553A (en) * | 2020-05-28 | 2020-08-14 | 东北农业大学 | Application of plant lactobacillus capable of degrading tryptophan and tryptophan mixture |
| CN111529553B (en) * | 2020-05-28 | 2020-11-27 | 东北农业大学 | Application of mixture of lactobacillus plantarum KLDS1.0386 and tryptophan in preparation of medicine for preventing colitis |
| CN112553115A (en) * | 2020-12-23 | 2021-03-26 | 南昌大学 | Application of lactobacillus plantarum ZDY2013 in preparation of products for relieving kidney injury |
| CN113678895A (en) * | 2021-08-18 | 2021-11-23 | 南昌大学 | Synbiotic fermented milk with high antioxidant activity in cold storage period and preparation method thereof |
| CN115089619A (en) * | 2022-07-18 | 2022-09-23 | 南昌大学 | Application of lactobacillus plantarum ZDY2013 in preparation with function of relieving non-alcoholic fatty liver disease |
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