CN109439592A - Lactobacillus plantarum preparation and its application - Google Patents
Lactobacillus plantarum preparation and its application Download PDFInfo
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- CN109439592A CN109439592A CN201811490004.7A CN201811490004A CN109439592A CN 109439592 A CN109439592 A CN 109439592A CN 201811490004 A CN201811490004 A CN 201811490004A CN 109439592 A CN109439592 A CN 109439592A
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- lactobacillus plantarum
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- 235000013965 Lactobacillus plantarum Nutrition 0.000 title claims abstract description 58
- 229940072205 lactobacillus plantarum Drugs 0.000 title claims abstract description 58
- 238000002360 preparation method Methods 0.000 title claims abstract description 36
- 239000006041 probiotic Substances 0.000 claims abstract description 27
- 235000018291 probiotics Nutrition 0.000 claims abstract description 27
- 241000894006 Bacteria Species 0.000 claims description 20
- 241000588724 Escherichia coli Species 0.000 claims description 7
- 241000191967 Staphylococcus aureus Species 0.000 claims description 6
- 206010046914 Vaginal infection Diseases 0.000 claims description 5
- 201000008100 Vaginitis Diseases 0.000 claims description 5
- 239000007788 liquid Substances 0.000 claims description 5
- 241000222122 Candida albicans Species 0.000 claims description 4
- 229940095731 candida albicans Drugs 0.000 claims description 4
- 206010007134 Candida infections Diseases 0.000 claims description 3
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- 230000002265 prevention Effects 0.000 claims description 3
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- 208000004926 Bacterial Vaginosis Diseases 0.000 claims description 2
- 208000037009 Vaginitis bacterial Diseases 0.000 claims description 2
- 201000007096 Vulvovaginal Candidiasis Diseases 0.000 claims description 2
- UELITFHSCLAHKR-UHFFFAOYSA-N acibenzolar-S-methyl Chemical compound CSC(=O)C1=CC=CC2=C1SN=N2 UELITFHSCLAHKR-UHFFFAOYSA-N 0.000 claims description 2
- 239000002131 composite material Substances 0.000 claims description 2
- 239000003814 drug Substances 0.000 claims description 2
- 235000013305 food Nutrition 0.000 claims description 2
- 239000000825 pharmaceutical preparation Substances 0.000 claims description 2
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- 206010003693 Atrophic vulvovaginitis Diseases 0.000 claims 1
- 241001478240 Coccus Species 0.000 claims 1
- 241000196324 Embryophyta Species 0.000 claims 1
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- 201000010808 postmenopausal atrophic vaginitis Diseases 0.000 claims 1
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- UQZIYBXSHAGNOE-XNSRJBNMSA-N stachyose Chemical compound O[C@H]1[C@H](O)[C@@H](CO)O[C@@]1(CO)O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO[C@@H]2[C@@H]([C@@H](O)[C@@H](O)[C@@H](CO[C@@H]3[C@@H]([C@@H](O)[C@@H](O)[C@@H](CO)O3)O)O2)O)O1 UQZIYBXSHAGNOE-XNSRJBNMSA-N 0.000 description 1
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Classifications
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/20—Bacteria; Culture media therefor
-
- 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
- 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
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P15/00—Drugs for genital or sexual disorders; Contraceptives
- A61P15/02—Drugs for genital or sexual disorders; Contraceptives for disorders of the vagina
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
- A61P31/04—Antibacterial agents
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
- A61P31/10—Antimycotics
-
- 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
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
- Y02A50/30—Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change
Abstract
The present invention relates to a kind of probiotics and its applications, and in particular to main active includes that the probiotics of lactobacillus plantarum and said preparation are preventing or treating the application in related disease.
Description
Technical field
The present invention relates to a kind of probiotics and its applications, and in particular to main active includes lactobacillus plantarum
Probiotics and said preparation prevent or treat related disease in application.
Background technique
Lactobacillus plantarum is vagina beneficial bacterium, maintains vagina health, but due to originals such as abuses of antibiotics
Cause causes vagina lactobacillus plantarum to reduce, and Escherichia coli, staphylococcus aureus or white occur for vaginal dysbacteriosis
Monilial infection causes various vaginal diseases to occur.Separating plant lactobacillus exploitation probiotics become the heat studied at present
Point, but the screening for the lactobacillus plantarum that growth activity is high, inhibition pathogenic bacteria effect is good is current difficult point.
The present inventor has found that lactobacillus plantarum DH230 has high bioactivity, main active packet after study
The probiotics of the DH230 containing lactobacillus plantarum can effectively adjust body microecological balance, prevention or treatment related disease,
Safety has huge economic value and social value without any toxic side effect, and has no that correlative study is reported, spy's application
The invention patent.
Summary of the invention
The object of the present invention is to provide a kind of probiotics, the main active of said preparation includes DH230, preservation
Number CGMCC No.16194, and said preparation is provided and is preventing or treating the application in related disease.
Result is analyzed according to bacterium cell form, physiological and biochemical property, 16S rRNA gene order etc. to confirm that bacterial strain exists
Taxonomy, the bacterial strain of discovery number DH230 are lactobacillus plantarum；Feature of this plant of bacterium on morphology and general aspects
It is as follows:
Cellular morphology and physical and chemical experiment result:
Lactobacillus plantarum DH230 of the present invention inhibits Escherichia coli, staphylococcus aureus or Candida albicans
Effect is significant, has the application for preventing or treating Escherichia coli, staphylococcus aureus or candida albicans infection, has pre-
Anti- or treatment vaginitis application, vaginitis includes bacterial vaginosis BV, mycotic, senile vahinitis, colpomycosis
It further comprise monilial vaginitis.
Probiotics of the invention, which also can be used as effect components and apply, to be used in compositions related prevent or control
Treat above-mentioned disease.
The present inventor is repeatedly screened and is cultivated, and isolated lactobacillus plantarum strain DH230, the bacterial strain is compared with it
He has higher activity, superior adjusting body microecological balance ability and prebiotic effect by bacterial strain.Plant of the present invention
Lactobacillus further includes the bacterial strain derived from lactobacillus plantarum DH230, especially has high activity, or with excellent adjusting machine
The derivative strain of body microecological balance ability and prebiotic effect.Main active includes lactobacillus plantarum DH230 and its derivative
The probiotics of bacterial strain each fall within protection scope of the present invention.
Probiotics of the present invention, main active include but is not limited to lactobacillus plantarum DH230, and preservation is compiled
Number CGMCC No.16194, can also be added the oligosaccharide such as other viable bacterias, stachyose, oligofructose or other active components.
Probiotics of the invention are using the lactobacillus plantarum as main active or main active
One of, according to certain preparation process, be added conventional excipient, flavoring agent, disintegrating agent, preservative, lubricant, wetting agent,
The auxiliary materials such as binder, solvent, thickener, solubilizer, the dosage form that manufactured any one is adapted for use with, as tablet, suppository,
The dosage forms such as capsule, granule, powder, liquid preparation, pulvis.
Probiotics of the present invention include but is not limited to composite medicine, health care product, food or sterile products, are disappeared
Malicious product further comprises inhibiting-bacteria preparation.
Lactobacillus plantarum described in this probiotics refers to bion living.
Lactobacillus plantarum described in this probiotics has high bioactivity.
This probiotics, the total viable count of the lactobacillus plantarum that solid pharmaceutical preparation includes are not less than 1 × 106CFU/g, generally
1 × 107CFU/g or more can reach 1 × 1012CFU/g or 1 × 1012CFU/g or more；Or the plant that liquid preparation includes
The total viable count of object lactobacillus is not less than 1 × 106CFU/mL, generally 1 × 107CFU/mL or more, it can reach 1 ×
1012CFU/mL or 1 × 1012CFU/mL or more.
Specific embodiment
Preparation example explanation: it is above-mentioned that the preparation of lactobacillus plantarum preparation is illustrated, pass through lactobacillus plantarum here
It is specifically described for DH230, preparation method those skilled in the art of other plant lactobacillus strain preparation pass through this reality
It applies example to be easy to grasp, preparation method those skilled in the art of other dosage forms are easy to grasp by this implementation, herein no longer
Explanation is described one by one.Preparation method is not limited to described in the embodiment of the present invention, the well known method that can reach preparation purpose
It can be with it is not limiting the scope of the invention that the preparation explanation of embodiment, which is the description of the invention,.
Prepare the preparation of 1 lactobacillus plantarum pulvis of embodiment
The preparation of 1 bacterium powder and the identification of strain
The samples such as vagina secretion are taken, then sample is placed in sterilizing bottle, 2 grams of samples is therefrom taken to be added
It in the dilution of 18mL sterilizing, mixes well, in aseptic operating platform, carries out 10-1、10-2、10-3、 10-4, 10-5, 10-6, 10-7
Gradient dilution takes 10-5, 10-6, 10-7Three dilution gradients, are coated on MRS solid medium, are placed in incubator, at 37 DEG C
Culture 48 hours, selects the single colonie to grow fine to be inoculated into MRS fluid nutrient medium, amplification cultivation 48 hours at 37 DEG C.It will
After gained medium centrifugal (12000rpm) isolates thallus, by thallus vacuum freezedrying, dry bacterium powder, viable bacteria are modulated
Number is 1 × 1010Then CFU/g or more carries out strain idenfication.It is identified through Physiology and biochemistry and 16S rRNA sequence comparative analysis
It is lactobacillus plantarum DH230, deposit number CGMCC No.16194 for lactobacillus plantarum.
2 toxicity tests
2.1 animals and grouping take 20 SPF rank mouse, 6-8 week old, and weight 14-18g is randomly divided into lactobacillus plantarum
Group and non-administered group, every group 10.
Above-mentioned lactobacillus plantarum bacterium powder is modulated to containing bacterium number be 1 × 10 by 2.2 preparation bacterium solutions with purified water9CFU/mL
Bacterium solution.
2.3 method lactobacillus plantarum groups and non-administered group give identical basal feed, and rearing conditions are consistent,
Lactobacillus plantarum group gavages lactobacillus plantarum bacterium solution 0.5mL daily, and non-administered group gavages purified water 0.5mL, feeding 14 daily
It, observes weight and toxic reaction.
2.4 result
Each group mouse does not occur abnormal conditions, and chatter, spasm, ataxia, posture exception do not occur, and no eyeball is prominent
Out, urination is normal, and skin, breathing are normal, and no death condition has no toxic reaction.
3 are prepared into tablets and other formulations
It is separated and identified according to above-mentioned steps and method, through experimental check nontoxicity, so that it may by lactobacillus plantarum strain system
At bacterium powder, then it is added as required relevant auxiliary materials and various dosage forms is made, preferably according to the viable count of lactobacillus plantarum bacterium powder,
Add microcrystalline cellulose, sodium carboxymethyl starch, magnesium stearate etc. in proportion and tablet be made, make viable count not less than 1 ×
107Then CFU/g bottles.
The present inventor uses above-mentioned separation and preparation method, while also separation obtains lactobacillus plantarum DH104 and plant
Lactobacillus DH124.
Application effect embodiment explanation:
For the present invention with lactobacillus plantarum DH230, probiotics made of deposit number CGMCC No.16194 are representative
Illustrate the application effect of lactobacillus plantarum preparation.It should be understood that these embodiments are merely to illustrate the present invention rather than limitation originally
The range of invention.To better illustrate application effect of the invention, the present inventor also uses lactobacillus plantarum DH104, plant cream
Probiotics made of bacillus DH124 and commercial plant lactobacillus compare explanation.Commercially available plant used in the present invention
Active plant lactobacillus product of the object lactobacillus from market sale.Combined with specific embodiments below, this hair is further described
It is bright.
Application effect embodiment 1: the bioactivity of lactobacillus plantarum
The preparation of 1 bacterium powder has prepared lactobacillus plantarum DH230, plant cream bar using the preparation method of above-mentioned bacterium powder
Bacterium DH104, lactobacillus plantarum DH124 and commercial plant lactobacillus bacterium powder.
It is 1 × 10 that above-mentioned bacterium powder is diluted to containing bacterium number by 2 fermented and cultureds6The bacterium solution of CFU/mL uses liquid amplification cultivation
Base, amplification cultivation 48 hours (3 parallel) at 37 DEG C.Detect the lactobacillus plantarum viable count in culture solution.
3 results are through 48h amplification cultivation, and viable count is significantly higher than lactobacillus plantarum in lactobacillus plantarum DH230 culture solution
Viable count (P < 0.05) in DH104, lactobacillus plantarum DH124 and commercial plant lactobacillus culture solution, is shown in Table 1.
1 lactobacillus plantarum viable count (Log of table10CFU·mL-1, n=3)
4 discuss above-mentioned the experimental results showed that lactobacillus plantarum DH230 is obviously compared with other bacterial strains and commercially available under equal conditions
Strain growth is vigorous.Probiotic active is high, breeding is its basis for playing prebiotic effect and guarantee fastly.The experimental results showed that planting
Object lactobacillus DH230 has higher bioactivity compared with other plant lactobacillus strain, has very high prebiotic value.
Application effect embodiment 2: suppression of the lactobacillus plantarum to Escherichia coli, staphylococcus aureus or Candida albicans
Production is used
1 materials and methods
Melt MH culture medium, when the culture medium wait melt is cooled to 50 DEG C or so, sensitive indicator bacterium is added in every 200mL
Liquid 0.5mL is poured into after mixing in 9cm culture dish, is placed in plate after solidification with tweezers clamping Oxford cup, in each Oxford cup
250 μ L lactobacillus plantarum culture solutions (1 × 10 of middle addition7CFU/mL), insulating box culture for 24 hours, measures antibacterial circle diameter (mm).
2 results
The antibacterial circle diameter of lactobacillus plantarum DH230 be noticeably greater than lactobacillus plantarum DH104, lactobacillus plantarum DH124 and
Viable count (P < 0.05), is shown in Table 2 in commercial plant lactobacillus culture solution.
2 lactobacillus plantarum antibacterial situation of table
3 discuss
This is the experimental results showed that lactobacillus plantarum DH230 inhibits Escherichia coli, staphylococcus aureus or Candida albicans
Effect be better than other plant lactobacillus, show that effect of the lactobacillus plantarum DH230 in terms of the treatment diseases such as vaginitis is also shown
It writes and is better than other plant lactobacillus.
Microorganism fungus kind lactobacillus plantarum DH230 of the present invention used in implementation process is in 08 month 2018 01
Day is in China Committee for Culture Collection of Microorganisms's common micro-organisms center's (Yard 1, BeiChen xi Road, Chaoyang District, Beijing City 3
Institute of Microorganism, Academia Sinica's postcode 100101) preservation.Classification naming: lactobacillus plantarum Lactobacillus
Plantarum, deposit number CGMCC No.16194.For above-mentioned strain through the microorganism Spot detection, testing result is survival.
Claims (10)
1. a kind of probiotics, it is characterised in that said preparation main active includes lactobacillus plantarum, the plant cream bar
Bacterium bag includes lactobacillus plantarum DH230, deposit number CGMCC No.16194.
2. probiotics according to claim 1, it is characterised in that lactobacillus plantarum refers to bion living.
3. probiotics according to claim 1, it is characterised in that the preparation includes composite medicine, health care product, food
Product or sterile products, sterile products further comprise inhibiting-bacteria preparation.
4. probiotics according to claim 1, it is characterised in that lactobacillus plantarum has high bioactivity.
5. probiotics described in claim 1 are in preparation prevention or treatment Escherichia coli, staphylococcus aureus or white
Application in monilial infection preparation.
6. being applied by described in claim 5, it is characterised in that the probiotics inhibit Escherichia coli, golden yellow Portugal in preparation
Application in grape coccus or Candida albicans bacteria preparation.
7. application of the probiotics described in claim 1 in preparation prevention or treatment vaginitis preparation.
8. being applied by described in claim 7, it is characterised in that the vaginitis includes bacterial vaginosis BV, mycotic, senile
Vaginitis, colpomycosis further comprise monilial vaginitis.
9. probiotics according to claim 1, it is characterised in that the lactobacillus plantarum includes lactobacillus plantarum DH230
And its derivative strain.
10. probiotics according to claim 1, it is characterised in that the total viable count of the lactobacillus plantarum that solid pharmaceutical preparation includes
Not less than 1 × 106CFU/g, generally 1 × 107CFU/g or more can reach 1 × 1012CFU/g or 1 × 1012CFU/g with
On；Or the total viable count of lactobacillus plantarum that liquid preparation includes is not less than 1 × 106CFU/mL, generally 1 × 107CFU/mL with
On, it can reach 1 × 1012CFU/mL or 1 × 1012CFU/mL or more.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
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| CN201811490004.7A CN109439592A (en) | 2018-12-06 | 2018-12-06 | Lactobacillus plantarum preparation and its application |
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201811490004.7A CN109439592A (en) | 2018-12-06 | 2018-12-06 | Lactobacillus plantarum preparation and its application |
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| Publication Number | Publication Date |
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| CN109439592A true CN109439592A (en) | 2019-03-08 |
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| CN201811490004.7A Pending CN109439592A (en) | 2018-12-06 | 2018-12-06 | Lactobacillus plantarum preparation and its application |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN112175867A (en) * | 2020-09-30 | 2021-01-05 | 山东环亿生物科技有限公司 | Functional non-woven fabric based on inactivated lactic acid bacteria |
| CN114869917A (en) * | 2022-07-07 | 2022-08-09 | 微康益生菌(苏州)股份有限公司 | Application of lactobacillus plantarum N13 in preparation of medicine for preventing and treating vaginitis or inhibiting pathogenic bacteria |
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| CN108295096A (en) * | 2017-12-21 | 2018-07-20 | 天津科睿恒生物科技有限公司 | Application of the bacterial strain in prevention and treatment mycotic, bacterial vaginitis preparation |
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN112175867A (en) * | 2020-09-30 | 2021-01-05 | 山东环亿生物科技有限公司 | Functional non-woven fabric based on inactivated lactic acid bacteria |
| CN114869917A (en) * | 2022-07-07 | 2022-08-09 | 微康益生菌(苏州)股份有限公司 | Application of lactobacillus plantarum N13 in preparation of medicine for preventing and treating vaginitis or inhibiting pathogenic bacteria |
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