CN108057118A - Multiple hepatitis b vaccine and preparation method thereof - Google Patents
Multiple hepatitis b vaccine and preparation method thereof Download PDFInfo
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- CN108057118A CN108057118A CN201810123532.2A CN201810123532A CN108057118A CN 108057118 A CN108057118 A CN 108057118A CN 201810123532 A CN201810123532 A CN 201810123532A CN 108057118 A CN108057118 A CN 108057118A
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K39/00—Medicinal preparations containing antigens or antibodies
- A61K39/12—Viral antigens
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K39/00—Medicinal preparations containing antigens or antibodies
- A61K39/395—Antibodies; Immunoglobulins; Immune serum, e.g. antilymphocytic serum
- A61K39/42—Antibodies; Immunoglobulins; Immune serum, e.g. antilymphocytic serum viral
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K39/00—Medicinal preparations containing antigens or antibodies
- A61K2039/555—Medicinal preparations containing antigens or antibodies characterised by a specific combination antigen/adjuvant
- A61K2039/55511—Organic adjuvants
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- 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
- C12N2730/00—Reverse transcribing DNA viruses
- C12N2730/00011—Details
- C12N2730/10011—Hepadnaviridae
- C12N2730/10111—Orthohepadnavirus, e.g. hepatitis B virus
- C12N2730/10134—Use of virus or viral component as vaccine, e.g. live-attenuated or inactivated virus, VLP, viral protein
Abstract
The present invention relates to the preparation methods of multiple hepatitis B vaccine, comprise the following steps：1) liposome is first prepared using investment, then prepares the plasmid liposome containing HBsAg, HBsAg plasmid lipidosome freeze-dried powders are then made；2) by the lyophilized plasmid liposome and adjuvant, oligopeptides of gained, it is placed in sodium alginate aqueous solution, is sufficiently mixed uniformly, it is slowly dropped into the chitosan solution containing calcium chloride, magnetic agitation, supernatant is abandoned in centrifugation, distill water washing for several times, it is dry to get chitosan microball；3) crosslinking agent glutaraldehyde is added in chitosan microball and Anti-HBsAg antibody, 40 DEG C of water bath with thermostatic control conditions is kept, is sufficiently stirred reaction, after reaction, stand to complete layering, abandon supernatant, lower floor's substance with isopropanol and petroleum ether, is dried to get crosslinked microsphere respectively；Multiple hepatitis B vaccine produced by the present invention has been obviously improved the immune response ability of vaccine, and stability is preferable.
Description
[technical field]
The present invention relates to hepatitis B vaccine preparing technical field, specifically a kind of preparation method of multiple hepatitis B vaccine.
[background technology]
Hepatitis B is the communicable disease as caused by hepatitis B (HBV), there is prevalence in countries in the world, and China belongs to high
Endemic Area.The chronicity of infection often leads to hepatic sclerosis even liver cancer, seriously endangers health.It there is no the B-mode liver of radical cure at present
Scorching specific medicament, HB vaccination are prevention and the important measures for controlling spreading.
Recombinant hepatitis b vaccine is widely used vaccine at this stage, and current China's hepatitis B vaccine is with recombination yeast type
Based on, utilize modern genetic engineering technology, recombinant plasmid of the structure containing hepatitis B virus surface antigen (HBsAg) gene, implantation
Yeast, yeast generate HBsAg in reproductive process, purified through Breaking Yeast thalline, are made after inactivation.The second of genetic recombination
Liver vaccine safety is preferably improved, but its immunogenicity and stability reduction just can induce effectively, it is necessary to add adjuvant
Immune response.Aluminium salt is the main adjuvant of current vaccine for man, but the cellullar immunologic response ability of its promotion vaccine is limited.Closely
The study found that microballoon can promote antigen to be absorbed by presenting cells and offer through MHCI approach, induction is higher levels of thin within several years
Born of the same parents and humoral immune response.
[content of the invention]
A kind of preparation method of multiple hepatitis B vaccine is provided present invention aim to solve above-mentioned deficiency, is made
Multiple hepatitis B vaccine be obviously improved the immune response ability of vaccine, stability is preferable, and preparation method is simple, is easy to control
System has good operability.
A kind of preparation method of multiple hepatitis B vaccine is designed to achieve the above object, is comprised the following steps：
1) preparation of HBsAg plasmid liposomes is freezed：Lecithin, cholesterol are pressed 3:1-5:4 molar ratios are placed in round bottom burning
In bottle, a certain amount of organic solvent dissolving is added in, then the rotary evaporation in 37 DEG C of water-baths, removes organic solvent, forms phosphatide
Film；Then a certain amount of 0.01-0.05M pH7.4PBS-HBsAg plasmid solutions are added in, the fat in PBS-HBsAg plasmid solutions
The volume ratio of plastid and HBsAg plasmids is 5:1-20:1, continue rotary evaporation to forming milky, uniform Liposomal suspensions,
Slight whirlpool mixing 1min after 45 DEG C of water-bath 6h, then with filtering with microporous membrane, obtain plasmid liposomal samples；In plasmid liposome sample
Antifreeze is added in product, after mixing immediately after -80 DEG C of quick pre-freezes, then is freeze-dried, -20 DEG C save backup, warp
55000r/min Superfreezings centrifuge 1.5h to get lyophilized plasmid liposome；
2) preparation of chitosan microball：By the lyophilized plasmid liposome obtained by step 1) and a certain amount of adjuvant, widow
Peptide is placed in the sodium alginate aqueous solution of 1-1.5%, is sufficiently mixed uniformly, is slowly dropped into the shell containing 5-10g/L calcium chloride
In glycan solution, supernatant is abandoned in more than magnetic agitation 30min, centrifugation, and distillation water washing is for several times, dry to get chitosan microball；
3) preparation of crosslinked microsphere：A certain amount of crosslinking agent is added in the chitosan microball and Anti-HBs antibody obtained by step 2)
Glutaraldehyde keeps 40 DEG C of water bath with thermostatic control conditions, is sufficiently stirred reaction, after reaction, stands to complete layering, abandons supernatant,
Lower floor's substance is respectively with isopropanol and petroleum ether 3 times or more, and 50 DEG C of drying are overnight to get crosslinked microsphere.
Further, in step 1), the organic solvent is chloroform or dichloromethane.
Further, in step 1), after immobilized artificial membrane is formed, add in PBS-HBsAg plasmid solutions before, further include with
Lower step：Lower immobilized artificial membrane is washed with the same amount of ether of organic solvent so that organic phase：Inorganic phase=3:1-5:1 volume ratio adds
Enter PBS-HBsAg plasmid solutions, slight whirlpool mixing, rotary evaporation to gel-forming simultaneously comes off.
Further, in step 1), plasmid liposomal samples form rounding obtained, grain size 50-300nm.
Further, in step 1), the antifreeze is mannitol, sucrose or trehalose.
Further, in step 1), plasmid liposomal samples obtained are divided into quarter, wherein three parts be separately added into it is sweet
Reveal alcohol, sucrose, trehalose, after mixing immediately after -80 DEG C of quick pre-freezes, then be freeze-dried, -20 DEG C save backup, separately
A 4 DEG C save backup, and then aforementioned four sample are taken to be centrifuged through Superfreezing respectively.
Further, in step 1), when lecithin, cholesterol are placed in round-bottomed flask, octadecylamine also has been inserted simultaneously,
The lecithin, cholesterol, the molar ratio of octadecylamine are 5:4:1.
Further, in step 2), the adjuvant is PLGA or PELA.
The present invention provides the multiple hepatitis B vaccine that a kind of above-mentioned preparation method obtains.
The present invention compared with the existing technology, by by hepatitis B virus surface antigen and Antibody preparation in identical carrier, system
It is relatively isolated, does not react, antigen is wrapped in liposome, and liposome is wrapped in micro- with adjuvant and oligopeptides again under agent state
In ball, and it is antibody linked in microsphere surface, wherein, adjuvant can control the rate of release of antigen, reach sustained release purpose, and nothing itself
Toxic side effect, after reaching liver cell so as to carrier, released antigen antibody, induction immune response, and microballoon/liposome and PLGA etc.
Adjuvant is could act as, has been obviously improved the immune response ability of vaccine；The dosage form be verified by experiments envelop rate height, stability compared with
It is good, there are potential advantages of the exploitation into clinical preparation.In addition, preparation method technological process of the present invention is simple, experiment condition is mild,
React easily controllable, cost of investment is low, has good operability, is worthy of popularization.
[specific embodiment]
The influences of the factors to immune response such as the material and preparation method of the invention for having investigated microballoon comprehensively, particle size,
And it is creative selected liposome hepatitis B virus surface antigen plasmid, using liposome and adjuvant by the use of microballoon as carrier,
By antibody linked in microsphere surface, antigen and antibody is made to obtain larger in the state that is relatively isolated, stability under formulated state
It improves；It after liver cell is reached, is released from carrier, induces immune response.Its preparation method is as follows：
1st, the recombinant plasmid liposome containing hepatitis B virus surface antigen (HBsAg) gene is prepared
1.1 prepare liposome using investment (reverse evaporation)：Take lecithin：Cholesterol=3：1~5：4, by the two
It is placed in round-bottomed flask, adds in a certain amount of organic solvent (chloroform or dichloromethane) dissolving, rotary evaporation in 37 DEG C of water-baths removes
Organic solvent forms immobilized artificial membrane；Lower immobilized artificial membrane is washed with ether, by organic phase：Inorganic phase=3：1~5：1 (volume ratio) adds in
PH7.4PBS solution (0.01~0.05M), slight whirlpool mixing, rotary evaporation to gel-forming simultaneously come off, add in a certain amount of
PBS liquid continues rotary evaporation to forming milky, uniform Liposomal suspensions, slight whirlpool mixing 1min after 45 DEG C of water-bath 6h,
Again with filtering with microporous membrane to get；After plasmid liposome obtained is dyed, by projecting electron microscopic observation form；Fat obtained
Plastide morphology rounding, grain size is between 50-300nm.
1.2 prepare the plasmid liposome containing HBsAg：In above-mentioned 1.1, plasmid is added in pH7.4PBS solution, remaining
Step is identical；Liposome：HBsAg plasmid=5：1~20：1.
1.3 prepare HBsAg plasmid lipidosome freeze-dried powders：Sample obtained in above-mentioned 1.2 is divided into quarter, wherein three parts
Antifreeze-mannitol, sucrose, trehalose are separately added into, after mixing immediately after -80 DEG C of quick pre-freezes, then freezing is carried out and does
Dry, -20 DEG C save backup；Another 4 DEG C save backup；Aforementioned four sample is taken to be centrifuged through Superfreezing respectively, takes supernatant
In right amount, agarose gel electrophoresis, gel imaging system observation tentatively judge encapsulating effect with its band light levels；As a result
Show the lyophilized encapsulating stability that can significantly increase plasmid.
2nd, the preparation of chitosan microball
By HBsAg plasmids lipidosome freeze-dried powder made from above-mentioned 1.3 and a certain amount of adjuvant (PLGA or PELA) and
Oligopeptides is placed in 1~1.5% sodium alginate aqueous solution, is sufficiently mixed uniformly, is slowly dropped into containing a certain amount of calcium chloride
In chitosan solution, supernatant is abandoned in magnetic agitation certain time, centrifugation；Distill water washing for several times, it is dry, to obtain the final product.
3rd, glutaraldehyde cross-linking chitosan microball and Anti-HBs antibody
A certain amount of crosslinking agent (glutaraldehyde) is added in chitosan microball and antibody, keeps 40 DEG C of water bath with thermostatic control conditions,
Reaction is sufficiently stirred, after reaction, stands to complete layering, abandons supernatant, lower floor's substance uses isopropanol and petroleum ether respectively
Washing 3 times or more, 50 DEG C of drying overnight, obtain crosslinked microsphere.
The present invention is made with reference to specific embodiment further explained below：
Embodiment 1
Lecithin, cholesterol, octadecylamine, by 5:4:1 (molar ratio) weighs a certain amount of each substance and is placed in 25mL eggplants shape circle
In the glass flask of bottom, a certain amount of chloroform dissolving is added in, then on the rotary evaporator, 37 DEG C of water-baths, 190 revs/min, decompression rotation
Turn evaporating organic solvent chloroform, lipid film forming.Lower film is washed with the same amount of ether of chloroform, by organic phase：Inorganic phase=
3：1 adds in 0.01M pH7.4PBS- plasmid solution (liposomes：HBsAg plasmid=10：1), slight whirlpool mixing 5min or so,
Rotary evaporation is to gel-forming and comes off, and adds in a certain amount of PBS liquid and continues rotary evaporation to forming milky, uniform lipid
Body suspension, slight whirlpool mixing 1min after 45 DEG C of water-bath 6h, then with filtering with microporous membrane, 4 DEG C save backup.It is added in sample sweet
Reveal alcohol (mannitol：Liposome=3:1), after mixing immediately after -80 DEG C of quick pre-freezes, then be freeze-dried, -20 DEG C of preservations
It is spare；Another 4 DEG C save backup.Above-mentioned sample is taken to centrifuge 1.5h through 55000r/min Superfreezings to get lyophilized plasmid fat
Plastid.
Freeze-dried powder, PLGA and oligopeptides are placed in 1.5% sodium alginate aqueous solution, are sufficiently mixed uniformly, are slowly dropped into
Into 5% chitosan solution containing 5g/L calcium chloride, supernatant is abandoned in magnetic agitation 30min, 4000r/0min centrifugation.Distilled water
Washing for several times, 50 DEG C of oven drying 6h to get.
2% glutaraldehyde is added in chitosan microball and antibody, 40 DEG C of water bath with thermostatic control conditions is kept, is sufficiently stirred reaction
3h after reaction, is stood to complete layering, abandons supernatant, lower floor's substance respectively with isopropanol and petroleum ether 3 times with
On.50 DEG C of drying overnight, obtain crosslinked microsphere.
Embodiment 2
Lecithin, cholesterol, by 3:1 (molar ratio) weighs a certain amount of each substance and is placed in 25mL eggplant shape glass round bottom flasks
In, a certain amount of chloroform dissolving is added in, then on the rotary evaporator, 37 DEG C of water-baths, 200 revs/min, decompression rotary evaporation removes
Organic solvents, chloroform, lipid film forming.Add in a certain amount of 0.01M pH7.4PBS- plasmid solution (liposomes：HBsAg plasmid=5：
1), continue rotation and wash film, under small part is not washed, slightly shake to film and completely fall off on whirlpool oscillator, rotary evaporation is extremely
Milky, uniform Liposomal suspensions are formed, slight whirlpool mixing 1min after 45 DEG C of water-bath 6h, then with filtering with microporous membrane, 4 DEG C
It saves backup.Trehalose (trehalose is added in sample：Liposome=4:1), after mixing immediately after -80 DEG C of quick pre-freezes, then
It is freeze-dried, -20 DEG C save backup；Another 4 DEG C save backup.Take above-mentioned sample through 55000r/min Superfreezings from
Heart 1.5h is to get lyophilized plasmid liposome.
By HBsAg plasmids lipidosome freeze-dried powder obtained, PELA and oligopeptides, be placed in 1% sodium alginate aqueous solution
In, it is sufficiently mixed uniformly, is slowly dropped into 5% chitosan solution containing 10g/L calcium chloride, more than magnetic agitation 30min,
Supernatant is abandoned in centrifugation.Distill water washing for several times, it is dry to get.
5% crosslinking agent (glutaraldehyde) is added in chitosan microball and Anti-HBs antibody, 40 DEG C of water bath with thermostatic control conditions is kept, fills
Divide and be stirred to react, after reaction, stand to complete layering, abandon supernatant, lower floor's substance is washed respectively with isopropanol and petroleum ether
It washs 3 times or more.50 DEG C of drying overnight, obtain crosslinked microsphere.
The present invention is simultaneously not limited to the embodiments described above, other any Spirit Essences and principle without departing from the present invention
Lower made change, modification, replacement, combination, simplification, should be equivalent substitute mode, be included in the protection model of the present invention
Within enclosing.
Claims (9)
1. a kind of preparation method of multiple hepatitis B vaccine, which is characterized in that comprise the following steps：
1) preparation of HBsAg plasmid liposomes is freezed
Lecithin, cholesterol are pressed 3:1-5:4 molar ratios, are placed in round-bottomed flask, add in a certain amount of organic solvent dissolving, so
The rotary evaporation in 37 DEG C of water-baths afterwards removes organic solvent, forms immobilized artificial membrane；Then a certain amount of 0.01-0.05M is added in
PH7.4PBS-HBsAg plasmid solutions, the volume ratio of liposome and HBsAg plasmids in PBS-HBsAg plasmid solutions is 5:1-
20:1, continue rotary evaporation to forming milky, uniform Liposomal suspensions, slight whirlpool mixing 1min after 45 DEG C of water-bath 6h,
Again with filtering with microporous membrane, plasmid liposomal samples are obtained；Antifreeze is added in plasmid liposomal samples, after mixing immediately-
After 80 DEG C of quick pre-freezes, then be freeze-dried, -20 DEG C save backup, through 55000r/min Superfreezings centrifugation 1.5h to get
Lyophilized plasmid liposome；
2) preparation of chitosan microball
By the lyophilized plasmid liposome and a certain amount of adjuvant, oligopeptides obtained by step 1), the sodium alginate of 1-1.5% is placed in
In aqueous solution, it is sufficiently mixed uniformly, is slowly dropped into the chitosan solution containing 5-10g/L calcium chloride, magnetic agitation 30min
More than, supernatant is abandoned in centrifugation, and distillation water washing is for several times, dry to get chitosan microball；
3) preparation of crosslinked microsphere
A certain amount of crosslinking agent glutaraldehyde is added in the chitosan microball and Anti-HBs antibody obtained by step 2), keeps 40 DEG C of thermostatted waters
Bath condition is sufficiently stirred reaction, after reaction, stands to complete layering, abandons supernatant, lower floor's substance respectively with isopropanol and
Petroleum ether 3 times or more, 50 DEG C of drying are overnight to get crosslinked microsphere.
2. the preparation method of multiple hepatitis B vaccine as described in claim 1, it is characterised in that：It is described organic molten in step 1)
Agent is chloroform or dichloromethane.
3. the preparation method of multiple hepatitis B vaccine as described in claim 1, it is characterised in that：In step 1), phosphatide is being formed
After film, add in before PBS-HBsAg plasmid solutions, it is further comprising the steps of：It is washed down with the same amount of ether of organic solvent
Immobilized artificial membrane so that organic phase：Inorganic phase=3:1-5:1 volume ratio adds in PBS-HBsAg plasmid solutions, slight whirlpool mixing, rotation
Turn to be evaporated to gel-forming and come off.
4. the preparation method of multiple hepatitis B vaccine as described in claim 1, it is characterised in that：In step 1), plasmid obtained
Liposomal samples form rounding, grain size 50-300nm.
5. the preparation method of multiple hepatitis B vaccine as described in claim 1, it is characterised in that：In step 1), the antifreeze
For mannitol, sucrose or trehalose.
6. the preparation method of multiple hepatitis B vaccine as described in claim 1, it is characterised in that：In step 1), by matter obtained
Grain liposomal samples are divided into quarter, wherein three parts are separately added into mannitol, sucrose, trehalose, it is fast at -80 DEG C immediately after mixing
It after fast pre-freeze, then is freeze-dried, -20 DEG C save backup, another 4 DEG C save backup, and then take aforementioned four sample respectively
Product are centrifuged through Superfreezing.
7. the preparation method of multiple hepatitis B vaccine as described in claim 1, it is characterised in that：In step 1), by lecithin, courage
When sterol is placed in round-bottomed flask, octadecylamine has also been inserted simultaneously, the lecithin, cholesterol, the molar ratio of octadecylamine are 5:
4:1。
8. the preparation method of multiple hepatitis B vaccine as described in claim 1, it is characterised in that：In step 2), the adjuvant is
PLGA or PELA.
9. a kind of multiple hepatitis B vaccine that preparation method according to any one of claim 1 to 8 obtains.
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| Application Number | Priority Date | Filing Date | Title |
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| CN201810123532.2A CN108057118A (en) | 2018-02-07 | 2018-02-07 | Multiple hepatitis b vaccine and preparation method thereof |
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| CN201810123532.2A CN108057118A (en) | 2018-02-07 | 2018-02-07 | Multiple hepatitis b vaccine and preparation method thereof |
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Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101062015A (en) * | 2007-05-22 | 2007-10-31 | 中国药科大学 | Urease epitope fusion peptide liposome bacterin for preventing the helicobacter pylori infecting |
| CN102008719A (en) * | 2010-12-02 | 2011-04-13 | 上海微球生物科技有限公司 | Immunomicrosphere for overcoming B cell immunological tolerance and application thereof |
| CN102578110A (en) * | 2011-12-23 | 2012-07-18 | 河海大学 | Preparation method of artemisinin slow-release body |
| CN102796721A (en) * | 2012-05-17 | 2012-11-28 | 东北农业大学 | Method for immobilizing phospholipase A2 by using sodium alginate-chitosan |
| CN104983591A (en) * | 2015-07-09 | 2015-10-21 | 西安艾尔菲生物科技有限公司 | Double modified beta-carotene lipidosome and preparation method thereof |
-
2018
- 2018-02-07 CN CN201810123532.2A patent/CN108057118A/en active Pending
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101062015A (en) * | 2007-05-22 | 2007-10-31 | 中国药科大学 | Urease epitope fusion peptide liposome bacterin for preventing the helicobacter pylori infecting |
| CN102008719A (en) * | 2010-12-02 | 2011-04-13 | 上海微球生物科技有限公司 | Immunomicrosphere for overcoming B cell immunological tolerance and application thereof |
| CN102578110A (en) * | 2011-12-23 | 2012-07-18 | 河海大学 | Preparation method of artemisinin slow-release body |
| CN102796721A (en) * | 2012-05-17 | 2012-11-28 | 东北农业大学 | Method for immobilizing phospholipase A2 by using sodium alginate-chitosan |
| CN104983591A (en) * | 2015-07-09 | 2015-10-21 | 西安艾尔菲生物科技有限公司 | Double modified beta-carotene lipidosome and preparation method thereof |
Non-Patent Citations (4)
| Title |
|---|
| JAGANATHAN KS等: "Strong systemic and mucosal immune responses to surface-modified PLGA microspheres containing recombinant hepatitis B antigen administered intranasally", 《VACCINE》 * |
| 张亚停: "《张亚停 儿科医生说》", 30 June 2017, 长春：吉林科学技术出版社 * |
| 樊建勇等: "pVAX-Hsp70-HSV2gD DNA疫苗阳离子脂质体的制备", 《中国皮肤性病学杂志》 * |
| 汪多仁: "《现代高分子材料生产及应用手册》", 31 May 2002, 北京：中国石化出版社 * |
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