CN106334001A - Application of cistanche phenylethanoid glycosides effective part in preparation of bone formation accelerating drug and drug composition - Google Patents
Application of cistanche phenylethanoid glycosides effective part in preparation of bone formation accelerating drug and drug composition Download PDFInfo
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Abstract
The invention discloses an application of a cistanche phenylethanoid glycosides effective part in preparation of a bone formation accelerating drug, wherein the cistanche phenylethanoid glycosides effective part comprises echinacoside and verbascoside, and the sum total of the content of phenethyl alcohol glycoside ingredients in the cistanche phenylethanoid glycosides effective part is more than 50%. The invention further provides a drug composition comprising the cistanche phenylethanoid glycosides effective part to promote bone formation.
Description
Technical field
The present invention relates to a kind of new application of the total benzyl carbinol glycosides active component from saline cistanche medicinal plant, especially relate to
And a kind of cistanche benzyl carbinol glycosides active component is as the application preparing promoting bone growing medicine and pharmaceutical composition.
Background technology
Osteoporosis is the elderly's common disease and frequently-occurring disease, is all that a kind of stealth killer threatens in China or even the whole world
The health of people.The intensity of bone relies on bone information and osteoplastic dynamic equilibrium to realize, and bone information increases or bon e formation reduces
All this dysequilibrium may be led to, thus leading to osteoporosis.Osteoporosis belongs to generalized bone metabolic disease, at present
There is no specific drug.On market, existing anti-osteoporotic is broadly divided into three major types: bone resorption inhibitor, bone mineralizer and bone
Form accelerator.Although synthetic drug and hormonal medicaments in terms for the treatment of osteoporosis achieved with very big breakthrough, due to it
The presence of side effect and expensive and be unable to long term usage, finds treatment bone loss disorders and Small side effects from natural plants
Effective extract, active component and monomeric compound have become study hotspot.
Saline cistanche is China's tradition kidney tonifying class Chinese medicine, and version " Chinese Pharmacopoeia " record in 2015 is Orobanchaceae Cistanche deserticola plant
Saline cistanchecistanche deserticolaY. c.ma and Cistanche tubulosacistanche tubulosa(schenk)
The fleshy stem of wight.Saline cistanche has high medical value, and it enters kidney, large intestine channel, has kidney-replenishing, benefiting essence-blood, ease constipation is led to
Just the effect of；Can be used for the insufficiency of the kidney yang, the disease such as essence and blood side is empty, impotence is infertile, soreness and weakness of waist and knees, muscles and bones are powerless.The chemistry of saline cistanche
Composition Study shows, it mainly contains wheat-based diet, benzyl carbinol glycoside compound, iridoid and its glycosides compound, wood
Lipid and its glycosides compound etc., at present both at home and abroad just for saline cistanche crude extract and echinacoside monomeric compound therein
Carry out inside and outside anti-osteoporosis research, had no and have been reported that the total benzyl carbinol glycosides active component in saline cistanche is promoted
Osteoplastic experimental study.In addition, according to the Overall View of traditional Chinese medical theory and synergy viewpoint, any one composition in Chinese medicine
Drug action can not replace the element of the first species or the drug action of whole Chinese medicine, therefore, saline cistanche crude extract and pine nut therein
A kind of effect of chrysanthemum glycosides composition anti-osteoporosis drug effect can not replace the total benzyl carbinol glycosides active component anti-osteoporosis in saline cistanche
Drug action.Meanwhile, the drug action of anti-osteoporosis can be divided into suppression bone information and promoting bone growing two big class, and it acts on machine
System is entirely different.Based on the result of study of document above investigation and our early stages, propose the present invention a kind of from saline cistanche
Total benzyl carbinol glycosides active component is as the application preparing promoting bone growing medicine and pharmaceutical composition.
Content of the invention
The invention provides a kind of new opplication of cistanche benzyl carbinol glycosides active component, that is, saline cistanche benzyl carbinol glycosides are effective
Position is as the application preparing promoting bone growing medicine.
Present invention also offers a kind of drug regimen of the promoting bone growing containing cistanche benzyl carbinol glycosides active component
Thing.
A kind of cistanche benzyl carbinol glycosides active component is as the application preparing promoting bone growing medicine, wherein cistanche
Benzyl carbinol glycosides active component contains echinacoside, acteoside, and contained benzene in cistanche benzyl carbinol glycosides active component
The summation of ethanol methods of glycosides content is more than 50%.
A kind of pharmaceutical composition of the promoting bone growing containing cistanche benzyl carbinol glycosides active component, including active component
Major ingredient and auxiliary material, the weight of described active component major ingredient and described auxiliary material is than for 20:80～80:20, described active component major ingredient
For cistanche benzyl carbinol glycosides active component, described auxiliary material is drug excipient, wherein utilizes cistanche benzyl carbinol glycosides effective
Promoting bone growing is carried out at position, and wherein cistanche benzyl carbinol glycosides active component contains echinacoside, acteoside, and meat desert
In Rong's total benzyl carbinol glycosides active component, the summation of contained ingredient content of phenylethanoid glycosides is more than 50%.
From saline cistanche cistanche benzyl carbinol glycosides active component as the application preparing promoting bone growing medicine and
The pharmaceutical composition of the promoting bone growing containing cistanche benzyl carbinol glycosides active component, is inventor in saline cistanche pharmacological action
During composition Study, separation and concentration total benzyl carbinol glycosides active component of q.s from saline cistanche, cause big through removal ovary
Mouse osteoporosis animal experiment is it was confirmed cistanche benzyl carbinol glycosides active component has the effect of promoting bone growing, and prepares
Go out the pharmaceutical composition of the promoting bone growing containing cistanche benzyl carbinol glycosides active component.
Brief description
Fig. 1 is rat total bone density (t-bmd) figure of each administration group.
Fig. 2 is femur micro- ct scanning 3d figure on the right side of the rat of each administration group.
Fig. 3 is that each group rat blood serum alp schemes.
Fig. 4 is that each group rat blood serum oc schemes.
Specific embodiment
Inventor's separation and concentration total benzyl carbinol glycosides active component of q.s from saline cistanche, causes rat bone through removal ovary
The loose animal experiment of matter is it was demonstrated that cistanche benzyl carbinol glycosides active component has the effect of promoting bone growing, wherein, cistanche
Benzyl carbinol glycosides active component contains echinacoside, acteoside, and contained benzene in cistanche benzyl carbinol glycosides active component
The summation of ethanol methods of glycosides content is more than 50%.
Further, above-mentioned cistanche benzyl carbinol glycosides active component can make the pharmaceutical composition of promoting bone growing,
Particularly as follows:
A kind of pharmaceutical composition of the promoting bone growing containing cistanche benzyl carbinol glycosides active component, this pharmaceutical composition includes
Active component major ingredient and auxiliary material, the weight of described active component major ingredient and described auxiliary material is than for 20:80～80:20, described activity
Composition major ingredient is cistanche benzyl carbinol glycosides active component, and described auxiliary material is drug excipient, wherein utilizes cistanche benzene second
Alcohol glycosides effective part carrys out promoting bone growing, and wherein cistanche benzyl carbinol glycosides active component contains echinacoside, acteoside,
And the summation of contained ingredient content of phenylethanoid glycosides is more than 50% in cistanche benzyl carbinol glycosides active component.Other auxiliary materials are
Any acceptable excipient on medicament, such as starch, lactose, microcrystalline cellulose, dextrin, hydroxypropylcellulose, crosslinked poly- dimension
Ketone, pregelatinized starch, talcum powder, magnesium stearate, superfine silica gel powder, lauryl sodium sulfate, Tween 80, hydrogenated vegetable oil etc..
Inventor passes through the total benzyl carbinol glycosides active component preparing from saline cistanche is carried out with removal ovary cause rat bone
The loose animal experiment of matter screens, and confirms the effect that this active component has promoting bone growing, concrete experiment sieving process is as follows:
Total benzyl carbinol glycosides active component is prepared from saline cistanche:
Mode one, prepares total benzyl carbinol glycosides active component: take the meat of Yongning Ningxia saline cistanche planting base cultivation from saline cistanche
Desert cistanche (cistanche deserticolaY. c.ma), take 30 kg after drying at room temperature, with the leaching of 80% ethanol 100 l room temperature
Steep 48 h, refluxing extraction 3 times, 3 h every time, merge extract, drying under reduced pressure obtains concentrate.Concentrate divides through macroporous absorbent resin
From, successively use water, 20% ethanol, 30% ethanol, 40% ethanol, 50% ethanol elution, 60% ethanol elution, discard water, 20% ethanol,
30% ethanol and 60% alcohol elution, collect 40% and 50% ethanol eluate merge concentrate after secondary loading, more successively use water,
20% ethanol, 30% ethanol, 40% ethanol, 50% ethanol elution, discard water, 20% ethanol eluate, collect 40% ethanol eluate and obtain
Cistanche benzyl carbinol glycosides active component, measures through hplc and wherein contains echinacoside, acteoside, 2- acetyl hair stamen simultaneously
Flower glucosides, 6- acetyl acteoside and five kinds of phenylethanoid glycosides of boschnaloside a, and above five kinds of component content summations
For cistanche benzyl carbinol glycosides active component gross mass 55.32%.
Mode two, prepares cistanche benzyl carbinol glycosides active component: take the meat of Yongning Ningxia saline cistanche planting base cultivation
Desert cistanche (cistanche deserticolaY. c.ma), take 10 kg after drying at room temperature, with 60% ethanol 60 l soaking at room temperature
24 h, refluxing extraction 3 times, 2 h every time, merge extract, be evaporated to liquid extract, then separate through macroporous absorbent resin: stream
After medicinal extract loading, use water, 20% ethanol, 40% ethanol, 50% ethanol, 80% ethanol elution successively, discard water, 20% ethanol eluate,
Collect 40% and 50% ethanol eluate, secondary loading afterwards and after concentrating, then use successively water, 20% ethanol, 40% ethanol, 60% ethanol,
80% ethanol elution, collects 60% ethanol eluate and obtains cistanche benzyl carbinol glycosides active component, measure through hplc and wherein contain simultaneously
There are echinacoside, acteoside, 2- acetyl acteoside and tetra- kinds of phenylethanoid glycosides of boschnaloside a, and four contain
Amount summation is the 51.14% of cistanche benzyl carbinol glycosides active component gross mass.
Mode three, prepares cistanche benzyl carbinol glycosides active component: take Cistanche tubulosa (cistanche tubulosa
(schenk) wight) dry product 10 kg, with 70% ethanol, 60 l soaking at room temperature 24 h, refluxing extraction 3 times, 2 h every time, merge
Extract, is evaporated to liquid extract, then through macroporous absorbent resin separate: after liquid extract loading, successively use water, 20% ethanol,
40% ethanol, 50% ethanol, 80% ethanol elution, discard water, 20% ethanol eluate, collect 40% and 50% ethanol eluate, afterwards simultaneously
Secondary loading after concentration, then use water, 20% ethanol, 40% ethanol, 60% ethanol, 80% ethanol elution successively, collect 40% ethanol elution
Liquid obtains cistanche benzyl carbinol glycosides active component, measures through hplc and wherein contains echinacoside and two kinds of benzene of acteoside simultaneously
Ethanol methods of glycosides, and the two content summation is the 52.28% of cistanche benzyl carbinol glycosides active component gross mass.
Mode four, prepares cistanche benzyl carbinol glycosides active component: take Cistanche tubulosa (cistanche tubulosa
(schenk) wight) dry product 10 kg, with 70% ethanol, 60 l soaking at room temperature 24 h, refluxing extraction 3 times, 2 h every time, merge
Extract, is evaporated to liquid extract, then through macroporous absorbent resin separate: after liquid extract loading, successively use water, 20% ethanol,
50% ethanol, 80% ethanol elution, discard water, 20% ethanol eluate, collect 50% ethanol eluate, afterwards and secondary after concentrating on
Sample, then use water, 20% ethanol, 40% ethanol, 60% ethanol, 80% ethanol elution successively, collect 40% ethanol eluate and obtain cistanche
Benzyl carbinol glycosides active component, measures through hplc and wherein contains echinacoside, acteoside and three kinds of 2- acetyl acteoside
Phenylethanoid glycoside, and three's content summation is the 56.74% of cistanche benzyl carbinol glycosides active component gross mass.
Mode five, prepares cistanche benzyl carbinol glycosides active component: take Cistanche tubulosa (cistanche tubulosa
(schenk) wight) dry product 8 kg, with 75% ethanol, 48 l soaking at room temperature 24 h, refluxing extraction 3 times, 2 h every time, merge
Extract, is evaporated to liquid extract, then through macroporous absorbent resin separate: after liquid extract loading, successively use water, 40% ethanol,
60% ethanol, 95% ethanol elution, discard water elution, collect 60% ethanol eluate, secondary loading afterwards and after concentrating, more successively
With water, 20% ethanol, 40% ethanol, 80% ethanol elution, collect 40% ethanol eluate and obtain cistanche benzyl carbinol glycosides active component,
Measure through hplc and wherein contain echinacoside, acteoside and tri- kinds of phenylethanoid glycosides of boschnaloside a, and three's content
Summation is the 61.32% of cistanche benzyl carbinol glycosides active component gross mass.
Cistanche benzyl carbinol glycosides active component causes the drug action of osteoporosis rat to removal ovary, and experimentation is such as
Under:
(1) animal, reagent and instrument
1st, animal: 3 month female sd rats 64, cleaning grade, body weight 250 ± 20 g, purchased from Ningxia Medical University's zoopery
Center.
2nd, medicine and reagent: positive control drug: Estradiol Valerate (Progynova), merck sharp ＆ dohme italia
Spa produces.Test medicine: cistanche benzyl carbinol glycosides active component 1,2,3,4 and 5 is prepared by embodiment 1,2,3,4 and 5 respectively
Obtain.BGP (oc) kit (abundant bio tech ltd of upper Hisense).
3rd, instrument: the micro- ct of explore locus sp type (ge company of the U.S.；Analysis software is micview v2.1.2
Three-dimensional reconstruction processes software；The special bone analysis software of aba)；Dual energy x-ray absorptiometry (lunar company dpx type)；
Assay balance (al104, mettler toledo, plum Teller-support benefit instrument Shanghai Co., Ltd)；Desk type high speed refrigerated centrifuge
Machine (1-15k type, Town in Shanghai booth instrument plant)；Ultra low temperature freezer (forma -86c ult freezer, bio-rad company).
(2) animal experiment method
1st, experiment packet and gastric infusion dosage: female 3 monthly age sd rats are randomly divided into 8 groups by body weight, every group 8, packet
As follows with dosage:
Sham-operation group (sham is orally administered to equal-volume 0.5% cmc-na)；
Model control group (ovx is orally administered to equal-volume 0.5% cmc-na)；
Positive drug group (ev, is orally administered to Estradiol Valerate, 1 mg/kg/d)；
1 group of cistanche benzyl carbinol glycosides active component (cdp1, oral dose 120 mg/kg/d respectively)；
2 groups of cistanche benzyl carbinol glycosides active component (cdp2, oral dose 120 mg/kg/d respectively)；
3 groups of cistanche benzyl carbinol glycosides active component (cdp3, oral dose 120 mg/kg/d respectively).
4 groups of cistanche benzyl carbinol glycosides active component (cdp4, oral dose 120 mg/kg/d respectively).
5 groups of cistanche benzyl carbinol glycosides active component (cdp5, oral dose 120 mg/kg/d respectively).
Above positive control drug and test medicine are joined as solvent with 0.5% CMC (0.5%cmc-na) respectively
Become the concentration needing.
2nd, zoopery
By above 64 female rats temperature be 24 ± 0.5 ° of c, humidity is 45-50%, adapts to one week in the environment of well-ventilated
Afterwards, carry out removal ovary experimental study.According to above animal packet, take rat, lumbar injection 10% chloraldurate (0.3 ml/100
G) anaesthetized.After confirming animal holonarcosis, aseptically, cut off about 1-2 cm otch in bilateral back rapidly: false
Operation group rat only take out back bilateral fat a little after sew up the incision, other each group rats are all taken out bilateral ovaries and tighten defeated ovum
Excise ovary after pipe, sew up the incision.Each group rat prevents infected wound in the postoperative penicillin of lumbar injection for three days on end.
Each group animal starts to be administered after performing the operation 3 days.Every group of rat presses body weight 10 ml/kg gastric infusion, sham-operation group
Sham group and model ovx group give same volume 0.5% cmc-na, and other each administration groups press the daily gavage of each administration group dosage above
It is administered once, 6 times a week.Successive administration 12 weeks.Claim 1 body weight every two weeks, adjust dosage.
After last 1 administration, every rat is respectively placed in metabolic cage, water is can't help in fasting, collects urine, and in anesthesia
Right common femoral artery takes blood to collect serum afterwards, and on the right side of rapid stripping hind leg, femur and shin bone masking foil are wrapped；Each sample standard deviation above
It is placed in 80 DEG C of Refrigerator stores standby.
3rd, bone index determining
Measure total bone density (t-bmd) of femur on the right side of every rat using dual energy x-ray borne densitometers.
4th, femur bone tissue Senile Mouse
Take right side femur, reject muscle and soft tissue, vertically will be cut along at femur about 1 cm between near end of thighbone tuberosity with cutting machine
Cut, put in micro-ct and be scanned.After the completion of scanning, software is processed using micview v2.1.2 three-dimensional reconstruction and carries out often
The reconstruction of individual sample 3-D view；The special bone analysis software of aba carries out Data Analysis Services.
5th, bon e formation index determining
Take each group rat blood serum, bon e formation index alkaline phosphatase (alp) activity is measured with literature method, according to kit detection
BGP (oc) activity.
6th, data analysis
Experimental data is all usedRepresent, statistical analysis carries out one-way analysis of variance with spss 16.0 software, with p <
0.05 is to have significant difference.
7th, interpretation of result
Each group rat femur total bone density t-bmd testing result is shown in accompanying drawing 1, compared with model group,* p< 0.05,** p<
0.01,*** p< 0.001；Compared with sham-operation group,### p< 0.001.After rat is administered 12 weeks, model ovx group rat bone is close
Compared with sham-operation group sham, bone density significantly reduces (p < 0.001) to degree.Compared with model group rats, cistanche benzene second
It is close that 1,2,3,4 and 5 groups of alcohol glycosides equal (cdp1, cdp2, cdp3, cdp4 and cdp5) can significantly improve the rat bone that removal ovary causes
Degree reduces (p < 0.01), and effect is suitable with sham-operation group sham.Therefore, bone density result shows, ovariectomized rats cause sclerotin to dredge
Loose modeling success, and cistanche benzyl carbinol glycosides can significantly improve the bone density of ovariectomized female rats.
As shown in accompanying drawing 2, wherein, a is sham-operation group to the micro- ct scanning figure of the bone tissue form of each group rat femur；b
For model group；C is Estradiol Valerate positive drug group；D is cistanche benzyl carbinol glycosides 1 dosage group；E is cistanche benzyl carbinol glycosides
2 dosage groups；F is cistanche benzyl carbinol glycosides 3 dosage group；G is cistanche benzyl carbinol glycosides 4 dosage group；H is cistanche benzene second
Alcohol glycosides 5 dosage group.Micro- ct scanning figure is to be represented with bone trabecula selection area (roi) the 3d figure of each group rat femur.By attached
Fig. 2 result shows, sham-operation group sham rat roi region is comparatively dense, and bone trabecula number is more, model group ovx group roi region
Very loose, bone trabecula number substantially reduces, and gap significantly increases, and cavity；Positive control drug group (ev), cistanche
The bone tissue form of 1,2,3,4 and 5 groups of (cdp1, cdp2, cdp3, cdp4 and cdp5) rats of benzyl carbinol glycosides is bright compared with model group ovx
Aobvious be greatly improved, the obvious packing volume in roi region increases, bone trabecula number showed increased and being completely embedded.
Each group rat blood serum alp and oc Activity determination result are as shown in accompanying drawing 3 and Fig. 4.Cistanche benzyl carbinol glycosides 1,2,
3rd, 4 and 5 groups (cdp1, cdp2, cdp3, cdp4 and cdp5) can significantly improve bon e formation index alp and the activity of oc, thus promoting
Enter bon e formation.
8th, conclusion
Cause osteoporosis model animal test results can draw cistanche benzyl carbinol glycosides active component by removal ovary
Can promoting bone growing, thus reaching the purpose of anti-curing osteoporosis.
Claims (10)
1. a kind of cistanche benzyl carbinol glycosides active component is as the application preparing promoting bone growing medicine, wherein cistanche benzene
Ethanol glycosides effective part contains echinacoside, acteoside, and contained benzene second in cistanche benzyl carbinol glycosides active component
The summation of alcohol glycoside component content is more than 50%.
2. cistanche benzyl carbinol glycosides active component according to claim 1 as prepare promoting bone growing medicine should
With it is characterised in that: cistanche benzyl carbinol glycosides active component also contains 2- acetyl acteoside, and cistanche benzene second
In alcohol glycosides effective part, the summation of contained ingredient content of phenylethanoid glycosides is more than 50%.
3. cistanche benzyl carbinol glycosides active component according to claim 1 as prepare promoting bone growing medicine should
With it is characterised in that: cistanche benzyl carbinol glycosides active component also contains boschnaloside a, and cistanche benzyl carbinol glycosides have
In effect position, the summation of contained ingredient content of phenylethanoid glycosides is more than 50%.
4. cistanche benzyl carbinol glycosides active component according to claim 1 as prepare promoting bone growing medicine should
With it is characterised in that: cistanche benzyl carbinol glycosides active component also contains 2- acetyl acteoside, boschnaloside a and 6- second
In acyl acteoside, and cistanche benzyl carbinol glycosides active component, the summation of contained ingredient content of phenylethanoid glycosides is more than
50%.
5. a kind of pharmaceutical composition of the promoting bone growing containing cistanche benzyl carbinol glycosides active component, including active component master
Material and auxiliary material, for 20:80～80:20, described active component major ingredient is the weight of described active component major ingredient and described auxiliary material ratio
Cistanche benzyl carbinol glycosides active component, described auxiliary material is drug excipient, wherein utilizes the effective portion of cistanche benzyl carbinol glycosides
Promoting bone growing is carried out in position, and wherein cistanche benzyl carbinol glycosides active component contains echinacoside, acteoside, and saline cistanche
In total benzyl carbinol glycosides active component, the summation of contained ingredient content of phenylethanoid glycosides is more than 50%.
6. the drug regimen of the promoting bone growing containing cistanche benzyl carbinol glycosides active component according to claim 5
Thing it is characterised in that: in cistanche benzyl carbinol glycosides active component the summation of contained ingredient content of phenylethanoid glycosides be more than 50%.
7. the drug regimen of the promoting bone growing containing cistanche benzyl carbinol glycosides active component according to claim 5
Thing it is characterised in that: cistanche benzyl carbinol glycosides active component also contains boschnaloside a, and cistanche benzyl carbinol glycosides have
In effect position, the summation of contained ingredient content of phenylethanoid glycosides is more than 50%.
8. the drug regimen of the promoting bone growing containing cistanche benzyl carbinol glycosides active component according to claim 5
Thing it is characterised in that: cistanche benzyl carbinol glycosides active component also contains 2- acetyl acteoside, and cistanche benzene second
In alcohol glycosides effective part, the summation of contained ingredient content of phenylethanoid glycosides is more than 50%.
9. the drug regimen of the promoting bone growing containing cistanche benzyl carbinol glycosides active component according to claim 5
Thing it is characterised in that: cistanche benzyl carbinol glycosides active component also contains 2- acetyl acteoside and boschnaloside a, and
In cistanche benzyl carbinol glycosides active component, the summation of contained ingredient content of phenylethanoid glycosides is more than 50%.
10. the drug regimen of the promoting bone growing containing cistanche benzyl carbinol glycosides active component according to claim 5
Thing it is characterised in that: cistanche benzyl carbinol glycosides active component also contains 2- acetyl acteoside, boschnaloside a and 6- second
In acyl acteoside, and cistanche benzyl carbinol glycosides active component, the summation of contained ingredient content of phenylethanoid glycosides is more than
50%.
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109674805A (en) * | 2019-02-12 | 2019-04-26 | 大连大学 | Application of the benzyl carbinol glycoside compound in the drug for promoting periodontosis Bone Defect Repari |
| CN111187309A (en) * | 2020-02-24 | 2020-05-22 | 中国药科大学 | Preparation process and application of four components in cistanche |
| CN116270810A (en) * | 2023-02-22 | 2023-06-23 | 河南中医药大学 | Application of cistanche phenylethanol total glycosides in preparation of postmenopausal osteoporosis drugs |
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