CN1544429A - Bee glue flavone extract preparation method, pharmaceutical preparation and its new medical uses - Google Patents
Bee glue flavone extract preparation method, pharmaceutical preparation and its new medical uses Download PDFInfo
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- CN1544429A CN1544429A CNA2003101100850A CN200310110085A CN1544429A CN 1544429 A CN1544429 A CN 1544429A CN A2003101100850 A CNA2003101100850 A CN A2003101100850A CN 200310110085 A CN200310110085 A CN 200310110085A CN 1544429 A CN1544429 A CN 1544429A
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Abstract
The invention provides a process for preparing bee glue flavone extract, its medicinal preparation and novel medical use, wherein the preparation process comprises, freezing and crushing the bee glue into coarse powder, charging in alcohol proportionally for lixiviating, filtering and combining the filter liquor, reclaiming the solvent by depression, condensing into thick grease, decompressing and drying, disintegrating into fines, dissolving by chloroform-ethanol miscible liquid, and extracting with sodium-hydroxide solution of equal volume, adjusting pH to 4-8 by charging in hydrochloric acid into alkaline extract liquor, obtaining deposition, filtering by suction, rinshing to neutral, decompressing and drying, disintegrating into fines, thus the product according to the invention can be obtained.
Description
Technical field
The invention relates to the invention of novel method, pharmaceutical preparation and the new purposes thereof of the extraction purifying of propolis flavone,, belong to Chinese medicine or natural medicine field comprising effects such as propolis flavone treatment coronary heart disease, stenocardia, hyperlipidaemias.
Background technology
Propolis is that honeybee gives birth to resene material and the endocrine complex compound of honeybee that the branch axillalry bud is gathered from the collagen plant.The chemical ingredients of propolis is very complicated, and by chromatogram and mass spectroscopy, the flavones that has separated affirmation at present just has kind more than 20.Conventional propolis product is to take off cured back with propolis to add other auxiliary material compacting in flakes, and every contains flavones and must not calculate with anhydrous rutin and be less than 3 milligrams (being equivalent to 100 milligrams of former propolis), and promptly its product is the crude extract of flavones in the propolis.Propolis is as a kind of natural drug, and its significant curative effect and magical multiple function have caused the great attention of the world of medicine.
The present invention that is reported in about methods such as propolis flavone treatment coronary heart disease, stenocardia, hyperlipidaemias finishes before not report.
Summary of the invention
One of the object of the invention has provided a kind of technology that the propolis flavone extract is produced in industrialization that is suitable for.
It is medicine material development dripping pill, tablet, capsule, granule etc. that two of purpose of the present invention has provided with the propolis flavone extract.
It is that main medicine is used for prevention and treats diseases such as coronary heart disease, stenocardia, hyperlipidaemia that three of purpose of the present invention has provided with active propolis flavone and phenolic acid component.
The said propolis flavone extract of the present invention is the mixture of a kind of total flavones and total phenolic acid, total flavones and total phenolic acid component with treatment coronary heart disease, stenocardia, hyperlipidaemia, also be to separate first to obtain, the total flavones of extract is the 30-50% and active strong, the purity high (more than 50%) of total phenolic acid 20-40% part of total amount, general flavone content is no less than 30.0% in rutin, total phenolic content is no less than 20.0% in Protocatechuic Acid, and chrysin content must not be lower than 3.45%.
Technical solution of the present invention is as follows:
Get propolis medicinal material, freezing (10 ℃-4 ℃) 1～5 hour are ground into meal, add 50-95% ethanol lixiviate 1～4 time in the propolis ratio of 5-25ml/g, each 6～24 hours, filter merging filtrate, decompression and solvent recovery, be condensed into thick paste, at 40 ℃-80 ℃, drying under reduced pressure under the 0.05Mpa condition is ground into fine powder.Every gram dry powder adds 50-200ml chloroform-ethanol (10: 1-3: 1) dissolving, with equal-volume 0.5-5% sodium hydroxide solution extraction 1～3 time, the alkali extract adds hydrochloric acid and transfers pH to 4-8, produces precipitation, leaves standstill, suction filtration, be washed to neutrality, at 40 ℃-80 ℃, drying under reduced pressure under the 0.05Mpa condition, be ground into fine powder, promptly get extract of the present invention.
The propolis flavone extract of method preparation of the present invention is that yellowish brown is to brown ceramic powder; Special fragrance is arranged, puckery, wherein general flavone content is the 30-50% of total amount, total phenolic content is the 20-40% of total amount.
Foundation is the thin-layer chromatography discrimination method of reference substance with propolis control medicinal material and chrysin.This product weight loss on drying, residue on ignition, microbial limit are checked.Propolis total flavones, total phenolic content adopt spectrophotometry to measure, and chrysin content adopts high effective liquid chromatography for measuring.Regulation this product weight loss on drying must not be higher than 9.0%, and residue on ignition must not be higher than 1.0%, and general flavone content is no less than 30.0% in rutin, and total phenolic content is no less than 20.0% in Protocatechuic Acid, and chrysin content must not be lower than 3.45%.Microbial limit is up to specification.
Compound of the present invention and composition can be used for preparing the medicine of diseases such as treatment coronary heart disease, stenocardia, hyperlipidaemia.
Press practice of pharmacy, propolis flavone of the present invention can be prepared into the various clinical pharmaceutical dosage form as treatment of diseases medicines such as coronary heart disease, stenocardia, hyperlipidaemias, oral preparations is selected from any in dripping pill, tablet, capsule, pill, granule, suspensoid, the oral liquid.
Auxiliary material in the medicine of the present invention is meant conventional vehicle, as solvent, disintegrating agent, correctives, sanitas, tinting material, tackiness agent etc.
Pharmaceutical composition of the present invention contains one or more acceptable carriers pharmaceutically of treatment significant quantity.
Carrier above is meant the pharmaceutical carrier of pharmaceutical field routine, comprises thinner, vehicle, weighting agent, tackiness agent, wetting agent, disintegrating agent, absorption enhancer, tensio-active agent, absorption carrier.
Pharmaceutical composition preferred weight ratio of the present invention is 0.1%～99.5% activeconstituents, preferably contains 10-90%, and more preferably 20-80% is preferably 70% The compounds of this invention.
Formulation rate of the present invention can be according to variations such as route of administration, patient age, body weight, disease type and severity, and per daily dose is 0.01～10mg/kg.
Propolis flavone pharmaceutical preparation of the present invention has diseases such as treatment coronary heart disease, stenocardia, hyperlipidaemia, is to be confirmed by following effect experiment.
Propolis total flavones dripping pill function of resisting myocardial ischemia
Summary propolis total flavones dripping pill can obviously improve the Acute Myocardial Infarction that ligation dog anterior descending coronary (LAD) causes through duodenal administration; Obviously dwindle through the shown myocardial infarction area (MIS) of N-ST dyeing; Rising to glutamic-oxal(o)acetic transaminase in the dog serum (AST) vigor has the obvious suppression effect, to serum lactic dehydrogenase (LDH) and creatine phosphokinase (CPK) though the rising of vigor has the trend of reduction, no difference of science of statistics.The propolis total flavones dripping pill can also obviously reduce hypercholesterolemia mouse Tch content.
Purpose is observed the pharmacological effect of propolis total flavones dripping pill to cardiovascular systems, for the heart, vascular disease such as clinical treatment coronary heart disease and stenocardia provide theoretical foundation by the pharmacodynamic study of system.
The animal Kunming mouse, body weight 18-22g, the male and female dual-purpose, available from the institute for drug control, Jilin Province, the animal quality conformity certification is numbered 10-1008; Healthy adult hybrid dog, body weight 12-16kg, the male and female dual-purpose, portion provides by the Norman Bethune Medical University laboratory animal.
Medicine propolis total flavones dripping pill provides lot number by dosage form research chamber, the court: 991028, and specification: 36mg/ ball (9mg propolis total flavones/ball), the time spent is made into desired concn with suitable quantity of water.FUFANG DANSHEN DIWAN, produce lot number by Tianjin Tasly Pharmaceutical Co., Ltd: 990116, specification: the 25mg/ grain, because of its function cures mainly closely with this product, and its formulation is dripping pill, and is identical with this product formulation, so be used as positive control drug.Zhibituo is produced by Chengdu Diao 9 Wang pharmaceutical factory, lot number 9809054.
Instrument RM-6000 type polygraph, Japanese photoelectricity company produces; The LIANG-100 microcomputer shows record blood plasma viscometer automatically; The CORMNG-178 Bloodgas Analyzer.
Experimental data is learned " t " check by statistics, and statistics is all represented with X ± SD.
Test-results
One, the propolis total flavones dripping pill causes the influence of Acute Myocardial Infarction to ligation dog anterior descending coronary
25 of hybrid dogs are divided into 5 groups at random, 5 every group, anaesthetize with Sodital 30mg/kg iv.Back of the body position is fixing, and skin of neck cuts, and trachea cannula connects WM-2 type breathing apparatus, separates 1/3 place under the left anterior descending coronary artery, and threading causes myocardial infarction in order to ligation, executes the duodenum art in order to administration.
Art finishes, and stablizes 15 minutes.Get blood from femoral vein then, survey AST, CPK and LDH value, as being worth before the administration, the ligation coronary artery is after 15 minutes, through duodenal administration, control group waits capacity physiological saline, and the positive drug control group gives FUFANG DANSHEN DIWAN 200mg/kg, propolis total flavones dripping pill 80,40, three dosage groups of 20mg/kg (being equivalent to propolis total flavones 20,10 and 5mg/kg), write down after 300 minutes, get blood thought-read flesh three enzymes for the second time.At last, take off heart, claim heavyly whole-heartedly, cut off great vessels root and atrium along coronary sulcus, claim left ventricle heavy, be cut into 5 with left ventricle is cross-section equably, place nitro tetrazole orchid (N-BT) dye liquor, dyeing is 15 minutes in 37 ℃ of constant water bath box.(36 points/cm2) are measured every myocardium bilateral infarct (the non-dyeing of N-BT district) and non-infarct (N-BT dye district) with the drop point planimetry, every cardiac muscle is weighed, calculate every myocardial area, the total area of the ventricle total area and infarct calculates infarct and ventricle and cardiac weight per-cent.
1, to the influence of MIS
Positive drug group, middle dosage group, heavy dose of group all can obviously reduce infraction/reach whole-heartedly infraction/left heart area (P＜0.05).See Table 1.
Table 1 propolis total flavones dripping pill is to the influence (n=5) of myocardial infarction dog myocardial infarction area
Dosage
(%) infraction/left heart (%) of group infraction/whole-heartedly
mg/kg
Salt solution group-13.23 ± 1.555 20.40 ± 1.734
Positive group 200 9.88 ± 0.576
*15.84 ± 1.237
*
Propolis total flavones dripping pill 20 13.04 ± 0.983 18.56 ± 1.401
Propolis total flavones dripping pill 40 10.84 ± 1.405
*17.16 ± 1.744
*
Propolis total flavones drips 80 8.79 ± 0.628
* *13.94 ± 0.833
* *
Annotate: compare with the physiological saline control group,
*P＜0.05,
*P＜0.01,
* *P＜0.001
2, to the influence of myocardium three enzymes (AST, CPK, LDH)
The positive drug group is to the percentile reduction of serum AST content obvious (P＜0.05); Small dose group reduces obviously (P＜0.05) to the variation percentage of serum AST content; Small dose group is more then reduced to 89.40 ± 38.901 (P＜0.01) to the measured value and the control group of serum AST content.Middle dosage group is more then reduced to 104.78 ± 22.05 (P＜0.01) to serum AST content measured value and control group.Heavy dose of group is more then reduced to 74.80 ± 21.856 (P＜0.001) to serum AST content measured value and control group.The positive drug group, though small dose group, middle dosage group, heavy dose of group also can to serum CK, LDH, content reduction trend is arranged, with the physiological saline control group relatively, there was no significant difference.See Table 2,3,4.
Table 2 propolis total flavones dripping pill is to the influence of AST (U/L) in the myocardial infarction dog serum
After the preceding administration of group dosage (mg/kg) administration (360min)
Salt solution group-36.00 ± 12.530 170.60 ± 33.126
％ 420.92±209.85
Positive group 200 47.60 ± 12.700 100.60 ± 28.112
％ 131.91±102.58#
Propolis total flavones dripping pill 20 36.00 ± 7.314 89.40 ± 38.901**
％ 149.58±109.89#
Propolis total flavones dripping pill 40 28.80 ± 12.736 104.78 ± 22.050**
％ 357.90±341.31
Propolis total flavones dripping pill 80 27.60 ± 7.570 74.80 ± 21.856***
％ 193.44±110.88
Table 3 propolis total flavones dripping pill is to the influence of CK (U/L) in the myocardial infarction dog serum
After the preceding administration of group dosage (mg/kg) administration (360min)
Salt solution group-67.60 ± 25.540 507.20 ± 226.61
％ 694.37±398.10
Positive group 200 99.40 ± 40.303 549.40 ± 175.19
％ 498.05±240.06
Propolis total flavones dripping pill 20 66.20 ± 15.595 312.40 ± 95.482
％ 389.85±178.32
Propolis total flavones dripping pill 40 48.80 ± 11.541 336.40 ± 118.38
％ 580.48±143.87
Propolis total flavones dripping pill 80 78.20 ± 10.450 313.20 ± 150.12
％ 309.06±206.92
Table 4 propolis total flavones dripping pill is to the influence of LDH (U/L) in the myocardial infarction dog serum
After the preceding administration of group dosage (mg/kg) administration (360min)
Salt solution group-80.40 ± 31.501 157.60 ± 45.643
％ 168.20±70.247
Positive group 200 105.0 ± 26.991 168.20 ± 70.247
％ 73.12±96.522
Propolis total flavones dripping pill 20 81.20 ± 22.532 168.00 ± 71.376
％ 112.75±104.20
Propolis total flavones dripping pill 40 81.20 ± 27.842 187.20 ± 39.354
％ 154.79±95.664
Propolis total flavones dripping pill 80 81.00 ± 29.445 117.40 ± 13.576
％ 59.33±56.369
Annotate: compare * * P＜0.01, * * *＜0.001 with the salt solution group.Change percentage and salt solution group relatively, #P＜0.05
Two, the propolis total flavones dripping pill is to the influence of hypercholesterolemia mice serum total cholesterol
Get 70 of Kunming mouses, male, body weight 20 ± 2g, be divided into 7 groups by body weight, and pressed the successive administration of dosage shown in the table 57, in fasting in the 6th day, behind the 16h except that the blank group, all the other respectively organize equal abdominal injection 75% fresh egg-nog 0.5ml/20g, respectively organize mouse socket of the eye vein behind the 16h and get blood, and separation of serum is total cholesterol Tch and is measured.The result shows the rising of propolis total flavones dripping pill energy antagonism yolk emulsion induced mice serum T ch content, and significant difference is arranged, and sees Table 5.
Table 5. propolis total flavones dripping pill is to the influence of hypercholesterolemia mice serum total cholesterol
Group dosage mg/kg Tch mmol/L
Blank group---2.48 ± 0.40***
Model control group---14.37 ± 3.77
Propolis total flavones dripping pill 320 9.34 ± 2.27**
160 11.29±2.11*
80 11.84±3.99
Compare * P＜0.05, * * P＜0.01, * * * P＜0.001 with model control group
Discuss
This experiment adopts ligation anterior descending coronary (LAD) to cause acute myocardial infarction model.Variation with seroenzyme after quantitative tissue mensuration myocardial infarct size and the myocardial infarction.Experimental result shows that the propolis total flavones dripping pill can dwindle myocardial infarct size (MIS) through duodenal administration, reduces glutamic-oxal(o)acetic transaminase (AST) activity, and prompting propolis total flavones dripping pill has function of resisting myocardial ischemia.The propolis total flavones dripping pill can also significantly reduce the content of hypercholesterolemia mouse Tch; In sum, the propolis total flavones dripping pill has tangible function of resisting myocardial ischemia.
Embodiment
The following example is intended to further describe for example the present invention, rather than limit the present invention by any way, under the prerequisite that does not deviate from the spirit and principles in the present invention, any change or change that those of ordinary skills that the present invention did are realized easily all will fall within the claim scope of the present invention.
Embodiment 1
Get propolis medicinal material, freezing (10 ℃-4 ℃) 5 hours are ground into meal, add 95% ethanol lixiviate twice in the ratio of every gram propolis 20ml, each 24 hours, filter merging filtrate, decompression and solvent recovery, be condensed into thick paste, 80 ℃, drying under reduced pressure under the 0.05Mpa condition is ground into fine powder.Every gram dry powder adds 100ml chloroform-ethanol (10: 1-3: 1) dissolving, with equal-volume 5% sodium hydroxide solution extraction secondary, the alkali extract adds hydrochloric acid and transfers pH to 4-8, produces precipitation, leaves standstill 24 hours, suction filtration, be washed to neutrality, 60 ℃, drying under reduced pressure under the 0.05Mpa condition, be ground into fine powder, promptly get propolis flavone extract of the present invention.
Embodiment 2
Get propolis medicinal material, freezing (10 ℃-4 ℃) 5 hours are ground into meal, in twice of the ratio 65% ethanol lixiviate of every gram propolis 10ml, each 10 hours, filter merging filtrate, decompression and solvent recovery, be condensed into thick paste, 55 ℃, drying under reduced pressure under the 0.05Mpa condition is ground into fine powder.Every gram dry powder adds 200ml chloroform-ethanol (10: 1-3: 1) dissolving, with equal-volume 1.5% sodium hydroxide solution extraction secondary, the alkali extract adds hydrochloric acid and transfers pH to 4-8, produces precipitation, leaves standstill 24 hours, suction filtration, be washed to neutrality, and drying under reduced pressure (40 ℃-80 ℃, 0.05MPa), be ground into fine powder, promptly get the propolis flavone extract.
Embodiment 3
The preparation of dripping pill
Get propolis flavone extract 83g and 3 times amount polyethylene glycol 6000 through mixing, fusing, drip and make, cool off, blot and make dripping pill.Other project should meet 2000 editions dripping pill projects of Pharmacopoeia of People's Republic of China relevant requirements.
Embodiment 4
The preparation of tablet
Get propolis flavone extract 1000g, medical starch 500g mixes, and uses an amount of alcohol granulation, through the whole grain of pelletizing machine, and compressing tablet, every 0.25g, oral, each 1-2 sheet, twice of every day.Other project should meet 2000 editions tablet projects of Pharmacopoeia of People's Republic of China relevant requirements.
Embodiment 5
The preparation of capsule
Propolis flavone extract 500g, medical starch 500g mixes, the 0# capsule of packing into, every 0.30g, each oral 1-2 grain, twice of every day.Other project should meet 2000 editions capsule projects of Pharmacopoeia of People's Republic of China relevant requirements.
Embodiment 6
The preparation of granule
Propolis flavone extract 100g, starch 1000g, Icing Sugar 400g mixes, and uses an amount of alcohol granulation, and dry, whole grain, packing are promptly.Other project should meet 2000 editions granule projects of Pharmacopoeia of People's Republic of China relevant requirements.Oral, a 5g, twice on the one.
Claims (7)
1, a kind of propolis flavone preparation method of extract is characterized in that may further comprise the steps:
Get propolis medicinal material,-10 ℃-4 ℃ freezing 1～5 hour, be ground into meal, add the ratio 50-95% alcohol lixiviate 1～4 time of 5-25ml in the every gram of propolis, each 6～24 hours, filter merging filtrate, decompression and solvent recovery, be condensed into thick paste, 40 ℃-80 ℃, drying under reduced pressure under the 0.05Mpa condition is ground into fine powder; Every gram dry powder adds 50-200ml chloroform-alcohol mixeding liquid dissolving, chloroform-ethanol blending ratio is 10: 1-3: 1, with equal-volume 0.5-5% sodium hydroxide solution extraction 1～2 time, the alkali extract adds hydrochloric acid and transfers pH to 4-8, produces precipitation, leave standstill, suction filtration is washed to neutrality, drying under reduced pressure under 40 ℃ of-80 ℃ of conditions, be ground into fine powder, promptly get product of the present invention.
2, be medicine and the pharmaceutical composition that main component is used to prevent and treat diseases such as coronary heart disease, stenocardia, hyperlipidaemia with active propolis flavone extract.
3, the propolis flavone extract is used for prevention and treats diseases such as coronary heart disease, stenocardia, hyperlipidaemia.
4, pharmaceutical composition according to claim 2 is characterized in that containing propolis total flavones and total phenolic acid and one or more pharmaceutically admissible vehicle for the treatment of effective dose.
5, according to claim 2 or 3 described medicines, it is characterized in that: the vehicle of the propolis flavone of 1%-99% and phenolic acid and 99%-1%.
6, pharmaceutical preparation according to claim 2 is characterized in that said medicine is the formulation of oral preparations.
7, pharmaceutical preparation according to claim 2 is characterized in that said oral preparations is selected from a kind of in the middle of the dripping pill, tablet, pill, capsule, granule, suspensoid, oral liquid.
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Cited By (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN100368408C (en) * | 2005-11-16 | 2008-02-13 | 李秉� | Method of extracting licorice flavone from licorice slag |
| CN101049320B (en) * | 2007-05-11 | 2010-05-19 | 山东大学 | Buccal tablets of extractive of propolis alcohol |
| CN101081233B (en) * | 2007-02-02 | 2010-09-22 | 北京蜜香村蜂胶有限责任公司 | Method for preparing water-soluble propolis extract liquid |
| CN101099547B (en) * | 2007-06-18 | 2011-01-05 | 中国农业科学院蜜蜂研究所 | Water soluble propolis extract with bioactivity and light-colored propolis extract |
| CN106420833A (en) * | 2016-08-15 | 2017-02-22 | 上海沪郊蜂业联合社有限公司 | Propolis flavone, propolis flavone refining method and application of propolis flavone |
| CN106942551A (en) * | 2017-03-24 | 2017-07-14 | 湖南文理学院 | A kind of threeleaf akebia pectin beverage and preparation method thereof |
| CN107125515A (en) * | 2017-03-22 | 2017-09-05 | 济南大学 | It is a kind of to prepare the method with anti-oxidation function bee peptide drink |
| CN109528843A (en) * | 2018-11-28 | 2019-03-29 | 河北科技大学 | A kind of extracting method of flavone of hawthorn fruit |
| CN106420833B (en) * | 2016-08-15 | 2019-07-16 | 上海沪郊蜂业联合社有限公司 | The refining methd and propolis flavone of a kind of propolis flavone and application |
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2003
- 2003-11-21 CN CN 200310110085 patent/CN1274684C/en not_active Expired - Fee Related
Cited By (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN100368408C (en) * | 2005-11-16 | 2008-02-13 | 李秉� | Method of extracting licorice flavone from licorice slag |
| CN101081233B (en) * | 2007-02-02 | 2010-09-22 | 北京蜜香村蜂胶有限责任公司 | Method for preparing water-soluble propolis extract liquid |
| CN101049320B (en) * | 2007-05-11 | 2010-05-19 | 山东大学 | Buccal tablets of extractive of propolis alcohol |
| CN101099547B (en) * | 2007-06-18 | 2011-01-05 | 中国农业科学院蜜蜂研究所 | Water soluble propolis extract with bioactivity and light-colored propolis extract |
| CN106420833A (en) * | 2016-08-15 | 2017-02-22 | 上海沪郊蜂业联合社有限公司 | Propolis flavone, propolis flavone refining method and application of propolis flavone |
| CN106420833B (en) * | 2016-08-15 | 2019-07-16 | 上海沪郊蜂业联合社有限公司 | The refining methd and propolis flavone of a kind of propolis flavone and application |
| CN107125515A (en) * | 2017-03-22 | 2017-09-05 | 济南大学 | It is a kind of to prepare the method with anti-oxidation function bee peptide drink |
| CN106942551A (en) * | 2017-03-24 | 2017-07-14 | 湖南文理学院 | A kind of threeleaf akebia pectin beverage and preparation method thereof |
| CN109528843A (en) * | 2018-11-28 | 2019-03-29 | 河北科技大学 | A kind of extracting method of flavone of hawthorn fruit |
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|---|---|
| CN1274684C (en) | 2006-09-13 |
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