CN103301290A - Compound longan polysaccharide immune foundation-strengthening prescription and preparation method thereof - Google Patents
Compound longan polysaccharide immune foundation-strengthening prescription and preparation method thereof Download PDFInfo
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Abstract
The invention discloses a compound longan polysaccharide immune foundation-strengthening prescription which is obtained by adding longan polysaccharide into 36g/kg qi-replenishing and blood-enriching longan soup and allowing the concentration of the longan polysaccharide to reach 80mg/kg. The qi-replenishing and blood-enriching longan soup is prepared from the following raw materials: 20g of longan aril, 20g of astragalus, 10g of Chinese angelica, 15g of Chinese yam and 6g of liquorice. The method comprises the following steps: separating and extracting the longan polysaccharide, removing proteins, pigments and small molecules, and mixing with the qi-replenishing and blood-enriching longan soup, thereby obtaining the compound longan polysaccharide immune foundation-strengthening prescription. The compound longan polysaccharide immune foundation-strengthening prescription has the effects of replenishing qi and enriching blood and soothing the nerves and tonifying spleen; the immune cells can be activated, and the immunologic function of a body is improved.
Description
Technical field
The present invention relates to the immunity of a kind of compound recipe longan polysaccharide and consolidate prescription and preparation method thereof, belong to the health medicine technical field.
Background technology
Contain abundant polysaccharide in the Arillus Longan, polysaccharide is a kind of active substance, is known as effects such as having the immunity of raising, antitumor, blood sugar lowering and antiviral by international medical community.
Chinese patent 03100545.4 discloses a kind of Arillus Longan, lycium barbarum polysaccharide health promoting liquid, formed by longan polysaccharide 10-90 weight portion and lycium barbarum polysaccharide 10-90 weight portion, also openly can add Radix Ginseng, the Radix Astragali, Radix Codonopsis, olive oil and other Chinese crude drugs in the health promoting liquid, this polysaccharide health-care liquid has higher biological activity and immunity, belong to trophism and functional food, be conducive to keep the health of human body.But this health promoting liquid does not disclose the rational proportion of longan polysaccharide and other Chinese crude drugs.
Summary of the invention
The object of the present invention is to provide the immunity of a kind of compound recipe longan polysaccharide to consolidate prescription and preparation method thereof, the ratio optimization by longan polysaccharide and other Chinese crude drugs improves its immunoloregulation function.
The present invention realizes that the technical scheme that above-mentioned purpose is taked is: the immunity of a kind of compound recipe longan polysaccharide consolidates prescription, is that 80mg/kg gets by adding longan polysaccharide in the 36g/kg Arillus Longan vigorate qi and replenish the blood soup and making the concentration of longan polysaccharide; Described Arillus Longan vigorate qi and replenish the blood soup is prepared from by following raw material: Arillus Longan 20g, Radix Astragali 20g, Radix Angelicae Sinensis 10g, Rhizoma Dioscoreae 15g and Radix Glycyrrhizae 6g.
The immunity of described compound recipe longan polysaccharide consolidates the preparation method of prescription, may further comprise the steps:
(1) separation and Extraction of Arillus Longan crude polysaccharides: commercial Arillus Longan dry fruit is peeled off after the enucleation, takes out sarcocarp and dries in the shade, and weighs, be Arillus Longan: distilled water=1:4 by mass volume ratio, adopt hot water extraction, the control temperature is 90 ℃ of lower extractions 4 hours, filter, filtering residue repeats to extract 3 times, merging filtrate, and 68 ℃ of lower Rotary Evaporators are concentrated, liquid to be extracted is concentrated into thick, slowly pour into while hot in 95% ethanol of 4 times of volumes, sealing and standing, pure hypostasis is the Arillus Longan crude polysaccharides;
(2) the Arillus Longan crude polysaccharides is removed albumen and pigment: take by weighing the Arillus Longan crude polysaccharides and be mixed with 5.00g/ml solution with distilled water, in solution, add papain, making its final concentration is 1g/ml, then place 50 ℃ of thermostat water baths to be incubated 3h, in 100 ℃ of deactivation 5min, the centrifugal 15min of 4000r/min removes precipitate at last, merge supernatant, namely get and remove proteoglycan solution; The Arillus Longan crude polysaccharides of removing albumen is made into the aqueous solution of 5.00g/ml, drips 30%H
2O
2Remove pigment, consumption is 15.00% of polysaccharide solution volume, the pH value of regulating polysaccharide liquid with the NaOH solution of 0.10mol/L is 8.0, in 45 ℃ of water-baths, react, detect a pH value every 30min, guarantee that the pH value of reactant liquor maintains 8.0 in the course of reaction, cessation reaction behind the 4h, be chilled to room temperature, recall to neutrality with 0.10mol/L hydrochloric acid, add sodium chloride, making its concentration is 5.00g/ml, then slowly add dehydrated alcohol, make its content reach 35～40%, 4 ℃ of placements are spent the night in refrigerator, next day centrifugal collecting precipitation, through dehydrated alcohol, behind the acetone cyclic washing, 45 ℃ of drying under reduced pressure obtain removing the Arillus Longan crude polysaccharides behind albumen and the pigment;
(3) except micromolecule: pack into after the Arillus Longan crude polysaccharides that will remove albumen and pigment dissolves and remove in the dialysis cellophane of impurity with distilled water immersion 30min, be sandwiched in the beaker edge, add distilled water and extremely do not have cellophane fully; Distilled water is placed the 5000ml wide mouthed bottle, splash into by glass tubing in the beaker that the dialysis cellophane is housed with the speed of 1/s, dialysis 48h removes small molecular weight impurity, concentrating under reduced pressure then, and 95% ethanol precipitate with ethanol namely gets the longan polysaccharide of purification;
(4) preparation of Arillus Longan vigorate qi and replenish the blood soup: take by weighing Arillus Longan 20g, Radix Astragali 20g, Radix Angelicae Sinensis 10g, Rhizoma Dioscoreae 15g and Radix Glycyrrhizae 6g, soak 30min, add 12 times of water gagings, intense fire boils uses slow fire continuation decoction 36min instead, decoct 2 times, merge decocting liquid, the centrifugal 10min remove impurity of 3000rpm/min, it is 36g/kg that decocting liquid is concentrated into crude drug concentration;
(5) be that to add the longan polysaccharide of purification in the Arillus Longan vigorate qi and replenish the blood soup of 36g/kg and make the concentration of longan polysaccharide be 80mg/kg in crude drug concentration, namely get the immunity of described compound recipe longan polysaccharide and consolidate prescription.
The beneficial effect that the present invention possesses is: form through proved recipe Arillus Longan vigorate qi and replenish the blood soup and longan polysaccharide compatibility with tcm clinical practice, wherein Arillus Longan vigorate qi and replenish the blood soup is comprised of Arillus Longan, the Radix Astragali, Rhizoma Dioscoreae, Radix Angelicae Sinensis and Radix Glycyrrhizae, Arillus Longan sweet in the mouth warm in nature is monarch drug in the side, the master enters heart spleen two warps, invigorating the heart and spleen is arranged, the effect of nourishing blood to tranquillize the mind; The Radix Astragali and Rhizoma Dioscoreae are minister, all enter spleen lung two warps, but qi-supplementing, blood-engendering, and Radix Angelicae Sinensis is all ministerial drug and helps the Arillus Longan reinforcing the heart of nourishing blood, Radix Glycyrrhizae sweet in the mouth, coordinating the actions of various ingredients in a prescription.Full side has vigorate qi and replenish the blood, the effect of the spleen invigorating of calming the nerves.Polysaccharide compound is a kind of desirable immunomodulator, and the energy immune cell activated improves the immunologic function of body, and normal cell is had no side effect.With for the first time use in conjunction of polysaccharide and Chinese medicinal formulae, embody the new characteristic that Chinese medicine is inherited innovation.
The specific embodiment
Below in conjunction with embodiment technical scheme of the present invention is described further.
Embodiment 1
The immunity of a kind of compound recipe longan polysaccharide consolidates prescription, is that 80mg/kg gets by adding longan polysaccharide in the 36g/kg Arillus Longan vigorate qi and replenish the blood soup and making the concentration of longan polysaccharide; Described Arillus Longan vigorate qi and replenish the blood soup is prepared from by following raw material: Arillus Longan 20g, Radix Astragali 20g, Radix Angelicae Sinensis 10g, Rhizoma Dioscoreae 15g and Radix Glycyrrhizae 6g.
Embodiment 2
Embodiment 1 described compound recipe longan polysaccharide immunity consolidates the preparation method of prescription, may further comprise the steps:
(1) separation and Extraction of Arillus Longan crude polysaccharides: commercial Arillus Longan dry fruit is peeled off after the enucleation, takes out sarcocarp and dries in the shade, and weighs, be Arillus Longan: distilled water=1:4 by mass volume ratio, adopt hot water extraction, the control temperature is 90 ℃ of lower extractions 4 hours, filter, filtering residue repeats to extract 3 times, merging filtrate, and 68 ℃ of lower Rotary Evaporators are concentrated, liquid to be extracted is concentrated into thick, slowly pour into while hot in 95% ethanol of 4 times of volumes, sealing and standing, pure hypostasis is the Arillus Longan crude polysaccharides;
(2) the Arillus Longan crude polysaccharides is removed albumen and pigment: take by weighing the Arillus Longan crude polysaccharides and be mixed with 5.00g/ml solution with distilled water, in solution, add papain, making its final concentration is 1g/ml, then place 50 ℃ of thermostat water baths to be incubated 3h, in 100 ℃ of deactivation 5min, the centrifugal 15min of 4000r/min removes precipitate at last, merge supernatant, namely get and remove proteoglycan solution; The Arillus Longan crude polysaccharides of removing albumen is made into the aqueous solution of 5.00g/ml, drips 30%H
2O
2Remove pigment, consumption is 15.00% of polysaccharide solution volume, the pH value of regulating polysaccharide liquid with the NaOH solution of 0.10mol/L is 8.0, in 45 ℃ of water-baths, react, detect a pH value every 30min, guarantee that the pH value of reactant liquor maintains 8.0 in the course of reaction, cessation reaction behind the 4h, be chilled to room temperature, recall to neutrality with 0.10mol/L hydrochloric acid, add sodium chloride, making its concentration is 5.00g/ml, then slowly add dehydrated alcohol, make its content reach 35～40%, 4 ℃ of placements are spent the night in refrigerator, next day centrifugal collecting precipitation, through dehydrated alcohol, behind the acetone cyclic washing, 45 ℃ of drying under reduced pressure obtain removing the Arillus Longan crude polysaccharides behind albumen and the pigment;
(3) except micromolecule: pack into after the Arillus Longan crude polysaccharides that will remove albumen and pigment dissolves and remove in the dialysis cellophane of impurity with distilled water immersion 30min, be sandwiched in the beaker edge, add distilled water and extremely do not have cellophane fully; Distilled water is placed the 5000ml wide mouthed bottle, splash into by glass tubing in the beaker that the dialysis cellophane is housed with the speed of 1/s, dialysis 48h removes small molecular weight impurity, concentrating under reduced pressure then, and 95% ethanol precipitate with ethanol namely gets the longan polysaccharide of purification;
(4) preparation of Arillus Longan vigorate qi and replenish the blood soup: take by weighing Arillus Longan 20g, Radix Astragali 20g, Radix Angelicae Sinensis 10g, Rhizoma Dioscoreae 15g and Radix Glycyrrhizae 6g, soak 30min, add 12 times of water gagings, intense fire boils uses slow fire continuation decoction 36min instead, decoct 2 times, merge decocting liquid, the centrifugal 10min remove impurity of 3000rpm/min, it is 36g/kg that decocting liquid is concentrated into crude drug concentration;
(5) be that to add the longan polysaccharide of purification in the Arillus Longan vigorate qi and replenish the blood soup of 36g/kg and make the concentration of longan polysaccharide be 80mg/kg in crude drug concentration, namely get the immunity of described compound recipe longan polysaccharide and consolidate prescription.
Embodiment 3
Embodiment 1 and embodiment 2 described compound recipe longan polysaccharide immunity consolidate the activity research of prescription
The inventor is that the Arillus Longan vigorate qi and replenish the blood soup of 36g/kg carries out compatibility with the high, medium and low various dose of longan polysaccharide respectively with crude drug concentration, behind the research compatibility on the impact of immunosuppressed mice immunologic function.
(1) animal grouping and modeling
70 of single sex Kunming mouses are divided into 7 groups at random by body weight, are respectively: blank group, negative control group, alone medicine treatment group and share the medicine treatment group.The experiment time-histories was 4 weeks.Cyclophosphamide is dissolved in normal saline, and except blank group intraperitoneal injection of saline, all the other every group every a cyclophosphamide of 3d lumbar injection (Cy), and 20mg/kg, capacity are 10ml/kg, makes immunodeficiency models.Blank and negative control group gavage every day normal saline is treated, and the administration volume is 10ml/kg; Individually dosed group gives respectively vigorate qi and replenish the blood soup 36g/kg or longan polysaccharide 160mg/kg; Share the administration group and give respectively vigorate qi and replenish the blood soup and longan polysaccharide 320mg/kg, 160mg/kg, 80mg/kg.
(2) leading indicator detection method
The mensuration of A, mouse immune organ index
Pluck eyeball sacrificed by exsanguination mice, strip spleen, and weigh after peeling off surrounding tissue totally, be calculated as follows immune organ index:
Index and spleen index=spleen average weight (mg)/average weight (g)
B, peritoneal macrophage are engulfed the dimethyl diaminophenazine chloride experiment
Each organizes the front lh of mice execution, injection 4%SRBC0.5ml in every mouse peritoneal, and eyeball of mouse is extractd and is got blood execution behind the 1h, after 75% soak with ethanol, takes under the aseptic operating platform and carries out the sterile working.Inject 2ml RPMI1640 culture fluid for every mouse peritoneal with syringe, gently rub mouse web portion lmin, draw peritoneal fluid with sharp suction pipe, the centrifugal 10min of 3000rpm abandons supernatant, adds the RPMI1640 culture fluid that 1ml contains hyclone in the precipitate, re-suspended cell, counting, adjusting cell concentration is 2 * 10
6Individual/ml.Get peritoneal macrophage suspension 100 μ l and add in the 96 porocyte culture plates, each sample is established three multiple holes, puts into 37 ℃, 5%CO
2Hatch 2h in the incubator, remove culture fluid, attached cell is the peritoneal macrophage monolayer, and every hole adds 100 μ l0.072% neutral red solution, 37 ℃, 5%CO again
2Continue in the incubator to cultivate after the 3h, clean 3 times with the RPMI1640 culture fluid that does not contain hyclone, every hole adding cytolysate (dehydrated alcohol: the 0.1mol/L glacial acetic acid=l:l) place and spend the night, and surveys absorbance OD value next day at microplate reader wavelength 570nm place by 100 μ l4 ℃ of refrigerators again.
The assay method of C, mouse spleen T cell and B cell proliferation ability
The extracorporeal culturing method of mouse spleen cell: mice is put to death after getting blood, in gnotobasis, take out spleen, with the disposable syringe inner core discrete cell of on stainless steel filtering net, milling gently, add trypsinization, and with suction pipe piping and druming for several times, the centrifugal 10min of 1200rpm (contains 5%FBS with RPMI-1640,50uM2-mercaptoethanol, 100 μ/ml penicillin and 100mg/ml streptomycin) to transfer cell number be 1 * 10
6Individual/the ml suspension.
The assay method of mouse spleen T cell and B cell proliferation ability: the every hole of the mouse spleen cell after stable adds LPS (final concentration 30 μ g/ml) or ConA (final concentration 20 μ g/ml) 100 μ l, and matched group gives blank culture fluid.After hatching 72h in the cell culture incubator, mtt assay is surveyed cell survival rate.
The mensuration of IL-2, IL-4 and INF-γ in D, the mice serum
Measure according to ELISA test kit method.
(3) measurement result
Arillus Longan vigorate qi and replenish the blood soup and various dose longan polysaccharide compatibility are on the impact of immunosuppressed mice immunologic function
The result is as shown in table 1.
Table 1:
Annotate: * represents to compare P＜0.05 with negative control group, and # represents to compare P＜0.05 with the blank group.
Arillus Longan vigorate qi and replenish the blood soup and various dose longan polysaccharide compatibility are on the impact of immunosuppressed mice cytokine
The result is as shown in table 2.
Table 2:
Annotate: * represents to compare P＜0.05 with negative control group, and # represents to compare P＜0.05 with the blank group
(4) conclusion
The result shows, Arillus Longan vigorate qi and replenish the blood soup administration group and longan polysaccharide immunity consolidate prescription administration group all significantly Promote immunity suppress mouse spleen lymphocyte conversion capability and macrophage phagocytic ability (P＜0.05).Arillus Longan vigorate qi and replenish the blood soup administration group and longan polysaccharide immunity consolidate prescription administration group all can improve the immunosuppressed mice Serum IL-2, points out them to adjust and has improved the immunosuppressive condition that Cy causes.The longan polysaccharide immunity consolidates INF-γ/IL-4 ratio and Normal group no difference of science of statistics in the prescription administration group mice serum, and the ratio of pointing out it can correct immunosuppressed mice Thl/Th2 cytokine is disorderly, thereby regulates immunologic function.Analysis-by-synthesis, longan polysaccharide immunity consolidate prescription administration group immunoregulation effect and are better than Arillus Longan vigorate qi and replenish the blood soup administration group, and action effect is best when Arillus Longan vigorate qi and replenish the blood soup and low dosage longan polysaccharide compatibility.Therefore the compound recipe longan polysaccharide immunity optimum formula that consolidates prescription is defined as: Arillus Longan vigorate qi and replenish the blood soup 36g/kg and longan polysaccharide 80mg/kg compatibility.
Claims (2)
1. compound recipe longan polysaccharide immunity consolidates prescription, it is characterized in that, and be that 80mg/kg gets by adding longan polysaccharide in the 36g/kg Arillus Longan vigorate qi and replenish the blood soup and making the concentration of longan polysaccharide; Described Arillus Longan vigorate qi and replenish the blood soup is prepared from by following raw material: Arillus Longan 20g, Radix Astragali 20g, Radix Angelicae Sinensis 10g, Rhizoma Dioscoreae 15g and Radix Glycyrrhizae 6g.
2. one kind is applicable to the preparation method that compound recipe longan polysaccharide claimed in claim 1 immunity consolidates prescription, it is characterized in that, may further comprise the steps:
(1) separation and Extraction of Arillus Longan crude polysaccharides: commercial Arillus Longan dry fruit is peeled off after the enucleation, takes out sarcocarp and dries in the shade, and weighs, be Arillus Longan: distilled water=1:4 by mass volume ratio, adopt hot water extraction, the control temperature is 90 ℃ of lower extractions 4 hours, filter, filtering residue repeats to extract 3 times, merging filtrate, and 68 ℃ of lower Rotary Evaporators are concentrated, liquid to be extracted is concentrated into thick, slowly pour into while hot in 95% ethanol of 4 times of volumes, sealing and standing, pure hypostasis is the Arillus Longan crude polysaccharides;
(2) the Arillus Longan crude polysaccharides is removed albumen and pigment: take by weighing the Arillus Longan crude polysaccharides and be mixed with 5.00g/ml solution with distilled water, in solution, add papain, making its final concentration is 1g/ml, then place 50 ℃ of thermostat water baths to be incubated 3h, in 100 ℃ of deactivation 5min, the centrifugal 15min of 4000r/min removes precipitate at last, merge supernatant, namely get and remove proteoglycan solution; The Arillus Longan crude polysaccharides of removing albumen is made into the aqueous solution of 5.00g/ml, drips 30%H
2O
2Remove pigment, consumption is 15.00% of polysaccharide solution volume, the pH value of regulating polysaccharide liquid with the NaOH solution of 0.10mol/L is 8.0, in 45 ℃ of water-baths, react, detect a pH value every 30min, guarantee that the pH value of reactant liquor maintains 8.0 in the course of reaction, cessation reaction behind the 4h, be chilled to room temperature, recall to neutrality with 0.10mol/L hydrochloric acid, add sodium chloride, making its concentration is 5.00g/ml, then slowly add dehydrated alcohol, make its content reach 35～40%, 4 ℃ of placements are spent the night in refrigerator, next day centrifugal collecting precipitation, through dehydrated alcohol, behind the acetone cyclic washing, 45 ℃ of drying under reduced pressure obtain removing the Arillus Longan crude polysaccharides behind albumen and the pigment;
(3) except micromolecule: pack into after the Arillus Longan crude polysaccharides that will remove albumen and pigment dissolves and remove in the dialysis cellophane of impurity with distilled water immersion 30min, be sandwiched in the beaker edge, add distilled water and extremely do not have cellophane fully; Distilled water is placed the 5000ml wide mouthed bottle, splash into by glass tubing in the beaker that the dialysis cellophane is housed with the speed of 1/s, dialysis 48h removes small molecular weight impurity, concentrating under reduced pressure then, and 95% ethanol precipitate with ethanol namely gets the longan polysaccharide of purification;
(4) preparation of Arillus Longan vigorate qi and replenish the blood soup: take by weighing Arillus Longan 20g, Radix Astragali 20g, Radix Angelicae Sinensis 10g, Rhizoma Dioscoreae 15g and Radix Glycyrrhizae 6g, soak 30min, add 12 times of water gagings, intense fire boils uses slow fire continuation decoction 36min instead, decoct 2 times, merge decocting liquid, the centrifugal 10min remove impurity of 3000rpm/min, it is 36g/kg that decocting liquid is concentrated into crude drug concentration;
(5) be that to add the longan polysaccharide of purification in the Arillus Longan vigorate qi and replenish the blood soup of 36g/kg and make the concentration of longan polysaccharide be 80mg/kg in crude drug concentration, namely get the immunity of described compound recipe longan polysaccharide and consolidate prescription.
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| CN104031155A (en) * | 2014-05-06 | 2014-09-10 | 广州医科大学附属第一医院 | Industrial extraction and purification method of Quanzhenyiqi decoction polysaccharides |
| CN104865333A (en) * | 2015-05-14 | 2015-08-26 | 武汉轻工大学 | Detection method of longan meat protein polysaccharide |
| CN104872656A (en) * | 2015-03-06 | 2015-09-02 | 天津江天同创酒业有限公司 | Composition with health function and preparation method thereof |
| CN105030626A (en) * | 2015-09-17 | 2015-11-11 | 天峨县平昌生态农业有限公司 | Anti-aging mask |
| CN106167531A (en) * | 2016-08-16 | 2016-11-30 | 集美大学 | A kind of method of enzyme process assisted extraction Arillus Longan polysaccharide |
| CN110604769A (en) * | 2018-06-17 | 2019-12-24 | 罗建平 | Multifunctional composite plant polysaccharide |
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Cited By (11)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103992415A (en) * | 2014-05-06 | 2014-08-20 | 广州医科大学附属第一医院 | Method for extracting and purifying Quanzhenyiqi decoction polysaccharide |
| CN104031155A (en) * | 2014-05-06 | 2014-09-10 | 广州医科大学附属第一医院 | Industrial extraction and purification method of Quanzhenyiqi decoction polysaccharides |
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| CN104031155B (en) * | 2014-05-06 | 2016-06-01 | 广州医科大学附属第一医院 | The method of the industrial extraction and purification of an authentic gas soup polysaccharide |
| CN104872656A (en) * | 2015-03-06 | 2015-09-02 | 天津江天同创酒业有限公司 | Composition with health function and preparation method thereof |
| CN104865333A (en) * | 2015-05-14 | 2015-08-26 | 武汉轻工大学 | Detection method of longan meat protein polysaccharide |
| CN104865333B (en) * | 2015-05-14 | 2016-08-24 | 武汉轻工大学 | A kind of detection method of Arillus Longan proteoglycan |
| CN105030626A (en) * | 2015-09-17 | 2015-11-11 | 天峨县平昌生态农业有限公司 | Anti-aging mask |
| CN106167531A (en) * | 2016-08-16 | 2016-11-30 | 集美大学 | A kind of method of enzyme process assisted extraction Arillus Longan polysaccharide |
| CN106167531B (en) * | 2016-08-16 | 2018-09-21 | 集美大学 | A kind of method of enzyme process assisted extraction longan polysaccharide |
| CN110604769A (en) * | 2018-06-17 | 2019-12-24 | 罗建平 | Multifunctional composite plant polysaccharide |
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