CN114903934A - Preparation method and application of mulberry leaf extract with bowel relaxing activity - Google Patents
Preparation method and application of mulberry leaf extract with bowel relaxing activity Download PDFInfo
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Classifications
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
- A61K36/18—Magnoliophyta (angiosperms)
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- A61K36/60—Moraceae (Mulberry family), e.g. breadfruit or fig
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
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- A61P1/10—Laxatives
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P1/00—Drugs for disorders of the alimentary tract or the digestive system
- A61P1/14—Prodigestives, e.g. acids, enzymes, appetite stimulants, antidyspeptics, tonics, antiflatulents
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
- A61K2236/30—Extraction of the material
- A61K2236/33—Extraction of the material involving extraction with hydrophilic solvents, e.g. lower alcohols, esters or ketones
- A61K2236/333—Extraction of the material involving extraction with hydrophilic solvents, e.g. lower alcohols, esters or ketones using mixed solvents, e.g. 70% EtOH
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
- A61K2236/50—Methods involving additional extraction steps
- A61K2236/51—Concentration or drying of the extract, e.g. Lyophilisation, freeze-drying or spray-drying
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
- A61K2236/50—Methods involving additional extraction steps
- A61K2236/53—Liquid-solid separation, e.g. centrifugation, sedimentation or crystallization
Abstract
The invention relates to preparation and application of a mulberry leaf extract, in particular to a preparation method and application of a mulberry leaf extract with bowel relaxing activity. A preparation method of a mulberry leaf extract with bowel relaxing activity comprises the step of carrying out hydrothermal reflux extraction on residues of mulberry leaves extracted by adopting an ethanol solution to obtain a liquid mulberry leaf extract with bowel relaxing activity. The prepared mulberry leaf extract has the function of relaxing bowel, which is obviously superior to the prior extract directly extracted from mulberry leaves by water, and the mulberry leaf waste residue is fully utilized, thereby effectively improving the comprehensive utilization rate of mulberry leaf resources.
Description
Technical Field
The invention relates to preparation and application of a mulberry leaf extract, in particular to a preparation method and application of a mulberry leaf extract with bowel relaxing activity.
Background
The mulberry leaves, also known as the Shenxiancao, are traditional Chinese medicine food materials, have various effects of relaxing bowel, reducing blood sugar, reducing blood fat and the like, and have high edible and medicinal values. Folium Mori contains multiple active ingredients such as alkaloid, polyphenol, flavone, and polysaccharide, and has wide pharmacological effects of lowering blood sugar, reducing blood lipid, relaxing bowels, resisting inflammation, resisting oxidation, and resisting virus. The mulberry leaves serving as one of medicinal and edible plants have great public foundation and industrial advantages in the field of functional food development, but the main pharmacological action and action mechanism of the mulberry leaves are not further verified, so that the development of the mulberry leaves is still in a grope state. The mulberry leaves are wide in source and low in price, wherein alkaloid component 1-deoxynojirimycin is a well-known active component with the function of reducing blood sugar, so that the market demand of mulberry leaf alkaloid extract is large, and a plurality of manufacturers exist. During production, 70% ethanol is used for extracting and preparing a mulberry leaf alkaloid crude extract, and a large amount of waste residues after ethanol extraction are generated. At present, the waste residue is rarely utilized, and resources are in a waste state.
The inventor team of the invention adopts the following method to prepare the mulberry leaf water extract in the existing research: weighing a proper amount of dry mulberry leaves, placing the dry mulberry leaves in a 60 ℃ oven for drying for 1 hour, taking distilled water with the volume 10-12 times of that of the mulberry leaves according to the pre-experimental experience, soaking, decocting until boiling, continuing to decoct for 60 minutes with soft fire, adding water for twice decoction, combining the filtrates of the two times, and concentrating with soft fire to obtain the mulberry leaf water extract with the crude drug concentration of 1 g/ml (the component content: 14.6 mg/g of total flavonoids and 17.8 mg/g of polysaccharides). The existing research (the function of mulberry leaf water extract in improving intestinal muscle movement and relieving constipation, food industry science and technology, network initials, published 3 months at 2022) shows that the experimental results show that the mulberry leaf water extract can play the roles of relieving constipation and relaxing bowel by increasing defecation, prolonging small intestine propulsion, thickening intestinal muscle layers, enhancing intestinal contraction tension, reducing AQP3, inhibiting excessive water absorption of colon and the like for different disease courses and administration periods.
The applicant does not find the same or similar literature reports in the prior art.
Disclosure of Invention
The invention discloses a preparation method and application of a mulberry leaf extract with bowel relaxing activity, which can utilize residues of mulberry leaves after alcohol extraction as raw materials, extract the mulberry leaf extract with bowel relaxing activity in the residues by a water extraction mode, have the bowel relaxing function obviously superior to that of the existing extract directly extracted from mulberry leaves, change waste into valuable, effectively improve the utilization rate of the residues and fully utilize mulberry leaf resources.
The overall technical concept of the invention is as follows:
a preparation method of a mulberry leaf extract with bowel relaxing activity comprises the step of carrying out hydrothermal reflux extraction on residues of mulberry leaves extracted by adopting an ethanol solution to obtain a liquid mulberry leaf extract with bowel relaxing activity.
Application of folium Mori extract in preparing medicine for loosening bowel to relieve constipation and promoting gastrointestinal peristalsis is provided.
The extract of the invention can be prepared into clinically acceptable formulations according to requirements, including but not limited to oral liquid, granules, water injection, tablets and the like, without departing from the technical essence of the invention.
The specific technical concept of the invention is as follows:
in order to obtain a better water extraction effect, the preferable technical implementation means is that the mass ratio of water to residues of the folium mori after alcohol extraction in hydrothermal reflux extraction is 10-12: 1, and the time of the hot reflux extraction is 1-2 hours.
For the convenience of storage, transportation and subsequent processing, the preferable technical implementation means is that the extract obtained after hydrothermal reflux extraction is dried to obtain mulberry leaf extract powder.
In order to keep the effective components in the extract not easy to damage, the preferable technical implementation means is that the drying adopts spray drying.
Further, the concentration and drying conditions are that the relative density is 1.07-1.12 when the mulberry leaves are concentrated to 60 ℃, the air inlet temperature is 155-165 ℃, and the air outlet temperature is 85-95 ℃, so that the mulberry leaf extract powder is obtained.
In order to obtain the mulberry leaf extract with more obvious effect, the more preferable technical implementation means is that the mulberry leaf extract powder is washed by ethanol with the mass percentage concentration of 60-80% and dried to obtain the mulberry leaf extract with obvious bowel relaxing activity.
Furthermore, the washing times of the ethanol are 1-2 times, and the drying adopts reduced pressure drying.
The invention has the substantive characteristics and the technical progress that are obtained:
1. a large number of experiments prove and creatively provide the following conclusion that the content of components which are easily dissolved in 60-80% ethanol solution in the folium mori ethanol extraction residue is reduced, so that the bowel relaxing effect of the residue hot reflux water extract is remarkably improved, and the effect is obviously superior to the bowel relaxing effect of the existing folium mori water extract.
2. The method fully utilizes the wastes of the alcohol extraction residues of the mulberry leaves, effectively improves the resource utilization rate of the mulberry leaves, has wide raw material source and low price, reduces the environmental protection cost, has simple preparation method, is easy to operate, and is easy to realize industrial production.
Drawings
The attached drawings of the invention are as follows:
FIG. 1 is a graph showing the comparison of small intestine propulsion rates of rats in each group according to the present invention.
In FIG. 1, # denotes P < 0.05 compared to the model group; + denotes P < 0.05 and N-5-6 compared to control sample 1.
FIG. 2 is a graph showing the results of the isolated ileum mean muscle tone of the rats in each group of the present invention.
FIG. 3 is a schematic representation of HE staining (x 100) of colon tissue of various groups of rats in accordance with the present invention.
In FIG. 3, A: blank group, B: model group, C: control sample 1 group, D: example 1 panel, Bars 100 μm, N5-6.
FIG. 4A is a schematic representation of mucin immunohistochemical staining (magnification:. times.400) of colon tissue in various groups of rats according to the present invention.
In fig. 4A: blank group, b: model group, c: control sample 1 group, d: example 1 panel, Bars 100 μm.
FIG. 4B is a statistical graph of mucin expression in colon tissues of rats in various groups according to the present invention.
In fig. 4B + indicates that P < 0.05 and N-5-6 compared to control sample 1.
FIG. 5A is a schematic representation of immunohistochemical staining (magnification 400) of 5-HT4R in colon tissue from various groups of rats according to the invention.
In fig. 5A: blank group, b: model group, c: control sample 1 group, d: example 1 panel, Bars 100 μm.
FIG. 5B is a statistical graph showing the expression of 5-HT4R in colon tissue of rats in each group according to the present invention.
In fig. 5B + indicates that P < 0.05 and N-5-6 compared to control sample 1.
FIG. 6 is a graphical representation of colon tissue 5-HT levels for various groups of rats in accordance with the present invention.
FIG. 7 is a schematic diagram of the analysis of intestinal flora PCoA of rats in each group according to the present invention.
K in FIG. 7: blank group, M7: model group, S7: control sample group 1, B7: example 1 group.
FIG. 8 is a histogram of the relative abundance of species at the level of each group of rats in the present invention.
K in fig. 8: blank group, M7: model group, S7: control sample group 1, B7: example 1 group.
Detailed Description
The present invention is further described with reference to the following examples, which should not be construed as limiting the scope of the present invention, but the scope of the present invention is defined by the appended claims, and any equivalent technical means may be substituted according to the present specification without departing from the scope of the present invention.
Example 1
The preparation method of the mulberry leaf extract with the activity of relaxing bowel comprises the following process steps:
(1) taking residues of mulberry leaves extracted by adopting an ethanol solution as a raw material, adding water for hot reflux extraction, wherein the mass ratio of water to the residues of the mulberry leaves after ethanol extraction in the hot reflux extraction is 10:1, the hot reflux extraction time is 1.5 hours, and filtering to obtain a mulberry leaf residue extracting solution;
(2) concentrating the mulberry leaf residue extracting solution obtained in the step (1), and performing spray drying; wherein the process parameters are that the relative density is 1.07-1.12 when the mulberry leaves are concentrated to 60 ℃, the air inlet temperature is 155-165 ℃, and the air outlet temperature is 85-95 ℃, so that the mulberry leaf extract powder is obtained.
(3) Washing the mulberry leaf extract powder obtained in the step (2) with 70% ethanol for 1 time, drying the insoluble substances under reduced pressure, setting the drying temperature at 50 ℃, and drying for 24 hours to obtain the mulberry leaf extract in the embodiment.
Application of folium Mori extract with effects of loosening bowel to relieve constipation in preparing medicine for loosening bowel to relieve constipation and promoting gastrointestinal peristalsis is provided.
Example 2
The preparation method of the mulberry leaf extract with the activity of relaxing bowel comprises the following process steps:
(1) taking residues of mulberry leaves extracted by adopting an ethanol solution as a raw material, adding water for hot reflux extraction, wherein the mass ratio of water to the residues of the mulberry leaves after ethanol extraction in the hot reflux extraction is 12:1, carrying out hot reflux for 1 hour, and filtering to obtain a mulberry leaf residue extracting solution;
(2) concentrating the mulberry leaf residue extracting solution obtained in the step (1), and performing spray drying; wherein the process parameters are that the relative density is 1.07-1.12 when the mulberry leaves are concentrated to 60 ℃, the air inlet temperature is 155-165 ℃, and the air outlet temperature is 85-95 ℃, so that the mulberry leaf extract powder is obtained.
(3) Washing the mulberry leaf extract powder obtained in the step (2) with 80% ethanol for 2 times, drying insoluble substances under reduced pressure, setting the drying temperature at 50 ℃, and drying for 24 hours to obtain the mulberry leaf extract in the embodiment.
The rest is the same as in example 1.
Example 3
The preparation method of the mulberry leaf extract with the activity of relaxing bowel comprises the following process steps:
(1) taking residues of mulberry leaves extracted by adopting an ethanol solution as a raw material, adding water for hot reflux extraction, wherein the mass ratio of water to the residues of the mulberry leaves after ethanol extraction in the hot reflux extraction is 10:1, carrying out hot reflux for 2 hours, and filtering to obtain a mulberry leaf residue extracting solution;
(2) concentrating the mulberry leaf residue extracting solution obtained in the step (1), and performing spray drying; wherein the process parameters are that the relative density is 1.07-1.12 when the mulberry leaves are concentrated to 60 ℃, the air inlet temperature is 155-165 ℃, and the air outlet temperature is 85-95 ℃, so that the mulberry leaf extract powder is obtained.
(3) Washing the mulberry leaf extract powder obtained in the step (2) with 60% ethanol for 1 time, drying the insoluble substances under reduced pressure, setting the drying temperature at 50 ℃, and drying for 24 hours to obtain the mulberry leaf extract in the embodiment.
The rest is the same as in example 1.
Example 4
The preparation method of the mulberry leaf extract with the activity of relaxing bowel comprises the following process steps:
(1) taking residues of mulberry leaves extracted by adopting an ethanol solution as a raw material, adding water for hot reflux extraction, wherein the mass ratio of water to the residues of the mulberry leaves after ethanol extraction in the hot reflux extraction is 10.5: 1, refluxing for 1.2 hours under heating, and filtering to obtain a mulberry leaf residue extracting solution;
(2) concentrating the mulberry leaf residue extracting solution obtained in the step (1), and performing spray drying; wherein the process parameters are that the relative density is 1.07-1.12 when the mulberry leaves are concentrated to 60 ℃, the air inlet temperature is 155-165 ℃, and the air outlet temperature is 85-95 ℃, so that the mulberry leaf extract powder is obtained.
(3) Washing the mulberry leaf extract powder obtained in the step (2) with 65% ethanol for 1 time, drying the insoluble substances under reduced pressure, setting the drying temperature at 50 ℃, and drying for 24 hours to obtain the mulberry leaf extract in the embodiment.
The rest is the same as in example 1.
Example 5
The preparation method of the mulberry leaf extract with the activity of relaxing bowel comprises the following process steps:
(1) taking residues obtained after folium mori is extracted by adopting an ethanol solution as a raw material, adding water for hot reflux extraction, wherein the mass ratio of water in the hot reflux extraction to the residues obtained after folium mori is extracted by ethanol is 11.5: 1, refluxing for 1.8 hours under heating, and filtering to obtain a mulberry leaf residue extracting solution;
(2) concentrating the mulberry leaf residue extracting solution obtained in the step (1), and performing spray drying; wherein the process parameters are that the relative density is 1.07-1.12 when the mulberry leaves are concentrated to 60 ℃, the air inlet temperature is 155-165 ℃, and the air outlet temperature is 85-95 ℃, so that the mulberry leaf extract powder is obtained.
(3) Washing the mulberry leaf extract powder obtained in the step (2) with 75% ethanol for 2 times, drying the insoluble substances under reduced pressure, setting the drying temperature at 50 ℃, and drying for 24 hours to obtain the mulberry leaf extract in the embodiment.
The rest is the same as in example 1.
To further illustrate the beneficial effects achieved by the present invention, applicants have verified the pharmacological activity as follows.
To further illustrate the effect of the present invention, a loperamide constipation model rat model was established, the small intestine propulsion rate was measured, and the improvement effect of the samples of control sample 1, example 2, example 3, example 4 and example 5 on the intestinal tract movement of the constipation model rat was compared, the control sample 1 was mulberry leaf water extract, and the results of the study were as follows:
referring to FIG. 1, as can be seen from the results of small intestine propulsion rate, the small intestine propulsion rate of the rats in the model group is reduced compared with that in the blank group; compared with the model group, the small intestine propulsion rate of rats in the groups of example 1, example 2, example 3, example 4 and example 5 is obviously increased (P < 0.05), and the small intestine propulsion rate difference of rats in the group of the control sample 1 is not obvious; compared with the control sample 1 group, the small intestine propulsion rate of rats in the groups of example 1, example 2, example 3, example 4 and example 5 is obviously increased (P is less than 0.05), which shows that the 5 extracts prepared by the invention can promote intestinal peristalsis, and the effect is better than that of a direct water extract of mulberry leaves.
Based on the comparison of control sample 1 as reference and the rat model of loperamide constipation model for the sample of example 1, the further pharmacological activity test was performed to compare the effects of the isolated ileum average muscle tension (gram), colon tissue HE staining, aliskiren blue staining, 5-HT4R immunohistochemical staining, 5-HT level and intestinal flora composition change of the rats given the control sample 1 and the sample of example 1, so as to illustrate the advantages of the present invention, and the results are as follows:
(1) referring to fig. 2, as can be seen from the isolated ileum mean muscle tone results, the model group rats mean muscle tone was decreased compared to the blank group; the mean muscle tone of the rats of the control sample 1 group and the example 1 group was increased compared to the model group, indicating that the effect of the extract of example 1 of the present invention was not inferior to that of the comparative sample 1.
(2) Referring to the attached figure 3, the HE staining result shows that the thickness of the colon muscle layer of the rat in the model group is obviously reduced compared with that in the blank group, the thickness of the colon muscle layer of the rat in the control sample 1 group and the rat in the example 1 group is thickened compared with that in the model group, and the muscle layer thickening effect of the rat in the example 1 group is more obvious, which indicates that the extract in the example 1 of the invention can obviously improve the pathological change of the intestinal muscle layer, and the effect is better than that of the direct water extract of mulberry leaves.
(3) Referring to FIG. 4, the results of Alisin blue staining revealed that the expression of mucin in colon tissues of rats in the model group was reduced compared with that in the blank group; compared with the model group, the colon tissue mucin of rats in the control sample 1 group and the example 1 group is increased; compared with the control sample 1 group, the rat colon tissue mucin expression of the example 1 group is obviously increased (P is less than 0.05), which shows that the extract of the example 1 can increase the expression of the colon mucin, protect the intestinal mucosa barrier and has better effect than the direct water extract of mulberry leaves.
(4) Referring to FIG. 5, as seen from the results of 5-HT4R staining, the expression of 5-HT4R was reduced in rat colon tissue of the model group as compared with that of the blank group; the colon tissue 5-HT4R of rats in the control sample group 1 and the group of example 1 was increased compared to the model group; compared with the control sample 1 group, the rat colon tissue 5-HT4R expression of the example 1 group is increased (P < 0.05), which shows that the extract of the example 1 of the invention can improve the colon 5-HT4R expression and promote the intestinal movement, and the effect is better than that of the direct water extract of mulberry leaves.
(5) Referring to FIG. 6, as seen from the results of the 5-HT levels in colon tissue, the 5-HT levels in colon were decreased in the rats in the model group compared to the rats in the blank group; in the control sample 1 group and the example 1 group, the level of 5-HT in the colon tissue of rats tends to be increased compared with the model group.
(6) Referring to fig. 7, it can be seen from the results of the intestinal flora PCoA that the blank group of flora is concentrated on the left side of the first principal component axis, the model group of flora is concentrated on the right side of the axis, the control sample group 1 and the example group 1 of flora both have a tendency to move to the blank group, and the example group 1 of flora is closer to the blank group.
(7) Referring to fig. 8, as can be seen from the results of the relative abundance of the intestinal flora species, when the groups of the flora with the relative abundance ranking first three are analyzed, the abundance of the model group is reduced compared with that of the blank group and is increased compared with that of the group of the control sample 1 and the group of the example 1; for the flora Ruminococcus _ UCG-005, the abundance of the model group is increased compared with that of a blank group, and the abundance of the control sample 1 group and the example 1 group is closer to that of the blank group; in addition, when the probiotic bacteria, namely Lactobacillus (Lactobacillus), are analyzed, the abundance of the model group is reduced compared with that of the blank group, the control sample 1 group and the example 1 group are increased, and the effect of the example 1 group is more remarkable.
Claims (8)
1. A preparation method of a mulberry leaf extract with bowel relaxing activity is characterized in that residue after mulberry leaves are extracted by adopting an ethanol solution is subjected to hydrothermal reflux extraction to obtain a liquid mulberry leaf extract with bowel relaxing activity.
2. The method for preparing mulberry leaf extract with bowel relaxing activity according to claim 1, wherein the mass ratio of water to residue after ethanol solution extraction of mulberry leaves in hydrothermal reflux extraction is 10-12: 1, and the time of thermal reflux extraction is 1-2 hours.
3. The method for preparing mulberry leaf extract with bowel relaxing activity as claimed in claim 1, wherein the extract after hydrothermal reflux extraction is concentrated and dried to obtain mulberry leaf extract powder.
4. The method for preparing mulberry leaf extract with bowel relaxing activity as claimed in claim 3, wherein the drying is spray drying.
5. The method for preparing mulberry leaf extract with bowel relaxing activity according to claim 3 or 4, wherein the conditions of concentration and drying are that the relative density is 1.07-1.12 when the concentration is carried out to 60 ℃, the air inlet temperature is 155-165 ℃, and the air outlet temperature is 85-95 ℃, so as to obtain the mulberry leaf extract powder.
6. The preparation method of the mulberry leaf extract with intestine-moistening and bowel-relaxing activities according to claim 3, wherein the mulberry leaf extract powder is washed by ethanol with the mass percent concentration of 60% -80%, and dried to obtain the mulberry leaf extract with remarkable intestine-moistening and bowel-relaxing activities.
7. The method for preparing mulberry leaf extract with bowel relaxing activity according to claim 6, wherein the number of times of ethanol washing is 1-2, and drying is performed under reduced pressure.
8. The use of the mulberry leaf extract according to any one of claims 1 to 7 in the preparation of a medicament for relaxing bowel and promoting gastrointestinal motility.
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Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101209284A (en) * | 2007-12-24 | 2008-07-02 | 天津大学 | Method for continuously extracting alkaloids, flavone and polysaccharide active components from mulberry leaves |
| CN102370707A (en) * | 2010-08-25 | 2012-03-14 | 上海辉文生物技术有限公司 | Method for preparing mulberry leaf and/or mulberry twig extract, obtained product thereof and application thereof |
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Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101209284A (en) * | 2007-12-24 | 2008-07-02 | 天津大学 | Method for continuously extracting alkaloids, flavone and polysaccharide active components from mulberry leaves |
| CN102370707A (en) * | 2010-08-25 | 2012-03-14 | 上海辉文生物技术有限公司 | Method for preparing mulberry leaf and/or mulberry twig extract, obtained product thereof and application thereof |
Non-Patent Citations (1)
| Title |
|---|
| 段晓峰: "桑叶水提物改善肠肌运动及缓解便秘的作用", 食品工业科技, vol. 43, no. 15, 3 March 2022 (2022-03-03), pages 344 - 351 * |
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