CN104163874A - Phellinus linteus polysaccharide ZDT and preparation method thereof - Google Patents
Phellinus linteus polysaccharide ZDT and preparation method thereof Download PDFInfo
- Publication number
- CN104163874A CN104163874A CN201410355608.6A CN201410355608A CN104163874A CN 104163874 A CN104163874 A CN 104163874A CN 201410355608 A CN201410355608 A CN 201410355608A CN 104163874 A CN104163874 A CN 104163874A
- Authority
- CN
- China
- Prior art keywords
- polysaccharide
- phellinus
- zdt
- precipitation
- milliliters
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Granted
Links
Landscapes
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
Abstract
The invention relates to a phellinus linteus polysaccharide ZDT component and a preparation method thereof. The phellinus linteus polysaccharide ZDT component is a beta glucomannan ZDT separated from phellinus linteus sporocarp as a raw material by crushing, extraction and deposition, and has weight-average molecular weight of 880000-1020000 Da and polysaccharide mass percentage content more than 90%. The extraction technology utilizes polyvinylpyrrolidone K30 to replace ethanol and based on the same volume of extract, a use amount of the polyvinylpyrrolidone K30 is fiftieth that of ethanol so that a production cost is greatly reduced. The phellinus linteus polysaccharide ZDT has substantial antineoplastic activity.
Description
(1) technical field
The present invention relates to a kind of Phellinus polysaccharide ZDT and preparation method thereof.Phellinus sporophore, through pulverizing, is extracted, and the steps such as separation obtain a kind of Phellinus polysaccharide ZDT, and this polysaccharide is mainly made up of glucose and seminose, can be used for making foodstuff additive, various pharmaceutical preparations.The invention belongs to food-processing, medicine industry association area.
(2) background technology
Phellinus is traditional medicinal fungi of China, is the sporophore of Phellinus fungi phelliuns igniarius, and this fungi is born on the trunks such as mulberry tree, willow, birch, robur, is distributed in the ground such as China North China, northeast, northwest and Sichuan, Yunnan.In China's traditional medicine, Phellinus is mainly used in treating metrorrhagia, and blood drenches, and prolapse of the anus is rushed down blood, is with inferior disease, is a kind of medicinal fungi of preciousness.Nearly decades are along with finding that to the further research of Phellinus Phellinus has higher inhibition tumour and antiviral activity, at present the ancillary drug mainly as immunostimulant and cancer therapy in Japan and Korea S.The main active ingredient of Phellinus is polysaccharide, and foreign study thinks, Phellinus polysaccharide can reach the object to tumor suppression by improving immunity of organisms.
The Wen Ke of Jilin University etc. find (Jilin University's journal after deliberation; the 28th the 3rd phase of volume in 2002) at Phellinus, glossy ganoderma, A Jiali Si young pilose antler, PL-2; in several materials of PL-5 (MeSima); Phellinus has better function of tumor inhibition; further in research, find, animal takes after Phellinus water body thing, the cytophagous phagocytic index k of monokaryon one and engulf coefficient n and obviously increase; and can improve spleen coefficient and thymus gland coefficient, promote the T lymphproliferation response of ConA induction.Korea S Sang-BaeHan etc. published an article in International Immunopharmacology magazine the 6th phase in 2006; result of study shows a kind of polysaccharide with treatment cancer activity of separating from Phellinus, and this polysaccharide can suppress the transfer of tumour.Gi-Young Kim etc. publishes an article at Journal of Ethnopharmacology magazine 95 volumes for 2004; find that a kind of polysaccharide extracting from Phellinus has anti-tumor activity, the antitumor mechanism of this polysaccharide may be relevant with TNF-α concentration with improving nitrogen protoxide.(the Food science such as Ge Qing; the 09th phase in 2008) Phellinus sporophore water extraction, alcohol precipitation, lyophilize are obtained to water-soluble polysaccharide; use gel column to be further purified to obtain holosaccharide PIFI, its monose consists of seminose, glucose and semi-lactosi, and mol ratio is 0.64: 1: 1.94.
Although the industrialization of extraction purifying of Phellinus polysaccharide; there are many companies on market, to sell Phellinus polysaccharide; but extraction process is more complicated mostly; too busy to get away ethanol precipitates this step; also there is people to carry out the extraction of Phellinus polysaccharide by the way of film processing; but because the cost of film is higher, reproducibility is bad, extracts the not high reason of polysaccharide content and cause the production cost of Phellinus polysaccharide significantly to decline.The Phellinus polysaccharide quality percentage composition of selling in the market mostly, below 60%, lacks the high-end product that exceedes 90% content.
(3) summary of the invention
Phellinus polysaccharide ZDT component of the present invention is taking Phellinus sporophore as raw material, through pulverizing, extracts, and the techniques such as precipitation separate a kind of β konjac glucomanna ZDT obtaining, and this extracting method and polysaccharide fraction have no report in document before.Extract this component and use PVP K30 to replace ethanol, the extracting solution PVP K30 consumption of same volume only has 1/50th of ethanol, and this greatly reduces production cost, and gained polysaccharide ZDT has significant anti-tumor activity.PVP K30 is a kind of macromole solubility promoter, is also used as the impurity of flocculation agent in purifying waste water, and 2010 editions second of Chinese Pharmacopoeia included as medicinal raw material.Use it as and extract the reagent of Phellinus polysaccharide not only cost is low, and security is also secure, is conducive to industrial-scale production from now on.The principle why PVP K30 can extract Phellinus polysaccharide as precipitation agent not yet has bibliographical information at present; supposition may be relevant with its throwing out; the Phellinus polysaccharide ZDT weight-average molecular weight that extraction obtains is between 880,000～1,020,000 dalton; specific aim is stronger, and this point is different from ethanol precipitation gained Phellinus polysaccharide.Ethanol precipitation gained polysaccharide component complexity, need just can obtain molecular weight ratio compared with the relative sterling of homogeneous through a series of chromatographic column purifying.Jia Jianbo etc. utilize ethanol precipitation to obtain Phellinus polysaccharide, further separate and obtain two kinds of significant polysaccharide fractions of molecular weight difference of 2,840,000 and 5.33 ten thousand dalton (Phellinus separation of polysaccharides purifying and structure preliminary evaluation Food science Vol.27No.12 in 2006 thereof).The polysaccharide quality percentage composition that PVP K30 extracts as precipitation agent the Phellinus polysaccharide ZDT obtaining is higher, extracts 3 batches of content and all exceedes 92%, and this has reduced cost for being further purified Phellinus polysaccharide.
The preferred extraction process concrete steps of preparation Phellinus polysaccharide ZDT are:
One, dry, pulverize
Phellinus sporophore dry product uses powder beater to smash, and sieves and obtains Phellinus powder with No. four screen clothes.
Two, extract
1 kilogram, Phellinus powder, adds 100 DEG C of 10 liters of hot water and extracts 2 hours, uses whizzer to carry out solid-liquid separation, and residue discards, and totally 7200 milliliters of the extracting solutions of clarification are for subsequent use.
Three, precipitation
The extracting solution 7200ml in " two, extract " that gets step adds 720 grams of PVP K30s in extracting solution, is stirred well to PVP K30 and all dissolves.Hold over night, centrifugal being precipitated, precipitation is washed 2 times with the aqueous ethanolic solution of 50% mass concentration, and each 2000 milliliters, then use absolute ethanol washing 2 times, each 2000 milliliters, after 80 DEG C of oven dry, be weighed as 230 grams.Precipitation is pulverized with No. four screen cloth sieve precipitation powder, and precipitation powder adds 100 DEG C of 6900 ml waters and extracts 2 hours, centrifugal, above resets and add 690 grams of PVP K30s, is stirred well to PVP K30 and all dissolves.Hold over night, centrifugal being precipitated, precipitation is washed 2 times with the aqueous ethanolic solution of 50% mass concentration; each 2000 milliliters, then use absolute ethanol washing 2 times, each 2000 milliliters; dry postprecipitations for 80 DEG C and pulverize, repeatedly sieved to obtain 182 grams of Powdered Phellinus polysaccharide ZDT with No. four screen clothes.
The structural information of Phellinus polysaccharide ZDT component is:
1, assay
The Phellinus polysaccharide ZDT sample of 3 batches is measured through phenol sulfuric acid method, and polysaccharide quality percentage composition is all more than 90%.
2, monose compositional analysis
Reference literature (Dai Jun etc.; analytical test journal; the 26th volume the 2nd phase P206～216) Phellinus polysaccharide ZDT component is carried out to monose compositional analysis, result shows that the monose composition of ZDT is mainly glucose and seminose, the mole ratio of glucose and seminose is 1: 2.
3, weight-average molecular weight
Taking Dextran series dextran as reference substance, measure the weight-average molecular weight of 3 crowdes of ZDT, result shows that the weight-average molecular weight of 3 crowdes of ZDT is respectively 880,000,930,000,1,020,000 dalton.
4, infrared scan (configuration)
Infrared scan result shows that ZDT sample is at 890cm
-1there is weak absorption peak, 840 cm
-1without absorbing, illustrate that ZDT component is beta comfiguration.
Described Phellinus polysaccharide ZDT can be applicable to prepare antitumor drug, also can be used for the additive of food prepared therefrom, protective foods, is applied to the patient who suffers from cancer.
Phellinus polysaccharide ZDT component abdominal injection (20mg/kg, 50mg/kg and 100mg/kg) administration 10 days; lentinan (Lentinan) is contrast, and result shows that 3 kinds of dosage of Phellinus polysaccharide ZDT component all have remarkable restraining effect to mouse S-180 knurl.Wherein 50mg/kg and Lentinan 20mg/kg dosage tumour inhibiting rate are without marked difference, all higher than 80%.Prompting Phellinus polysaccharide ZDT component can be for the preparation of antitumor drug.
(4) Figure of description
Nothing
(5) embodiment
Below in conjunction with specific embodiment, the present invention is described further, but protection scope of the present invention is not limited in this:
The preparation of embodiment 1 Phellinus polysaccharide ZDT
1 kilogram of Phellinus sporophore dry product, pulverizes with powder beater, and No. four screen cloth sieves Phellinus powder.Add 10 liters of hot water to Phellinus powder, stir, 100 DEG C are extracted 2 hours, use whizzer to carry out solid-liquid separation, and residue discards, and obtains 7200 milliliters of extracting solutions.In extracting solution, add 720 grams of PVP K30s, be stirred well to PVP K30 and all dissolve.Hold over night, centrifugal being precipitated, precipitation is washed 2 times with the aqueous ethanolic solution of 50% mass concentration, and each 2000 milliliters, then use absolute ethanol washing 2 times, each 2000 milliliters, after 80 DEG C of oven dry, be weighed as 230 grams.Precipitation is pulverized with No. four screen cloth sieve precipitation powder, and precipitation powder adds 100 DEG C of 6900 ml waters and extracts 2 hours, centrifugal, above resets and add 690 grams of PVP K30s, is stirred well to PVP K30 and all dissolves.Hold over night, centrifugal being precipitated, precipitation is washed 2 times with the aqueous ethanolic solution of 50% mass concentration; each 2000 milliliters, then use absolute ethanol washing 2 times, each 2000 milliliters; after 80 DEG C of oven dry, pulverize, repeatedly sieved to obtain 182 grams of Powdered Phellinus polysaccharide ZDT with No. four screen clothes.
The physics and chemistry of embodiment 2 Phellinus polysaccharide ZDT detects
Prepare 3 batches of ZDT polysaccharide (lot number 20120601,20120602,20120603) according to the preparation method in embodiment 1 and carry out structural analysis.Monose compositional analysis adopts PMP (1-phenyl-3-methyl-5-pyrazolones ketone) derivatization method; Polysaccharide configuration adopts infrared analysis; Weight-average molecular weight takes high performance gel permeation chromatography (GPC) to measure, and detector uses differential detector.Above test result is as follows:
The monose composition measuring result of 3 batches of ZDT polysaccharide is as following table 1:
Table 1
The quality percentage composition of 3 batches of ZDT polysaccharide and weight-average molecular weight measurement result are as following table 2
Table 2
| Lot number | 20120601 | 20120602 | 20120603 |
| Weight-average molecular weight (Mw, dalton) | 880000 | 930000 | 1020000 |
| Polysaccharide quality percentage composition | 93.1％ | 92.6％ | 94.2％ |
?infrared scan result shows that 3 batches of ZDT samples are at 890cm
-1all there is weak absorption peak, 840cm
-1all, without absorbing, interpret sample is beta comfiguration, not containing α configuration.
Comprehensive above various physical and chemical determination results, the polysaccharide quality percentage composition in Phellinus polysaccharide ZDT all exceedes 90%, belongs to high purity polysaccharide.The monose formation of this polysaccharide is mainly glucose and seminose, and two kinds of monose mol ratios are 1: 2, and weight-average molecular weight is between 88～1,020,000 dalton.
The preparation of embodiment 3 Phellinus polysaccharide ZDT sheets
Get the Phellinus polysaccharide ZDT20 gram in embodiment 1, add 200 grams of auxiliary materials, mix, granulate, dry, be pressed into 1000, to obtain final product.
The preparation of embodiment 4 Phellinus polysaccharide ZDT capsules
The Phellinus polysaccharide ZDT20 gram that gets embodiment 1, adds 300 grams of auxiliary materials, mixes, and granulates, dry, is filled to 1000 of capsules, to obtain final product.
The preparation of embodiment 5 Phellinus polysaccharide ZDT injection liquids
The Phellinus polysaccharide ZDT20 gram that gets embodiment 1, adds 200 grams of auxiliary materials, dissolves by 2L purified water, and sterile filtration, filling, freeze-drying and get final product.
The restraining effect of embodiment 6 Phellinus polysaccharide ZDT to S-180 knurl
Sample: Phellinus polysaccharide ZDT (polysaccharide quality percentage composition is greater than 92%), lentinan (Lentinan) is commercially available.
Animal: 50 of healthy Kunming mouses, male and female half and half.
Model preparation: get in the S180 of logarithmic phase cell strain oncocyte, become 1 × 10 with normal saline dilution
6the tumor cell suspension of mL, is inoculated in mouse right fore armpit subcutaneous, and the every mouse 0.2mL of inoculum size, starts the administration of dividing into groups next day.
Animal grouping and administering mode: divide 5 groups, 10 every group, male and female half and half.Phellinus polysaccharide ZDT by low (20mg/kg), in (50mg/kg); high (100mg/kg) dosage is divided into 3 groups, 1 group of lentinan (20mg/kg), model group injecting normal saline; each intraperitoneal injection 10 days of organizing all continuously.
Indicator-specific statistics calculates: after last administration, next day, eye socket venous plexus was got blood, centrifugal, got supernatant, adopts enzyme mark double antibody sandwich ELISA to detect TNF-α and IL 2 content.Put to death mouse, the complete knurl piece of peeling off, weighs and record, calculates tumor control rate.
The average knurl of tumor control rate=(the average knurl weight of the average knurl weight-administration of model group group)/model group heavy × 100%
Experimental result: lentinan and Phellinus polysaccharide ZDT senior middle school low dose group all have significant tumor-inhibiting action, concrete data see the following form 3:
The tumor-inhibiting action of table 3 Phellinus polysaccharide ZDT component (
n=10)
Statistical method: T-test, with relatively * P < 0.05 of model group, * * P < 0.01.
| Group | Average knurl heavy (g) | Tumour inhibiting rate | TNF-α(ng/mL) | IL?2(ng/mL) |
| Model group | 1.55±0.12 | - | 5.05±0.20 | 0.18±0.04 |
| ZDT high dose group | 0.37±0.08* | 76％ | 6.81±0.32* | 0.28±0.02* |
| Dosage group in ZDT | 0.35±0.09* | 77％ | 6.92±0.26* | 0.28±0.03* |
| ZDT low dose group | 0.38±0.05* | 75％ | 6.88±0.25* | 0.29±0.03* |
| Lentinan group | 0.41±0.07* | 72％ | 6.49±0.26* | 0.25±0.02* |
The TNF-α of each administration group and IL2 content all increase than model group as can be seen from Table 3, and this may be that they have one of reason of tumor-inhibiting action.
Claims (3)
1. a Phellinus polysaccharide ZDT; it is characterized in that described Phellinus polysaccharide ZDT is the β-konjac glucomanna of glucose and seminose composition; the molar ratio of two kinds of monose is 1: 2, and weight-average molecular weight is between 88～1,020,000 dalton, and the quality percentage composition of polysaccharide is higher than 90%.
2. the method for preparation Phellinus polysaccharide ZDT as claimed in claim 1, it is characterized in that 1 kilogram of Phellinus sporophore dry product, pulverize with powder beater, No. four screen cloth sieve Phellinus powder, add 10 liters of hot water to Phellinus powder, stir, 100 DEG C are extracted 2 hours, use whizzer to carry out solid-liquid separation, residue discards, obtain 7200 milliliters of extracting solutions, in extracting solution, add 720 grams of PVP K30s, being stirred well to PVP K30 all dissolves, hold over night, centrifugal being precipitated, precipitation is washed 2 times with the aqueous ethanolic solution of 50% mass concentration, each 2000 milliliters, use again absolute ethanol washing 2 times, each 2000 milliliters, after 80 DEG C of oven dry, be weighed as 230 grams, precipitation is pulverized, with No. four screen cloth sieve precipitation powder, precipitation powder adds 100 DEG C of 6900 ml waters and extracts 2 hours, centrifugal, on reset and add 690 grams of PVP K30s, being stirred well to PVP K30 all dissolves, hold over night, centrifugal being precipitated, precipitation is washed 2 times with the aqueous ethanolic solution of 50% mass concentration, each 2000 milliliters, use again absolute ethanol washing 2 times, each 2000 milliliters, after 80 DEG C of oven dry, pulverize, repeatedly sieved to obtain Powdered Phellinus polysaccharide ZDT182 gram with No. four screen clothes.
3. the Phellinus polysaccharide ZDT as described in claim 1～2 is in the application of preparing in antitumor drug.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201410355608.6A CN104163874B (en) | 2014-07-21 | 2014-07-21 | A kind of Phellinus polysaccharide ZDT and its preparation method |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201410355608.6A CN104163874B (en) | 2014-07-21 | 2014-07-21 | A kind of Phellinus polysaccharide ZDT and its preparation method |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN104163874A true CN104163874A (en) | 2014-11-26 |
| CN104163874B CN104163874B (en) | 2016-06-01 |
Family
ID=51907869
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201410355608.6A Active CN104163874B (en) | 2014-07-21 | 2014-07-21 | A kind of Phellinus polysaccharide ZDT and its preparation method |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN104163874B (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110038034A (en) * | 2019-04-11 | 2019-07-23 | 安徽省森湶谷药业股份有限公司 | A kind of small molecule water prepares the method and application of phellinus linteus extract |
Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPS4924211A (en) * | 1972-07-01 | 1974-03-04 | ||
| JPH0924211A (en) * | 1995-07-11 | 1997-01-28 | Takane Kitao | Filter device for waste water |
| CN101323648A (en) * | 2008-07-29 | 2008-12-17 | 上海璞诚生物科技有限公司 | Extraction method and and purification method of Sanghuang mushroom polysaccharide |
| CN101463093A (en) * | 2007-12-17 | 2009-06-24 | 天津市华泰森淼生物工程技术有限公司 | Technique for extracting polysaccharide from Phellinus linteus |
| CN101474211A (en) * | 2008-10-27 | 2009-07-08 | 上海市农业科学院 | Phellinus linteus extract and preparation method thereof |
| CN103059155A (en) * | 2012-12-28 | 2013-04-24 | 崇左市江州区生产力促进中心 | A method of extracting Phellinus igniarius polysaccharides |
-
2014
- 2014-07-21 CN CN201410355608.6A patent/CN104163874B/en active Active
Patent Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPS4924211A (en) * | 1972-07-01 | 1974-03-04 | ||
| JPH0924211A (en) * | 1995-07-11 | 1997-01-28 | Takane Kitao | Filter device for waste water |
| CN101463093A (en) * | 2007-12-17 | 2009-06-24 | 天津市华泰森淼生物工程技术有限公司 | Technique for extracting polysaccharide from Phellinus linteus |
| CN101323648A (en) * | 2008-07-29 | 2008-12-17 | 上海璞诚生物科技有限公司 | Extraction method and and purification method of Sanghuang mushroom polysaccharide |
| CN101474211A (en) * | 2008-10-27 | 2009-07-08 | 上海市农业科学院 | Phellinus linteus extract and preparation method thereof |
| CN103059155A (en) * | 2012-12-28 | 2013-04-24 | 崇左市江州区生产力促进中心 | A method of extracting Phellinus igniarius polysaccharides |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110038034A (en) * | 2019-04-11 | 2019-07-23 | 安徽省森湶谷药业股份有限公司 | A kind of small molecule water prepares the method and application of phellinus linteus extract |
Also Published As
| Publication number | Publication date |
|---|---|
| CN104163874B (en) | 2016-06-01 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Yang et al. | Antitumor activity of a polysaccharide from Pleurotus eryngii on mice bearing renal cancer | |
| Xia et al. | Partial characterization and immunomodulatory activity of polysaccharides from the stem of Dendrobium officinale (Tiepishihu) in vitro | |
| Sun et al. | Physicochemical properties and immunological activities of polysaccharides from both crude and wine-processed Polygonatum sibiricum | |
| Yu et al. | Immunomodulatory activities of sulfated Cyclocarya paliurus polysaccharides with different degrees of substitution on mouse spleen lymphocytes | |
| CN103463531B (en) | Use of rhizoma smilacis glabrae extract in preparation of lipid-lowering and fat-reducing medicines and health-care foods | |
| CN109276576A (en) | The purposes of plain boiled pork ganoderma lucidum polysaccharide in the preparation of antitumor drugs | |
| Pu et al. | Immunomodulatory effects of a polysaccharide from Solanum nigrum Linne through TLR4-MyD88 signaling pathway | |
| CN103479683A (en) | North cordyceps sinensis sporophores cultivated in large scale and health care and pharmaceutical application thereof | |
| Wang et al. | Preparation and anti-tumor activity of selenium nanoparticles based on a polysaccharide from Paeonia lactiflora | |
| CN102408494A (en) | Grifola frondosa polysaccharide ZZK component and preparation method thereof | |
| CN106749729B (en) | A kind of Smilacina japonica polysaccharide and its preparation method and application | |
| CN105663444A (en) | Compound immunity-enhancing and aging-resisting agent and preparation method thereof | |
| CN101023976B (en) | Medicine composition with anti-virus function | |
| CN102526219A (en) | Extract for Relinqing Granule, as well as preparation method and application of same | |
| CN102600368A (en) | Pharmaceutical composition, as well as preparation method and application thereof | |
| CN104163874B (en) | A kind of Phellinus polysaccharide ZDT and its preparation method | |
| CN105125573A (en) | Application of Borojo polysaccharides to enhancement of immunity of human bodies and animal bodies | |
| CN104877037B (en) | Separation and purification method, products and application of Christia vespertilionis polysaccharides | |
| CN101423558B (en) | Eupatorium adenophorum spreng polysaccharide and preparation method and use | |
| CN103610048B (en) | To chemical damage health food having assistant protection function and preparation method thereof | |
| CN103330211A (en) | Composition with immunoregulation function and preparation method and application of composition | |
| CN102078368B (en) | Chinese traditional medicine extract preparation for treating cancer and preparation method thereof | |
| CN101518556B (en) | Polysaccharide with antitumor and immunologic adjuvant function as well as preparation method and purpose thereof | |
| CN103554289A (en) | Rhizoma atractylodis sinensis polysaccharide and extraction method and applications thereof in preparing anti-tumor medicaments | |
| CN107556401A (en) | A kind of kuh-seng polysaccharide, its preparation method and liver protection and immunomodulation applications |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant |