CN107090049B - Arborvitae total starches and its preparing the purposes in immunosuppressive drug - Google Patents
Arborvitae total starches and its preparing the purposes in immunosuppressive drug Download PDFInfo
- Publication number
- CN107090049B CN107090049B CN201610088573.3A CN201610088573A CN107090049B CN 107090049 B CN107090049 B CN 107090049B CN 201610088573 A CN201610088573 A CN 201610088573A CN 107090049 B CN107090049 B CN 107090049B
- Authority
- CN
- China
- Prior art keywords
- arborvitae
- total starches
- polysaccharide
- preparing
- total
- 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.)
- Active
Links
Classifications
-
- C—CHEMISTRY; METALLURGY
- C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
- C08B—POLYSACCHARIDES; DERIVATIVES THEREOF
- C08B37/00—Preparation of polysaccharides not provided for in groups C08B1/00 - C08B35/00; Derivatives thereof
- C08B37/006—Heteroglycans, i.e. polysaccharides having more than one sugar residue in the main chain in either alternating or less regular sequence; Gellans; Succinoglycans; Arabinogalactans; Tragacanth or gum tragacanth or traganth from Astragalus; Gum Karaya from Sterculia urens; Gum Ghatti from Anogeissus latifolia; Derivatives thereof
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/70—Carbohydrates; Sugars; Derivatives thereof
- A61K31/715—Polysaccharides, i.e. having more than five saccharide radicals attached to each other by glycosidic linkages; Derivatives thereof, e.g. ethers, esters
-
- C—CHEMISTRY; METALLURGY
- C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
- C08B—POLYSACCHARIDES; DERIVATIVES THEREOF
- C08B37/00—Preparation of polysaccharides not provided for in groups C08B1/00 - C08B35/00; Derivatives thereof
- C08B37/0003—General processes for their isolation or fractionation, e.g. purification or extraction from biomass
-
- 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
- A61K36/13—Coniferophyta (gymnosperms)
- A61K36/14—Cupressaceae (Cypress family), e.g. juniper or cypress
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Medicinal Chemistry (AREA)
- General Health & Medical Sciences (AREA)
- Molecular Biology (AREA)
- Polymers & Plastics (AREA)
- Materials Engineering (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Biochemistry (AREA)
- Engineering & Computer Science (AREA)
- Organic Chemistry (AREA)
- Epidemiology (AREA)
- Pharmacology & Pharmacy (AREA)
- Animal Behavior & Ethology (AREA)
- Public Health (AREA)
- Veterinary Medicine (AREA)
- Sustainable Development (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
- Polysaccharides And Polysaccharide Derivatives (AREA)
- Medicines Containing Plant Substances (AREA)
Abstract
The invention belongs to the field of Chinese medicines, be related to a kind of arborvitae total starches, the polysaccharide be from Tibetan medicine Xu Bayuan plant arborvitae (Sabina pingii var.wilsonii) Water-soluble polysccharide of preparation is extracted in branches and leaves, the present invention characterizes the structure features such as its sugared content, glucuronic acid content, protein content, molecular weight distribution, monosaccharide composition；Through determination of activity, the results showed that the arborvitae polysaccharide has apparent anticomplementary action, reaches 23~35 μ g/ml to the half effective inhibition concentration of complement activation classical pathway, suitable with positive control medicine heparin sodium activity；LPS can be significantly inhibited simultaneously stimulates the level of lower Turnover of Mouse Peritoneal Macrophages secretion NO, shows it with good anti-inflammatory effect；Arborvitae total starches of the invention can be used for preparing the immunosupress related drugs such as anticomplement, anti-inflammatory.
Description
Technical field
The invention belongs to the field of Chinese medicines, it is related to a kind of plant polyose and its medicinal usage, and in particular to a kind of from arborvitae
(Sabina pingii var. wilsonii) in extract the total starches of preparation and its preparing anticomplement, in anti-inflammatory drug
Purposes.
Background technique
Prior art discloses the normal Activates of complement system to eliminate external microorganism, remove damage or death in human body
Cell and tissue, maintain body balance in play an important role.However the improper activation of the system can cause human body
The overreaction of immune system causes the damage of human body itself normal tissue, participates in rheumatoid arthritis, systemic red yabbi
The pathologic process of a variety of diseases such as sore, acute respiratory distress syndrome.Current clinically widely used glucocorticoid, ring phosphorus
Although the immunosuppressor such as amide, the cry of certain animals of methylamine butterfly have certain therapeutic effect to certain diseases relevant to excess complement activation,
Since such drug is not single-minded complement inhibitor, poor selectivity, prolonged application can reduce the defensive enginery of body, cause to resist
Infection ability decline, easy secondary infection and spreads hidden focus, generation multiple complications and side effect.Therefore clinically it is badly in need of
Efficiently, low toxicity, single-minded novel complement inhibitor.
Natural polysaccharide is the main active monoid of Chinese herbal medicine, such as panaxan, ganoderma lucidum polysaccharide, pachymaran, fructus lycii
Polysaccharide etc. is often reported with the important activities such as Immune-enhancing effect, antitumor, hypoglycemic, increasingly causes natural drug or health care
The attention of field of food researcher.And part Traditional Chinese Medicine of Qin gre lei polysaccharide, it is shown if cordate houttuynia polysaccharide, lithosperman etc. bright
Aobvious anticomplement, anti-inflammatory activity, the table in terms of preventing and treating the severes inflammatory diseases such as acute lung injury relevant to excess complement activation
Reveal good curative effect (ZL201010022880.4, ZL201110284642.5).
Arborvitae (Sabina pingii var. wilsonii) be Tibetan medicine " Xu bar " one of former plant, be mainly distributed on
Height above sea level 2600-4900 meters of the provinces and regions such as the Hubei northwestward, Southern Shaanxi, SOUTH OF GANSU, Sichuan, Yunnan and Tibet alpine belt, with
Branches and leaves are used as medicine, and have heat-clearing, clearing damp, removing toxic substances and other effects, for treating pneumonia, rheumatic arthritis, rheumatoid arthritis etc.
Disease.Containing the ingredients such as volatile oil, flavones, Phenylpropanoid Glycosides, steroidal in document report arborvitae, but so far there is not yet work to its polysaccharide
Journal of Sex Research report.
Summary of the invention
The object of the present invention is to provide a kind of water-soluble plant total starches and its preparing the purposes in immunosuppressive drug.
Especially arborvitae total starches and its prepare anticomplement, the purposes in anti-inflammatory drug.
Arborvitae total starches of the present invention be from Tibetan medicine Xu Bayuan plant arborvitae (Sabina pingii var.wilsonii) a kind of Water-soluble polysccharide that preparation is extracted in branches and leaves, it is made by following steps: arborvitae branches and leaves being crushed, are used
Ethyl alcohol grease removal carries out water and mentions, and alcohol precipitation after Aqueous extracts concentration, precipitating is freeze-dried after redissolving.
Arborvitae total starches produced by the present invention have following structure feature:
Molecular weight distribution: less than 25000 Da；Monosaccharide composition: with arabinose, galacturonic acid, galactolipin, glucose
Based on, and contain a small amount of rhamnose, mannose, glucuronic acid；Sugared content is greater than 55%, and glucuronic acid content is greater than 8%, and albumen contains
Amount is less than 2%.
It is measured in the present invention using cell in vitro haemolysis method, the results show that the arborvitae total starches have apparent suppression
Excess complement activation effect processed, the IC of three batches batches of arborvitae total starches of measurement50Value is the 23 μ g/ml of μ g/ml~35, with positive control
Medicine heparin sodium (27.81 μ g/ml) activity is quite；
It is measured in the present invention using macrophages secrete NO method, the results show that the arborvitae total starches can significantly inhibit
The peritoneal macrophage NO secretion of LPS induction, has stronger antiphlogistic effects；
The present invention through determination of activity the result shows that, the arborvitae total starches can be used as good innate immunity inhibitor,
Further preparation prepares anticomplement, anti-inflammatory drug, the inflammatory disease caused for treating excess complement activation or pathogenic microorganism.
Detailed description of the invention
Fig. 1 is arborvitae polysaccharide efficient gel electrophoretogram.
Fig. 2 is influence of arborvitae total starches SP1, the SP2 to macrophages secrete NO.
Specific embodiment
Embodiment 1 prepares arborvitae total starches
Arborvitae branches and leaves are crushed, with 95% ethyl alcohol by the solid-liquid ratio of 1:30 carry out heating under reduced pressure reflux (55 DEG C, points 3 times, often
Secondary 4h) dregs of a decoction after grease removal volatilize, with water by the solid-liquid ratio of 1:50 carry out heating under reduced pressure refluxing extraction (70 DEG C, points 3 times, every time
Aqueous extracts 5h) are concentrated into 0.4g crude drug/ml, add 95% ethyl alcohol of 4 times of volumes, alcohol precipitation filters afterwards for 24 hours, is redissolved precipitating with water
After be freeze-dried, obtain total starches；Technique according to this prepares three crowdes of arborvitae total starches SP1, SP2 and SP3 respectively.
The structural characterization of 2 arborvitae total starches of embodiment
(1) molecular weight distribution:
Arborvitae total starches SP3 is configured to 0.3mg/ml aqueous solution, carries out HPLC analysis: using TSK-GEL GMPWXL
Gel column (300 × 7.6 mm), mobile phase are deionized water, 0.8 mL/min of flow velocity, 25 DEG C of column temperature, Detection wavelength 210nm；
Polysaccharide standard items are analyzed with same procedure, show that the molecular weight of arborvitae polysaccharide is approximately less than 25000 Da by comparing spectrogram；
(2) monosaccharide forms:
Take 8 mg polysaccharide samples that 1M TFA 2ml, 110 DEG C of heating 5h, centrifugation are added, supernatant is evaporated, and precipitating washes three with methanol
It is secondary；Then carry out PMP derivatization: acid hydrolysis products are dissolved with 2mldd-H2O, take 1ml that 1ml PMP solution and 1ml 0.3 is added
M NaOH, 70 DEG C of heating 2h hydrolysis.1ml 0.3M HCl is added after cooling to neutralize, is added after chloroform extracts 3 times and discards chloroform layer,
Then it carries out HPLC analysis: using Agilent extend C18 column (250*4.6mm, 5 μm), with PBS(phosphate-buffered
Liquid): acetonitrile is eluent gradient elution, flow velocity 0.7ml/min, Detection wavelength 245nm；By being compareed with monosaccharide standard, obtain
Monosaccharide composition and its relative amount (as shown in table 1), the composition of three batches of arborvitae total monosaccharides is almost the same, mainly contains Arab
Sugar, galacturonic acid, galactolipin, glucose, and containing a small amount of mannose, rhamnose and glucuronic acid；
The monosaccharide composition HPLC of 1 three batches of arborvitae total starches of table analyzes result
(3) sugared content:
It establishes standard curve: weighing arabinose monosaccharide reference substance 3.8mg, be dissolved in water, be settled to 25.0ml as mark
Quasi- solution draws 0,0.5,0.25,0.15,0.1,0.05,0.02 ml standard solution in rub oral examination tube respectively, adds distilled water
Then water is added 5% phenol solution 0.5ml, adds concentrated sulfuric acid 2.5ml and (be preferably applied directly to liquid in the present embodiment to 0.5ml
On face) acutely oscillation 5min, placement 15min measure its absorption value with spectrophotometric at 490nm, draw standard curve afterwards；
Measurement sample solution: arborvitae polysaccharide 3.0mg is weighed, is placed in 10ml volumetric flask, scale, pipette samples liquid are added water to
0.2 ml volume surveys its absorption value in rub oral examination tube, by the above operating procedure, and calculating content by standard curve is 59.96%；
(4) glucuronic acid content:
It establishes standard curve: weighing galacturonic acid 1.9mg, be dissolved in water, be settled to 25.0ml as standard solution, divide
Not Xi Qu 0,0.04,0.08,0.12,0.24,0.40ml standard solution in rub oral examination tube, add distilled water to 0.50 ml, in ice
It is pre-chilled, is added 3.0 ml sodium tetraborates-sulphate reagent (sulfuric acid solution of 0.0125 M sodium tetraborate) in bath, oscillation mixing,
5 min are heated in boiling water-bath.After being cooled to room temperature with ice bath plus 50 μ l, 0.15% hydroxyl biphenyl solution is (with 0.5% sodium hydroxide
Prepare, refrigerator saves, a month stabilization), it shakes up, measures its absorption value with spectrophotometric at 520nm, draw standard curve；
Measure sample solution: precision weighs 1.2 mg of arborvitae polysaccharide, is placed in 10 ml volumetric flasks, adds water to scale, draws
0.5 ml volume of sample liquid surveys its absorption value by the above operating procedure in rub oral examination tube, calculates content by standard curve and is
9.43%；
(5) protein content:
The preparation of Coomassie brilliant G-250 solution: weighing 50mg Coomassie brilliant G-250, is dissolved in 90% ethyl alcohol of 25ml,
85%(W/V is added) phosphoric acid 50ml, finally with distilled water constant volume to 500ml；
It establishes regression equation: taking 25mg bovine serum albumin(BSA), be dissolved in 25ml distilled water, as the stoste of 1.0mg/ml,
The stoste is taken to obtain the bovine serum albumin solution of 0.5,0.25,0.125,0.0625,0.0312 mg/ml with water two-fold dilution, from
0.1 ml of draw solution is put into 10 ml tool plug test tubes respectively in each test tube, using 0.1 ml water as blank, 5 ml is added and examine horse
This brilliant blue G-250 protein reagent, Gai Sai, mixing survey absorbance under 595 nm after placing 5 min, draw standard curve；
Measurement sample solution: weighing arborvitae polysaccharide 3.0mg, be placed in 10 ml volumetric flasks, adds water to scale, draws 0.1
For ml in rub oral examination tube, surveying its absorption value to calculate content by standard curve by the above operating procedure is 1.05%.
3 cell haemolysis method of embodiment measures arborvitae total starches anti-complement activity
About 2.0mg arborvitae total starches are taken, it is accurately weighed, it is dissolved in 0.8ml BBS(barbiturate buffer) in, to dilute again
Release, obtain 2.5,1.25,0.625,0.3125,0.1563,0.0781,0.0391,0.0195mg/ml sample solution it is each
0.4ml draws in 0.2ml to 0.4ml BBS do blank control respectively；It is separately added into 0.2ml's in remaining sample solution
The sheep red blood cell solution of complement, the hemolysin of 0.1ml and 0.1ml 2% takes supernatant to survey absorption value at 405nm after centrifugation, button
Except the IC for the haemolysis inhibiting rate for calculating three batches of arborvitae polysaccharide after the absorption value of blank control50Respectively SP1:25.04 μ g/ml,
SP2: 35.40μg/ml,SP3: 23.82μg/ml；The IC of positive control drug heparin sodium50For 27.81 μ g/ml.
Embodiment 4 measures influence of the arborvitae total starches to macrophages secrete NO
It is thin to collect mouse peritoneal macrophage for 4 days 5% sulphur glycollate culture mediums of intraperitoneal injection before Balb/c mouse experiment after 4 days
Cell concentration is adjusted to 5 × 10 by born of the same parents5/ ml kind enters in 96 orifice plates, and the hole 200ul/ is placed in adhere-wall culture 2h in cell incubator
(37 DEG C, 5% CO2)；It is loaded according to following grouping: (1) blank control group；(2) the independent processing group of polysaccharide；(3) LPS is handled
Group (the final concentration of 1 μ g/ml of LPS)；(4) polysaccharide and LPS Combined Treatment group；(5) positive drug dexamethasone and LPS Combined Treatment
Group (DEX)；Every group of four multiple holes are placed in incubator and cultivate 24 h, collect supernatant, using the level of Griess method measurement NO, take
50 μ l Griess reagents are added in 50 μ l of supernatant, and 10 min are placed at room temperature for after mixing, react it sufficiently, 540 nm detection
Absorbance value establishes NO standard curve (0-200 μM of measurement range) by standard solution of sodium nitrite solution, determines each experiment
NO in group cells and supernatant is horizontal；
The experimental results showed that can significantly raise macrophage NO secretion level when the mono- processing of SP1, SP2, there is Immune-enhancing effect
Activity；Meanwhile SP1, SP2 can significantly inhibit the peritoneal macrophage NO secretion of LPS induction, have stronger antiphlogistic effects；
The result shows that SP1, SP2 can have dual regulation to function of peritoneal macrophages.
Claims (3)
1. arborvitae total starches, which is characterized in that the arborvitae total starches are water-soluble plant total starches, derive from arborvitae Sabina
Pingii var. wilsonii, has the feature that
Molecular weight is less than 25000 Da；Total sugar content is not less than 55%, and glucuronic acid content is higher than 8%, and protein content is lower than 2%；Monosaccharide
Composition contains rhamnose, mannose and glucuronic acid based on arabinose, galacturonic acid, galactolipin and glucose.
2. arborvitae total starches as described in claim 1 are preparing the purposes in Complement inhibition drug.
3. arborvitae total starches purposes in preparing anti-inflammatory drugs as described in claim 1.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610088573.3A CN107090049B (en) | 2016-02-17 | 2016-02-17 | Arborvitae total starches and its preparing the purposes in immunosuppressive drug |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610088573.3A CN107090049B (en) | 2016-02-17 | 2016-02-17 | Arborvitae total starches and its preparing the purposes in immunosuppressive drug |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN107090049A CN107090049A (en) | 2017-08-25 |
| CN107090049B true CN107090049B (en) | 2019-10-08 |
Family
ID=59646009
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201610088573.3A Active CN107090049B (en) | 2016-02-17 | 2016-02-17 | Arborvitae total starches and its preparing the purposes in immunosuppressive drug |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN107090049B (en) |
Families Citing this family (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110051682B (en) * | 2018-01-18 | 2022-01-18 | 复旦大学 | Application of polysaccharide of thuja in preparation of medicine for preventing and treating viral acute lung injury |
| CN110498864B (en) * | 2018-05-17 | 2021-09-07 | 复旦大学 | Selaginella polysaccharide, preparation method thereof and application thereof in preparation of anticomplement medicines |
| CN111825775B (en) * | 2019-04-18 | 2021-11-02 | 复旦大学 | Cupressus sempervirens polysaccharide, preparation method thereof and application thereof in preparing antitumor drugs |
| CN116425897B (en) * | 2023-04-23 | 2024-03-29 | 青岛农业大学 | Rosa roxburghii polysaccharide, and preparation method and application thereof |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US5773005A (en) * | 1995-06-09 | 1998-06-30 | Takahashi; Hidehiko | Purified flavonoid and diterpene 5α-reductase inhibitors from thuja orientalis for androgen-related diseases |
| CN102090989A (en) * | 2011-01-27 | 2011-06-15 | 中国人民解放军北京军区总医院 | Application of phellodendron polysaccharides for resisting ultraviolet radiation and preparation method of anti-ultraviolet sun-screening agent |
| CN105061615A (en) * | 2015-07-28 | 2015-11-18 | 华南理工大学 | Chinese arborvitae twig polysaccharide with antiviral and immunity-enhancing activity, and preparation method and application thereof |
-
2016
- 2016-02-17 CN CN201610088573.3A patent/CN107090049B/en active Active
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US5773005A (en) * | 1995-06-09 | 1998-06-30 | Takahashi; Hidehiko | Purified flavonoid and diterpene 5α-reductase inhibitors from thuja orientalis for androgen-related diseases |
| CN102090989A (en) * | 2011-01-27 | 2011-06-15 | 中国人民解放军北京军区总医院 | Application of phellodendron polysaccharides for resisting ultraviolet radiation and preparation method of anti-ultraviolet sun-screening agent |
| CN105061615A (en) * | 2015-07-28 | 2015-11-18 | 华南理工大学 | Chinese arborvitae twig polysaccharide with antiviral and immunity-enhancing activity, and preparation method and application thereof |
Also Published As
| Publication number | Publication date |
|---|---|
| CN107090049A (en) | 2017-08-25 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN107090049B (en) | Arborvitae total starches and its preparing the purposes in immunosuppressive drug | |
| Xie et al. | Polysaccharide from alfalfa activates RAW 264.7 macrophages through MAPK and NF-κB signaling pathways | |
| CN102366432B (en) | Saussurea involucrata culture extract and novel purpose thereof | |
| Zhang et al. | Extraction, purification, hypoglycemic and antioxidant activities of red clover (Trifolium pratense L.) polysaccharides | |
| CN102417544A (en) | Ziyang selenium-rich green tea selenium-containing polysaccharide, and preparation method and application thereof | |
| Aher et al. | Pharmacological study of Tinospora cordifolia as an immunomodulator | |
| EP3178482A1 (en) | Applications of phillyrin/phillygenin composition in preparing drugs or health care products for relieving and/or treating viral diseases | |
| CN100457139C (en) | Method for preparing a Shuanhuanglian injection and the component detecting method | |
| CN101684163B (en) | Gastrodia elata polysaccharide sulphated derivative and preparation method and antitumor application thereof | |
| Wu et al. | Immunoregulatory effects of Tetrastigma hemsleyanum polysaccharide via TLR4-mediated NF-κB and MAPK signaling pathways in Raw264. 7 macrophages | |
| CN101843667B (en) | Shuanghuanglian medicinal composition and preparation method thereof | |
| CN106749729B (en) | A kind of Smilacina japonica polysaccharide and its preparation method and application | |
| CN101890084B (en) | Semen nigellae total glycoside extract and preparation method and application thereof | |
| CN100443498C (en) | Use of anti-inflammatory medicine for scheelite total saponin and its saponin compound | |
| CN100408594C (en) | Lonicera macranthoides active total saponin extract and its preparing method and use | |
| CN1879688B (en) | Preparation for treating wind-heat type cold, its preparation process and quality control method | |
| CN104586831A (en) | Application of high-content salvianolic acid B to prepare medicines for resisting cervical carcinoma and preparation method thereof | |
| Yang et al. | Study onin vitro anti-tumor activity of Bidens bipinnata L. extract | |
| CN107213239B (en) | Traditional Chinese medicine composition and preparation method and application thereof | |
| CN102362877B (en) | Application of Pouzolzia extract | |
| CN103558303A (en) | Method for measuring content of oleanolic acid in glossy privet fruit and astragalus membranaceus healthy energy-strengthening preparation | |
| CN102028722B (en) | Extract of effective parts of total saponins in pterocephalus hookeri as well as preparation method and application thereof | |
| CN103405375B (en) | Poecilobdella manillensis active extract and preparation method and application thereof | |
| CN107090050B (en) | Screwtree root polysaccharide and preparation method thereof and preparing the purposes in anticomplement medicament | |
| CN1927309B (en) | Traditional Chinese medicine composition detection method |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |