CN105030838A - Preparing method of sea anemone crude extract and anti-tumor application of sea anemone crude extract - Google Patents
Preparing method of sea anemone crude extract and anti-tumor application of sea anemone crude extract Download PDFInfo
- Publication number
- CN105030838A CN105030838A CN201510292554.8A CN201510292554A CN105030838A CN 105030838 A CN105030838 A CN 105030838A CN 201510292554 A CN201510292554 A CN 201510292554A CN 105030838 A CN105030838 A CN 105030838A
- Authority
- CN
- China
- Prior art keywords
- sea anemone
- sea
- crude extract
- pure water
- preparation
- 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.)
- Pending
Links
Landscapes
- Medicines Containing Material From Animals Or Micro-Organisms (AREA)
Abstract
The invention relates to a preparing method of sea anemone crude extract. The preparing method includes the steps of 1, washing bred live sea anemones in pure water, placing the washed sea anemones in a beaker, adding pure water into the beaker according to a sea anemone to pure water ratio being 1:1.5 to 1:2, and performing freeze thawing twice to three times at -60 DEG C; 2, moderately unfreezing the sea anemones obtained in the step 1, and performing coarse filtration; 3, centrifuging filtrate obtained in the step 2 at 4 DEG C to 6 DEG C at the speed of 12000-14000 r/min for 10 to 15 min, and collecting supernate to obtain sea anemone crude venom; 4, subjecting the sea anemone crude venom obtained in the step 3 to fractional precipitation with acetone, centrifuging precipitate obtained in each step for 15min, collecting final precipitate, and freeze-drying the final precipitate to obtain sea anemone crude toxin. The invention further provides application of the sea anemone crude toxin in the preparation of anti-lung cancer medicaments.
Description
Technical field
The present invention relates to marine prods field of deep, particularly a kind of preparation method of sea anemone crude extract and the application of anti-tumor thereof.
Background technology
Sea anemone (SeaAnemone) has another name called extra large Flos Chrysanthemi, it is a kind of Marine coelenterata, belonging to Coelenterata (Coelenterata), Anthozoa (Anthozoa), Actiniaria (actiniaria) animal, is animal more original in ocean.Sea anemone has 6 section 37 kinds, is extensively distributed in the torrid zone and warm Tropical Ocean Area, mainly anchors on marine rock or in silt.Sea anemone has the effects such as YIN nourishing and YANG strengthening, astringing to arrest diarrhea, removing dampness and killing parasites.20 century 70s are started to the research of sea anemone chemical composition, numerous cnidoblasts is contained in the tentacle of sea anemone and health, wherein there is the organelle-nematocyst storing venom, when sea anemone runs into physics or chemical stimulation, ecthoaeum can thrust prey health and discharges venom and benumb prey, grabs prey for catching or resists harmful animal.But because it contains various bioactivators, the concern of extremely scholars, mainly concentrates on the research of the aspects such as Actinia neurotoxin, sea anemone cytolysin and sea anemone inhibitors of ion channels in recent years.Also scholar is had to find to have antineoplastic active substance in sea anemone, as actinocongestin has apoptosis-induced effect to glioma U251 cell, apoptosis-induced by increase Fas expression.Sea anemone crude extract activates Caspases by mitochondria pathway and reduces mitochondrial membrane potential and induce A549 apoptosis, sea anemone synocytotoxin Gigantoxin-4 has the effect killing and wounding MCF-7 Breast Cancer Cell and cancer of pancreas SW1991 cell, such as StI, EqtII etc. have stronger inhibitory action to different tumor cells, because it has biologic activity widely, one of study hotspot becoming marine natural products.Zhoushan Anthopleura lanthogrammica (Berkly). (AnthopLeuraxanthograrnmica) belongs to sea anemone section, Anthopleura belongs to, multiple lps molecule is rich in its body, a kind of irritability polypeptide toxin and the multiple insecticidal peptide with strong insecticidal activity and low mammal activity are obtained, be the ideal object carrying out marine natural product exploitation, but there is not yet report about the research of Zhoushan Anthopleura lanthogrammica (Berkly). antitumor action.
Pulmonary carcinoma is a class disease of serious threat human health, also has very high sickness rate, and finds effective cancer therapy drug and method, thoroughly capture cancer in China, is the important research topic of world medical circle.Current clinical existing anti-pulmonary carcinoma, while performance antitumaous effect, has larger toxic and side effects more, and therefore active high, that determined curative effect, toxic and side effects the are low medicine of research and development becomes the key issue of anti-lung-cancer medicament research.
Summary of the invention
A technical problem to be solved by this invention is the preparation method providing a kind of sea anemone crude extract for the present situation of prior art.
Another technical problem to be solved by this invention is the anti-tumor application providing a kind of above-mentioned sea anemone crude extract for the present situation of prior art.
The present invention solves the problems of the technologies described above adopted technical scheme: the preparation method of this sea anemone crude extract, is characterized in that: comprise the following steps:
1) cleaned in pure water by the sea anemone alives after raising, remove its surface impurity, be positioned in beaker by the sea anemone after clean and add pure water, the solid-liquid ratio of sea anemone and pure water is that 1:1.5 ~ 1:2 carries out 2 ~ 3 freeze thawing at-60 DEG C;
2) by step 1) in sea anemone suitably thaw at 20 DEG C ~ 28 DEG C after carry out coarse filtration;
3) get step 2) in filtrate, in 4 DEG C ~ 6 DEG C, centrifugal 10 ~ 15min under 12000 ~ 14000r/min condition, collect supernatant and obtain sea anemone crude venom;
4) by step 3) in sea anemone crude venom with 30%, 60%, 90% acetone carry out fractional precipitation, and by the precipitate of each step gained in 4 DEG C ~ 6 DEG C, centrifugal 15min under 12000 ~ 14000r/min condition, after final gained precipitation is collected postlyophilization, obtain sea anemone slightly malicious, save backup in-20 DEG C.
Further, described step 1) in the solid-liquid ratio of sea anemone and pure water be 1:1.5.
Further, described step 4) in acetone is carried out pre-cooling at-20 DEG C.
Although bibliographical information sea anemone synocytotoxin has the effect killing and wounding MCF-7 Breast Cancer Cell and cancer of pancreas SW1991 cell, as StI, EqtII etc. have stronger inhibitory action to different tumor cells, but because pulmonary carcinoma and breast carcinoma and cancer of pancreas are dissimilar tumor, there is different indications, therefore, the thick poison of the sea anemone that can not push away also has the effect suppressing pulmonary carcinoma.
Therefore, the present invention also provides the thick poison of a kind of sea anemone utilizing the preparation method of sea anemone crude extract to prepare preparing the application in anti-lung-cancer medicament.
Compared with prior art, tool of the present invention has the following advantages: by sea anemone carry out multigelation, acetone fractional precipitation method extract obtain actinocongestin crude extract, adopt Freeze Drying Technique simultaneously, the original biological activity of sea anemone can be retained preferably, and through mtt assay detection display, actinocongestin crude extract has strong anti tumor activity in vitro, can be used for preparing tumor.
Accompanying drawing explanation
The concentration of the thick poison of Fig. 1 sea anemone is on the impact of SPC-A1 cell inhibitory rate;
The thick poison of Fig. 2 sea anemone action time on cell proliferation impact;
The thick poison of Fig. 3 sea anemone suppresses the microexamination figure of pulmonary carcinoma SPC-A1 Growth of Cells, and wherein, A figure contrasts, and in B figure, the concentration of the concentration of actinocongestin to be the concentration of actinocongestin in 0.625mg/mL, C figure be actinocongestin in 1.25mg/mL, D figure is 2.5mg/mL;
The thick poison of Fig. 4 sea anemone suppresses the HE colored graph of pulmonary carcinoma SPC-A1 Growth of Cells, and wherein, A figure contrasts, and in B figure, the concentration of the concentration of actinocongestin to be the concentration of actinocongestin in 0.625mg/mL, C figure be actinocongestin in 1.25mg/mL, D figure is 2.5mg/mL;
The thick poison of Fig. 5 sea anemone suppresses the AO/EB fluorescence staining figure of pulmonary carcinoma SPC-A1 Growth of Cells, wherein, A figure is contrast, and in B figure, the concentration of actinocongestin is 0.625mg/mL, in C figure, the concentration of actinocongestin is the concentration of actinocongestin in 1.25mg/mL, D figure is 2.5mg/mL.
Detailed description of the invention
Below by way of by reference to the accompanying drawings and embodiment the invention will be further described.
The preparation of embodiment 1 sea anemone crude extract
1) sea anemone alive after raising is cleaned in pure water, remove its surface impurity, sea anemone after cleaning is positioned over beaker, a certain amount of pure water is added by material ratio 1:1.5, and be placed in-60 DEG C of ultralow temperature freeze overnight, naturally thaw at taking out 20 DEG C next day, after all dissolving, be placed in refrigerator-freezer more freezing, freeze thawing step cycle like this 3 times;
2) by step 1) in the gauze parcel sea anemone of all melting suitably thaw at 20 DEG C after carry out coarse filtration, discard the impurity in sea anemone body;
3) get its filtrate, filtrate, at 4 DEG C, centrifugal 10min under 12000r/min condition, is collected supernatant and is obtained sea anemone crude venom;
4) by step 3) in sea anemone crude venom add the acetone fractional precipitation of-20 DEG C precooled 30%, 60%, 90% respectively, stir with Glass rod rapidly and make rapid deposition, the precipitate of each step gained is at 4 DEG C, the centrifugal 15min of 12000r/min.Namely final gained precipitation is collected postlyophilization is that gained sea anemone is slightly malicious, saves backup in-20 DEG C.
The preparation of embodiment 2 sea anemone crude extract
1) sea anemone alive after raising is cleaned in pure water, remove its surface impurity, sea anemone after cleaning is positioned over beaker, a certain amount of pure water is added by material ratio 1:2, and be placed in-60 DEG C of ultralow temperature freeze overnight, naturally thaw at taking out 22 DEG C next day, after all dissolving, be placed in refrigerator-freezer more freezing, freeze thawing step cycle like this 2 times;
2) by step 1) in the gauze parcel sea anemone of all melting suitably thaw at 28 DEG C after carry out coarse filtration, discard the impurity in sea anemone body;
3) get its filtrate, filtrate, at 6 DEG C, centrifugal 15min under 14000r/min condition, is collected supernatant and is obtained sea anemone crude venom;
4) by step 3) in sea anemone crude venom add the acetone fractional precipitation of-20 DEG C precooled 30%, 60%, 90% respectively, stir with Glass rod rapidly and make rapid deposition, the precipitate of each step gained is at 6 DEG C, the centrifugal 15min of 14000r/min.Namely final gained precipitation is collected postlyophilization is that gained sea anemone is slightly malicious, saves backup in-20 DEG C.
The preparation of embodiment 3 sea anemone crude extract
1) sea anemone alive after raising is cleaned in pure water, remove its surface impurity, sea anemone after cleaning is positioned over beaker, a certain amount of pure water is added by material ratio 1:1.8, and be placed in-60 DEG C of ultralow temperature freeze overnight, naturally thaw at taking out 20 DEG C next day, after all dissolving, be placed in refrigerator-freezer more freezing, freeze thawing step cycle like this 3 times;
2) by step 1) in the gauze parcel sea anemone of all melting suitably thaw at 25 DEG C after carry out coarse filtration, discard the impurity in sea anemone body;
3) get its filtrate, filtrate, at 5 DEG C, centrifugal 12min under 13000r/min condition, is collected supernatant and is obtained sea anemone crude venom;
4) by step 3) in sea anemone crude venom add the acetone fractional precipitation of-20 DEG C precooled 30%, 60%, 90% respectively, stir with Glass rod rapidly and make rapid deposition, the precipitate of each step gained is at 5 DEG C, the centrifugal 15min of 13000r/min.Namely final gained precipitation is collected postlyophilization is that gained sea anemone is slightly malicious, saves backup in-20 DEG C.
Embodiment 4 cell culture
Human lung adenocarcinoma SPC-A1 cell culture in containing streptomycin, penicillin and 10% hyclone 1640 nutritional solutions in, 37 DEG C, containing 5%CO
2incubator in cultivate, in time covering with more than 80% with containing 0.25% trypsin carry out had digestive transfer culture, get the cell being in exponential phase and test.
Embodiment 5 cell growth inhibition assay (mtt assay)
Get SPC-A1 cell and make single cell suspension, carry out counting and being diluted to 1 × 10
4/ mL concentration is seeded to 96 orifice plates, every hole 200 μ L.Discard culture fluid after cultivating 24h under normal condition, blank group, matched group and medicine group are set.Blank group does not add cell, and just 1640 nutritional solutions, give 1640 nutritional solutions in cellular control unit, medicine group add in 1640 nutritional solutions mass gradient be 0.625,1.25, the sea anemone of 2.5mg/mL is slightly malicious, often group establishes 3 multiple holes, after cultivating 24h, 48h, 72h respectively, abandons culture fluid.Every hole adds the 200 μ LPBS buffer containing 10%MTT, continues to cultivate 4h.Experiment terminates, and abandon liquid, every hole adds the DMSO of 150 μ L, and fully vibrate 10min.Survey its absorbance A value in microplate reader 490nm place, calculate cell inhibitory effect index (IR), above experiment repetition 3 times.IR (%)=[(control group A value-medicine group A value)/(control group A value-blank group A value)] × 100%.And calculate IC
50time drug level.From MTT experiment result as depicted in figs. 1 and 2, in certain sphere of action, along with increase and the prolongation of action time of sea anemone crude extract concentration, suppression ratio increases gradually, has obvious dosage and time dependence.As sea anemone crude extract concentration 2.5mg/mL, when action time is 72h, proliferation inhibition rate is 82.12%.The IC of effect 24h, 48h and 72h
50be respectively 2.52mg/mL, 2.05mg/mL and 1.89mg/mL.
In order to observe the living cells of cultivation, the observed result under inverted microscope is shown in Fig. 3-A, and matched group SPC-A1 Growth of Cells is good, and form is full, grows, clear border in Epithelial.As Fig. 3-B, 3-C, after medication, cell volume reduces, and cellular contraction becomes circle, and form is irregular, and cell attachment ability declines, and part cells float is in culture fluid.As Fig. 3-D, when drug level is 2.5mg/mL, cell loses normal morphology, part cell rupture, visible significantly cell debris and particulate matter in culture fluid, part necrocytosis.
Embodiment 6HE dyeing observation of cell form
The coverslip processed is put into six orifice plates, and inoculum density is 1 × 10
4the SPC-A1 cell suspension of/mL, cultivates after 24h and removes culture fluid, set up matched group and medicine group, the mass concentration of the thick poison of sea anemone is 0.625,1.25,2.5mg/mL, cultivate 48h and under inverted microscope, observe form and take pictures.After remove culture fluid, wash with PBS, 20min fixed by 95% ethanol, and PBS washes, haematoxylin dyeing 5min, tap water embathes and makes nucleus oil blackeite, 0.5min is redyed in Yihong, ethanol (50%, 75%, 95%, 100%) serial dehydration, transparent 2 times of dimethylbenzene, neutral gum mounting, optical microphotograph Microscopic observation is also taken pictures.From HE coloration result as shown in Fig. 4-A, matched group SPC-A1 cell size is even, and kernel number is many.Along with the increase of drug level, cellular morphology occurs irregular.When drug level is 0.625mg/mL, there is cavity in kytoplasm, kernel decreased number, as shown in Fig. 4-B; When drug level is 1.25mg/mL, cell volume obviously reduces, and occurs gemma, karyopycnosis, part hypochromatosis, as shown in Fig. 4-C; When drug level is 2.5mg/mL, cellular morphology is different, most cells karyolysis, disappears, and the nucleus also pyknosis further of existence, occurs the morphological change of apoptosis, as shown in Fig. 4-D.Illustrate that the thick poison of sea anemone can suppress the growth of SPC-A1 cell, and apoptosis-induced, and the apoptosis of lung carcinoma cell is the important evaluation index of anticancer lung-cancer medicament.
Embodiment 7AO/EB Fluorescent Staining Observation cellular morphology
Cell inoculation is divided into groups with embodiment 4 with experiment.After 24h, take out coverslip, PBS washes 2-3 time, carries out AO/EB dyeing.The compound method of AO/EB is as follows: get each 1mg of AO, EB, be dissolved in respectively in the PBS buffer of 10mLPH7.2, mixing shakes up, now with the current and lucifuge.On microscope slide, drip 40 μ LPBS and 10 μ LAO/EB mixed liquors before observing, carefully clean the PBS that coverslip remains, will one of cell be had to face down, take pictures in fluorescence microscopy Microscopic observation.As shown in fig. 5-A, cellular control unit size is even, and karyon presents uniform green fluorescence for AO/EB fluorescence staining result.After medication there is metamorphosis in cell all in various degree.0.625mg/mL group presents yellow-green fluorescence, as shown in fig. 5-b.1.25mg/mL group abnormal cell showed increased, core presents stronger yellow-green fluorescence or yellow fluorescence, as shown in fig. 5-c; 2.5mg/mL group cell volume obviously reduces, part hypochromatosis, and part nucleus deflection cell side, karyopycnosis is in " crescent moon " shape, and occur apoptotic body, core is dyed to orange-red fluorescence, as shown in Fig. 5-D.Similarly, the cellular morphology change after the dosing of AO/EB Fluorescent Staining Observation.SPC-A1 cell after dosing occurs that cell volume reduces, there is " gemma " phenomenon and apoptotic body in cell peripheral, cavity is there is in Cytoplasm, kernel decreased number, karyopyknosis or karyolysis disappear, these morphologic changes, illustrate that the thick poison of sea anemone can suppress the growth of SPC-A1 cell.
Claims (4)
1. a preparation method for sea anemone crude extract, is characterized in that: comprise the following steps:
1) cleaned in pure water by the sea anemone alives after raising, remove its surface impurity, be positioned in beaker by the sea anemone after clean and add pure water, the solid-liquid ratio of sea anemone and pure water is that 1:1.5 ~ 1:2 carries out 2 ~ 3 freeze thawing at-60 DEG C;
2) by step 1) in sea anemone suitably thaw at 20 DEG C ~ 28 DEG C after carry out coarse filtration;
3) get step 2) in filtrate, in 4 DEG C ~ 6 DEG C, centrifugal 10 ~ 15min under 12000 ~ 14000r/min condition, collect supernatant and obtain sea anemone crude venom;
4) by step 3) in sea anemone crude venom with 30%, 60%, 90% acetone carry out fractional precipitation, and by the precipitate of each step gained in 4 DEG C ~ 6 DEG C, centrifugal 15min under 12000 ~ 14000r/min condition, after final gained precipitation is collected postlyophilization, obtain sea anemone slightly malicious, save backup in-20 DEG C.
2. the preparation method of sea anemone crude extract according to claim 1, is characterized in that described step 1) in the solid-liquid ratio of sea anemone and pure water be 1:1.5.
3. the preparation method of sea anemone crude extract according to claim 1, is characterized in that described step 4) in acetone is carried out pre-cooling at-20 DEG C.
4. the application in anti-lung-cancer medicament prepared by the thick poison of sea anemone prepared by the preparation method of the sea anemone crude extract according to claims 1 to 3 any one claim.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201510292554.8A CN105030838A (en) | 2015-05-31 | 2015-05-31 | Preparing method of sea anemone crude extract and anti-tumor application of sea anemone crude extract |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201510292554.8A CN105030838A (en) | 2015-05-31 | 2015-05-31 | Preparing method of sea anemone crude extract and anti-tumor application of sea anemone crude extract |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN105030838A true CN105030838A (en) | 2015-11-11 |
Family
ID=54438202
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201510292554.8A Pending CN105030838A (en) | 2015-05-31 | 2015-05-31 | Preparing method of sea anemone crude extract and anti-tumor application of sea anemone crude extract |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN105030838A (en) |
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106560518A (en) * | 2016-11-24 | 2017-04-12 | 浙江海洋大学 | Preparing method of authopleura midori uchida muramatsu resisting prostatic cancer oligopeptides |
| CN107375338A (en) * | 2017-07-31 | 2017-11-24 | 中国医学科学院药用植物研究所海南分所 | A kind of preparation method and application of Onchidium struma extract |
| CN112458138A (en) * | 2020-11-30 | 2021-03-09 | 海南医学院 | Preparation method and application of actinia violaceus enzymatic hydrolysis polypeptide |
| CN113663052A (en) * | 2021-07-30 | 2021-11-19 | 海南康盾生物制药有限公司 | Pharmaceutical composition for treating brain glioma and preparation method thereof |
| CN113683671A (en) * | 2021-05-31 | 2021-11-23 | 海南医学院 | Preparation method and anti-tumor application of actinia violaceus polypeptide toxin |
Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102399280A (en) * | 2011-11-07 | 2012-04-04 | 浙江海洋学院 | Sea anemone cardiotonic peptide and extraction method and application thereof |
| CN103408650A (en) * | 2013-07-26 | 2013-11-27 | 张朝晖 | Cereus sinensis verrill soluble cell peptide CCT-I and separating and purifying method and application thereof |
| CN103421869A (en) * | 2012-05-14 | 2013-12-04 | 宁波博丰生物科技有限公司 | Preparation method of sea anemone low molecular polypeptides |
| CN103694374A (en) * | 2013-12-18 | 2014-04-02 | 青岛福创环境科技有限公司 | Method for extracting agar, fucoidin and protein from gracilaria as raw material by using enzyme process |
| CN104231105A (en) * | 2014-09-28 | 2014-12-24 | 浙江工业大学 | Preparation method for sea anemone polysaccharide and application thereof to tumor resistance |
-
2015
- 2015-05-31 CN CN201510292554.8A patent/CN105030838A/en active Pending
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102399280A (en) * | 2011-11-07 | 2012-04-04 | 浙江海洋学院 | Sea anemone cardiotonic peptide and extraction method and application thereof |
| CN103421869A (en) * | 2012-05-14 | 2013-12-04 | 宁波博丰生物科技有限公司 | Preparation method of sea anemone low molecular polypeptides |
| CN103408650A (en) * | 2013-07-26 | 2013-11-27 | 张朝晖 | Cereus sinensis verrill soluble cell peptide CCT-I and separating and purifying method and application thereof |
| CN103694374A (en) * | 2013-12-18 | 2014-04-02 | 青岛福创环境科技有限公司 | Method for extracting agar, fucoidin and protein from gracilaria as raw material by using enzyme process |
| CN104231105A (en) * | 2014-09-28 | 2014-12-24 | 浙江工业大学 | Preparation method for sea anemone polysaccharide and application thereof to tumor resistance |
Cited By (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106560518A (en) * | 2016-11-24 | 2017-04-12 | 浙江海洋大学 | Preparing method of authopleura midori uchida muramatsu resisting prostatic cancer oligopeptides |
| CN106560518B (en) * | 2016-11-24 | 2020-02-07 | 浙江海洋大学 | Preparation method of anti-prostate cancer oligopeptide from actinia viridis |
| CN107375338A (en) * | 2017-07-31 | 2017-11-24 | 中国医学科学院药用植物研究所海南分所 | A kind of preparation method and application of Onchidium struma extract |
| CN112458138A (en) * | 2020-11-30 | 2021-03-09 | 海南医学院 | Preparation method and application of actinia violaceus enzymatic hydrolysis polypeptide |
| CN112458138B (en) * | 2020-11-30 | 2023-05-05 | 海南医学院 | Preparation method and application of sea anemone enzymatic polypeptide |
| CN113683671A (en) * | 2021-05-31 | 2021-11-23 | 海南医学院 | Preparation method and anti-tumor application of actinia violaceus polypeptide toxin |
| CN113683671B (en) * | 2021-05-31 | 2023-08-25 | 海南医学院 | Preparation method of echinacea purpurea polypeptide toxin and anti-tumor application thereof |
| CN113663052A (en) * | 2021-07-30 | 2021-11-19 | 海南康盾生物制药有限公司 | Pharmaceutical composition for treating brain glioma and preparation method thereof |
| CN113663052B (en) * | 2021-07-30 | 2024-01-16 | 广东康盾制药有限公司 | Pharmaceutical composition for treating brain glioma and preparation method thereof |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN105030838A (en) | Preparing method of sea anemone crude extract and anti-tumor application of sea anemone crude extract | |
| Field et al. | A Hematodinium-like dinoflagellate infection of the Norway lobster Nephrops norvegicus: observations on pathology and progression of infection | |
| CN102018759B (en) | Rosmarinic acid, rosmarinic acid-containing common selfheal fruit-spike active ingredient and preparation methods and application thereof to prevention and treatment of cancer postoperative metastasis | |
| CN104140953B (en) | Breast cancer multidrug-resistant cell strain constructed by virtue of epirubicin induction as well as construction method and application thereof | |
| Pietsch et al. | Freshwater fishes of the Kuril Islands and adjacent regions | |
| CN102038690A (en) | Application of bufarenogin to preparing antitumor medicinal preparation | |
| CN101391961A (en) | Phyllanthus emblica leaf or fruit anticancer Chinese medicine and preparation method and use thereof | |
| Murugami et al. | Helminth parasites of farmed fish and water birds in Kirinyaga County, Kenya | |
| CN104940239A (en) | Cockroach extract, and preparation method and use thereof | |
| CN104083522B (en) | The processing method and extract particles and purposes of a kind of black nightshade fresh fruit | |
| Ke et al. | Impacts of two biomanipulation fishes stocked in a large pen on the plankton abundance and water quality during a period of phytoplankton seasonal succession | |
| CN102319425A (en) | Wound healing agent and radix astragali composite freeze-dried platelet gel kit for preparing wound healing agent | |
| Sudatri et al. | Kidney histology and broiler serum creatinine levels supplemented with a mixture of water extract of turmeric and tamarind fruit | |
| CN102078317A (en) | Dihydroartemisinine-phospholipid complex and preparation and application thereof | |
| Chen et al. | Immunology and diseases | |
| Chen et al. | Study on the effect of polysaccharides from Solanum nigrum Linne on cellular immune function in tumour-bearing mice | |
| CN103599162A (en) | Application of Tibet rhodiola crenulata extract in preparing medicament for resisting intestinal inflammation | |
| CN106167491A (en) | The compound of a kind of effective suppression lung cancer metastasis and application thereof | |
| CN103966357B (en) | A kind of fast quantitative measurement method for detecting of Apostichopus japonicus coelomic fluid virus | |
| Russell | Epidemiology of malaria in the Philippines | |
| CN103583589A (en) | Karenia mikimotoi inhibitor, preparation method and application thereof | |
| CN103316062A (en) | Novel application of mulgedium tataricum | |
| CN102973633B (en) | Method for screening anti-bacterial adhesion inhibitors and preparation of anti-adhesion oral liquid | |
| CN102631446B (en) | Grapefruit peel flavone extract, preparation and application thereof | |
| CN103638068A (en) | Grateloupia livida extracts, and preparing methods and applications thereof |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| RJ01 | Rejection of invention patent application after publication | ||
| RJ01 | Rejection of invention patent application after publication |
Application publication date: 20151111 |