CN101306020A - Use of clostridium butyricum in preparing medicine composition for preventing and curing intestinal cancer - Google Patents
Use of clostridium butyricum in preparing medicine composition for preventing and curing intestinal cancer Download PDFInfo
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- CN101306020A CN101306020A CNA2007101033977A CN200710103397A CN101306020A CN 101306020 A CN101306020 A CN 101306020A CN A2007101033977 A CNA2007101033977 A CN A2007101033977A CN 200710103397 A CN200710103397 A CN 200710103397A CN 101306020 A CN101306020 A CN 101306020A
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- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
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Abstract
The invention discloses the novel application of clostridium butyricum to drugs, in particular to the application of the clostridium butyricum to the preparation of a drug combination used for preventing and treating intestinal cancer. The novel application comprises the prevention of chronic intestinal disease canceration such as intestinal polyp, Crohn disease, ulcerative colitis, etc., the adjuvant therapy on advanced inoperable intestinal cancer, the adjuvant therapy after the intestinal cancer operation, etc.; the clostridium butyricum is selected from (but is not limited to) clostridium butyricum, CGMCC, and storage serial number 0313.1. In the forms of live bacteria and pharmic receivable different carriers, the clostridium butyricum is made into tablets, capsules, powders, liquid preparations, suppositories, and enemas; the clostridium butyricum can be used for preventing and treating the intestinal cancer for a long time through the methods of oral administration, clysis, injection in the tumor tissue; therefore, the life of a patient suffering from the intestinal cancer is prolonged, and the life quality of the patient suffering from the intestinal cancer is increased.
Description
Technical field
The present invention relates to the new medicine use of clostridium butyricum, be specifically related to the application of clostridium butyricum in preparation prevention and treatment bowelcancer medicine compositions, wherein said clostridium butyricum is made active bacteria formulation as main medicament active composition, belongs to biological technical field.
Background technology
Recent research shows, descends on the contrary year by year with most of cancer morbidity, and the sickness rate of intestinal cancer just rises with average annual 2% speed in the world, and the investigation in Shanghai shows, the speedup of China's intestinal cancer sickness rate is the twice of world average level, reaches average annual 4%.Colorectal cancer has been one of primary killer in the cancer, has every year 140000 people to be made a definite diagnosis.Also have investigation to show that the age of onset of colorectal cancer is tending towards rejuvenation.Colorectal cancer is one of malignant tumor that threatens health of people, worldwide belongs to one of 4 big common cancers, become the malignant tumor that mortality rate occupies the 2nd at developed country's colorectal cancer, and the sickness rate of colorectal cancer is the trend that raises year by year.Incidence trend prediction according to western countries' colorectal cancer, each probability generation large bowel neoplasm of 50% of having per capita of living more than 70 years old (comprises early stage benign polypus, estimate 10% benign polypus canceration), everyone has 6% chance trouble colorectal cancer, there is 1 people may suffer from colorectal cancer for per 17 philtrums of women, there is obvious ascendant trend in China's colorectal cancer in recent years, especially economical than flourishing coastal cities and eastern region, and colorectal cancer has become one of the fastest malignant tumor of sickness rate rising in the most of area of China at present.Colorectal cancer is one of China's kinds of tumor, accounts for the 5th of the national malignant tumor cause of the death.Therefore, searching can prevent intestinal cancer to take place and be extremely urgent without any medicine, health food and the beverage etc. of toxic and side effects, easily develops into the medicine of the intestinal chronic disease canceration of intestinal cancer as early prevention knot or rectal polyp, schistosomicide, bacillary dysentery, ameba, Crohn disease and ulcerative colitis etc.
Colorectal cancer is generally all wanted operative treatment except that late period, cooperate radiotherapy, chemotherapy, immunization therapy and treatment by Chinese herbs etc. after the operation again.Though radiotherapy, chemotherapy, immunization therapy effect are obvious, side effect is very big, and the patient is difficult to tolerance, the patient who has even die from side effect.Though the Chinese medicine side effect is little, effect is undesirable.
Pharmaceutical composition prevention of the present invention, treatment or auxiliary treatment intestinal cancer are evident in efficacy, and especially intestinal cancer operation back auxiliary treatment has unique effect, and without any side effect, can prolonged application prevent and treatment intestinal cancer special this patent of invention of application.
Summary of the invention
The inventor is that main medicament active composition is made medicine, health food or beverage by deep basis and clinical research invention with clostridium butyricum, and intestinal cancer can effectively be prevented and treat to prolonged application.Pre-anti-caking or rectal polyp, schistosomicide, bacillary dysentery, ameba, Crohn disease and ulcerative colitis etc. easily develop into the intestinal chronic disease canceration of intestinal cancer; Inoperable intestinal cancer patient was put, when changing, was used clostridium butyricum simultaneously and can effectively suppress tumor growth and delay further diffusion transfer late period, and human body immunity improving power is alleviated symptoms such as stomachache, diarrhoea, improves the quality of living life-saving; The clostridium butyricum auxiliary treatment is used in intestinal cancer patient with operation operation back, can promote the healing of surgical wound surface, the infection that prevents surgical wound surface and antiinflammatory, the recurrence of prevention intestinal cancer and diffusion transfer, raising intestinal cancer patient immunity of organisms.Because clostridium butyricum belongs to probiotic bacteria, without any toxic and side effects, can prolonged application.
Be mainly the following aspects by the mechanism of action of discovering clostridium butyricum prevention and treatment intestinal cancer:
1, secretion butyric acid, regulate gene expression
Clostridium butyricum can be secreted a large amount of butyric acid after entering intestinal, and butyric acid is a kind of deacetylase (HDAC) inhibitor, mainly changes chromatin Structure by the degree of acetylation that changes histone, regulation and control P
21Etc. the expression of several genes, stop the colon-cancer cell growth, induce the ripe differentiation of colon-cancer cell, induce the colon-cancer cell apoptosis, can effectively prevent canceration, prevent intestinal cancer recurrence and diffusion transfer.
2, secretion butyric acid, the nutrition intestinal is repaired intestinal mucosa
70% of intestinal mucosa institute energy requirement derives from butyric acid, and butyric acid is the main energy source and the nutrient substance of intestinal mucosa cells, can provide metabolisable energy for intestinal, promotes enterocyte propagation and mucosa growth.Simultaneously, butyric acid can stimulate the generation of the gut hormone of mediation enterocyte propagation and intestinal mucosa growth, as gastrin, enteroglucagon, cheese cheese peptide etc.Thereby promote the healing of intestinal cancer operation back wound, promote the reparation of chronic intestinal tract disease injured intestinal mucosa to reduce the generation of canceration.Studies show that clostridium butyricum can obviously dwindle the ulcer area of ulcerative colitis rat intestine mucosa.
3, produce kininase, the dissolving cancerous tissue
Clostridium butyricum can produce kininase and decompose Kallidin I, the blood capillary circulation in the cancerous tissue is destroyed, thereby make the cancerous tissue necrosis, reaches oncolysis.
4, suppress harmful intestinal tract bacteria, eliminate enterotoxin, prevention of postoperative infection
Clostridium butyricum can be secreted the butyric acid rhzomorph, suppress harmful intestinal tract bacteria such as enterohemorrhagic Escherichia coli, Shigella dysenteriae, Salmonella typhi and staphylococcus aureus, eliminate the malicious source of producing, reduce of the further infringement of enterotoxin such as intestinal ammonia, amine, indole, scatol, can prevent infection and other intestinal infections of intestinal cancer postoperative wound simultaneously intestinal.
5, diminish inflammation
Clostridium butyricum can be reduced the overexpression that IL-2, IL-8 etc. cause inflammatory factor, promotes the healing of inflammation or postoperative wound.
6, human body immunity improving power
Clostridium butyricum can promote the phagocytic function of monokaryon-macrophage system, increase the generation of serum hemolysin, leukocyte increasing etc., human body immunity improving power, intestinal cancer patient uses clostridium butyricum can human body immunity improving power, the growth of anticancer and diffusion transfer, chemicotherapy cause immunity of organisms to descend, and taking clostridium butyricum during chemicotherapy can human body immunity improving power.
7, recover the intestinal microbial population balance, set up the intestinal biological barrier
Clostridium butyricum enters intestinal secretion butyric acid rhzomorph and suppresses harmful bacterium, decomposing polysaccharide is that oligosaccharide promotes the intestinal beneficial bacterium growth, and fast quick-recovery intestinal microbial population balance is set up biological barrier, safeguard intestinal health, intestinal cancer patient is owing to all there is alteration of intestinal flora in reasons such as chemicotherapy.
Easily develop into the chronic pathological changes of intestinal of intestinal cancer for knot or rectal polyp, schistosomicide, bacillary dysentery, ameba, Crohn disease and ulcerative colitis etc., generation that can prolonged application clostridium butyricum preparation prevention canceration.
Many intestinal cancer patients die from recurrence and diffusion transfer, for the intestinal cancer patient, comprise that can not perform the operation late period or postoperative patient, but equal prolonged application clostridium butyricum auxiliary treatment, promote the surgical wound healing, suppress intestinal cancer recurrence and diffusion transfer, prevent from or delay intestinal cancer to further develop deterioration.
Clostridium butyricum of the present invention is selected from but is not limited to clostridium butyricum (Clostridium butyricum), CGMCC, preservation numbering 0313.1.
Clostridium butyricum of the present invention refers to the living organism individuality.
Application of the present invention is meant the clostridium butyricum preparation and is used for prevention and treatment intestinal cancer by oral, coloclysis or mode such as inject in the cancerous tissue.
The present invention is the preparation that main medicament active composition is made with clostridium butyricum, can be to be separately that active ingredient is made relevant dosage form with the clostridium butyricum, also can make pharmaceutical composition with other active ingredient combinations, improves therapeutic effect.
The present invention is that clostridium butyricum with effective dose is according to top described as main medicament active composition, according to certain preparation process, add excipient substances such as conventional excipient, flavoring agent, disintegrating agent, antiseptic, lubricant, wetting agent, adhesive, solvent, thickening agent, solubilizing agent, make any dosage form that is suitable for using clinically, as clinical required dosage forms such as tablet, capsule, granule, oral liquid, other liquid preparations or enemas.
Indication effective dose of the present invention is meant with clostridium butyricum can not be lower than 1 * 10 according to total viable count that the top described final solid live bacteria preparation of making as main medicament active composition comprises
5CFU/g is generally 1 * 10
7More than the CFU/g, can reach 1 * 10
12CFU/g or 1 * 10
12More than the CFU/g.
Indication effective dose of the present invention is meant with clostridium butyricum can not be lower than 1 * 10 according to total viable count that the top described final liquid active bacteria formulation of making as main medicament active composition comprises
5CFU/mL is generally 1 * 10
7More than the CFU/mL, can reach 1 * 10
12CFU/mL or 1 * 10
12More than the CFU/mL.
The preferred preparation process of the present invention is, inoculates strain in the first liquid medium within, cultivates or multistage amplification cultivation, and is then that liquid culture is centrifugal, collects wet bacterium mud, and with described bacterium mud drying, pulverizing, obtains dry mycopowder.The dry mycopowder of gained is mixed with pharmaceutical carrier, make final dosage form.
Because it is the application of main medicament active composition in preparation prevention and treatment bowelcancer medicine compositions that the present invention discloses with clostridium butyricum first; therefore; make medicament with clostridium butyricum as medicament active composition and adjuvant combination; so long as this medicament is used for prevention and treatment intestinal cancer, all belong to protection scope of the present invention.
Clostridium butyricum of the present invention all has the effect of prevention and treatment intestinal cancer when making any dosage form.Any medicament prepares patent medicine if contain the clostridium butyricum composition in its component, as long as indicate or point out the effect with prevention and treatment intestinal cancer, then falls within protection scope of the present invention on signs such as its packing or description or on other any propaganda materials.
Clostridium butyricum of the present invention belongs to probiotic bacteria.Therefore, also clostridium butyricum can be made health food or beverage.Health food or beverage with clostridium butyricum is made indicate or point out the effect with prevention and treatment intestinal cancer if need only on signs such as its packing or description or on other any propaganda materials, then fall within protection scope of the present invention.
The specific embodiment
Medication preparation example explanation: with the clostridium butyricum be active constituents of medicine to make capsule be example, preparation method is described.Preparation method is not only limited to following method, and known method all can.
Medication preparation embodiment 1, the preparation of clostridium butyricum viable bacteria mycopowder:
Get clostridium butyricum viable bacteria strain Guan Yizhi, being dissolved in sterilizes is equipped with in the 100ml triangular flask of 10ml normal saline and bead, activate 10 minutes, drawing the 1ml bacterial suspension inoculation with the 1ml aseptic straw is equipped with in the 250ml triangular flask of 50ml amplification culture medium, putting the interior 37 ℃ of constant temperature vibrations of rocking bed (190rpm) cultivated 24 hours, switching is equipped with in the 450ml amplification culture medium 2500ml plate washer triangular flask, 37 ℃ of constant-temperature shaking culture 24 hours, microscopy is not transferred in the seed tank that the 4.5L amplification culture medium is housed after having assorted bacterium again, anaerobism was cultivated (aeration quantity 3: 1) 24 hours, microscopy does not have to transfer behind the assorted bacterium and has in the fermentation tank of 45L fermentation medium, 37 ℃ of anaerobism were cultivated 24 hours, microscopy spore rate reaches more than 80%, stops to cultivate.Use continuous centrifuge, 12000rpm is centrifugal.Collect wet bacterium mud, weigh, after protective agents such as an amount of defatted milk powder of 1: 1 (w/v) interpolation, drying is pulverized, and room temperature is preserved standby.
Medication preparation embodiment 2, the preparation of clostridium butyricum viable capsule
Composition % (weight)
20.00 parts of clostridium butyricum viable bacteria mycopowder
40.00 parts of microcrystalline Cellulose
40.00 parts of glucoses
In in the above-mentioned component 1～3 group, fully mix thoroughly, utilize conventional fill capsule technique to make capsule then by unit dose.
The experimentation of observation of curative effect experimental example 1, clostridium butyricum prevention intestinal cancer
1, experimental technique
Selecting body weight is 40 of the female Kunming mouses of 18-20g, is divided into administration group and matched group (20 every group) at random, and experimental period was 20 weeks.
Carcinogen is 1, and (symmetrical 1, and 1-dimethylhydrazine DMH), is white powdery crystallization, is made into 0.4% solution with physiological saline solution before facing injection at every turn, and uses NaHCO for the 2-Dimethylhydrazine
3Its pH is transferred to 6.5.Weekly once, continuous 20 weeks to mouse peritoneal injection DMH 20mg/kg (being 0.4%DMH solution 0.05ml/10g).After 1 week, add dextrorotation glucosan glycosides sodium (DSS) in lumbar injection DMH in the drinking-water, concentration is 20g/L, drinks continuously 7 days.
The administration group, in the time of lumbar injection DMH, clostridium butyricum CGMCC0313-1 preparation (10
7CFU/ml) irritate stomach, each 0.5mL, every day 1 time, continuous 20 weeks.
Matched group, in the time of lumbar injection DMH, normal saline is irritated stomach, each 0.5mL, every day 1 time, continuous 20 weeks.
2, experimental result
After the 20th week that experiment finishes, after with ether mouse anesthesia being caused death, open the abdominal cavity, observe intestinal and change.The result shows 20 mices of matched group, and lesser tubercle appears in the large intestine intestinal wall, and the intestinal gauffer reduces or disappears, and the intestinal quality is hard, and 100% has generated intestinal cancer.The administration group has 8 mice large intestine intestinal wall lesser tubercle to occur, and the intestinal gauffer reduces or disappears, and the intestinal quality is hard, and 40% has generated intestinal cancer.
3, discuss
This experiment shows, clostridium butyricum is the canceration of prevention of ulcerative colitis obviously, prevention rate has reached 60%, therefore can make relevant dosage form to clostridium butyricum, be used to prevent intestinal cancer, be particularly useful for intestinal chronic disease generation cancerations such as pre-anti-caking or rectal polyp, schistosomicide, bacillary dysentery, ameba, Crohn disease and ulcerative colitis, safeguard human intestine's health.Most of intestinal cancer patient dies from postoperative recurrence and transfer, therefore clostridium butyricum also can be used for intestinal cancer operation back prevention of recurrence and transfer, take the healing that the clostridium butyricum preparation can promote wound after the operation simultaneously, can suppress harmful bacterium, antiinflammatory prevention infection, can human body immunity improving power.
The experimentation of observation of curative effect experimental example 2, clostridium butyricum treatment intestinal cancer
1, experimental technique
20 of BALB/c mouse are got the mouse junction cancer C26 tumor tissue that goes down to posterity under the animal skins, become suspension with normal saline by 1: 3 dilution proportion, and it is subcutaneous only to be inoculated in mouse armpit by 0.2ml/, then mice is divided into administration group and matched group (10 every group).
The administration group is injected clostridium butyricum solution (10 in inoculation respectively in tumor tissue after 72 hours, 144 hours
7CFU/ml) 0.1mL/ only.Injecting normal saline 0.1mL/ is only in tumor tissue after 72 hours, 144 hours in inoculation respectively for matched group.Test and measured the gross tumor volume of each treated animal on the 12nd day, and press gross tumor volume and calculate tumour inhibiting rate.
2, experimental result
Administration group tumour inhibiting rate and matched group be P＜0.01 relatively
3, discuss
Experimental result shows that clostridium butyricum has tangible anti-intestinal cancer effect, and late period, inoperable intestinal cancer patient or postoperative intestinal cancer patient can use all that the clostridium butyricum preparation is treated or auxiliary treatment and prevention of recurrence and diffusion transfer.
Observation of curative effect experimental example 3, intestinal cancer operation back patient take oral clostridium butyricum viable capsule observation of curative effect
Liu * *, the man, 68 years old, occurred having blood in stool before 3 years, suffer from diarrhoea, symptom such as stomachache, (postoperative continues stomachache for Dukes ' A), row surgical resection therapy to be diagnosed as the intestinal cancer I phase.Operation back begins to take oral clostridium butyricum viable capsule (420mg/ grain, 1.0 * 10 1 week
7CFU/g, Qingdao DongHai Pharmacy Co., Ltd provides), 3/time, 3 times/day, take the 3rd day pain relief, the back pain disappearance of 1 week.Took continuously 1 year, 3 years so far, do not see stomachache, abdominal discomfort, have blood in stool, symptom such as diarrhoea occurs, to examination in hospital do not find any unusually, intestinal cancer does not recur and shifts.
The present invention in implementation process employed microorganism fungus kind respectively on July 28th, 1997 in (No. 13, North No.1 Row, Zhongguancun, Haidian District, Beijing City, China Committee for Culture Collection of Microorganisms common micro-organisms center, Institute of Microorganism, Academia Sinica's postcode 100080) preservation, but clostridium butyricum of the present invention is not limited to this microbial strains.
The classification name: clostridium butyricum Clostridium butyricum, preserve numbering 0313.1.
The mentioned microorganism bacterial strain detects through this microorganism center, and testing result is survival.
Claims (8)
1, the application of clostridium butyricum in preparation prevention and treatment bowelcancer medicine compositions, wherein said clostridium butyricum is made active bacteria formulation as main medicament active composition.
2, by the described application of claim 1, it is characterized in that the application of clostridium butyricum in preparation prevention intestinal chronic disease canceration pharmaceutical composition, the intestinal chronic disease comprises knot or rectal polyp, schistosomicide, bacillary dysentery, ameba, Crohn disease, ulcerative colitis.
3,, it is characterized in that the application of clostridium butyricum in preparation auxiliary treatment bowelcancer medicine in late period compositions by the described application of claim 1.
4, by the described application of claim 1, it is characterized in that the application of clostridium butyricum in the auxiliary therapeutic agent compositions of preparation intestinal cancer operation back, comprise the healing that promotes surgical wound surface, the infection that prevents surgical wound surface and antiinflammatory, the recurrence of prevention intestinal cancer and diffusion transfer, raising intestinal cancer patient immunity of organisms.
5, by the described application of claim 1, it is characterized in that clostridium butyricum comprises clostridium butyricum CGMCCO313.1 strain.
6, by the described application of claim 1, it is characterized in that described clostridium butyricum of the claim 1 of effective dose and pharmaceutically acceptable different carriers are made tablet, capsule, powder, liquid preparation, suppository, enema.
7,, it is characterized in that in clostridium butyricum active bacteria formulation oral administration, anum administration or the injection tumor tissues by the described application of claim 1.
8, by the described application of claim 1, it is characterized in that pharmaceutical composition comprises medicine, health food, beverage.
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Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101849969A (en) * | 2009-03-31 | 2010-10-06 | 青岛东海药业有限公司 | Application of butyric acid producing beneficial bacterium in preparing preparation for preventing and treating severe disease gut barrier injury and post-injury complication |
| CN103202861A (en) * | 2013-04-15 | 2013-07-17 | 山东科兴生物制品有限公司 | Application of composition of clostridium butyricum and bifidobacterium dual live bacteria preparation and anticancer medicament in preparation of medicament for treating intestinal cancer |
| CN104857026A (en) * | 2015-05-18 | 2015-08-26 | 新乡医学院 | Method for researching preventive effect of combined clostridium butyricum and bifidobacterium powder on NEC (neonatal necrotizing enterocolitis) |
| CN109771445A (en) * | 2019-03-20 | 2019-05-21 | 青岛东海药业有限公司 | Clostridium butyricum is preparing the application in inducing antitumor immunity and immunologic test point inhibitor enhanced sensitivity preparation |
| CN110090231A (en) * | 2018-01-30 | 2019-08-06 | 青岛东海药业有限公司 | Application of the clostridium butyricum in preparation prevention or treatment cholangiocarcinoma preparation |
| CN110652528A (en) * | 2010-06-04 | 2020-01-07 | 国立大学法人东京大学 | Composition for inducing proliferation or accumulation of regulatory T cells |
| CN112843096A (en) * | 2020-02-11 | 2021-05-28 | 青岛东海药业有限公司 | Application of clostridium butyricum in preparation of preparation for treating coronavirus infection |
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- 2007-05-20 CN CNA2007101033977A patent/CN101306020A/en active Pending
Cited By (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101849969A (en) * | 2009-03-31 | 2010-10-06 | 青岛东海药业有限公司 | Application of butyric acid producing beneficial bacterium in preparing preparation for preventing and treating severe disease gut barrier injury and post-injury complication |
| CN110652528A (en) * | 2010-06-04 | 2020-01-07 | 国立大学法人东京大学 | Composition for inducing proliferation or accumulation of regulatory T cells |
| CN103202861A (en) * | 2013-04-15 | 2013-07-17 | 山东科兴生物制品有限公司 | Application of composition of clostridium butyricum and bifidobacterium dual live bacteria preparation and anticancer medicament in preparation of medicament for treating intestinal cancer |
| CN104857026A (en) * | 2015-05-18 | 2015-08-26 | 新乡医学院 | Method for researching preventive effect of combined clostridium butyricum and bifidobacterium powder on NEC (neonatal necrotizing enterocolitis) |
| CN110090231A (en) * | 2018-01-30 | 2019-08-06 | 青岛东海药业有限公司 | Application of the clostridium butyricum in preparation prevention or treatment cholangiocarcinoma preparation |
| CN109771445A (en) * | 2019-03-20 | 2019-05-21 | 青岛东海药业有限公司 | Clostridium butyricum is preparing the application in inducing antitumor immunity and immunologic test point inhibitor enhanced sensitivity preparation |
| CN109771445B (en) * | 2019-03-20 | 2022-08-05 | 青岛东海药业有限公司 | Application of clostridium butyricum in preparing sensitization preparation for inducing anti-tumor immunity and immune checkpoint inhibitor |
| CN112843096A (en) * | 2020-02-11 | 2021-05-28 | 青岛东海药业有限公司 | Application of clostridium butyricum in preparation of preparation for treating coronavirus infection |
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Effective date of registration: 20091218 Address after: No. 8, Shanghai Road, Jiaonan, Qingdao, Shandong Applicant after: Qingdao Donghai Pharmaceutical Co., Ltd. Co-applicant after: Beijing Puerkang Medical High Technology Co., Ltd. Address before: No. 8, Shanghai Road, Jiaonan, Qingdao, Shandong Applicant before: Qingdao Donghai Pharmaceutical Co., Ltd. Co-applicant before: Dongfanghaixin Biological Technology Co., Ltdd., Beijing |
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Open date: 20081119 |