CN1044468A - Cell growth stimulant and manufacture method thereof - Google Patents
Cell growth stimulant and manufacture method thereof Download PDFInfo
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- CN1044468A CN1044468A CN 89105016 CN89105016A CN1044468A CN 1044468 A CN1044468 A CN 1044468A CN 89105016 CN89105016 CN 89105016 CN 89105016 A CN89105016 A CN 89105016A CN 1044468 A CN1044468 A CN 1044468A
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- cell growth
- growth stimulant
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- streptococcus aureus
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Abstract
The invention provides a kind of cell growth stimulant, it is characterized in that: it is the extracting solution of a kind of Xie Shi streptococcus aureus (Staphy lococcusaureus Xie) meta-bolites, is main component and includes protein, polypeptide and multiple amino acids such as 2,5-diaminovaleric acid, Gelucystine with aseptic plasma-coagulase.The present invention also provides a kind of manufacture method of cell growth stimulant, it is characterized in that adopting pig myocardium, and the aqueous solution of peptone and sodium-chlor is made substratum, bacterial classification is cultivated down at 37 ℃ directly extracted aseptic meta-bolites in 48 hours.This cell growth stimulant is to delayed union of fracture, and diseases such as disunion and ulcer have special efficacy.
Description
The invention belongs to the technical field of microorganism biological goods class medicine, promptly provide a kind of reparation for the treatment of wound healing and human body particular tissues or regenerated and the medicine made and the method for making this medicine treated by microbial metabolites.
Handle various wounds and be a frequent and important job in the surgical clinical practice.The purpose of handling wound is that wound is in time healed perfectly.Yet the healing of some wound for example union of fracture is the comparison difficulty, and at first, the normal healing of fracture takes longer; Secondly, be prone to the slow concrescence phenomenon that even do not heal for open fracture.At present both at home and abroad the method for treatment delayed union of fracture and disunion disease mainly contains two kinds of operation bone grafting (comprising microsurgery band blood bone grafting) and electricity irritation (comprising electric pulse stimulation) methods, they all are that surgical treatment is a non-drug therapy, and all have: the course of treatment is long, treatment rate is low, and (the former only reaches about 50% by a bone grafting; The latter also only reaches 70%), shortcoming such as painful big, the medical expense height of patient.Wu Lisite (Urist) has invented a kind of Delicious peptide (abbreviating BMP as) in recent years, though experimental study shows that it has certain good influence to union of fracture, Shang Weijian has its clinical application and obtains the case report of produce effects.We can say, do not find a kind of medicine that can obviously directly promote union of fracture up to now both at home and abroad as yet.In addition, processing for ulcer (comprising sinus and fistula) also is a relatively stubborn problem of Clinical Surgery, owing to there is not a kind of medicine that can directly promote the human body cell growth, usually the surgeon can only adopt cleaning ulcer spot, skin-grafting, strengthen nutrition, blood transfusion, replenishes various VITAMIN to improve " indirectly " therapy such as its whole body situation to the patient, and its course of treatment is long, patient is painful well imagines greatly.
The objective of the invention is to develop a kind of specifics that can be used for treating diseases such as above-mentioned delayed union of fracture, disunion and ulcer, a kind of injection liquid that can promote the cell growth promptly is provided, only it need be expelled to patient's the fracture site or the position of festering, just can impel fracture or wound healing effectively.
The invention provides a kind of cell growth stimulant, it is characterized in that it is the extracting solution of a kind of Xie Shi streptococcus aureus (Staphylococcus aureus Xie) meta-bolites, it is major ingredient with aseptic plasma-coagulase and includes protein, polypeptide and multiple amino acids such as 2,5-diaminovaleric acid, Gelucystine.
Xie Shi streptococcus aureus (Staphylococcus aureus Xie) is a kind of special bacterial classification provided by the present invention.The Xie Shi streptococcus aureus is compared with common streptococcus aureus (Staphylococcus aureus Rosenbach), and its feature has obvious difference.At first, our both difference in view of outward appearance, Xie Shi streptococcus aureus and common streptococcus aureus are observed contrast after electron microscope amplifies 54800 times, from the visual field obviously as seen: Xie Shi streptococcus aureus volume is big, cell walls is abundant and more tough and tensile, sclerotium is big, karyomit(e) is thick and obvious, be complete symmetry when dividing; Then volume is less, cell walls approaches cracky, sclerotium is less even obviously, karyomit(e) is little or it is unclear to show, be the asymmetrical type division for common streptococcus aureus.The feature of above-mentioned two kinds of streptococcus aureuses is done further relatively, and its data information is listed in the table 1.It is (many that the original strain of Xie Shi streptococcus aureus is taken from a trouble femur bone marrow inflammation
Table 1
| The bacterial classification test subject | The Xie Shi streptococcus aureus | Common streptococcus aureus |
| The mannose ferment test | Be the light red reaction | Be purple color reaction |
| Clotting of plasma enzymatic determination | + | + |
| The zone of hemolysis test | +?+ | + |
| Scleroproein haemolysis degraded product (FDP) | 1：16 | 1：1024 |
The property sent out sinus companion hyperplasia bony shell disease) patient's in year lesions position surplus in the of 20, this bacterial classification institute for drug control, Zhejiang Province is identified, puts on record in pharmaceutical biological product institute of Ministry of Health of the People's Republic of China.
Cell growth stimulant provided by the present invention is used for delayed union of fracture and disunion disease is effective especially, the 345 routine delayed union of fracture of accepting for medical treatment through No.3 People's Hospital, Hangzhou City., the case of disunion, curative ratio reaches 95.1%, and the clinical healing time (opposing than the test statistics with the fresh fracture patient) can shorten 1/3～1/2.Compare with existing operation bone grafting and electrical stimulating therapy, it has: short treating period, curative ratio height, applied widely, patient does not have obvious misery, medical expense is low, sequela is few and series of advantages such as easy to use, safe.Cell growth stimulant provided by the present invention is lost ulcer even illly through the cureless ulcer of lower limb of various therapies (being commonly called as " old-rotten-leg ") obvious curative effects is arranged also for many years for soft tissue.
The present invention also provides a kind of method of making above-mentioned cell growth stimulant, and it comprises the preparation and six procedures such as processing, inoculation, bacterium colony, cultivation, soup extraction, sterilization and packing of substratum, it is characterized in that:
A) adopt the Xie Shi streptococcus aureus to make bacterial classification;
B) substratum is made raw material with pig myocardium, peptone, sodium-chlor and water, and the proportioning of former three (calculating with 10,000 milliliters of substratum) is:
Pig myocardium 5,000 grams
Peptone 100 grams
Sodium-chlor 50 grams
Water adds to 10,000 milliliters
C) preparation of substratum and processing comprise boil, remove slag, filtration and four steps of autoclaving, its pH value is 7.5;
D) raise craft condition of Xie Shi streptococcus aureus is that adding concentration is 2 * 10 in per 500 milliliters of substratum
91.0 milliliters of the bacterium liquid of/milliliter were cultivated 48 hours at 37 ℃;
E) soup extracts by multiple tracks filtration and two steps of degerming and forms.
In order to ensure the quality of cell growth stimulant of the present invention, prevent that the variation of Xie Shi streptococcus aureus bacterial classification from being very necessary.So before the inoculation operation, preferably earlier the golden yellow staphylococcus original seed of Xie Shi bacterial strain is placed on the inclined-plane (or palm) with 0.25 milliliter of dissolving of physiological saline, put into substratum and cultivated 24 hours down at 37 ℃, do variation inspection.Bacterium colony after the variation passed examination can get off with a small amount of normal saline flushing, and preparation becomes the production that is suitable for raise craft condition desired concn bacterium liquid.
Cell growth stimulant provided by the present invention is faint yellow transparent injection liquid.Usually can preserve under 4～10 ℃ envrionment temperature, also the vacuum available desiccating method is made for the pulvis prolonged preservation.In order to improve the effective storage life limit of injection liquid, can in injection liquid, add the phenol of some amount and make sanitas, the add-on of phenol (calculating by weight percentage) is advisable with 0.2～0.3.
Adopt the cell growth stimulant of above-mentioned manufacture method manufacturing, carry out toxicological test, acute and a series of checks such as subacute toxicity test, hypersensitive test, pyrogen testing, local excitation test, hemolytic test, limulus test, sterility test and enterotoxin test, depressor substance test, its assay all meets the Ministry of Health and the " relevant regulations of Chinese pharmacopoeia.The acute medium lethal dose L of small white mouse wherein
D50Be 344.8 ± 16.6mg/kg, the cell growth stimulant that the confirmation manufacturing is come out is safe in utilization, nontoxic.Medullary cell nuclear test, Salmonella reversion test, U.D.S test-results are all negative, teratogenesis, carcinogenic observation were followed up a case by regular visits to 1～5 year, the result all belongs to feminine gender, cell growth stimulant and manufacture method thereof that these further illustrate the present invention closely provides possess practicality, nontoxicity of medicine own and apparent side effect can be applied clinically.
Cell growth stimulant provided by the present invention and manufacture method thereof, embodiment is as follows:
(1) culture medium preparation and processing.Substratum is made raw material with pig myocardium, peptone, sodium-chlor and water, and the actual proportioning of each component (calculating by weight percentage) is as follows:
Pigs Hearts (clean and drain) 5000 grams
Peptone 100 grams
Sodium chloride for injection 50 grams
Distilled water adds to 10,000 milliliters
Earlier Pigs Hearts is dehematized pipe, clot, fat and manadesma are cleaned the back and are dashed to drench with distilled water and drain, and weigh, and add 2 times distilled water, boil 2 hours, cool off in the rearmounted refrigerator refrigeration 12 hours, this slag of elimination, filtrate; Add sodium chloride for injection and peptone in filtrate, boil the dissolving after-filtration, transferring to pH value with 10% Hcl again is 3～4.Add injection gac 50 grams in case of necessity, continued heated and boiled 15 minutes, use the paper pulp filtering carbon removal while hot.If adding distil water is to full dose, and transfers to pH value 7.5(after transferring pH value with NaOH liquid, and solution becomes gets and muddyly should add filtration with clarification), in 500 mL of saline bottles, seal at last with aluminium lid.Need behind the substratum envelope bottle through 10 kilograms, 30 minutes or autoclaving processing in 15 kilograms, 20 minutes, and after being cooled to room temperature, preserve standby.
(2) inoculation and cultivation.Get one of original seed bacterial strain, after 0.25 milliliter of dissolving of physiological saline, culture transferring is on inclined-plane (or dull and stereotyped), through 37 ℃ of culture medium culturing 24 hours, after the variation passed examination, lawn on inclined-plane (or dull and stereotyped) is got off with a small amount of normal saline flushing, is made into the concentration of 2,000,000,000 bacteria/milliliters, with bacterium than turbid standard pipe coarse adjustment (10
-8Dilution back enumeration core) in per 500 milliliters of substratum, adds 1.0 milliliters bacterium liquid, cultivated 48 hours down, observed in preferably per 2 hours and also to proofread and correct a subculture temperature at 37 ℃.
(3) extract soup.With in the last process through the bacterium liquid of 37 ℃, 48 hours culture medium culturing, filter twice at least, faint yellow clarifying filtrate.With filtrate degerming and filtration again, and sterility test is made in sampling.The bacteria-free filtrate of passed examination is makes good cell growth stimulant injection liquid.Consider the needs of prolonged preservation, can adopt boulton process to be made into pulvis; The phenol that perhaps adds (calculating by weight percentage) 0.2～0.3 in above-mentioned injection liquid is made sanitas, divides in 2 milliliters the peace bottle of packing into, seals, and refrigeration is stand-by down at 4～10 ℃.
Claims (4)
1, a kind of cell growth stimulant, it is characterized in that it is the extracting solution of a kind of Xie Shi streptococcus aureus (Staphy lococcus aurus Xie) meta-bolites, is main component and includes protein, polypeptide and multiple amino acids such as 2,5-diaminovaleric acid, Gelucystine with aseptic plasma-coagulase.
2, a kind of method that is exclusively used in manufacturing by the described cell growth stimulant of claim 1, it comprises the preparation and six procedures such as processing, inoculation, bacterium colony cultivation, soup extraction, sterilization and packing of substratum, it is characterized in that:
A) adopt the Xie Shi streptococcus aureus to make bacterial classification;
B) substratum is made raw material with pig myocardium, peptone, sodium-chlor and water, and the proportioning of former three (calculating with 10,000 milliliters of substratum) is: pig myocardium 5000 grams, and peptone 100 grams, sodium-chlor 50 grams, water adds to 10,000 milliliters;
C) preparation of substratum and processing comprise boil, remove slag, filtration and four steps of autoclaving, the substratum pH value is 7.5;
D) raise craft condition of Xie Shi streptococcus aureus is that wherein to add concentration be 2 * 10 in every milliliter of cultivation
91.0 milliliters of the bacterium liquid of milliliter were cultivated 48 hours down at 37 ℃;
E) soup extracts by multiple tracks filtration and two steps of degerming and forms, and preserves down at 4～10 ℃.
3,, it is characterized in that soup vacuum available desiccating method makes pulvis and preserve by the described cell growth stimulant of claim 2 manufacture method.
4,, it is characterized in that the phenol that soup can add (calculating by weight percentage) 0.2～0.3 makes sanitas by the described cell growth stimulant of claim 2 manufacture method.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 89105016 CN1025118C (en) | 1989-01-28 | 1989-01-28 | Cell growth hormone and prepn. thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 89105016 CN1025118C (en) | 1989-01-28 | 1989-01-28 | Cell growth hormone and prepn. thereof |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN1044468A true CN1044468A (en) | 1990-08-08 |
| CN1025118C CN1025118C (en) | 1994-06-22 |
Family
ID=4855788
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN 89105016 Expired - Fee Related CN1025118C (en) | 1989-01-28 | 1989-01-28 | Cell growth hormone and prepn. thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN1025118C (en) |
Cited By (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1043350C (en) * | 1994-03-15 | 1999-05-12 | 中国人民解放军第458医院 | Method for preparation of cardiac muscle cell growth stimulus peptide |
| CN1108170C (en) * | 1996-10-11 | 2003-05-14 | 李绍光 | Buffering liquid for staphylococcus Aureus Injection and stephylococcus Aureus Injection with buffering liquid |
| WO2006011137A3 (en) * | 2004-07-26 | 2006-03-30 | State Of Israel Dept Of Agricu | Novel bacteria and pharmaceutically active products obtained therefrom |
| CN1293865C (en) * | 2002-12-06 | 2007-01-10 | 谢旭明 | Cell growth regulating factor R and its prepn process |
| CN1293875C (en) * | 2002-12-06 | 2007-01-10 | 谢旭明 | Cell growth regulatnig factor S and its prepn process |
| CN1293866C (en) * | 2002-12-06 | 2007-01-10 | 谢旭明 | Cell growth regulating factor II and its prepn process |
| CN1305465C (en) * | 2002-12-06 | 2007-03-21 | 谢旭明 | Cell growth regulating factor I and its prepn process |
| CN1323669C (en) * | 2002-12-06 | 2007-07-04 | 谢旭明 | Cell growth regulating factor P and its prepn process |
-
1989
- 1989-01-28 CN CN 89105016 patent/CN1025118C/en not_active Expired - Fee Related
Cited By (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1043350C (en) * | 1994-03-15 | 1999-05-12 | 中国人民解放军第458医院 | Method for preparation of cardiac muscle cell growth stimulus peptide |
| CN1108170C (en) * | 1996-10-11 | 2003-05-14 | 李绍光 | Buffering liquid for staphylococcus Aureus Injection and stephylococcus Aureus Injection with buffering liquid |
| CN1293865C (en) * | 2002-12-06 | 2007-01-10 | 谢旭明 | Cell growth regulating factor R and its prepn process |
| CN1293875C (en) * | 2002-12-06 | 2007-01-10 | 谢旭明 | Cell growth regulatnig factor S and its prepn process |
| CN1293866C (en) * | 2002-12-06 | 2007-01-10 | 谢旭明 | Cell growth regulating factor II and its prepn process |
| CN1305465C (en) * | 2002-12-06 | 2007-03-21 | 谢旭明 | Cell growth regulating factor I and its prepn process |
| CN1323669C (en) * | 2002-12-06 | 2007-07-04 | 谢旭明 | Cell growth regulating factor P and its prepn process |
| WO2006011137A3 (en) * | 2004-07-26 | 2006-03-30 | State Of Israel Dept Of Agricu | Novel bacteria and pharmaceutically active products obtained therefrom |
Also Published As
| Publication number | Publication date |
|---|---|
| CN1025118C (en) | 1994-06-22 |
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