CN101612392A - A kind ofly be used to prevent and treat preparation of mammalian endometritis and preparation method thereof - Google Patents
A kind ofly be used to prevent and treat preparation of mammalian endometritis and preparation method thereof Download PDFInfo
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- CN101612392A CN101612392A CN200810043569A CN200810043569A CN101612392A CN 101612392 A CN101612392 A CN 101612392A CN 200810043569 A CN200810043569 A CN 200810043569A CN 200810043569 A CN200810043569 A CN 200810043569A CN 101612392 A CN101612392 A CN 101612392A
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Abstract
The invention discloses a kind of suppository for the treatment of mammalian endometritis and preparation method thereof.In the gross weight of preparation, component comprises: lysozyme 0.5-8%, gelatin 1-30%, glycerol 1-40%, sodium alginate 0.1-20%, water 40-95%.Said preparation adopts the topical mode, can effectively treat mammalian endometritis, bactericidal effect is strong, good stability, and in the process of processing, lysozyme is wrapped up through sodium alginate owing to first, make that activity of lysozyme can keep effectively in the process of preparation suppository, can in uterus slowly discharge, reach better therapeutic effect.
Description
Technical field
The present invention relates to a kind of biotype biocide preparation, specifically, relate to a kind of biological preparation that is used to prevent and treat mammalian endometritis.
Background technology
Endometritis is a common genital organ disease such as mammal such as milch cow, pig etc., is the one of the main reasons that causes female animal infertile.Seriously influence breeding and the economic benefit of female animal.Intrusion antibacterials such as the sterilization when being artificial insemination, childbirth or practise midwifery of its main diseases is not tight, and postpartum care is improper cause infection, this epivaginitis, the prolapsus uteri, placenta retention etc., but secondary endometritis all.Its cardinal symptom has: acute when generally betiding post-abortion or producing placenta retention, and fervescence, loss of appetite, spirit is depressed.When serious, hogback is exerted duty, often makes to urinate shape, and chronic by acute secondary, the estrus cycle is undesired.
At present both at home and abroad to the treatment of female animal endometritis still based on antibiotic, but there are many unfavorable factors in antibiotic therapy.Make antibacterial produce drug resistance easily as (1); (2) the single medicine antimicrobial spectrum is wideless; (3) cause antibiotic remains easily; (4) other side effect.For example, the milch cow that suffers from endometritis can cause Determination of antibiotic in milk residual through after the antibiotic therapy, and it is extremely serious that these milk that used antibiotic milch cow to extrude are sneaked into the consequence that causes in the edible milk.FAO (Food and Agriculture Organization of the United Nation) (FAO) and The World Health Organization's suggestion, the milk of extruding in (preferably drug withdrawal 5-7 days) at least 3 days after milch cow is accepted the antibiotic therapy drug withdrawal is not directly as edible milk material.Be present in the milk or meat in residue, can cause people's anaphylaxis, also can make Crinis Carbonisatus rawhide rash etc.Antibiotic residual in milk as the important social public hazards, forbidden listing already in American-European countries, and implements the management method that milk that residual period extrudes all abandons.As the food and bureau of drug (F.D.A) regulation of the U.S., the antibiotic content in every ml milk must not surpass 0.006 iu, uses antibiotic milk discarded 96 hours planted agents." food hygiene law " of various countries' promulgation at present all stipulated, uses the milk of antibiotic therapy mastitis, must not be as food selling in 5 days.Developed country also to the selective examination in every month of each product fresh milk once, penalizes as finding the residual antibiotic person, stipulates simultaneously in a single day the poultry master that must accuse when the veterinary uses antibiotic must not sell the edible time limit with fresh milk, is to blame otherwise find the veterinary.
The traditional Chinese medical science, traditional Chinese veterinary medicine are used the disease that natural drug (Chinese herbal medicine) is treated the human and animal, now are described as natural medical science.As Chinese patent CN1302634A (medicine of treatment inflammation of uterus), but because the drug action of Chinese medicine is slow, so that state of an illness protracted course of disease, or outbreak repeatedly, thereby clinical result of use and bad.
U.S. Pat 4692452 (Method for treatment of endometritis in mammalianfemales), reported with chemicals and treated mammiferous endometritis, but chemicals often has zest to endometrium, can cause the endometrium fibrosis, so that the formation cicatrix, be unfavorable for milch cow further breeding become pregnant.
Lysozyme is one or more in bacterial cell wall lyase, yeast cell wall lyase, the mycete cell wall lyase; Used lysozyme is the active component that extracts from Ovum Gallus domesticus album, is the crude drug that meets the Chinese Pharmacopoeia standard, and lysozyme is by acting on β-1,4 glycosidic bond in the cell wall or β-1,3 glucosan and killing bacteria.Because the sterilization mechanism of lysozyme uniqueness makes it be different from general antibiotic.Antibiotic effect generally is a bacteria growing inhibiting, impels the R-plasmid of Production by Bacteria tissue regeneration promoting or the effect that new enzyme stops antibiotics.This enzyme then is cracking and killing bacteria, and action time is short, and antibacterial is not easy to produce drug resistance, and bactericidal effect is good simultaneously.Because lysozyme is a kind of protein of extensive existence, nontoxic, harmless, the narrow spectrum purpose cells of microorganisms wall that acts on of energy, gram positive bacteria, good gas spermatium formation bacterium, bacillus subtilis, Bacillus licheniformis etc. all there is antibacterial action, and can not have a negative impact to the animal somatic cell that does not have cell wall, thereby safety is very high.Because the food-safety problem that causes because of antibiotic abuse in the animal feeding process in recent years is more and more outstanding, how not only to ensure the safety of animal foodstuff but also the fertility performance of animal can normally be brought into play, as a kind of to humans and animals all safety antibiotic preparation, lysozyme is subject to people's attention gradually, and is applied to animal production.
Yang Weirong (lysozyme treatment milch cow puerperal endometritis test, the dairy industry science and technology, 2001 (4): 44-46) adopt lysozyme injection for curing cow endometritis, the result show lysozyme to the total cure rate of endometritis, treatment back recover of uterus degree equal effective than antibiotic therapy.Total cure rate to endometritis reaches 82.35%～88.24%, apparently higher than the cure rate (56.25%) of chlortetracycline.Because of administering mode is that lysozyme directly is dissolved in normal saline, intrauterine perfusion administration, total cure rate is also not too high, and the use amount of lysozyme is bigger than normal, and uses comparatively inconvenience.(the composite lysozyme preparation is to the bacteriostatic test of mammitis of cow pathogen for Liu Wen virtues etc., China's Dairy Industry, 2006 (6): 33-35) with the compound formulation coupling of hen's egg-white lysozyme and Chinese herbal medicine the pathogenic bacterium that cause mastitis are made bacteriostatic test, the lysozyme experimental group has obtained good fungistatic effect.
Chinese patent ZL03142299.3 (a kind of be used to prevent and treat preparation of cow endometritis and preparation method thereof), disclose the preparation made from staphylococcus lysozyme, lysozyme, bacteriostatic peptide and medically acceptable carrier, can prevent and treat cow endometritis.
Summary of the invention
One of technical problem to be solved by this invention is to provide a kind of biological antibiotic suppository that is used for mammalian endometritis, with overcome the bactericidal effect that prior art exists undesirable, in uterus retention time short, with the interaction of mucosa synovial fluid a little less than, be unfavorable for penetrating into the shortcoming of vaginal mucosa.
Two of the technical issues that need to address of the present invention provide the preparation method of this kind suppository.
Inventive concept of the present invention is such:
The suppository of treatment mammalian endometritis of the present invention, its composition and weight percentage are: lysozyme 0.5-8%; Gelatin 1-30%; Glycerol 1-40%; Sodium alginate 0.1-20%, water 40-95%.
The suppository of the treatment mammalian endometritis of being addressed also can comprise the staphylococcus lysozyme that percentage by weight is 0.01-0.1%.
The suppository of the treatment mammalian endometritis of being addressed also can comprise the hibitane that percentage by weight is 0-1%.
The suppository of the treatment mammalian endometritis of being addressed, the composition of its optimization formula and percentage by weight are: lysozyme 2-8%; Gelatin 5-15%; Glycerol 5-20%; Sodium alginate 0.1-5%; Hibitane 0.1-0.5%, water 50-85%.
The preparation method of the suppository of the treatment mammalian endometritis of being addressed comprises the steps:
(1) contains the preparation of lysozyme microgranule: lysozyme (containing or do not contain staphylococcus lysozyme) is joined mix homogeneously in the sodium alginate soln, joining volume ratio is ethyl acetate: in the solution of dichloromethane=3: 1, add polyvinyl alcohol, powerful stirring (rotating speed is 100-300 rev/min), dropping calcium chloride solution dropwises the back and continues to stir then, forms the microgranule of diameter at 10um～1mm, filtration obtains containing the microsphere of lysozyme, lyophilization.
(2) preparation of suppository formulations: take by weighing gelatin, glycerol according to the above ratio, add water and be heated to 60 ℃ of dissolvings, then the microgranule for preparing is joined in the gelatin glycerol that dissolves according to the above ratio, pour the suppository die for molding rapidly into and get final product.
Lysozyme can effectively be killed the various pathogen that cause endometritis for pressing down sterilization component; Gelatin is an excipient; Glycerol is wetting agent; Sodium alginate is an excipient, and has slow-release function, makes lysozyme slowly to discharge in uterus and reaches better therapeutic effect.This suppository is at first gelatin liquefaction in uterus, the sodium alginate granule in uterus slowly liquefies and progressively discharges lysozyme then, has the advantage that slowly discharges in uterus, while owing to earlier lysozyme is wrapped up through sodium alginate, makes that activity of lysozyme can keep effectively in the process of preparation suppository in the process of processing.
Lysozyme suppository disclosed by the invention has following characteristics:
1, time of in animal uterus, existing of this preparation long, after administration 48 hours, also kept very high bactericidal activity in the mucus of uterus, administration just can effectively prevent and treat animal endometritis 1-2 time basically.
2, bactericidal effect is strong.Said preparation has very strong bactericidal action to mammal uterus pathogenic bacterium.Experiment in vitro can effectively be killed gram positive bacteria and negative bacterium.
2, stability is strong.This product was preserved 2 years at 25 ℃ of room temperature, and biological activity does not have to change substantially.Be better than lyophilized powder aspect the storage of preparation and the transportation.
3, the present invention is a biological preparation, has overcome traditional chemical class preparation uterine mucosa is had zest, and antibacterial is easy to generate chemical sproof shortcoming.Easily accepted by body, and without any toxic and side effects.
4, this preparation adopts the mode of local application, convenient drug administration.Comparing with lyophilized powder does not need other dissolving, and administration is more convenient.
Description of drawings
Fig. 1 is the release in vitro curve chart of lysozyme suppository.
The specific embodiment
Embodiment 1 staphylococcus lysozyme suppository preparation method 1 (prescription 1)
1, composition and proportioning
Lysozyme 0.5%, 0.01% staphylococcus lysozyme, gelatin 5%, glycerol 5%, sodium alginate 5%, hibitane 0.5%, water 83.99%.
2, preparation
(1) at first 0.5 gram lysozyme and 0.01g staphylococcus lysozyme joined mix homogeneously in the sodium alginate soln of 100mL 5%, join the 200mL ethyl acetate: in dichloromethane=3: 1 solution such as grade, add 0.5 gram polyvinyl alcohol, powerful stirring (rotating speed is 100-300 rev/min), the calcium chloride solution 5-10mL of Dropwise 5 % then, dropwising the back continues to stir 30 minutes, form the microgranule of diameter at 10um～1mm, filtration obtains containing the sodium alginate micro ball of lysozyme, lyophilization then.
(2) take by weighing gelatin 5g, glycerol 5g, 0.5 gram hibitane adds 83.99mL water and is heated to 60 ℃ of dissolvings.
(3) sodium alginate micro ball that contains lysozyme that step (1) is made joins in the gelatin glycerol dissolving liquid that step (2) makes, and pours the suppository die for molding rapidly into and gets final product.
Embodiment 2 staphylococcus lysozyme suppository preparation methoies 2 (prescription 2)
1, composition and proportioning
Lysozyme 2%, gelatin 10%, glycerol 10%, sodium alginate 2%, hibitane 0.3%, water 75.7%.
2, preparation
(1) at first 2 gram lysozyme is joined mix homogeneously in the sodium alginate soln of 100mL4%, join the 200mL ethyl acetate: in dichloromethane=3: 1 solution such as grade, add 0.5 gram polyvinyl alcohol, powerful stirring (rotating speed is 100-300 rev/min), the calcium chloride solution 5-10mL of Dropwise 5 % dropwises the back and continues to stir 30 minutes then, forms the microgranule of diameter at 10um～1mm, filtration obtains containing the microsphere of lysozyme, lyophilization then.
(2) take by weighing gelatin 10g, glycerol 10g, hibitane 0.3g adds 75.7mL water and is heated to 60 ℃ of dissolvings.
(3) sodium alginate micro ball that contains lysozyme that step (1) is made joins in the gelatin glycerol dissolving liquid that step (2) makes, and pours the suppository die for molding rapidly into and gets final product.
Embodiment 3 staphylococcus lysozyme suppository preparation methoies 2 (prescription 3)
1, composition and proportioning
Lysozyme 8%, gelatin 15%, glycerol 20%, sodium alginate 0.5%, hibitane 0.1%, water 56.4%.
2, preparation
(1) at first 8 gram lysozyme is joined mix homogeneously in the sodium alginate soln of 100mL 0.5%, join the 200mL ethyl acetate: in dichloromethane=3: 1 solution such as grade, add 0.5 gram polyvinyl alcohol, powerful stirring (rotating speed is 100-300 rev/min), the calcium chloride solution 5-10mL of Dropwise 5 % dropwises the back and continues to stir 30 minutes then, forms the microgranule of diameter at 10um～1mm, filtration obtains containing the microsphere of lysozyme, lyophilization then.
(2) take by weighing gelatin 15g, glycerol 20g, hibitane 0.1g adds 56.4mL water and is heated to 60 ℃ of dissolvings.
(3) sodium alginate micro ball that contains lysozyme that step (1) is made joins in the gelatin glycerol dissolving liquid that step (2) makes, and pours the suppository die for molding rapidly into and gets final product.
The stripping release test of embodiment 4 suppositorys
1, experiment material
Suppositorys by above-mentioned prescription 1, prescription 2, prescription 3 preparations (contain gelatin 20%, glycerol 20%, lysozyme 2%) in contrast with the lysozyme gelatin glycerin suppository of routine.Fill a prescription 4 ingredients with prescription 1, but the used sodium alginate mean particle dia that contains lysozyme is about 1-10um; Fill a prescription 5 ingredients with prescription 1, but the used sodium alginate mean particle dia that contains lysozyme is about 1-10mm.
2, test method
Dissolution determination method: get 1 piece of the suppository (about 5.0 grams) for preparing, with the 10mL0.9% normal saline solution is solvent, and temperature is 37 ℃, and rotating speed is 75r/min, take a sample respectively at 0, when 30min, 60min, 3h, 6h, 12h, 24h, 48h, measure antalzyme activity unit.And calculate the dissolution of each sample according to the unit of activity of the lysozyme in the suppository that records.
3, experimental result
From table 1, fill a prescription the as can be seen external slow release effect of 1, the 2 and 3 lysozyme suppositorys that prepare of Fig. 1 (the release in vitro curve chart of lysozyme suppository) is better than glycerinated gelatin suppository far away, at 48 hours, three kinds of just basic all releases of suppository, and conventional glycerol gelatin matrix just all discharged at 3 hours.Prescription 4 is because the particle diameter of microgranule is too little, and rate of release is obviously fast than preceding 3 prescriptions, and prescription 5 is because particle diameter is too big, and in the suppository preparation, the difficult control of the quality of suppository does not discharge so further detect its stripping.Result of the test shows its had good sustained release effect of suppository of the prescription 1-3 that the present invention prepares.
The comparison of the release in vitro effect of 3 kinds of lysozyme suppositorys of table 1 and glycerinated gelatin suppository
The external pharmacodynamics test of embodiment 5 suppositorys
1, test equipment
Test strain: staphylococcus aureus (ATCC6538); Escherichia coli (ATCC 11229) phosphate buffered solution (PBS0.03mol/l pH7.2)
2, test method
Test organisms 24h slant culture is washed with PBS, make bacteria suspension.
Get above-mentioned bacteria suspension, nutrient agar (antibacterial) or sabouraud's agar (yeast) 15mL with cold to 40～45 ℃ pour into, rotate plate, make it full and uniform, after agar solidifies, with being attached to the surface that solid medium gets after the suppository section (the about 3-5mm of thickness) to be measured, then flat board is placed 35 ℃ ± 2 ℃ cultivations.The effectiveness of suppository is estimated by the size that has or not inhibition zone and inhibition zone.Having or not by "+" of inhibition zone, "-" represent, " ++ " or " +++" represents that respectively inhibition zone is greater than 10mm or greater than 15mm in addition.
3, result of the test
1, the 2 and 3 lysozyme suppositorys that prepare of filling a prescription as can be seen from Table 2 can effectively suppress staphylococcus aureus and colibacillary growth.The external bactericidal effect that presses down that proves above-mentioned 3 kinds of suppositorys is good.
The external fungistatic effect of 3 kinds of lysozyme suppositorys of table 2 relatively
| Prescription | Staphylococcus aureus (ATCC6538) | Escherichia coli (ATCC 11229) |
| ????1 | ????+++ | ????++ |
| ??2 | ????+++ | ????++ |
| ??3 | ????+++ | ????++ |
| The blank formula that does not contain lysozyme | ????- | ????- |
The stability test of embodiment 6 lysozyme suppositorys
1, test equipment
Test strain: staphylococcus aureus (ATCC6538);
Phosphate buffered solution (PBS0.03mol/l pH7.2)
2, test method
Test organisms 24h slant culture is washed with PBS, make bacteria suspension.
Get by each 4 of test agent (suppository that contains lysozyme) and contrast prints (with the sample homogeneous material, equal size, but do not contain anti-biotic material, and), be divided into 4 groups and place 4 sterilization conical flasks through sterilization treatment.
Get above-mentioned bacteria suspension, through 103 dilution back countings, be inoculated into conical flask then, every bottle graft kind 20mL joins print in the flask respectively, places 35 ℃ ± 2 ℃ to cultivate 18-20 hour, and OD600 is detected in the back, calculates bacteriostasis rate.
X=(the blank group-OD600 experimental group of the OD600)/blank group * 100% of OD600
3, result of the test
Lyase suppository is killed activity to staphylococcus aureus at 25 ℃ after depositing 24 months and is not changed as can be seen from Table 3.
Table 3 is deposited after 24 months the staphylococcus aureus killing effect for 25 ℃
Annotate: negative control group does not have bacterial growth.
Embodiment 7 lysozyme suppositorys in the intrauterine metabolism of milch cow
1, test material
Lysozyme suppository
Test strain: staphylococcus aureus (ATCC6538);
Phosphate buffered solution (PBS0.03mol/l pH7.2)
2, test method
Test organisms 24h slant culture is washed with PBS, make bacteria suspension, standby through 103 dilution backs.After ill milch cow intrauterine is given lysozyme suppository, respectively at detecting mucous bacteriostatic activity in 1,3,6,12,24,48 hour.Different time is separated to each 0.5 gram of the excretory mucus of intrauterine after drawing medicine, joins 4.5mL and contains in the culture medium of test organisms, places 35 ℃ ± 2 ℃ to cultivate 18-20 hour, and OD600 is detected in the back, calculates bacteriostasis rate.
X=(the blank group-OD600 experimental group of the OD600)/blank group * 100% of OD600
3, result of the test
Experimental result shows that lysozyme suppository can keep the long time at the milch cow intrauterine after the administration of milk cattle uterus, also kept certain bactericidal activity in the mucus in 48 hours after the administration.
The comparison of the mucous bactericidal activity in different time uterus after the administration of table 4 lysozyme suppository
Embodiment 8 lysozyme suppositorys are to the action effect of cow endometritis
1, the sick cattle symptom of chronic endometritis
Discharging purulent secretion with vagina is main clinic symptoms, and sick cattle has canescence or the thin pus of yellowish-brown from vaginal orifice row, so be stained with vaginal discharge or dry crusts at root of the tail, vaginal orifice and Fei Jiechu.Visible vaginal mucosa of examination per vagina and uterus abdominal part hyperemia, cervix uteri swelling, lax opening, the cervix uteri mouth is with purulent secretion; Examination per rectum finds that cornua uteri increases, and contractile response is faint, lacks flexibility, and palace wall thickening hardening when the rheuminess thing is accumulated in the uterus, then has the slight fluctuations sense.
2, trial drug
Lysozyme suppository; Sharp native agent group.
3, test method
In cow reproduction year, select to divide 20～40 days puerperiums to make a definite diagnosis the milch cow that suffers from chronic endometritis and be for experiment, according to the treatment order, it is divided at random " lysozyme " group and " sharp native agent " group.
Lysozyme group: use 1 piece of lysozyme suppository directly to put into the milk cattle uterus at every turn, be administered once every one day.
Sharp native agent group: sharp native agent medicinal liquid 100mL (ethacridine 0.5g, Oxytetracycline Base 2g are dissolved in the 100mL normal saline) directly injects intrauterine, the next day 1 time, continuous 3～4 times is 1 course of treatment.
4, therapeutic effect criterion
Cure: the transparent free from extraneous odour of mucus that vagina is discharged, other clinical symptom disappearance are recovered the normal estrus cycle, and gestation is determined in the straight inspection in 2～3 months of breeding back.
Effectively: the uterus is obviously softening, and contractility is strong, other sxs, but not pregnant after the breeding of oestrusing.
Invalid: with the treatment before relatively do not have significant change.
5, result of the test
Table 5 lysozyme group and sharp native agent group therapeutic effect are relatively
As can be seen from Table 5, treat cow endometritis (based on chronic purulence and Catarrhal purulence) effect with lysozyme and be significantly improved than traditional sharp native agent therapy, the former improves cure rate more than 30 percentage points than the latter.
Claims (5)
1. a suppository for the treatment of mammalian endometritis is characterized in that, in the gross weight of preparation, component comprises: lysozyme 0.5-8%, gelatin 1-30%, glycerol 1-40%, sodium alginate 0.1-20%, water 40-95%.
2. the suppository of treatment mammalian endometritis according to claim 1 is characterized in that, also comprises the staphylococcus lysozyme of 0.01-0.1% percentage by weight in the formulation components.
3. the suppository of treatment mammalian endometritis according to claim 1 and 2 is characterized in that, also comprises the hibitane of 0-1% percentage by weight in the formulation components.
4. the suppository of treatment mammalian endometritis according to claim 1 is characterized in that, in the gross weight of preparation, component comprises lysozyme 0.5-8%; Gelatin 5-15%; Glycerol 5-20%; Sodium alginate 0.5-5%; Hibitane 0.1-0.5%, water 50%-85%.
5. the preparation method of the suppository of treatment mammalian endometritis according to claim 1 is characterized in that, comprises the steps:
(1) contain the preparation of lysozyme microgranule: lysozyme (containing or do not contain staphylococcus lysozyme) joins mix homogeneously in the sodium alginate soln, joining volume ratio is ethyl acetate: in the solution of dichloromethane=3: 1, add polyvinyl alcohol, powerful stirring (rotating speed is 100-300 rev/min), dropping calcium chloride solution dropwises the back and continues to stir then, forms the microgranule of diameter at 10um～1mm, filtration obtains containing the microsphere of lysozyme, lyophilization then.
(2) preparation of suppository: take by weighing gelatin, glycerol according to the above ratio, add the water heating for dissolving, the microgranule that step (1) is prepared joins in the gelatin glycerol that dissolves according to the above ratio then, pours the suppository die for molding rapidly into.
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102552888A (en) * | 2010-12-10 | 2012-07-11 | 上海高科联合生物技术研发有限公司 | Suppository for treating mammal endometritis |
| CN102552983A (en) * | 2010-12-27 | 2012-07-11 | 上海高科联合生物技术研发有限公司 | Biological enzyme-carrying calcium phosphate bone cement for preventing and treating bone infection and preparation method thereof |
| CN116617318A (en) * | 2022-02-14 | 2023-08-22 | 菲吉乐科(南京)生物科技有限公司 | Phage suppository for treating cow endometritis and preparation method thereof |
-
2008
- 2008-06-27 CN CN200810043569A patent/CN101612392A/en active Pending
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102552888A (en) * | 2010-12-10 | 2012-07-11 | 上海高科联合生物技术研发有限公司 | Suppository for treating mammal endometritis |
| CN102552983A (en) * | 2010-12-27 | 2012-07-11 | 上海高科联合生物技术研发有限公司 | Biological enzyme-carrying calcium phosphate bone cement for preventing and treating bone infection and preparation method thereof |
| CN102552983B (en) * | 2010-12-27 | 2015-10-07 | 上海高科联合生物技术研发有限公司 | Year enzyme calcium phosphate bone cement of prevention and therapy infection of bone and preparation method thereof |
| CN116617318A (en) * | 2022-02-14 | 2023-08-22 | 菲吉乐科(南京)生物科技有限公司 | Phage suppository for treating cow endometritis and preparation method thereof |
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Application publication date: 20091230 |