CN1164278C - Biological preparation for treating burn and preparation method thereof - Google Patents
Biological preparation for treating burn and preparation method thereof Download PDFInfo
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- CN1164278C CN1164278C CNB011235837A CN01123583A CN1164278C CN 1164278 C CN1164278 C CN 1164278C CN B011235837 A CNB011235837 A CN B011235837A CN 01123583 A CN01123583 A CN 01123583A CN 1164278 C CN1164278 C CN 1164278C
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Landscapes
- Medicines Containing Material From Animals Or Micro-Organisms (AREA)
- Medicines Containing Plant Substances (AREA)
Abstract
The invention discloses a biological preparation for treating burn and a method for producing the preparation, wherein the preparation contains bacillus cereus CGMCC0601 fermentation liquor. The biological preparation has good curative effects on pain, exudation, infection, deep accumulation, progressive necrosis of burn tissues and scar healing of deep II-degree wounds, can dry the surface of a scald, reduce exudation, accelerate the growth of new granulation tissues and promote the healing of the scald tissues by using the biological preparation, and has the advantages of small irritation, quick pain relieving, effective infection control, short treatment course, no scar and the like.
Description
Technical field
The present invention relates to medicinal preparation and preparation method thereof, specifically belong to the biological preparation of treatment burn and the method for producing said preparation.
Background technology
Burn is a kind of commonly encountered diseases, frequently-occurring disease, also is the higher trauma of disability rate.Show that according to the area sampling investigation annual burn crowd's sickness rate is 2% person-time, remained scar ratio is about 30%.The normal method that adopts of clinical treatment burn generally is after carrying out the wound surface early time treatment at present, or adopts occlusive dressing or adopt exposure method to treat (" burn treating " Li Ao chief editor, the People's Health Publisher publishes).Occlusive dressing is the thick dressing wrapping wound surface with the sterilization suction, makes it to be isolated from the outside, with the protection wound surface.The weak point of this method is, when wrapping is handled, the dressing of wrapping internal layer usually occur or with wound surface adhesion, exchange difficulty, or drainage effect is not good enough.In the bandaging method for treating process, also usually because the wound surface humidity is brought out infection.Exposure method is that burn wound is exposed in the dry air, covers or wrapping without dressing, makes the sepage of wound surface and slough be dried to crust, with the temporary protection wound surface.Though this method wound surface is unfavorable for the growth of antibacterial, can make the district's tissue that is retarded by silt withered downright bad, its damaged condition is not second to the damage of initial stage burn.Exposure method requires high to external environment condition, and the dry crusts time is long, generally can become crust at burn after 48 hours.Prior treatment method is fit to the medicine that burned skin uses because of lacking, thereby curative effect is not fully up to expectations.In addition, no matter be occlusive dressing or exposure method, during debridement, all to carry out the wound surface infection and handle in early days.The selected anti-infectives of debridement mainly contains three classes: the one, and Chinese herbal and crude drugs preparations is as Ilex purpurea Hassk.[I.chinensis Sims, Radix Ilicis Pubescentis, Rhizoma Coptidis, Cortex Phellodendri, eucalyptus leaves etc.; The 2nd, the chemicals preparation is as hydrogen peroxide, bromo geramine, mafenide etc.; The 3rd, antibiotic formulations such as penicillin, gentamycin etc.Because most drug all can absorb from wound surface, so large tracts of land uses these medicines, and especially concentration is excessive or usually can cause when using powder and poison or visceral lesion.Moreover these medicines generally all have certain zest, usually can increase misery to the patient in use.
Summary of the invention
Purpose of the present invention just provides a kind of suitable burned skin healing, little, the eutherapeutic biological preparation of zest, and a kind of method for preparing said preparation is provided simultaneously.
The object of the present invention is achieved like this, and prepared biological preparation contains Bacillus cercus CGMCC0601 fermentation liquid.
The microorganism L-72 (being microorganism used therefor of the present invention) that Bacillus cercus CGMCC0601 fermentation liquid is used, through Institute of Microorganism, Academia Sinica (foundation 1, Krieg N.R.et al.Bergey ' s Manual ofSystematic Bacteriology (Vol, 2) p.1105-1138.1986.2, Colin RHarwood, Bacillus, Plenum Press, 1989.3, " bacillus " Cai Miaoying etc. such as RE Gordon translates agriculture publishing house 1978.) be accredited as Bacillus cercus (Bacillus cereus).Microorganism used therefor of the present invention has been submitted China Microbial Culture Preservation Commission's common micro-organisms center preservation in the present patent application a few days ago, and preserving number is CGMCC0601, and preservation day is July 12 calendar year 2001.
It is shaft-like that microorganism used therefor of the present invention is Gram-positive, and size is 0.9-1.0 * 2.5-4.0 μ; Spore is oval, and near-end is given birth to, and does not expand; The particulate accumulation of poly-(PHB) is arranged in the born of the same parents; Catalase and oxidase are all positive, and the anaerobe growth is positive, and the V-P reacting positive produces acid to glucose, arabinose, xylose, mannitol are not produced acid, hydrolyzed starch, decomposing casein utilizes citrate and propionate, V-P reacts end of a period, and pH value is 5.0, does not grow in 7%NaCl.
Microorganism used therefor growth conditions of the present invention: aerobic environment, pH value 7.8-8.5; Temperature 28-32 ℃; Cultivation conditions: leave standstill, dynamically all can; Production cycle: left standstill dynamically 10-16 hour 60-72 hour.
Microorganism used therefor of the present invention has good stable.This microorganism is found in 1972, reserves seed for planting because of mycete there being stronger inhibitory action separate, and 4 ℃ of preservations are standby.Every through two months voltage regulator tube (going down to posterity) once; Each to screening in 2 years once, choosing colony is big, thickness, stretched filament, neat in edge, Jing Xiaguan are looked into the special pattern structure that this bacterium is arranged, and can choose, amplification culture is decided to be female the kind, is used for preparation.
The Physiology and biochemistry of doing in per 5 years once than system detects (seeing Table 1), uses through the cultivation of two more than ten years, does not find that this bacterium has variation phenomenon, the preparation that annual different times is produced, and drug effect is zero difference also.
This microorganism is difficult for producing variation phenomenon in use, preservation and the process that repeatedly goes down to posterity, and pretending is that a production strain is reliable and promising.
This microorganism had for two more than ten years from finding so far, always went down to posterity more than 100 time, in biochemical reactions and use, found no variation and decay.Its main biochemical reactions detects and sees Table 1.
Table 1: Bacillus cercus CGMCC0601 major physiological biochemical reaction detects table
Test item detection time (year, month)
72.5 77.5 82.8 88.5 93.7
Gram+++++
Bacterium colony thickness stretched filament+++++
The special decorative pattern of bacterium colony+++++
Liquid culture formation Mycoderma+++++
Amylase+++++
Casein+++++
Nitrate+++++
Acetate+++++
The V-P reaction+++++
The 7.5%NaCI growth-----
Anaerobic growth-----
The acid of glucose product+++++
The acid of lactose product-----
The acid of sucrose product+++++
The acid of maltose product+++++
The acid of mannitol product-----
The acid of xylose product-----
Peroxidase+++++
Its culture of strains method of the contained microorganism of the present invention is:
1, mother culture media: Carnis Bovis seu Bubali cream 10g, peptone 5g, sucrose 10g, agar 18g, sodium chloride 2.5g light water 1000ml, pH value 7.5,1.034 * 10
5Pa sterilization in 30 minutes.Be cooled to 60 ℃, pour plate.
2, inoculation: strain (CGMCC0601) is inserted in the inoculating loop adhesional wetting line on the flat board, 30 ℃ of constant temperature culture 24 hours, choose single, shape big, milky white, thickness, wire drawing length, neat in edge, mirror are observed the bacterium colony of the unique pattern structure of maintenance down as the production strain.
3, system is female plants: preferred bacterium colony is inserted in the inclined-plane 30 ℃ cultivated three days, Xiang Guanzhong adds sterilized water 3ml, and Mycoderma is washed, and hook goes out culture medium then, inserts the inclined-plane pipe after bacterium liquid is boiled three times, cultivated three days for 30 ℃, and 4 ℃ of preservations, standby.
4, system original seed: culture medium is Carnis Bovis seu Bubali cream 10g, peptone 5g, and sucrose 10g, sodium chloride 2.5g, water steams 200ml, 1.034 * 10
5Sterilization in Pa30 minute.
5, mother is planted connect five bottles of original seeds with one and be as the criterion, plant to mother and add 5.5ml original seed base in the base, the 2ml suction pipe of inhaling ball with band is pressure-vaccum repeatedly, and lawn is washed in the solution, and every milliliter connects one bottle in original seed, mixes back 30 ℃ and cultivates 24 hours, is original seed.
The preparation method of biological preparation of the present invention comprises the following steps:
A, preparation culture medium: culture medium contains Folium Populi Pseudo-simonii powder 25-35 part, Japanese pagodatree leaf powder 20-30 part, Semen Maydis powder 80-120 part, analysis for soybean powder 20-30 part, sodium carbonate 8-10 part, add water to paste, placed 0.5-1 hour, and added water 8000-10000 part, transferring pH value with sodium hydroxide is 8.0-8.5, stir, seal 1.034 * 10
5Sterilization in Pa20-30 minute;
B, culture medium temperature are reduced to 28-32 ℃, by the 5-12% access original seed of culture medium capacity; Cultivated 2-3 days for 30 ± 1 ℃, to white Mycoderma formation;
The phenol of c, adding 0.5-1% stirs, and adds the filter aid of 0.5-1%, filters, and collects limpid transparency liquid.
For guaranteeing that preparation has better therapeutic, wherein the Folium Populi Pseudo-simonii powder is preferably selected lobule Folium Populi Pseudo-simonii powder for use, and the Japanese pagodatree leaf powder is preferably selected Flos Robiniae Pseudoacaciae leaf powder for use.
That the reaction of biological preparation of the present invention and potassium oxalate can generate etc. is big, evenly, disperse, have the spherical particle of light folding endurance, prepared biological preparation contains spherical particle for every milliliter and adds up to 10-20 hundred million, its curative effect is good especially.
Biological preparation of the present invention can be guaranteed its stable curative effect when disease is burnt in treatment by quality control.
Its qualitative checking method is: get this product 5ml and add 10% potassium oxalate 0.2ml mixing and form white emulsion liquid, its precipitation is dissolved in 0.2% dilute sulfuric acid, is insoluble to 36% acetic acid, and dried powder is dissolved in dimethylbenzene.
The reaction of this product and potassium oxalate can generate the water-insoluble solid material, examine under a microscope be wait big, evenly, disperse, have the spherical particle of refractivity, its numbers of particles becomes positive to concern with active ingredient.
Quantitative analysis method is: get this product 5ml, add 10% potassium oxalate 0.2ml, get the 1ml adding distil water after the mixing and be mixed to 100ml, get a little and place on the blood counting chamber, the total number of particles of getting with 400 power microscopes in four medium squares (each medium square has 16 lattices) of four jiaos is calculated.Formula:
Example: 328 of granule numbers.
Calculate:
The specific granule number that is biological preparation of the present invention is 20.5 hundred million.
The specific granule of biological preparation of the present invention has identical charges, can form homodisperse individuality in solution, can count on blood counting chamber.
Biological preparation of the present invention to the pain of burn wound, ooze out, infect, long-pending dense, the necrosis progrediens of tissue of burn and the cicatrix healing of dark II ° of wound surface, good curative effect is arranged.Use biological preparation of the present invention, can make the scald dry tack free, ooze out minimizing, can accelerate new granulation tissue growth, promote to scald organization healing, have simultaneously that zest is little, pain relieving fast, effective advantages such as control infection, short treating period, preventing from scar.
CGMCC0601 microorganism of the present invention makes the chlorophyll in the culture medium become the red porphyrin of leaf during the fermentation.The carbonyl on the red porphyrin of leaf and the hydroxyl reaction of carboxyl and phenol generate corresponding fat or ether.This is similar to the lipid material that the normal skin surface exists, thereby can play emulsification with the moisture on trauma skin surface and produce the lipid film, can play a protective role to the skin trauma surface.
Excellent results of the present invention has obtained further confirmation by following experimentation.
Pharmacodynamic study:
1, experiment material
1.1 medicine: experimental group, biological preparation of the present invention, lot number 94025; Matched group, the burn spirit, lot number 940766, Le Rentang pharmaceutical factory in Shijiazhuang produces.
1.2 animal: Kunming mouse 24-25g, male and female half and half, effluent north medical college Experimental Animal Center provides.
1.3 strain: staphylococcus aureus (26003), escherichia coli (44102), streptococcus faecalis (32221), bacillus pyocyaneus (10104), Candida albicans (10231) provide by the calibrating of Beijing pharmaceutical biological product.
1.4 culture medium: sabouraud culture medium, plain agar culture medium and meat soup.
2, method and result
2.1 influence to the mice wound tissue healing:
Get healthy mice, the back depilation was used etherization with animal after 24 hours, placed on 80 ℃ of constant temperature aluminium sheets, continued 15 seconds, caused 10% II ° of burn.Burn wound's area is (2 * 3cm approximately
2) (this condition is to determine after trial test is by histopathologic examination).Animal is divided into four groups at random, the blank group, matched group, experimental group is divided into 1 group of experiment, 2 groups of experiments according to high and low two dosage.Every group 12, spray medicine after the scald immediately, blank group water spray, matched group is given clever 0.5ml/ time of burn, experimental group dosage 0.5ml/ time, each is organized 2 times/day, and successive administration 14 days is observed body weight change situation between incrustation time after each treated animal administration, each group of decrustation time and 4 days, 8 days, 12 days.The results are shown in Table 2, table 3.
Table 2: biological preparation of the present invention is to the influence of mice burn wound healing
The group incrustation time (my god) the decrustation time (my god)
Blank group 7.0 ± 1.3 15.6 ± 4.2
Test 1 group of (a large amount) 4.8 ± 0.4
*11.3 ± 2.5
*
Test 2 groups of (low amounts) 4.6 ± 0.8
*11.4 ± 1.3
*
Matched group 5.8 ± 1.1 13.4 ± 3.8
x±s，n＝12，
*P＞0.05
Table 3: biological preparation of the present invention is to the x ± s that influences of mice burn back body weight, and n=12 restrains
Before the group administration after the administration 4 days 8 days 12 days
Blank group 24.4 ± 0.5 22.6 ± 0.6 24.1 ± 0.8 27.2 ± 1.3
Test 1 group of (a large amount) 24.5 ± 0.6 25.8 ± 0.9
*25.7 ± 1.4
*27.2 ± 1.4
Test 2 groups of (low amounts) 24.3 ± 0.3 25.8 ± 0.5
*25.8 ± 0.5
*27.4 ± 0.8
Matched group 24.4 ± 0.4 24.6 ± 0.9 25.2 ± 1.0
*27.4 ± 1.4
*P＞0.05
**P＜0.01
Experiment shows: experimental group can be alleviated the phenomenons such as shouting, jump, tremble that the scald animal causes because of pain, scald wound drying, nothing infect behind the animal continuous use of experimental group, the time that grows the time of crust and take off crust relatively has significant difference all early than matched group with matched group.From experiment, can also observe, after mice is scalded, cause the minimizing of activity and food ration because of severe pain, make weight loss, after the administration, no matter experimental group is a large amount group (biological preparation contains spherical particle for every milliliter and adds up to 2,000,000,000) or low amount group (biological preparation contains spherical particle for every milliliter and adds up to 1,000,000,000), animal ingestion and activity are all very fast to have recovered normal, body weight promptly was significantly improved in administration in 4 days, compared with matched group, had significant difference.
2.2 extracorporeal bacteria inhibitor test:
The bacteriostatic test method is carried out according to Ministry of Public Health " drug inspection method ".
Result of the test sees Table 4.
Table 4: send out of the influence of this open-birth thing preparation to pathogenic bacterium
Golden yellow white
Time (branch) escherichia coli bacillus pyocyaneus contrast
The staphylococcus candidiasis
0 thirty thirty thirty thirty thirty
2 Lv Lv-Lv thirty
4++-Lv thirty
6---+thirty
8---+thirty
10----thirty
Experiment shows that biological preparation of the present invention can produce significant inhibitory effect to 5 kinds of higher pathogenic bacterium of burn wound's infection rate in 10 minutes.
This biological preparation shows through skin medication toxicity test: each treated animal that uses biological preparation of the present invention, at the medication viewing duration, each treated animal feed, drainage and activity are all no abnormal, and breathing and extremity are movable normal, do not have significant change between each group of body weight, do not have dead the generation.
This biological preparation shows through skin medication stimulation test: intact skin and damaged skin group there is no irritant reaction such as erythema and edema.
Its clinical research of biological preparation of the present invention obtains good effect, and clinical research is reported as follows.
Methods of clinical observation and project:
1, observation unit: Hebei Medical College the 3rd hospital, the 256 hospital of the Chinese People's Liberation Army, Shijiazhuang City Changan District hospital, the Shijiazhuang City pendant English biomedical developmental research institute etc. that burns.
2, observation item: the tissue reaction behind exudation, pain relieving, infection, antipruritic, granulation promoting, the termination burn etc.To difference burn factor, degree and consultation time sooner or later, observe its cure time and have or not cicatrix etc.
3, administrated method: I ° burn, wound surface are interrupted sprinkling biological preparation of the present invention to pain and disappear.The wound surface of II ° of burn cleaning, no dirt, the person that do not use other medicines can directly spray this medicine, every day 2-6 time, but if any the emptying of blister low level clip.The pain wound surface every 5 minutes spray medicines once changes positive usual amounts into behind painful the ending.Spray value is not advisable evenly not flow.General 160cm
2Area, consumption liquid 1ml.Dirt being arranged or be covered with other medicines as wound surface should be clean with normal saline flushing earlier, uses the same method then to spray this medicine.Generally take exposure method.The wound surface and the low temperature environment of commute friction can adopt the soak bandaging strapping.Infective wound surface with empyema or necrotic tissue cleaning, is used the soak method, change sprinkling into after wound surface is clean.
Clinical observation:
1, general clinical data: treated fire victim's 787 examples in October, 1993 altogether from 1987 to, and male 388 examples account for 49.3%; Woman's 399 examples account for 50.7%.Minimal ages 8 months, maximum 65 years old age.Burn surface area is 1～37%.II ° of burn 756 examples account for 96.06%, and dark II ° of burn 31 examples account for 3.94%.Wherein treat 3 examples of burning of burning.There are head, face, neck, nose, mouth, ear, shirtfront, back, extremity, buttocks, external genitalia, scrotum etc. in the burn position.
2, fully recovering of therapeutic effect: II ° of burn 84.5% at 5-10 days; 15.6% recovery from illness in 10-15 days, and 3% secondary infection (mostly be and accidentally injure crust) is wherein arranged.Dark II ° of area 10% is with the patient of interior burn, and 80.5% recovery from illness in 15-20 days, and 19.4% recovery from illness in 15-30 days.Can there be the cicatrix healing without skin-grafting yet.
The burn observation of factor, administration time, pain relieving and recovery from illness sees Table 5 to biological preparation of the present invention to difference.
Table 5: biological preparation of the present invention is observed the medication of burn
Pain is ended recovery from illness
The burn burn is caused injury to using burn % time time remarks
Factor position medicine time example number (branch) (my god)
Liquid extremity, the timely 200 25.4 5-10 2-5 decrustations in shirtfront, no cicatrix boil
Steam back, face etc.
The same 14～30 minutes 105 13.3 5～15 5～10 is the same
The same 1-7 days 456 57.9 5～15 5～20 is the same
The same with 6 0.8 10～20 15～30 the same more than first half of the month
Red ferrum hands, buttocks timely 2 0.3 5～15 2～9 are the same
Flame front, back 15 minutes 5 0.6 40～45 2～4 are the same
Nitric acid eye, hands detumescence in 10～15 minutes 2 0.3 10～15 2～6 20 minutes
Caustic soda scrotum 3 days 1 0.1 8 takes off thin skin one deck immediately
The same buttocks first quarter moon 1 0.1 is 29 no cicatrixes immediately
Electric arc the back of the hand, 20 minutes-1 day 3 0.6 15 2～4 no cicatrixes of face
Sulphuric acid instep, upper limb 6 minutes-1 day 3 0.6 be 3～6 no cicatrixes immediately
Colophonium hands, sufficient 1-4 days 3 0.6 30 10 no cicatrixes
From observed result as can be seen, pain relieving in 10-20 minute (flame burn pain relieving in 45 minutes) behind burn wound's usefulness biological preparation of the present invention.Recovery from illness is sooner or later relevant with medication, and medication is more early fully recovered soon more.
To II ° of burn, go to a doctor back 30 minutes of burn with interior person, generally can take off thin skin one deck recovery from illness in 5-10 days, leave no trace; Burn has reached 6-48 hour, and wound surface forms blister, and part skin lesion person is arranged, and treats recovery from illness in 5-12 days; Burn has surpassed a week, through other medicines therapeutic effect gastrointestinal disease patient, uses biological preparation treatment of the present invention instead, 40% recovery from illness in 10 days, and 45% recovery from illness in 20 days, 15% (genus residual wound) fully recovered in 30 days, and all do not have scar tissue.
To the treatment that chemical burn, electric arc are hindered, because burn degree and consultation time difference, treatment is generally at recovery from illness in 2-30 days, preventing from scar.
The observation of exudation: burn wound's spraying biological preparation of the present invention is in half an hour, and visible skin lesion wound surface serosity oozes out minimizing, gets rid of the blister of serosity about 1 hour and no longer heaves, and can see red and swollen obvious disappearance of I ° of wound surface in two hours.After the medication, wound surface does not have the intensification phenomenon, shows that biological preparation of the present invention has the effect of anti-tissue reaction.
Biological preparation of the present invention is applicable to: the skin and flesh damage that various chemical factors such as boiled water, the oil that boils, steam, high temp objects, flame, gunpowder, photoelectricity, strong acid, highly basic cause.
The using method of biological preparation of the present invention is:
1, debridement
(1) wound surface infection suppurates not, and do not used any medicine person, as belong to fresh wound surface, can directly use this medicine; As not belonging to fresh wound surface, use this medicine behind the available normal saline flushing.
(2) traumatic infection suppuration person re-uses this medicine after cleaning up with a small amount of 3% hydrogen peroxide earlier.
(3) the wound surface person that uses the other drug after must thoroughly removing, can use this medicine.
2, handle blister
Complete blister adopts the low level clip to get rid of sepage, but will protect the blister skin.Bigger blister can be cut several mouthfuls more, and the sepage pressurization is got rid of, and makes the bubble skin be close to wound surface.The bubble skin of shrinkage or pollution will be wiped out.
3, medication
(1) rubbing method: soak into medicinal liquid with absorbent cotton, softly be applied to equably wound surface, every day 3-4 time.This method is applicable to cleaning, pollution and incrustation face.
(2) spraying: with sprayer unit medicinal liquid is sprayed on wound surface equably, is advisable every day 3-6 time not flow.This method is applicable to the wound surface of cleaning and incrustation.
The specific embodiment
Embodiment 1:
Preparation lobule Folium Populi Pseudo-simonii powder, Flos Robiniae Pseudoacaciae leaf powder: select the leaves (band petiole) of PopulussimoniiCarr. in 5 to October for use, Flos Robiniae Pseudoacaciae leaf (band petiole) dries, and wears into 40-60 order powder with little steel, and is standby.
Take by weighing PopulussimoniiCarr. powder 3kg, Flos Robiniae Pseudoacaciae leaf powder 2kg, Semen Maydis powder 10kg, analysis for soybean powder 2kg, sodium carbonate 1kg places container, it is an amount of to add water, stir into paste, place half an hour, add water 1000kg, transfer pH value to reach 8.0 with 5N NaOH, under fully stirring, be sub-packed in the glass jar capacity 350ml, insert original seed 35ml, cultivated three days for 30 ℃, white Mycoderma forms, and bacterium liquid is limpid transparent, the thalline long-chain forms spore, count with the specific granule counting method, quantity reaches 2,000,000,000/milliliter, promptly makes the CGMCC0601 fermentation liquid.Pressing 1% of fermentation liquid total amount and add phenol, fully mix, is that radix adds 1% kieselguhr as filter aid with the mixed liquor total amount, stirs 1 minute, with 7
#The canvas normal pressure hangs filter, collects limpid transparency liquid and is finished product.
Embodiment 2:
Preparation Folium Populi Pseudo-simonii powder, Japanese pagodatree leaf powder: select Folium Populi Pseudo-simonii, the Folium sophorae in 5 to October for use, dry, wear into 40-60 order powder with little steel, standby.
Take by weighing Folium Populi Pseudo-simonii powder 2.5kg, Japanese pagodatree leaf powder 3kg, Semen Maydis powder 8kg, analysis for soybean powder 3kg, sodium carbonate 1kg, place container, it is an amount of to add water, stir into paste, place half an hour, add water 800kg, transfer pH value to reach 8.0 with 5N NaOH, under fully stirring, be sub-packed in the glass jar, capacity 350ml inserts original seed 30ml, cultivates three days for 30 ℃, the white Mycoderma forms, bacterium liquid is limpid transparent, and the thalline long-chain forms spore, counts with the specific granule counting method, quantity reaches 1,500,000,000/milliliter, promptly makes the CGMCC0601 fermentation liquid.Pressing 0.5% of fermentation liquid total amount and add phenol, fully mix, is that radix adds 1% kieselguhr as filter aid with the mixed liquor total amount, stirs 1 minute, with 7
#The canvas normal pressure hangs filter, collects limpid transparency liquid and is finished product.
Claims (7)
1, a kind of biological preparation that is used for the treatment of burn is characterized in that it contains Bacillus cercus CGMCC0601 fermentation liquid.
2, the biological preparation that is used for the treatment of burn according to claim 1 is characterized in that being added with in the said fermentation liquid phenol of 0.5-1.0%.
3, the biological preparation that is used for the treatment of burn according to claim 1, it is characterized in that the reaction of said biological preparation and potassium oxalate generate etc. big, evenly, disperse, have the spherical particle of refractivity.
4, the biological preparation that is used for the treatment of burn according to claim 3 is characterized in that every milliliter of described biological preparation contains spherical particle and adds up to 10-20 hundred million.
5, a kind of preparation method that is used for the treatment of the biological preparation of burn according to claim 2 is characterized in that it includes the following step:
A, preparation culture medium: culture medium contains Folium Populi Pseudo-simonii powder 25-35 part, Japanese pagodatree leaf powder 20-30 part, Semen Maydis powder 80-120 part, analysis for soybean powder 20-30 part, sodium carbonate 8-10 part, add water to paste, placed 0.5-1 hour, and added water 8000-10000 part, transferring pH value with sodium hydroxide is 8.0-8.5, stir, seal 1.034 * 10
5Sterilization in Pa20-30 minute;
B, culture medium temperature are reduced to 28-32 ℃, by the 5-12% access CGMCC0601 strain of culture medium capacity; Cultivated 2-3 days for 30 ± 1 ℃, to white Mycoderma formation;
The phenol of c, adding 0.5-1% stirs, and adds the filter aid of 0.5--1%, filters, and collects limpid transparency liquid.
6, the preparation method that is used for the treatment of the biological preparation of burn according to claim 5 is characterized in that said Folium Populi Pseudo-simonii powder is a lobule Folium Populi Pseudo-simonii powder.
7, the preparation method that is used for the treatment of the biological preparation of burn according to claim 5 is characterized in that said Japanese pagodatree leaf powder is a Flos Robiniae Pseudoacaciae leaf powder.
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