1059 results about "Streptomycin" patented technology

The invention belongs to the field of plant protection, and relates to an agricultural composition containing antibiotics and plant source materials, wherein antibiotic is one or several selected from abamectin, ivermectin, milbemectin, pleocidin, nikkomycin, liuyangmycin, streptomycin, polyoxins, chunleimeisu, jinggangmeisu, ningnanmycin and the like; the plant source material is one or several of smashing material, crude extract, or active ingredient of derris, thunder god vine, tobacco, pyrethrum, tagetes, crofton weed, radix stemonae, cayenne, macleaya cordata, radix sophorae flavescentis and the like. The composition provided by the invention expresses well synergism in a certain matching range, and has excellent control efficiency for crop diseases and pests.

The invention discloses a method of preparing a bombyx mori silk fibroin specific antibody by utilizing a characteristic polypeptide. The method comprises the following steps: synthesizing a "CGAGAGSGAGAGS" polypeptide sequence by utilizing an Fmoc method, coupling the polypeptide with keyhole limpet hemocyanin (KLH) through the cysteine on the N terminus of the polypeptide so as to obtain a complete antigen; diluting the complete antigen with normal saline, mixing the diluted complete antigen with a complete Freund's adjuvant, adding streptomycin and penicillin to carry out an emulsion treatment so as to obtain a primary immunized antigen emulsion, subjecting a rabbit to a primary immunization by using the primary immunized antigen emulsion, then subjecting the rabbit to a strengthened immunization, wherein the strengthened immunization uses a strengthened immunized antigen emulsion, which is prepared by the following steps: mixing the diluted complete antigen with an incomplete Freund's adjuvant, and then adding streptomycin and penicillin to carry out an emulsion treatment so as to obtain the target product; collecting the blood of the immunized rabbit, when the antibody titer in the rabbit blood sample reaches 1 / 10000; making the blood blocks fully contract to completely separate out the antiserum, then collecting the antiserum, and subjecting the antiserum to a centrifugation treatment so as to obtain a supernate.

The invention relates to a method for simultaneously detecting three antibiotic residues including streptomycin, chlorampenicol and tetracycline based on nucleic acid aptamers and quantum dots and belongs to the technical field of analytical chemistry. Three-section complementary cDNA 1 sequences are designed and synthesized and are complemented with three antibiotic aptamers DNA sequences and are further complemented with respective capturing probe DNA sequences of three antibiotics and complementary cDNA 2 sequence part. The three cDNAs 1 are first hybridized with aptamer DNAs supplemented with each other to form double-stranded DNAs. When target antibiotics exist in a sample, the aptamer DNAs are combined with corresponding antibiotics as a priority to cause the double-stranded DNAs to be de-hybridized and release cDNAs 1. The free cDNAs 1 are combined with capturing probe DNAs modified on the surface of a gold electrode, and cDNAs 2 modified with three quantum dots are hybridized with the other end of each cDNA 1. The three quantum dots on the surface of the gold electrode generate dissolution volt-ampere peaks corresponding to the target antibiotics. The three antibiotics are detected by utilizing the relations of changes of peak current values and antibiotic concentration. The method is good in repeatability and stability and high in sensitivity and can be used for directly measuring the three antibiotic residues in samples such as milk.

The invention provides an inducing method and inducing culture medium for differentiation of bone marrow mesenchymal stem cells into osteoblasts in vitro. The inducing culture medium is composed of 1*10<-8> mol / L of dexamethasone, 50 mu mol / L of ascorbic acid and 10 mmol / L of sodium beta-glycerophosphate; and solvent is a supernatant of a sclerite complete culture medium and comprises 10% of fetal calf serum, 100 U / mL of penicillin, 100 mg / L of streptomycin, a mixture of DMEM culture fluid and F12 culture fluid and multiple growth factors secreted by bone cells in the sclerite culture process. According to the invention, bone marrow mesenchymal stem cells of a mouse are purified by replacing the cell culture fluid through an adherent cell passage method, the obtained cells of the first generation are induced, and the supernatant of the sclerite complete culture medium cultured for 72-96 hours is used as the solvent of osteoblast differentiation inducer, thereby obviously improving the in vitro osteogenic differentiation efficiency of bone marrow mesenchymal stem cells.

The invention discloses a colon cancer protein mark parallel test liquid phase chip, which is mainly formed by: micro ball, capture antibody, test antibody and streptomycin-phycoerythrin, wherein the capture antibody with the corresponding micro balls form coupling conjugated, which uses red laser to active the red categorizing fluorescence of the sphere base material and ascertains the type by the different color of the sphere base material; the test antibody is a skin factor mark antibody; the capture antibody and the test antibody can combine with the colon cancer protein mark; the test antibody combines with the streptomycin-phycoerythrin and uses green laser to active the phycoerythrin to measure the report fluorescence molecular number of the sphere base material, which can indirect ascertain the colon cancer protein mark content combines with the sphere base material. The invention also discloses the colon cancer protein mark parallel test liquid phase chip applied in preparing the test agent.

The invention relates to a herbal medicine preparation method for the children bronchopneumonia treatment, belonging to the technical field of Chinese herbal medicine preparation methods; which adopts the technical proposal that: herbs like ginkgo seed, ephedra, apricot kernel, perilla, pinellia ternate, Swallowwort Rhizome (SRh), aster, Tussilago farfara, cortex mori, Chinese wolfberry root-bark, scutelarria, blackberry lily, dyers woad leaf, isatis root, bupleurum, dandelion, honeysuckle, houttuynia cordata, sargentgloryvine stem, sophora tonkinensis, APN (Andrographis Paniculta), Franchet Groundcherry Fruit, balloonflower and liquorice are immersed in 1000ml water for half an hour, and then decocted on mild fire; after decoctation, the herbal liquor is filtered to get 300ml herbal medicine for the treatment of children bronchopneumonia. In prior art, penicillin, streptomycin, and gentamicin are used for the treatment of children bronchopneumonia; the application of penicillin can cause allergic reaction, as a result, the life of the patient may be in danger if rescue is not timely. The preparation method has the advantages of simple manufacturing technology, small side effect and durable and stable therapeutic effect.

The invention provides a strain of Mycobacterium sp.16F for efficiently degrading polycyclic aromatic hydrocarbon and benzene organic matters, which has a preservation number of CGMCC No.6367. The mycobacterium 16F can efficiently, safely and rapidly degrade polycyclic aromatic hydrocarbons and benzene organic matters, can grow and degrade by using fluorene, naphthalene, anthracene, acenaphthene, phenanthrene, pyrene and benzopyrene as the sole carbon source and energy in aerobic condition, and can utilize benzene, m-xylene, toluene, salicylic acid, catechol and other multiple aromatic organic matters. The mycobacterium 16F is sensitive to streptomycin, rifampin, tetracycline, kanamycins and other antibiotics, has good degradation effects to mixed polycyclic aromatic hydrocarbons in aging soils and monocyclic benzene organic matters in water bodies, can be used for restoring and purifying the water-soil environment combinedly polluted by aromatic hydrocarbon organic matters, is important for promoting sustainable development, and has a wide application prospect.

The invention relates to a livestock semen freezing diluent as well as a preparation method and application of the livestock semen freezing diluent. The preparation method of the livestock semen freezing diluent comprises the following steps of: adding 2.71 grams of trihydroxymethyl aminomethane, 1,4 grams of citric acids, 1.0 gram of monosaccharides, 10 myriad IU (International Unit) of penicillium, 10 myriad IU of streptomycin and 0.29-6.97 grams of inositol compounds to a right amount of ultrapure water; optionally adding 5-20 milliliters of penetrability protectants; uniformly stirring; regulating a pH value to 6.8-7.2; adding 5-20 milliliters of fresh yolk subjected to inactivation treatment; uniformly mixing, and then fixing the volume to 100 milliliters; centrifugalizing at low temperature for 1 hour; and filtering supernate to prepare the livestock semen freezing diluent. Because the inositol compounds can effectively inhibit the generation of ice crystals, the livestock semen freezing diluent disclosed by the invention reduces the mechanical injury caused by the ice crystals on semens, achieves the survival rate of the defrozen semens at about 75%, the activity more than 50%, the acrosome integrality at about 60% and the plasmalemma integrality at about 50% and achieves the non-return rate more than 70% after artificial insemination; and the livestock semen freezing diluent has the advantages of no immune response generation, purity guarantee, low price, and the like by using the inositol compounds as the ice crystal inhibitors of the semen freezing diluent.

Disclosed is a cyclic peptide antibiotic in which an enzyme-sensitive bond is replaced by an enzyme-resistant bond.

The invention discloses a method for artificially culturing seawater anucleate pearl. The method comprises the following steps: taking out the mantle epithelium of a healthy nacre, removing the skirt band part, digesting the mantle epithelium in protease solution, then adding the culture solution with shell blood serum to stop the digestion, culturing the culture solution containing shell tissue fluid and a property amount of penicillin and streptomycin in a sterile environment for 5 to 30 hours, then injecting the cell suspension into a pearl breeding shell body, and putting the pearl breeding shell body in the seawater for suspended culture or into a culture pond box for feeding after a recreation period. The method has little damage to the pearl breeding shell, a high survival rate and a high pearl formation rate. The cultured pearl is suitable for medicinal use. The method has convenient operation and low cost.

The invention discloses a single-chain DNA aptamer specifically recognizing streptomycin and application of the DNA aptamer, belonging to the field of biochemistry and molecular biology, analytical chemistry and combinatorial chemistry. The invention provides a method for preferably selecting the streptomycin aptamer by using an improved magnetic bead SELEX technology and four single-chain DNA aptamers with high appetency with the streptomycin and sequences of A1, A3, A6 and A12, and provides a high-specificity detection recognition element with the advantages of better stability, high sensitivity, low cost, and easy preparation, modification and mark for the detection of residue of the streptomycin, especially streptomycin in foods.

The invention relates to a serum-free adipose tissue-derived mesenchymal stem cell culture medium, which consists of a basic culture medium and added ingredients, wherein the basic culture medium is DMEM-LG, and the added ingredients and the content of each added ingredient are shown as follows: 5 to 20ng / mL of alkaline fiberblast growth factors, 5 to 20ng / mL of epidermal growth factors, 100U / mL of penicillin, 100 micrograms / mL of streptomycin, 50 to 200 micrograms / mL of heparin, 2 to 8mM of L-glutamine, 100 to 300 microM of 2-mercaptoethanol, 500 to 2000U / mL of leukaemia inhibitory factors and 0.5 to 2mM of sodium pyruvate. The serum-free adipose tissue-derived mesenchymal stem cell culture medium does not contain the serum, so that the inter-batch difference and the influence of the serum component on the cell culture can be avoided; the exogenous pollution of the serum and the toxicity of the serum on the cells can be avoided; and the ingredients are clear, so that the research of the psychological regulation mechanism of the cells can be facilitated.

The invention discloses a normal temperature preservation diluent for sperm, comprising 120ml to 140ml double distilled water, 1.0g to 3.0g fructose, 3.5g to 5.5g Tris, 2.0g to 3.5g citric acid, 30mg to 50mg GSH, 140mg to 180mg BS, 30mg to 50mg ATP, 450mg to 550mg VC, 300,000 to 500,000 penicillin, 150,000 to 250,000 streptomycin and 1.0g to 2.0g trehalose; the optimum operating temperature ranges from 15 DEG C to 25 DEG C. The normal temperature preservation diluent for sperm has the advantages that, the diluent has good effect on the normal temperature preservation of the sperm of cattle, sheep and deer; the invention has simple production method, low production cost and convenient popularization and application.

The invention discloses a microbial fertilizer resistant to diseases and insects and a preparation method thereof. The microbial fertilizer resistant to diseases and insects comprises the following components in parts by mass: 100 parts of plant stalks, 40-100 parts of grain alcohol residue, 0.5-0.7 part of a bacillus inoculant, 0.2-0.4 part of a streptomycete inoculant, 0.4-0.6 part of a trichoderma inoculant, 0.06-0.08 part of a photosynthetic bacterium inoculant, 5-15 parts of calcium sulphate, 5-15 parts of magnesium chloride and 8-16 parts of potassium dihydrogen phosphate. The microbial fertilizer resistant to diseases and insects contains the plant stalks as well as the bacillus inoculant, streptomycete inoculant, trichoderma inoculant and photosynthetic bacterium inoculant, wherein the bacillus inoculant contains bacillus thuringiensis and can resist to diseases and kill insects; streptomycete can produce streptomycin capable of resisting to diseases and inhibiting proliferation of insects, and trichoderma can enhance stress resistance of crops; and the photosynthetic bacterium inoculant contains substances which are resistant to bacteria and viruses and can passivate pathogenicity of pathogens and inhibit growth of the pathogens. By adopting the technical scheme, disease and insect resistance of the microbial fertilizer is improved.

The invention provides carnation high-efficiency rootone which is characterized by containing the following materials according to weight part: 10 parts of rhodofix, 1-2 parts of seradix, 80-120 parts of mildothane, 15-30 parts of streptomycin and 6000-6400 parts of talcum powder. In the invention, aiming at the problems of root rot and stem rot of the carnation easily harmed by epiphyte and bacteria and being combined with the characteristics that roots produced by rhodofix NAA induction are less in quantity and thick and adventitious roots produced by seradix IBA action are thin and long, NAA and IBA are mixed in the rootone, and meanwhile, mildothane used for sterilizing and agricultural streptomycin are added; therefore, the rooting ratio of the carnation is high, the growing roots are long and thick, the antibiosis and disease-resisting capacity is enhanced, and the root rot ratio is greatly reduced, and the invention has the advantages of tidy rooting, high field production survival rate, convenient usage, low cost, good effect, and the like and is very convenient for the growth of the carnations.

The invention provides a human adiponectin enzyme-linked immunosorbent detection kit, a preparation method and application thereof. The kit comprises antihuman APN monoclonal antibody-coated ELISA plates, recombinant gAPN protein standard, antihuman APN monoclonal detection antibodies, HRP-streptomycin, 10 mM of phosphate buffer with a pH of 7.4, TMB enzyme-substrate chromogenic solution, 0.1 M of phosphate buffer with a pH of 7.4 and 2 M of H2SO4. The invention also comprises the preparation method of the kit and the application of the kit in the detection of human-blood APN content. The kit has the advantages that the kit is convenient and fast to use and operate, capable of getting detection results within 2 hours, high in specificity, low in cost and convenient for clinical large-scale use.

The invention discloses a tissue sample preservative solution used for preserving tissue samples, such as fat, umbilical cords, placentas and the like, after collection and before separation. The tissue sample preservative solution mainly comprises following components: high-glucose DMEM dry-powder culture medium, sodium bicarbonate, DMSO, dexamethasone, insulin, penicillin, streptomycin, amphotericin and a heparin sodium injection. The high-glucose DMEM dry-powder culture medium and the sodium bicarbonate are used for maintaining osmotic equilibrium among cells inside and outside tissue, maintaining a pH value, maintaining a moistening situation of the tissue and providing nutritional components. The DMSO is a freeze-storage protective agent and can prevent freeze-injuries on the tissue samples. The dexamethasone can inhibit immunization and protect activity of stem cells in the tissue sample; the insulin can promote absorption and utilization of the tissue sample to glucose. The penicillin, the streptomycin and the amphotericin can prevent pollution from bacteria and moulds and can eliminate pollution which has occurred. The heparin sodium injection can prevent blood solidification in the tissue samples and increase a yield of the stem cells. The tissue sample preservative solution is simple in components, is low in cost, is convenient to use, can maintain activities of the stem cells in the tissue samples, such as fat, umbilical cords, placentas and the like, and can greatly reduce a time limit from collection to preparation of the tissue samples.

The invention discloses a limbal stem cell serum-free medium, consisting of the following raw materials (calculated by weight percent): DMEM / F12 60ml-90ml, BSA 0.5g-4g, EGF 1Mug-4Mug, insulin 0.1mg-1mg, hydrogen 20ug-100ug, cholera toxin 1ug-10ug, transferrin 0.1mg-1mg, selenious acid 0.1ug-1ug, GCLCM 10ml-40ml, penicillin 5*103IU-2*104IU and streptomycin 5*103IU-2*104IU. The invention adopts the method that the culture medium is added with fibroblast conditioned medium, and the nutritive factors are supplemented with the hormone and cytokine combination, fibroblast conditioned medium and BSA. As a result, the nutritive needs for long-time in-vitro expansion and cultivation of the limbal stem cell serum-free medium are satisfied, and the medium is serum-free and suitable for commercial production.

The invention relates to a freezing diluent for a seminal fluid of livestock. Every 100 ml of the freezing diluent comprises components as follows: 2.71 g of trihydroxymethyl aminomethane, 1.4 g of citric acid, 1.0 g of monosaccharide, 5-20 ml of fresh yolk, 0-10 ml of a permeable protecting agent, 0.1 million IU of penicillin, 0.1 million IU of streptomycin, 0.05-5 g of polyvinyl alcohol and the balance of ultrapure water. The common high-molecular polymer polyvinyl alcohol serving as an ice crystal inhibitor replaces natural antifreeze protein and is applied to the cryopreservation research of the seminal fluid of livestock for the first time. The survival rate of unfrozen sperms is about 75%, the activity is 50% or higher, the acrosome completeness is 65% or higher, the plasma membrane completeness is about 50%, and the non-return rate after artificial insemination is 70% or higher.

The invention relates to an equus semen storage diluter and a preparing method and a use method thereof, belonging to the technical field of animal semen storage. The equus semen storage diluter is prepared from double distilled water, glucose, glycin, penicillin, streptomycin and yolk. The equus semen storage diluter has a good effect of preserving the equus semen under the normal or low temperature, has the advantages of simple operation, low cost, easy mastery, and is suitable for popularization and application, which is beneficial to the development of the equus breeding.

The invention relates to the technical field of biological control of breast cancer disease and discloses a polypeptide for promoting apoptosis of breast cancer cells by targeted uptake of siRNA. Thepolypeptide comprises a polypeptide 1; the sequence of the polypeptide 1 is H-Ile-Phe-D-Trp-Leu-Leu-Trp-Gln-Gly-Arg-Gly-Gly-Gly-Arg-Arg-Arg-Arg-Arg-Arg-Arg-OH. A detection method for the polypeptide for promoting the apoptosis of the breast cancer cells by targeted uptake of the siRNA comprises the following steps: firstly, culturing the breast cancer cells: selecting breast cancer cells MDA-MB-231, culturing the breast cancer cells MDA-MB-231 by adopting a DMEM culture medium, and sequentially adding 10 percent of fetal calf serum, 100 unit / ml of penicillin and 100g / mL of streptomycin in theculture medium. According to the polypeptide for promoting the apoptosis of the breast cancer cells by targeted uptake of the siRNA, the polypeptide 1 wraps the siRNA to form nanoparticles for targeting delivery of the siRNA to the breast cancer cells; the nanoparticles can realize the targeting delivery of the siRNA through a surface receptor of the breast cancer cells and have little damage to normal cells, and the practicality of the polypeptide is improved; meanwhile, the targeting delivery of TRPC1 siRNA by using the polypeptide 1 causes the apoptosis of the breast cancer cells, so that the effect of treating breast cancer is realized.

The invention relates to a diluent for storing the sperm of a pig under normal temperature, which is characterized by comprises the following components by weight ratio: 9-11 of glucose, 0.2-0.3 of ethylene diamine tetraacetic acid, 0.5-0.6 of trisodium citrate, 0.08-0.12 of 1570U / mg penicillin and 0.2-0.25 of 720U / mg streptomycin. The diluent is completely dissolved in distilled water with distilled water according to the dilution ratio that diluent of 1g is diluted with distilled water of 20ml, and dilution liquid with the pH of being 6.5-6.8 and the osmotic pressure of being 290-310mOsmo / L is obtained; the diluent of the sperm of the pig, produced by the formulation, can effectively prolong the holding time, has convenient use and low cost, is suitable for being used by an artificial insemination station of the pig and a large-scale pig farm, has good application effect in the long-term production, has wider application range than the imported sperm, and can lead the conception rate of a sow in the oestrous period when being applied to a foreign pig to enable conception rate of a sow to be up to 86.8 percent with the litter size of 11.2 heads.

A Bacillus strain characterized by (i): sensitivity for ampicillin, vancomycin, gentamicin, kanamycin, streptomycin, erythromycin, clindamycin, tetracycline and chloramphenicol; (ii) antimicrobial activity against E. coli and Clostridium perfringens; and (iii) a sporulation percentage of at least 80 when measured after 2 days of incubation. The invention further relates to a method for selecting such strains. Many of the identified strains according to the invention are of the species Bacillus amyloliquefaciens. Some of the Bacillus amyloliquefaciens were further identified as Bacillus amyloliquefaciens subsp. amyloliquefaciens whereas others were identified as amyloliquefaciens subsp. plantarum. A Bacillus strain of the invention may be used as a feed additive to animal feed where it has a probiotic effect.

The invention relates to an HUVEC (human umbilical vein endothelial cell) separation, culture and subculture method belonging to the technical field of biology. The method comprises the following steps: (1) separation of HUVEC: taking a fresh umbilical cord, cleaning with a sterile PBS (phosphate buffer solution) containing penicillin-streptomycin, adding collagenase to perform water bath digestion at 37 DEG C, centrifuging the digestion solution, then adding an M199 culture medium, transferring into a gelatin-coated culture bottle, and culturing in a CO2 incubator at 37 DEG C; (2) culture of HUVEC: after the HUVEC is cultured at 37 DEG C for 24 hours, pouring out the culture medium, cleaning with the PBS, adding a fresh M199 culture medium, and afterwards, changing the culture medium once every two days, wherein the HUVEC can be subcultured generally after being cultured for 5-7 days; and (3) subculture of HUVEC: pouring out the culture medium, cleaning with the PBS, adding the digestion solution to digest the cell, adding a DMEM (dulbecco's modified eagle medium) containing serum to terminate the reaction once the cell is rounded, centrifuging the digested cell, and adding a fresh culture medium, wherein the HUVEC subcultured for 2-3 generations is used for experiments. The separated HUVEC is economical and practical, is simple and easy to use, and is beneficial to obtaining required in-vitro experimental model cells.

The invention provides a sterilizing method of a spherical phaeocystis culture solution, and relates to a microalgae culture solution. The method comprises the following steps: centrifuging algae solution which grows to an exponential phase, and removing supernatant; carrying out gravity suspension on algae cells with an f / 2 culture medium, centrifuging, and removing supernatant, repeating twice, and carrying out gravity suspension on the algae cells with the f / 2 culture medium; adding SDS (sodium dodecyl sulfate) and antibiotics, wherein the antibiotics are clindamycin, azithromycin, gentamicin, kanamycin, streptomycin, cefotaxime, ampicillin and rifamycin; culturing under illumination, centrifuging the algae solution and removing supernatant during culturing, carrying out gravity suspension on the algae cells with the f / 2 culture medium, centrifuging and removing supernatant, removing residual SDS and the antibiotics, transferring into the f / 2 culture medium, and culturing under illumination; and selecting survival transferred algae solution which is treated with SDS and the antibiotics, and detecting whether bacteria exist or not after the transferred algae solution grows to the exponential phase. The method is convenient to operate, complex operations, such as bacterium separation and test on sensitivity to antibiotics and the like, are avoided, and long-term treatment of high-concentration antibiotics has good sterilization effect.

The invention discloses an engineering bacterium through anthracene nucleus antibiotic and appliance, which is characterized by the following: inserting or replacing restriction enzymes site of dnmV gene nucleic acid order of sky blue rufus streptomycete with antibiotic against property gene and surface cerubidin synthetic gene; blocking-up engineering bacterium of dnmV gene; or transforming expressing carrier of anthracene nucleus antibiotic synthetic gene to lead streptomycin (S.lividans)TK24; extracting expressing plasmid from S.lividans TK24; leading-in sky blue rufus streptomycete gene only with antibiotic against property gene to block dnmV gene; blocking-up abrupt change bacterium. This invention can produce cerubidin and / or analogue, such as anthracene nucleus antibiotic.

The invention discloses Chicken semen diluent as well as a preparation method and application thereof, and belongs to the technical field of animal breeding. Chicken semen is diluted for normal temperature preservation and artificial insemination. The Chicken semen diluent takes sterile distilled water as a solvent and contains fructose, citric acid, sodium glutamate, potassium chloride, dipotassium hydrogen phosphate, Tris, bovine serum albumin, VB12 (Vitamin B12), streptomycin and penicillin. The preparation method comprises the following steps of: dissolving the fructose, citric acid, sodium glutamate, potassium chloride, dipotassium hydrogen phosphate, Tris, bovine serum albumin and VB12 into a small amount of sterile distilled water under the sterile condition, adding the penicillin and the streptomycin, adding the sterile distilled water, mixing uniformly to obtain the Chicken semen diluent, and preserving at the temperature of 4 DEG C; and preheating the Chicken semen diluent to the temperature of 37 DEG C, and mixing the Chicken semen diluent and Chicken semen, wherein the mass ratio of the Chicken semen to the Chicken semen diluent is (1:1)-(1:2). The Chicken semen diluent has the advantages of simplicity and convenience in preparation, convenience in material acquisition, low cost and easiness in popularization.

The invention provides an expansion in vitro purification culture method of mesenchymal stem cells and a cell culture medium used in a culture process. According to the method, mesenchymal stem cells differentiate to osteogenic stem cells. The cell culture medium comprises the following components: 8-12mu g / L of bFGF (Basic Fibroblast Growth Factor), 8-12mu g / L of EGF (Epidermal Growth Factor), 8-12mu g / L of PDGF-BB (Platelet Derived Growth Factor-BB), 10 percent of fetal calf serum, 100U / mL of penicillin and 100mg of L of streptomycin; a solvent is a basic culture medium; and the basic culture medium is a DMEM (Dulbecco Modified Eagle Medium) culture solution, an F12 culture solution or a mixture of the DMEM culture solution and the F12 culture solution. The mesenchymal stem cells obtained by adopting a method comprising the steps of early replacement of the cell culture medium, cell adherence passage and combined addition of cell factors (bFGF, EGF and PDGF-BB) in the culture solution are high in purity and strong in vitro proliferation capacity, and have good capability of differentiation to osteogenic stem cells.

This invention relates to a method of treating otitis externa using a topical combination medication, including one or more antifungal agent such as, for example fluconazole, voriconazole, itraconazole, clotrimazole, amphotericin B, caspofungin, micafungin, terbinafine, naftifine, butenafine, amorolfine, ravuconazole, posaconazole, flucytosine, econazole, enilaconazole, miconazole, oxiconazole, saperconazole, sulconazole, terconazole, tioconazole, nikkomycin Z, anidulafungin (LY303366), nystatin, pimaricin, griseofulvin, ciclopirox, haloprogin, tolnaftate, and undecylenate and one or more antibacterial agent such as neomycin sulfate, polymyxin B sulfate, colistin sulfate, gentamycin, tobramycin, chloramphenicol, Ciprofloxacin, Ofloxacin, a penicillin compound, a cephalosporin compound, a macrolide compound, a fluoroquinolone compound, streptomycin, or kanamycin.

The present invention discloses an adipose mesenchymal stem cell preparation kit, the kit includes a fat storage solution, a fat washing liquid, a fat decomposition liquid, a cell washing liquid, a cell culture liquid, a cell digestion liquid and a cell cryopreservation liquid; the fat storage solution is a DMEM, DMEM / F12, MEM or RPMI-1640 medium including 100-200ug / ml of penicillin, 100-200ug / ml of streptomycin and 2.5-5 mug / ml of amphotericin; the fat washing liquid is normal saline, PBS, D-Hanks or HBS including 100-200ug / ml of penicillin, 100-200ug / ml of streptomycin and 2.5-5mug / ml of amphotericin; the fat decomposition liquid is a DMEM, DMEM / F12, MEM or RPMI-1640 medium including 0.75-2mg / ml of collagenase I; the cell washing liquid is PBS, D-Hanks or HBSS; the cell culture liquid is a mesenchymal stem cell serum-free growth medium, the cell digestion liquid is a PBS, D-Hanks or HBSS solution including 1.25-2.5mg / ml of Trypsin-EDTA; and the cell cryopreservation liquid is a mesenchymal stem cell serum-free growth medium including 60-200mg / ml of DMSO.