Method for separation and organotypic preservation of allogenic limb transplant
SUBSTANCE: invention refers to medicine, namely ophthalmology, and may be used in separation and organotypic preservation of an allogenic limb transplant. A method involves early surgical separation of the limb transplant from a whole corpse donor eyeball with eliminated perforation after donor's death. The transplant is preserved within 28 days in a medium containing insulin, dexamethasone, blood serum, penicillin, streptomycin, dextran 40000, HEPES M-solution, 7.5% bocarbonate, 40% glucose, L-glutamine, penicillin, streptomycin, amphotericin B and Hanks salt medium 199 under certain proportion of the components. The preservation is performed at temperature 37°C with the medium changed every 3 days. The same medium, but serum-free is used for washing. The method provides preserving transplant viability with proliferating progenitor cells that leads to achieving stable, long-term epithelisation, stroma defect recovery, prevention of corneal conjunstivisation.
EFFECT: prepared transplant may be used for surgical management of the patients with limb cell insufficiency syndrome of various aetiologies.
2 cl, 1 ex, 9 dwg
The invention relates to ophthalmology, and can be used in the surgical treatment of patients with the syndrome of limbus cell failure (LCN) of various etiologies.
To date, all proposed methods of treatment LCN are divided into two main groups.
1. Cytotherapy cultured limbus epithelial cells on different carriers. As the carrier can speak: amniotic membrane [Grueterich m, Scheffer C., Tseng G. Human Limbal Progenitor Cells Expanded on Intact Amniotic Membrane Ex Vivo // Arch Ophthalmol. 2002; 120:783-790; Koizumi N., Cooper, L.J., N.J. Fullwood et al. An evaluation of cultivated corneal limbal epithelial cells, using cell-suspension culture Invest. Ophthalmol. Vis. Sci. 2002. 43(7):2114-21], fibrin substrate [Sudha Century, H.N. Madhavan, Sitalakshmi g et al. Cultivation of human corneal limbal stem cells in Mebiol gel is A thermo-reversible gelation polymer // Indian. J. Med. Res. 2006. Dec; 124(6):655-64] and a soft hydrogel contact lens [Sangwan V.S. Limbal stem cells in health and disease // BioSci. Rep. 2001. Vol.21. P.385-405].
The disadvantage of this method is the technical complexity and high cost of implementation, and the need to use feeder layer under cultivation.
2. Limbus transplantation [Kenyon K.R., S.C. Tseng // Ophthalmology. - 1989. - Vol.96. N5. - P.709-722].
The known method autotransplantation limbus flap, taken with a healthy bilateral eye recipient [patent No. 2348384 from 10.03.2009].
The disadvantage of this method is the inability excision quite a large shopping Mall is the area of the limb, and the disagreement of many patients for this procedure.
The known method of allogeneic transplantation "fresh" limbus flaps isolated from cadaveric donor eyes received in ophthalmic institutions transplanting the cornea [Dua HS, Blanco AA. Allolimbal transplantation in patients with limbal stem cell defi ciency // Br J Opthalmol., 1999. 83: 414-19; RF Patent №2361550 from 20.07.2009].
The disadvantage of this method is high risk of transplant rejection due to the presence of a large number of Langerhans cells expressing HLA-DR antigens, which creates the need for immunosuppressive therapy.
The closest analogue is the allocation method and organo-typical cultivation limbus grafts described in the publication Zito-Abbad E, Borderie VM, Baudrimont M, et al. Corneal epithelial cultures generated from organ-cultured limbal tissue: factors influencing epithelial cell growth // Curr Eye Res. 2006 May; 31(5): 391-9.
How is that organo-typical cultivation are corneoscleral ring remaining after removal of corneal transplant for end-to-end or layer-by-layer keratoplasty, including: processing of 0.05%trypsin solution for 1 hour and disposal II (1,2 U/ml) for 15 minutes at 37°C, further cultivation using a feeder layer of human keratocytes in DMEM/F12 (Dulbecco''s Modified Eagle's Medium Nutrient Mixture F-12 US with the addition of fetal bovine serum, insulin, dexamethasone, cholera toxin, gentamicin within 28 days in CO2-incubator at a temperature of 31°C, with 5%content of CO2and 99% humidity, replacing medium every 3 days. The advantage of this method is the reduction of the expression of HLA-DR antigens by natural elimination of Langerhans cells from the limbus of the graft due to long-term (28 days) organo-typical cultivation [Holland E.J, DeRuyter D.N., Doughman D.J. Langerhans cells in organ-cultured corneas // Arch Ophthalmol 1987; 105: 542-5].
However, the described method has several disadvantages:
for organo-typical cultivation used donor corneoscleral ring, after removal of corneal transplant, in this regard, the length of time from the death of the donor before cutting out the limbus grafts is not less than 72 hours, which adversely affects survival and proliferative activity of limbus epithelial stem cells;
- the necessity of using special tools for cutting out the limbus allografts, as well as the inability to form the limbus transplant uniform thickness and the necessary forms with knives-rasseivatelei of corneoscleral rings;
- used during processing corneoscleral what about the ring of the high concentration of enzymes (trypsin and dispose II) leads to a large loss of limbus epithelial cells (Figure 1, 2).
- cultivation at 31°C promotes proliferation only epithelial cells of the limb, which leads to the necessity to use under cultivation feeder layer (Figure 3).
The task of the invention is to develop a method of preoperative preparation of the surgical selection and organo-typical conservation) allogeneic limbus transplant for transplant patients with LCN.
The technical result is to achieve stable, long-epithelialization, the restoration of defects stroma and prevention of conjunctivitis of the cornea at the limbus cell failure.
The technical result is achieved in that in the method of preoperative preparation of allogeneic limbus transplant for transplant patients with limbus cell failure, including surgical selection limbus grafts from cadaveric donor eyes, organo-typical conservation within 28 days in medium containing insulin, dexamethasone, serum, antibiotics, replacing the medium every 3 days, laundering, according to the invention the surgical selection of limbus transplants performed from a single cadaveric donor of the eyeball, except for its perforation, organo-typical conservation is carried out at a temperature of 37°C, and the environment for organo-typical conse the activate your product further comprises dextran 40000, HEPES odnokolernyh, a 7.5%solution of bicarbonate, 40%solution of glucose, L-glutamine, amphotericin b and medium 199 on the Hanks salts, and contains antibiotic penicillin and streptomycin in the following ratio of the components in 1000 ml solution:
|dextran 40000||50.0 g|
|HEPES odnokolernyh||15.0 ml|
|a 7.5%solution of bicarbonate||10.0 ml|
|40%glucose solution||5.0 ml|
|amphotericin b||1.4 mg|
|insulin "Humulin of regular"||0.8 ml|
|dexamethasone with both molarity 10-8||1.0 ml|
|serum human blood|
|or cattle||100,0 ml|
|medium 199 salts Hanks||rest|
while laundering from the whey produced the claimed medium not containing serum.
As serum can be used fetal bovine serum or human serum, cord blood or human serum IV blood group.
Catalog number medium 199 salts Hanks - S230 LLC Paneco".
The advantage of this method is the possibility of taking limbus of the graft in the early stages after the death of the donor, comprising not more than 18 hours, which contributes to maintaining reserves the viability of the cells (Figure 4).
The advantage of this method is the technical simplicity of cutting out the limbus of the graft with the entire circumference of the limb whole cadaveric donor eyes, when this is achieved a uniform thickness graft, not more than ½ of the thickness of the sclera, which prevents perforation of the donor's eyeball, to perform subsequent dissection of corneoscleral disc, preservation and planned through or layer-by-layer keratoplasty.
The proposed method organo-typical conservation limbus grafts can preserve the natural microenvironment of limbus epithelial progenitor cells, namely, mesenchymal stem cells, which are goods which owing to its paracrine properties fulfil the function of a feeder layer.
The preservation of the structure of limbus graft with viable, proliferating progenitor cells in the process of implementation of the method makes it possible to further cryopreservation limbus grafts and long-term storage in liquid nitrogen at a temperature of -196° to create a Bank of limbus grafts (Figure 5, 6).
The method is as follows. From cadaveric eyeball prepared for the donor cornea on medical technology "Algorithm procurement of cadaveric donor human corneas for transplantation" (of IRTC "eye microsurgery" named. Acad. Fyodorov") in sterile ward, after decapitalization of the cornea and remove the bulbar conjunctiva with the entire surface of the diamond or metal blade are 2 circular cut: one - sclera, retreating to 2 mm from the transparent part of the cornea to a depth of 0.5 mm; the other on the boundary between the limbus and transparent areas of the cornea. Next on the same depth, perpendicular incision is made in the direction of the cornea are 4 holes in the 3, 6, 9 and 12 hours, then a knife-rasseivatele tsaparevetsa tissue layer at a given depth, holding the resulting flap of corneal forceps, the result of limbus grafts stretch fabric with a width of 2 mm and thickness is Noah 0.5 mm over the entire length (Figure 7, 8, 9).
Received limbus grafts are placed in a sterile vial of 25 cm2and filled with 4 ml of medium for organo-typical conservation, which can be obtained by mixing the components according to the formula of the invention or may be obtained from the Solution for the storage of the cornea" (THE No. 9398-013-29039336-2008 produced LLC NEP "eye microsurgery", Moscow, Beskudnikovsky Boulevard, building 59A), including dextran, HEPES, bicarbonate, glucose, L-glutamine, penicillin, streptomycin, amphotericin b and medium 199 on the Hanks salts, which is optionally added insulin, dexamethasone and serum of humans or cattle, when the ratio of the components in 1000 ml solution: dextran 40000 - 50.0 g, HEPES odnokolernyh - 15.0 ml of 7.5%solution of bicarbonate - 10.0 ml, 40%glucose solution to 5.0 ml, L-glutamine and 1.5 g, penicillin - 250000 units, streptomycin - 200.0 mg, amphotericin b - 1.4 mg, insulin "Humulin of regular" - 0.8 ml, dexamethasone with both molarity 10-8to 1.0 ml serum human or cattle in 100.0 ml medium 199 salts Hanks - the rest.
Conservation is carried out for 28 days at 37°C by incubation in an atmosphere of air with 5% CO2with 99% humidity. Replacement conservation of the environment is provided every 3 days. Before using limbus transplant washed in the environment such as the level, but free from serum, namely when the ratio of the components in 1000 ml: dextran 40000 - 50.0 g, HEPES odnokolernyh - 15.0 ml of 7.5%solution of bicarbonate - 10.0 ml, 40%glucose solution to 5.0 ml, L-glutamine and 1.5 g, penicillin - 250000 units, streptomycin - 200.0 mg, amphotericin b - 1.4 mg, insulin "Humulin of regular" - 0.8 ml, dexamethasone with both molarity 10-8to 1.0 ml medium 199 salts Hanks - the rest.
Then limbus graft is transferred into a sterile tube with the medium of the same composition, in which laundered, that is free of serum and in the cool bag at a temperature of 4-6°C is delivered to the operating room.
Example. Patient K., 54 years old.
Diagnosis: Chronic recurrent erosion of cornea of both eyes.
Anamnesis morbi: suffering for 5 years, receive conservative therapy.
Anamnesis vitae: 6 years ago was held radiation and chemotherapy on breast cancer.
Status localis: visual acuity in both eyes (OU) of 0.1 not corrects.
OU - photophobia, in the center of the cornea decapitalization.
Under conjunctival limbus in the upper segment of the right eye produced transplant was prepared according to the proposed method limbus transplant, at the same time as the serum used human serum IV blood group.
The operation and the postoperative period was uneventful, was called ACANA standard local immunosuppressive therapy. After surgery on the 7th day was marked complete epithelialization of the cornea. A month later conducted a similar operation on my left eye. When control inspections throughout the entire observation period (6 months) signs of rejection limbus grafts were observed, cornea epithelisation completely transparent, visual acuity in both eyes of 0.6-corrected sph+1,0D=0,9.
Thus, the proposed method of treatment (surgical selection and organo-typical conservation) allogeneic limbus grafts allows to achieve stable and long-epithelialization of the cornea in patients with limbus cell failure.
1. Method of preoperative preparation of allogeneic limbus transplant for transplant patients with limbus cell failure, including surgical selection limbus grafts from cadaveric donor eyes, organo-typical conservation within 28 days in medium containing insulin, dexamethasone, serum, antibiotic, replacing the medium every 3 days, laundering, characterized in that the surgical selection of limbus transplants performed early after the death of the donor, from a single cadaveric donor of the eyeball, except for its perforation, organo-typical conservation is carried out at a temperature of 37°C, and the environment for organo-is epicheskoi conservation further comprises dextran 40000,
HEPES odnokolernyh, a 7.5%solution of bicarbonate, 40%solution of glucose, L-glutamine, amphotericin b, medium 199 on the Hanks salts, and contains antibiotic penicillin and streptomycin in the following ratio of the components in 1000 ml solution:
|dextran 40000||50.0 g|
|HEPES odnokolernyh||15.0 ml|
|a 7.5%solution of bicarbonate||10.0 ml|
|40%glucose solution||5.0 ml|
|amphotericin b||1.4 mg|
|insulin "Humulin of regular"||0.8 ml|
|dexamethasone with both molarity 10-8||1.0 ml|
|serum human blood|
|or cattle||100,0 ml|
while laundering produce medium containing all of the above components, except for the serum.
2. The method according to claim 1, characterized in that as the serum can be used fetal bovine serum or human serum, cord blood or human serum IV blood groups.
SUBSTANCE: there are described versions of humanized monoclonal anti-factor D antibodies and their functional fragments. There are offered: a coding nucleic acid, an expression vector, as well as a cell for preparing c the antibody containing the vector. What is described is a method for preparing the anti-factor D antibody by cell culture and expressed antibody purification. Also, there are offered: an antibody composition and use of the antibody for treating disorders mediated by a complement system.
EFFECT: higher clinical effectiveness in the diseases related to excessive or uncontrolled activation of the complement system.
32 cl, 9 dwg, 4 tbl, 6 ex
SUBSTANCE: group of inventions relates to medicine, particularly to ophthalmology. An agent for promotion of corneal endotheliocyte adhesion containing an Rho kinase inhibitor, a culture medium for corneal endotheliocytes containing the Rho kinase inhibitor, a solution of cornea preservation containing the Rho kinase inhibitor, and a method for making a preparation of corneal endothelium involving the stage of corneal endotheliocyte culture with the use of the above culture medium.
EFFECT: group of inventions provides an ability for effective growing of corneal endotheliocytes and stable delivery of the preparation of corneal endothelium.
43 cl, 17 dwg, 19 ex
SUBSTANCE: invention relates to medicine, namely to ophthalmology, and may be used for treating chalazion. That is ensured by surgical excision of chalazion. The surgery is terminated by the introduction of Kenalog 0.2-0.3 ml into an incisional wound of eyelid tissue after the removal of chalazion.
EFFECT: method reduces a possibility of recurrences within a site of chalazion thereby enabling removing greater chalazion without recurrent surgical interventions.
SUBSTANCE: cell population is recovered from peripheral blood or umbilical blood by a method involving positive cell secretion from peripheral blood or umbilical blood able for immune reaction with an antibody specified in a group consisting of aHTH-CD44, anti-CD11b and their combination. The recovered myeloid-like cell population contains the cells expressing CD44 antigen, CD11b antigen and hypoxia-induced factor lα (HIF-lα). The prepared population is used for recovery and stabilisation of the functional vasculature, stimulation of microgliacyte formation, and for stimulation of physiologic intra-retinal vascularisation of hypoxic retinal tissue in simultaneous suppression of abnormal preretinal vessel formation.
EFFECT: invention enables preparing the cells with vasculotrophic and neurotrophic activity which possess a considerable therapeutic potential in the intraocular introduction into a mammal suffering a degenerative ocular disease.
7 cl, 63 dwg, 2 tbl, 21 ex
FIELD: medicine, pharmaceutics.
SUBSTANCE: invention refers to medicine, particularly ophthalmology. Eye drops containing recombinant interferon stabilising the biological and physicochemical properties, stabilising resistance to microbial contamination, an antioxidant and a buffer mixture, contain antihistamine preparations specified in a group: antazoline, azelastin, tetryzoline, diphenhydramine hydrochloride as stabilising the biological and physicochemical properties, contains specified in the group: low-molecular polyvinylpyrrolidone, macrogol 400-12000, propylene glycol as stabilising resistance to microbial contamination contains specified in a group: Nipasol, sorbic acid, boric acid, besides it contains sodium chloride and/or sodium acetate in the following proportions in 1 ml of the solution. As an antioxidant, the eye drops contain disodium dihydrogen ethylenediaminetetraacetate and/or buthylhydroxytoluene in the amount of 0.0001-0.1 g in 1 ml of the solution. The eye drops contain recombinant interferon specified in a group: alpha-, beta-, gamma-interferon. As a buffer mixture, the eye drops contain borate-acetate or phosphate, or borate buffer.
EFFECT: invention provides higher antiviral activity of the eye drops.
4 cl, 5 ex
SUBSTANCE: invention relates to ophthalmology and is intended for conservative treatment of dacryostenosis. Furacilinum-based ophthalmic gel is introduced into lacrimal canaliculus. Furacilinum-based ophthalmic gel is introduced by means of syringe in dose 3 ml, every second day, 8-10 times in total.
EFFECT: method ensures recovery of lacrimal passages and prevention of stenosis recurrences in postoperative period after dacryorhinocystostomy, as well as possibility of application in patients with allergic predisposition to antibiotics.
SUBSTANCE: invention relates to compounds of formula (IX) wherein radicals and symbols have values given in the claim, and pharmaceutically acceptable salts or tautomers thereof. Said compounds are inhibitors of poly(ADP-ribose)polymerase (PARP) and can be used to treat cancer, inflammatory diseases, reperfusion injuries, ischaemic conditions, stroke, renal failure, cardiovascular diseases, vascular diseases other than cardiovascular diseases, diabetes mellitus, neurodegenerative diseases, retroviral infections, retinal damage, skin senescence and UV-induced skin damage, and as chemo- or radiosensitisers for cancer treatment. The invention also relates to a pharmaceutical composition containing said compounds, use of said compounds and a method of treating said diseases.
EFFECT: high efficiency of using the compounds.
10 cl, 18 ex
SUBSTANCE: invention refers to medicine. There are applied at least two proteases for preparing a drug for treating and/or preventing ocular diseases related to neoangiogenesis and specified in a group consisting of age-related macular degeneration (AMD), choroidal revascularization, Von Hippel-Lindau disease, iris revascularisation, ischemic proliferative retinopathy, corneal revascularisation, and proliferative crescent cell retinopathy wherein at least one protease is specified in a group consisting of a group consisting of herbal protease.
EFFECT: invention provides reducing angiogenesis in treating or preventing ocular diseases related to neoangiogenesis.
14 cl, 1 tbl, 7 dwg, 7 ex
SUBSTANCE: group of inventions refers to medicine, particularly to ophthalmology, namely: to antibacterial lenses containing metals, as well as to methods for producing them. The method for producing an antibacterial lens containing a metal salt involves the stages: (a) preparation of a hard lens by a salt precursor, as well as (b) preparation of the lens produced at the stage (a) by a disperse additive specified in a group consisting of polyvinylpyrrolidone, polyvinyl alcohol, glycerol and polyethylene oxide, and a metal-containing agent specified in a group consisting of silver tetrafluoroborate, silver sulphate, zinc acetate, zinc sulphate, copper acetate, copper sulphate, silver nitrate, manganese sulphide, zinc oxide, zinc sulphide, copper sulphide, copper phosphate, silver nitrate, silver sulphate, silver iodate, silver carbonate, silver phosphate, silver sulphide, silver chloride, silver chloride, silver bromide, silver iodide and silver oxide. The other method for producing involves the stages (a) of preparation of a hard lens by a metal-containing agent, and a disperse agent, (b) preparation of the lens prepared at the stage (a) by a salt precursor. The antibacterial lens containing the metal salt is produced by said methods.
EFFECT: group of inventions provides producing the lenses with a decreased degree of opacity.
16 cl, 1 tbl
SUBSTANCE: invention relates to medicine, namely to ophthalmology, and can be used for treatment of "dry" form of age-related macular degeneration. For this purpose at preliminary stage, magnetic implant in form of flat rectangular plate is implanted intrasclerally in projection of macular area. 2 weeks after that, mesenchymal stem cells (MSC) of patient's own bone marrow, labelled with magnetic microparticles, are introduced to patient intravenously during 30-40 minutes. 2 hours before MSC introduction macular area of retina is irradiated by low-intensive laser irradiation with wavelength 632-633 nm, voltage 10 mW, diameter of laser irradiation spot is 4 mm, total time of irradiation is 5 minutes.
EFFECT: invention ensures targeted delivery of MSC to pathological nidus and retaining said cells for the time, required for efficient improvement of microcirculation and trophism of tissues, which contributes to steady stabilisation of visual functions.
SUBSTANCE: invention relates to medicine, namely to ophthalmology and is intended for slowing down rough scarring of conjunctiva in carrying out photodynamic therapy in conditions of vascular proliferation of conjunctiva. First, not less than 2 mg of alasense are introduced subconjunctivally. After 1.5-2 hours impact with laser radiation with wavelength 632.8 nm, power density 47.7-55.7 mW/cm2, is performed for 10-14 minutes. 12-24 hours after intervention medication viscleron is introduced into conjunctival sac of affected eye 1 drop 3 times per day, during not fewer than 10 days.
EFFECT: method ensures efficient suppression of angiogenesis of conjunctiva vessels in zones of proliferating cells due to application of aggressive parameters of laser impact with simultaneous prevention of necrobiotic and destructive changes due to protective effect on irradiated tissues.
SUBSTANCE: invention refers to medicine, specifically dentistry and concerns treating temporomandibular joint arthrosis. For this purpose, mesenchymal cord and placental stem cells received after easy delivery. The cells 5-10·106 are injected in the temporomandibular joint in 1-2 ml of patient's blood plasma and intra-articular fluid. taken from a patient's healthy major joint.
EFFECT: while being low-traumatic, the method provides replacement of articular cartilage defects, creating the environments for optimising the repair processes in treating the most severe and frequent temporomandibular joint injures.
SUBSTANCE: invention relates to medicine, namely surgical stomatology and can be used for treating paradontium. Periodontal osteogenesis and neogenesis are ensured by removing pathologically changed structures and introducing the mesenchymal stem cells suspended in a carrier into the defect cavity. As the stem cells, the mesenchymal cord and placental stem cells received after easy delivery are used. The cells 5-10·106 are injected in 1-2 ml of patient's blood plasma.
EFFECT: method provides more effective replacement of the manifested periodontal defect associated with paradontium, reduced length of treatment with no side effects.
SUBSTANCE: method of obtaining medications from fetus umbilical cord lies in the following: from calf fetus in the first part of intrauterine development umbilical cord as amputated in area of umbilical ring and on border of amnion-chorionic part of fetus placenta, blood is discharged from umbilical veins and arteries, amputated end of umbilical cord are fixed, umbilical blood is separated, components are separated, passed through nano-filters, amputated umbilical cord is disinfected and Wharton's jelly is separated, umbilical arteries, veins, umbilical envelope are separated and fixed at specified conditions, obtained materials are frozen in liquid nitrogen.
EFFECT: method makes it possible to increase purity of obtained preparation without application of chemical reagents.
1 tbl, 3 dwg
SUBSTANCE: invention refers to medicine - transfusiology. Nuclear cells with hematopoietic stem cells (HSC) of cryoprotector - 55% of dimethyl sulfoxide solution with 5% dextran 40 arranged in a cryobag are added into a suspension. Mechanically mixed in a mixing device at +4°C, the cryobag is sealed and put into a shrink-wrap bag. Programmed freezing is carried out, at the first stage of which a sample is maintained for 10 min at +4°C, then cooled with the speed of 1°C/min down to the temperature of -12°C, then cooled down with the speed of 15°C/min down to the temperature of -60°C. After the sample thawing with the speed of 15°C/min down to the temperature of -18°C, it is cooled with the speed of 1°C/min down to -60°C . At the end of the freezing program the sample is cooled with the speed of 3°C/min down to -100°C. On completion of freezing the sample is put into a quarantine Dewar vessel with liquid nitrogen until results of tests for infection availability are ready. On completion of a quarantine period of storage, the sample is transferred for long-term storage at the temperature of at least -150°C into a Dewar vessel with liquid nitrogen provided the tests results are negative. In case the tests results are positive, the sample with HSC are transferred to a Dewar vessel for infection material for long-term storage.
EFFECT: invention makes it possible to increase HSC sterility during freezing, integrity and their viability.
SUBSTANCE: method related to field of medicine, namely to transfusiology. Method includes collecting umbilical cord blood (UCB) into container, weighing it with calculation of its volume, determining type of human leukocyte antigen for comparing hypocompatibility, presence or absence of infection, introduction into packet with UCB, anticoagulant and hydroxyethyl starch, mechanical mixing of packet content in various planes, incubation after mixing at room temperature with separation of erythrocyte mass (EM) by sedimentation, placement of centrifuge chamber in separator, connecting to it on symmetry axis main pipeline, the latter being connected via distribution unit via pipelines to sacks for collection of blood components and to packet with mixture of UCB, anticoagulant and hydroxyethyl starch. After that, chamber is filled with packet content, and mixture is subjected to separation, with its separation into EM, plasma and leukocyte concentrate with their distribution into corresponding sacks and into empty packet. After separation, viability and amount of SC are determined, they are checked on biological inoculation of sample of plasma and EM mixture, dose of plasma is tested for detection of antibodies, hemotransmissive infections, dose of EM is used to determine blood group and Rhesus factor.
EFFECT: method application makes it possible to increase quality and reliability of determining SM from various biological contaminations.
SUBSTANCE: invention refers to medicine, particularly - to ophthalmology. The method involves laser exposure and retinalamine administration For this purpose, retinalamine 5 mg is dissolved in 2% lidocaine 1.5 ml. Parabulbarly, the prepared solution is introduced by 75 ml into each eye. 5-10 minutes later, retina focused continuous noncontact exposure to helium-neon laser follows. The light guide diametre is 3 mm, the distance of a distal end of the light guide to a front surface of cornea is 2-4 cm. The power is 2.2 - 2.6 Wt. Said combined exposure is daily for ten days. The duration of first three sessions of laser exposure is 3 minutes, and the following seven sessions of laser exposure last for 5 minutes.
EFFECT: method improves visual functions due to enhancing visual acuity and extending fields of vision, provides continuous stable maintenance of the effect; it is atraumatic.
SUBSTANCE: invention refers to medicine, specifically to ophthalmology, and can be used in experimental ophthalmology for preparing proliferating crystalline lens cell culture with preferential epithelial cell content for future screening analysis of various methods for prevention of posterior capsule opacity in vitro. Substance of the invention implies mechanical release of a crystalline lens capsule with adjacent cortical mass of a nucleus, refinement to 1 mm3 and processing with mixed 0.05% collagenase first type and 0.25% trypsin; then cell culture is washed by centrifugation with dissolving precipitated cells in a serum-medium DMEM/Ham's F-12; prepared culture is sowed in culture cups in concentration 5×105 cells per cm3 and grown in a CO2 incubator.
EFFECT: advantage of the invention consists in development of a simple method for preparing crystalline lens cell culture preferentially consisting of epithelial cells.
SUBSTANCE: mixture of chondroitin-6-sulphate and dermatan sulphate extracted from umbilical cords is oxidised with 2,2,6,6-tetramethylpiperidine-1-oxyl-NaClO-NaBr at pH 10.2 and temperature 0-5°C for 1 hour with subsequent precipitation of oxidation products with three ethanol objects and their separation on a chromatographic column with DEAE-cellulose using eluent in form of aqueous solutions of NaCl with increasing concentration 0.2→0.6 M in increment of 0.05.
EFFECT: obtaining pure chondroitin-6-sulphate; the rest of the products are hybrid polysaccharides consisting of chondroitin-6-sulphate and dermatan sulphate oxidised on the primary groups.
SUBSTANCE: method consists in introducing of mesenchymal stem cells of umbilical cord or placenta, taken after normal childbirth. Endolymphatic introduction of cells is performed by injections of 1-2 ml of physiological solution, which contains 0.075-0.20×106 cells/kg of weight along spine or leg in single-step1-2 times per year.
EFFECT: method allows to improve animal's condition, increasing of its activity, normalises metabolism.
FIELD: food industry.
SUBSTANCE: invention relates to food for infants and/or young children. The food contains indigestible oligosaccharides with polymerisation degree equal to 2 - 200, unviable Bifidobacterium breve in an amount equivalent to 103 CFU - 1013 CFU of B. breve per g of the food dry weight, and viable Bifidobacterium breve in an amount of less than 103 CFU of B. breve per g of the food dry weight. The invention relates to the food application for production of a composition for feeding infants and/or young children.
EFFECT: indigestible oligosaccharides combination combined with unlivable Bifidobacterium breve promotes prevention and/or therapy of atopic illness and food allergy.
19 cl, 2 tbl, 3 ex