Method for simulating diabetic macular neovascularisation. RU patent 2504844.
 Method of decellularisation of small-calibre blood vessels / 2504334Invention relates to medicine, namely to regenerative medicine and tissue engineering, and can be applied for obtaining extracellular matrixes of small-caliber blood vessels. For this purpose at the first stage of tissues fragment of blood vessel is washed with distilled water for 1 hour at temperature +4°C. After that, fragment is placed into 0.05% solution of tripsin and 0.02% EDTA for 1 hour at temperature +37°C. At the third stage processing in 0.075% solution of sodium dodecylsulfate is performed for 24 hours at temperature 26°C. After that, fragment is placed into 0.25% solution of Triton X-100 for 24 hours at temperature 26°C. At the fourth stage said fragment is processed with solution which contains 20 mcg/ml of RNA-ase and 200 mcg/ml of DNA-ase I, for 6 hours at temperature +37°C. After each stage of processing, fragment of blood vessel is three times washed in phosphate-salt buffer solution, for 10 minutes each time. The entire process of processing is carried out with constant mixing of solutions and simultaneous vibration, created by vibromotor, located on the external wall of the reservoir. |
| Method for simulating experimental isolated optic neuritis / 2504020 Cultural fluid 0.1 ml containing the strain L2 of type I herpes simplex virus adapted to a pig's finite embryo renal cell line in a dose of 100000 TCD50 is introduced through a flat portion of a ciliary body into a chinchilla's vitreous body with a needle 33 G. That is followed by the histological analysis of the removed eyeball starting from the 21st post-infection day. |
| Method for simulating experimental adenoviral uveitis complicated by optic neuritis / 2504019 Cultural fluid 0.1 ml containing type 6 adenovirus adapted to a pig's finite embryo renal cell line in a dose of 10000 TCD50 is introduced through a flat portion of a ciliary body into a chinchilla's vitreous body with a needle 33 G. That is followed by the analysis of the removed eyeball starting from the 7th post-infection day. |
 Method for experimental finasteride simulation of induction of functional activity of glycoprotein-p / 2504018Invention aims at studying a membership of the analysed drug preparations among efflux carrier protein Pgp (glycoprotein P) substrates. That is ensured by the experimental simulation of the induction of functional activity of the same protein. Finasteride is used as an inducing preparation. The preparation is introduced into rabbits intragastrically in the form of a suspension in olive oil in a daily dose of 0.225 mg/kg of animal's body weight for 14 days. |
 Method for simulating enamel demineralisation centre / 2503067Dental bracket is fixed onto an extracted tooth. A centre surrounding the dental bracket on a vestibular surface is limited by a wax layer. The tooth is immersed into a separated container with a demineralising gel consisting of (wt %): calcium dihydrophosphate - 0.04-0.08, lactic acid - 0.8-1.0, praestol 2510 - 3.0-4.5, sodium hydroxide - 0.4, distilled water - the rest. Then, the container with the teeth is placed in a thermostat at pH=4.5 for 96 hours. |
 Method of simulating atherosclerosis / 2500041For the purpose of diagnosing, preventing and treating the same disease, a cholesterol powder in the amount of 1%, margarin 10%, mercazolilum 10 mg/kg and vitamin D 2.5 IU per kg of body weight is added to the laboratory rats' feed. That is combined with performing an operation of ligating a left renal pedicle with a non-absorbable suture and underrunning a right upper pole with 2/3 of the organ being preserved. |
| Method for combined tadalafil and l-norvaline correction of l-name induced nitrogen oxide deficiency in experiment / 2500040 Nitrogen oxide deficiency is simulated by the daily 7-day intraperitoneal introduction of N-nitro-L-arginine methyl ester 25 mg/kg into experimental Wistar male rats. The nitrogen oxide deficiency is corrected by the simultaneous 7-day intragastric introduction of a combination of tadalafil 0.09 mg/kg and L-norvaline 10 mg/kg once a day. |
| Method for simulating primary biliary cirrhosis / 2500039 Primary biliary cirrhosis is simulated by introducing 45-50% alcoholic solution of picryl sulfonic acid 0.08-0.12 ml into rat's bulbar duodenal lumen and terminal ileum every 5-10 minutes. The primary biliary cirrhosis is diagnosed 1-1.5 months later. |
| Method for simulating chronic infected bone wound / 2499295 Bone defect is formed along the bone; the museum strain Staphylococcus Aureus No.5 culture in the amount of 40-45 mln CFU per 1 kg of experimental animal's body weight mixed with 0.1 ml of sterile silica sand are placed therein. Three days later, necrotic mass and purulent matter are removed from the bone defect. The wound is re-infected with the same culture in a dose of 20-25 mln CFU per 1 kg of body weight. The wound is further preserved in the form of a fistula. |
| Method for simulating diabetic macular oedema / 2498415 Alloxan is introduced into rat's peritoneum in a dose of 15.0 mg/100 g of weight. Then, 6.5 weeks after alloxan has been introduced, and experimental diabetes has been developed, insulin-like growth factor 1 (IGF-1) 1 mcl is introduced into the vitreous body from an intravitreous approach. |
 Method for modeling acute pancreatitis / 2244345As experimental animals one should apply mongrel dogs of 12-17 kg body weight. Under general anesthesia one should conduct superior-median laparotomy, introduce 3.0 ml 70%-ethanol solution under pancreatic capsule and then laparotomic wound should be sutured up. Manipulation should be performed once. The method provides modeling adequate acute pancreatic inflammation at no side effects being very simple in implementation. |
 Method and device for studying transosseous osteosynthesis model stiffness / 2246139Method involves studying transverse longitudinal and rotation stiffness characteristics. The studies are carried out step-by-step from the first order units to complete external fixation apparatus structure. The device has frame and is provided with calibration loads, wire rope, displacement indicators, strip for fastening to loading end of bone imitator fragment, beam for fixing displacement indicators, beams having unit for modeling longitudinal and transverse loadings. The frame is manufactured as parallelepiped. The fixing panel has openings for bone imitator, for fixing external fixation apparatus and yoke connection union and is fixed in end face part of the frame. Beam for fixing displacement indicators has longitudinal slit for fixing the indicators and arranging them on lateral slots in frame base. The beams having unit for modeling rotational, longitudinal and transverse loadings are arranged on lateral frame sides on lateral slots in base. |
 Method for experimental modeling urinary calculosis / 2248045The present innovation deals with modeling urinary calculosis in rats due to injecting intraperitoneally 60%-glucose solution at 1 ml/100 g animal body weight twice daily for 2 mo. The method is very simple and enables to achieve lithogenesis in 25% experimental animals. |
| Method for applying wave action to pathogenic microorganisms, viruses and tumor cells in experiment / 2250788 Method involves using Hann diode crystal with proper frequencies of pathogenic microorganisms and cells during their death period or during the stimulating factors action period being applied. |
 Method for modeling hypoxia with hypercapnia in animals / 2251158Hypoxia with hypercapnia should be modeled due to creating a closed system of inhaled air circulation. Air enters lungs out of hermetically sealed reservoir and at expiration returns back. The process of recirculation is supplied with an apparatus of artificial pulmonary ventilation. The innovation suggested provides steadiness in development of hypoxia with hypercapnia excluding the development of stressor reaction. Conditions should be created to carry out any manipulations with an animal in the course of an experiment. |
| Method for obtaining parodontitis model / 2252009 The present innovation should be carried out for the purpose to study ethiology and pathogenesis of parodontitis. One should affect with emotional stress in experimental animals (mature rats) due to placing 10-11 experimental animals into the cage at area of 0.018 sq. cm/animal. Before placing into the cage one should create artificial dental plaque around the cervix of the upper and lower incisors with the help of stomatological cement for every experimental animal. In the course of modeling all experimental animals should eat paste-like food. The method enables to shorten terms for obtaining the model desired and increase its similarity with pathomorphological manifestations of human parodontitis. |
 Acupuncture point electric model device / 2252743Device has input first variable resistor, capacitor and permanent resistor. Permanent resistor is connected to arm second and third variable resistors. Second ends of variable resistors are connected with motionless contacts of polarized relay. Movable contact of relay is connected to common bus. Input of device is connected to amplifier which has output connected with control wiring of polarized relay. Second end of wiring is connected with common bus. Device is intended for electrical modeling of balanced and misbalanced conditions of acupuncture point at electropunctural action with unlike-poled signals due to liquidation mutual errors at any circuit of opposite arms of the device. Values of active resistances can be installed independently at any arm of device. |
| Method for modeling hypoxic encephalopathy / 2253152 Laboratory animals should be once injected intraperitoneally or intravenously with phenylhydrazine at the dosage of 100-150 mg/kg. |
| Method for modeling chronic toxic nephropathy / 2253153 At studying the mechanisms of heavy metals toxic action, in particular, cadmium upon renal function, it is suggested to introduce cadmium sulfate solution into stomach once daily for 2 mo at the dosage of 0.5 mg/kg, on conversion to metal, where cadmium corresponds to 0.5 mg per 1 ml solution. The present innovation enables to study the pathology in dynamics of development and elaborate and searching preparations for treating and preventing chronic toxic nephropathy. |
| Method for stopping carcinoma cells division in a biological object / 2253903 Method involves exposing cell or cell group to external power source. At least two electrodes are introduced before treating the cells. One of electrodes is set on cytoplasmatic external cell membrane surface and the other one cell membrane and membrane potential value is measured. External electric voltage source is connected to the introduced electrodes oppositely in polarity with cell membrane potential difference value being not less than cell membrane potential. |
FIELD: medicine.
SUBSTANCE: diabetes mellitus is simulated in rats by intraperitoneal introduction of alloxan 15.0 mg/100 g of body weight. Murine VEGF 164 is introduced into a vitreous body from an intravitreal approach 6.5 weeks later on the 1st, 3rd and 7th day in a dose of 1 mcg with a total dose of 3 mcg.
EFFECT: method provides macular neovascularisation specific for diabetes mellitus that enables the further study of the therapeutic effectiveness and applicability in the given disease.
1 ex
Invention refers to medicine, in particular to ophthalmology, and can be used to create a model of the pathological process neovascularization of the macular zone of the retina against diabetes in experimental ophthalmology.
There is a method of creation of experimental model of diabetes mellitus by a single intraperitoneal injection of hydrate experimental animals in a dose of 170 mg/kg used in the modeling of diabetes, generates toxic free radicals, damaging cells of the islets of Langerhans cells of the kidney, liver, lung, and adipocytes (Szkudelski T. The mechanism of alloxan end streptozotocin action in The cells of the rat pancreas // Physiol. Res. - 2001. - №50. - P.536-546.). When diabetes occurs oxidative stress in adipose tissue, characterized by increase of the content, products of lipid peroxidation, in violation of exchange. Activation of lipid peroxidation and decrease in antioxidant potential of glutathione system in adipose tissue of rats with experimental diabetes can lead to stimulation of spontaneous lipolysis in adipocytes and the emergence of insulin resistance (Ivanov V.V. and other lipid peroxidation and the glutathione system in adipose tissue of rats with diabetes /the Bulletin of the Russian Academy of medical Sciences. - 2010. - 30, №6. - P.101-104.). When modeling of diabetes mellitus in the experiment, pathological changes in the fundus not appear, because activates blood aqueous barrier. In addition, for the emergence of the process of proliferation in the vitreous body against diabetes requires up to 2 years, which makes it difficult to conduct experiments. These factors lead to the need for additional impact on the retina against the backdrop of systemic diabetes.
The closest analogue of the invention is a method of modelling of neovascularization of the retina and optic nerve in rabbits, which consists in the fact that rabbits spend intravitreal injections of water solution of recombinant human vascular endothelial growth factor in the amount of 0,05-0,1 ml, and the dose of the specified funds injected fractional: 1 and 3 days of the experiment dose of 3 mg, 7 and 11 of the day - 1.5 mcg, by day 15 - 1.0 mg and 24 day - to 5.0 mcg [patent RU 2408083, 2010]. The method provides a consequence of changes in the fundus with a clear depending on the input dose, which allows us to trace all the stages of the pathological process that leads to a neoplasm of blood vessels posterior of the eye. However, changes in the retina associated with damage on the local level. This model is the etiological for modelling of pathologic process, associated with the violation of General exchange, hyperglycemia.
Object of the invention is development of a reliable, technically simple in execution of the method of modeling the defeat of the macular area with retina against the background of experimental diabetes.
Technical result in the use of the invention is development of diabetic neovascularization, pathological changes in the fundus with common endocrine disorders.
This technical result is achieved by the method of modeling of diabetic neovascularization of the macular region of the retina, which includes introduction into the vitreous body of the eye experimental animal as a toxic agent Vascular endothelial growth factor (VEGF) fractional on the 1st, 3rd and 7th day, according to the invention as an experimental animal use a rat, and previously in the abdominal cavity dose of 15.0 mg/100 g weight through a 6.5 weeks after the introduction of and development of an experimental diabetes as VEGF enter 1 mcg rat VEGF 164 in a total dose of 3 mg.
The choice of this kind of animals is due to the fact that blood flow to the retina rats closer to people than the blood circulation in the eye of a rabbit and it is also more economical model. Introduction of rat VEGF 164 through 6.5 weeks after the development of an experimental diabetes provides formation of the model of diabetic neovascularization, pathological changes in the fundus with common endocrine disorders. Thus, the combination in a single model of two factors (experimental diabetes and intravitreal injection VEGF 164) gives the development of resistant diabetic neovascularization. Fragmentation introduction provides a consequence of changes in the fundus.
The proposed method of modelling of diabetic neovascularization is preliminary modeling of diabetes and subsequent introduction into the vitreous body of a rat VEGF (Vascular endothelial growth factor) 164 methodology intravitreal access. Diabetes reproduce after 24 hours of fasting by intraperitoneal injection of dose of 15.0 mg/100 g weight. Through 6.5 weeks after the introduction of and development of an experimental diabetes each animal under ether anaesthesia perform intravitreal injections: through the flat part of the ciliary body at a distance of 1 mm from the limbus in the lower outer sector impose rat VEGF 164 in a total dose of 3 mg, the fractional 1 microgram on the 1st, 3rd and 7th day. To control indirect ophthalmoscopy on the 3rd, 7th and 14th day after the injection under conditions of drug (installation S.Tropicamidi 1%) and histological studies of 1-, 3-, 7-, 14-th day. The method was tested on 112 rats-males breed Wistar weighing 200 g of Animals were taken from the experiment method of air embolism after the introduction of anaesthesia at the 3rd, 7th and 14th day, eyes (according to the order of Ministry of higher education of USSR №724 of 13.11.1984). For morphological study on microtome prepared serial sections of 7-10 mkm. Colouring of histological sections carried out -eosin.
As a result of combining models of diabetes and retinal neovascularization with edema development determines the appearance of a new pathogenetic parallel with the development of nosology, identical to the pathology of retina in case of diabetes. In contrast to the local influence of VEGF without systemic disorders, the proposed model defines macular region, typical for diabetes that allows further to determine the appropriateness of therapy.
The proposed method is illustrated by the following example: 3 rats-males breed Wistar, 200 gram play diabetes after 24 hours of fasting by intraperitoneal injection of dose of 15.0 mg/100 g weight. Through 6.5 weeks after the introduction of and development of an experimental diabetes animal under ether anaesthesia through the flat part of the ciliary body was administered rat VEGF 164 at a distance of 1 mm from the limbus in the lower outer sector. Conducted 3 injections of the drug on the 1-St, 3 and 7 days of experiment were injected with 1 mg of VEGF. The total dose of VEGF was 3 . data of instrumental and pathomorphological studies after the first injection of 1 mcg VEGF was observed expansion and tortuosity of the retinal vessels. Introduction 2 mcg VEGF (two injections of 1 microgram) caused emergence of multiple retinal hemorrhages, enrichment retinal vascular network by opening not normally functioning capillaries. On the 14th day after the beginning of the experiment the result of 3 injections of 1 microgram VEGF pointed to the emergence of massive retinal hemorrhages and newly formed vessels. The animals were sacrificed experiment method of air embolism after the introduction of anaesthesia on the 3rd, 7th and 14th day (according to the order of Ministry of higher education of USSR №724 of 13.11.1984). Histologically, it was revealed that the newly formed blood vessels appeared in the retina and part of the optic nerve, germinated on their inner surfaces and the contiguous zone of the vitreous body in the form of the kidneys, which consist of proliferating newly formed vessels. Was also determined plethora of vessels in the choroid with the regional state of the formed elements of blood. In the zone of photoreceptor layer visualized cysts, the Druze, the newly formed blood vessels that defines the appearance of neovascularization and diabetic macular edema simultaneously.
A method of modelling of diabetic neovascularization of the macular region of the retina, comprising an introduction into the vitreous body of the eye of an experimental animal as a toxic agent Vascular endothelial growth factor (VEGF) fractional on 1, 3 and 7 of the day, characterized in that in the capacity of an experimental animal use a rat, and previously in the abdominal cavity dose of 15.0 mg/100 g weight through a 6.5 weeks after the introduction of and development of an experimental diabetes as VEGF enter 1 mcg rat VEGF 164 in a total dose of 3 mg.