MD1179Z - Method for treating trophic ulcers - Google Patents

Method for treating trophic ulcers Download PDF

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MD1179Z
MD1179Z MDS20170043A MDS20170043A MD1179Z MD 1179 Z MD1179 Z MD 1179Z MD S20170043 A MDS20170043 A MD S20170043A MD S20170043 A MDS20170043 A MD S20170043A MD 1179 Z MD1179 Z MD 1179Z
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Moldova
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wound
blood
patient
suspension
fibrin clot
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MDS20170043A
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Romanian (ro)
Russian (ru)
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Серджиу ПИСАРЕНКО
Адриан КУШНИР
Георге АНГЕЛИЧ
Татьяна ЗУГРАВ
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Государственный Медицинский И Фармацевтический Университет "Nicolae Testemitanu" Республики Молдова
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Priority to MDS20170043A priority Critical patent/MD1179Z/en
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Publication of MD1179Z publication Critical patent/MD1179Z/en

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Abstract

The invention relates to medicine, in particular to purulent surgery and can be used for treating trophic ulcers.The method consists in that 24…48 hours before the treatment, a suspension of mononuclear cells from the patient's blood containing 3×107/mL cells is separated, 30…40 mL of blood is also taken from the patient 2…3 hours before the first procedure, which is centrifuged for 8…12 min with 3000…3500 rpm to produce a fibrin clot enriched with platelets, the suspension is administered subcutaneously around the wound, at a distance of 1 cm from its edges, in an amount of 5…15 mL, after which the fibrin clot is applied to the wound, the procedure is repeated every 4 days, and the course of treatment is 8…10 procedures.

Description

Invenţia se referă la medicină, în special la chirurgia septică, şi poate fi utilizată pentru tratamentul proceselor septico-purulente ale ulcerelor trofice. The invention relates to medicine, in particular to septic surgery, and can be used for the treatment of septic-purulent processes of trophic ulcers.

Este cunoscută metoda de tratament al ulcerelor trofice în piciorul diabetic, care constă în aceea că se efectuează necrectomia ţesuturilor necrotizate din regiunea ulcerului, apoi timp de 5...7 zile se efectuează lavajul plăgii cu soluţie antiseptică, după care se aplică un pansament îmbibat cu soluţie de 0,5% de iodopiron. După sanarea plăgii se aplică autotransplanturi dermale [1]. There is a known method of treating trophic ulcers in the diabetic foot, which consists in performing a necrotizing excision of necrotic tissues in the ulcer region, then for 5...7 days the wound is washed with an antiseptic solution, after which a dressing soaked in a 0.5% iodopyrone solution is applied. After the wound heals, dermal autotransplants are applied [1].

Este cunoscută, de asemenea, metoda de tratament al ulcerelor trofice în piciorul diabetic, care constă în aceea că se efectuează pansamente cu un amestec care conţine plasmă îmbogăţită cu trombocite, soluţie de 10% de CaCl2 în cantităţi egale şi un preparat antibacterian la care este sensibilă flora din plagă [2]. There is also a known method of treating trophic ulcers in the diabetic foot, which consists of applying dressings with a mixture containing platelet-rich plasma, 10% CaCl2 solution in equal amounts, and an antibacterial preparation to which the wound flora is sensitive [2].

Mai este cunoscută metoda de tratament al complicaţiilor septico-purulente cu o rezistenţă majoră a microorganismelor faţă de antibiotice în piciorul diabetic, care constă în aceea că se efectuează pansamente cu soluţie de 1% de acid clorhidric pe plagă după aplicarea antibioticelor pe plaga menţionată. Pansamentele se efectuează până la apariţia granulaţiilor [3]. Another known method of treating septic-purulent complications with major antibiotic resistance of microorganisms in the diabetic foot is to apply dressings with a 1% hydrochloric acid solution to the wound after applying antibiotics to the aforementioned wound. The dressings are applied until granulations appear [3].

Este cunoscută metoda de tratament al ulcerelor trofice în piciorul diabetic, care constă în aceea că se administrează i/v soluţie de Tiroxin-alanil-glicil-phenilalanil-leucil-arginină diacetat (Dapargin), şi anume 4 mg, doza pentru 24 ore, în 10 ml de ser fiziologic, care se administrează timp de 10 zile, apoi se repetă peste 2 săptămâni [4]. The method of treating trophic ulcers in the diabetic foot is known, which consists in administering i/v solution of Thyroxine-alanyl-glycyl-phenylalanyl-leucyl-arginine diacetate (Dapargin), namely 4 mg, the dose for 24 hours, in 10 ml of physiological saline, which is administered for 10 days, then repeated after 2 weeks [4].

Este cunoscută, de asemenea, metoda de tratament al ulcerelor trofice în piciorul diabetic, care constă în aceea că se administrează i/a în artera femurală a membrului afectat autolimfocite prelucrate cu glutoxim 20 mg timp de 45 min. Autolimfocitele sunt obţinute cu ajutorul unui plasmoseparator de celule sangvine AS-TEC 204 firma „Fresenius”. Procedura se efectuează de două ori cu un interval de 48...72 ore [5]. The method of treatment of trophic ulcers in the diabetic foot is also known, which consists in administering i/a into the femoral artery of the affected limb autolymphocytes processed with glutoxim 20 mg for 45 min. Autolymphocytes are obtained using a blood cell plasma separator AS-TEC 204 from the company "Fresenius". The procedure is performed twice with an interval of 48...72 hours [5].

Este cunoscută metoda de regenerare a ulcerelor trofice inclusiv de geneză diabetică, care constă în aceea că preliminar, cu 24...48 ore înainte de tratament, se separă o suspensie de celule mononucleare din sângele recipientului, care conţine 3×107/ml de celule, apoi suspensia separată, peste o zi, în cantitate de 5...15 ml, se administrează subcutanat în jurul plăgii, la o distanţă de 1 cm de la marginile ei, şi în muşchiul gastrocnemian afectat, seria de tratament constituie 12...14 zile [6]. The method of regeneration of trophic ulcers, including those of diabetic origin, is known, which consists in the preliminary separation, 24...48 hours before treatment, of a suspension of mononuclear cells from the recipient's blood, containing 3×107/ml of cells, then the separated suspension, after a day, in an amount of 5...15 ml, is administered subcutaneously around the wound, at a distance of 1 cm from its edges, and into the affected gastrocnemius muscle, the treatment series being 12...14 days [6].

Dezavantajele metodelor cunoscute constau în aceea că are loc o regenerare lentă a ţesuturilor afectate, în unele cazuri sunt necesare intervenţii chirurgicale de plastie a defectelor dermale, precum şi utilizarea antibioticelor timp îndelungat şi aplicarea repetată a metodelor menţionate pentru obţinerea unei regenerări complete. The disadvantages of known methods are that there is a slow regeneration of the affected tissues, in some cases surgical interventions are necessary to repair dermal defects, as well as the use of antibiotics for a long time and repeated application of the mentioned methods to achieve complete regeneration.

Problema pe care o rezolvă invenţia constă în sporirea eficienţei în combaterea complicaţiilor septico-purulente în ulcerele trofice, stimularea proceselor regenerative şi prevenirea dezvoltării proceselor degenerative şi necrotice ale ţesuturilor afectate. The problem solved by the invention consists in increasing the efficiency in combating septic-purulent complications in trophic ulcers, stimulating regenerative processes and preventing the development of degenerative and necrotic processes in the affected tissues.

Metoda constă în aceea că cu 24...48 ore înainte de tratament se separă o suspensie de celule mononucleare din sângele pacientului, care conţine 3×107/ml de celule, de asemenea de la pacient cu 2...3 ore înainte de prima procedură se prelevă 30...40 ml de sânge, care se centrifughează timp de 8...12 min cu 3000...3500 rot./min cu obţinerea unui cheag fibrinic bogat în trombocite, suspensia se administrează subcutanat în jurul plăgii, la o distanţă de 1 cm de la marginile ei, în cantitate de 5...15 ml, după care pe plagă se aplică cheagul fibrinic, procedura se repetă peste fiecare 4 zile, iar seria de tratament prevede 8...10 proceduri. The method consists in separating a suspension of mononuclear cells from the patient's blood 24...48 hours before treatment, containing 3×107/ml of cells. Also, 30...40 ml of blood is taken from the patient 2...3 hours before the first procedure, which is centrifuged for 8...12 min at 3000...3500 rpm to obtain a fibrin clot rich in platelets. The suspension is administered subcutaneously around the wound, at a distance of 1 cm from its edges, in an amount of 5...15 ml, after which the fibrin clot is applied to the wound. The procedure is repeated every 4 days, and the treatment series includes 8...10 procedures.

Rezultatul invenţiei constă în intensificarea proceselor regenerative, antiinflamatoare şi de resorbţie, ce contribuie la o cicatrizare per primum a plăgii cu preîntâmpinarea trecerii proceselor necrotice în ţesuturile profunde şi cu remisii îndelungate. The result of the invention consists in the intensification of regenerative, anti-inflammatory and resorption processes, which contribute to per primum wound healing, preventing the spread of necrotic processes into deep tissues and with long-term remissions.

Platelet Rich Fibrin (PRF) prezintă o matrice fibrinică bogată în trombocite, care include în sine citokine, factori de creştere şi leucocite, având posibilitatea de a elimina substanţele menţionate un timp îndelungat. El poate fi utilizat sub formă de cheag sau membrană. Trombocitele pot secreta factori de creştere numai după formarea cheagului fibrinic, care îi conferă un potenţial terapeutic. Platelet Rich Fibrin (PRF) presents a platelet-rich fibrin matrix, which includes cytokines, growth factors and leukocytes, having the ability to eliminate the mentioned substances for a long time. It can be used in the form of a clot or membrane. Platelets can secrete growth factors only after the formation of the fibrin clot, which gives it a therapeutic potential.

Produsul menţionat se pregăteşte în eprubete vacuumate cu un activator al plasmei, acestea pot fi eprubete din plastic cu un strat aplicat de SiO pe pereţii interni sau eprubete din sticlă fără adaos, deoarece sticla este un activator al plasmei sangvine. După care se centrifughează 30...40 ml de sânge, timp de 8...12 min cu 3000...3500 rot./min. Apoi cheagul se amplasează într-o boxă (PRF-BOX) pentru scurgerea cheagurilor de fibrină. Scopul centrifugării este sedimentarea eritrocitelor. Principala condiţie este factorul g (RCF, acceleraţia centrifugă), care depinde de viteza de rotaţie şi de distanţa de la eprubetă până la axa centrală a centrifugii. The mentioned product is prepared in vacuum tubes with a plasma activator, these can be plastic tubes with a SiO layer applied to the inner walls or glass tubes without addition, because glass is an activator of blood plasma. After that, 30...40 ml of blood is centrifuged for 8...12 min at 3000...3500 rpm. Then the clot is placed in a box (PRF-BOX) for draining fibrin clots. The purpose of centrifugation is sedimentation of erythrocytes. The main condition is the g-factor (RCF, centrifugal acceleration), which depends on the rotation speed and the distance from the tube to the central axis of the centrifuge.

Pentru obţinerea PRF se începe centrifugarea la cel mult 1 min după prelevarea sângelui. Până la începerea centrifugării se amestecă bine sângele în eprubetă. Activatorul plasmei activează factorul 12, cu cât mai mare este concentraţia activatorului cu atât mai bine se amestecă sângele şi cu atât mai multă şi mai repede se formează trombina, care transformă fibrinogenul în fibrină. To obtain PRF, centrifugation is started no later than 1 min after blood collection. The blood in the test tube is mixed well before centrifugation begins. The plasma activator activates factor 12, the higher the concentration of the activator, the better the blood is mixed and the more and faster thrombin is formed, which converts fibrinogen into fibrin.

În perioada de până la 7 zile după aplicarea cheagului fibrinic pe suprafaţa ulcerului din el se elimină următoarele: During the period of up to 7 days after applying the fibrin clot to the surface of the ulcer, the following is eliminated from it:

- leucocite şi monocite ce se transformă în macrofagi, celule care stimulează regenerarea ţesutului; - leukocytes and monocytes that transform into macrophages, cells that stimulate tissue regeneration;

- VEGF - factorul de creştere a endoteliului (Vascular endothelial growth factor), care este o proteină de semnalizare, se elimină de celule pentru stimularea vasculogenezei şi angiogenezei; - VEGF - vascular endothelial growth factor, which is a signaling protein, is secreted by cells to stimulate vasculogenesis and angiogenesis;

- PDGF - factorul de creştere a trombocitelor (Platelet-derived growth factor) - proteină ca factor de creştere şi are importanţă pentru angiogeneză; - PDGF - Platelet-derived growth factor - protein as a growth factor and is important for angiogenesis;

- TGF - beta - factor de creştere şi transformare (Transforming growth factor beta) - proteină care controlează proliferarea, diferenţierea celulară şi alte funcţii; - TGF - beta - transforming growth factor beta - protein that controls proliferation, cellular differentiation and other functions;

- proteine, care au importanţă în procesul de angiogeneză, stimulează creşterea ţesuturilor; - proteins, which are important in the angiogenesis process, stimulate tissue growth;

- TSP - trombospondina este un inhibitor al angiogenezei, acţionând asupra adgeziei şi creşterii celulelor endoteliale; - TSP - thrombospondin is an inhibitor of angiogenesis, acting on the adhesion and growth of endothelial cells;

- IGF-1 - factor de creştere de tip insulinic 1 - proteină din familia factorilor de creştere de tip insulinic. - IGF-1 - insulin-like growth factor 1 - protein from the insulin-like growth factor family.

Cultura de celule mononucleare conţine substanţe biologic active ce contribuie la activizarea reacţiilor imune, la intensificarea sintezei proteinelor şi a proceselor fermentative, precum şi la inhibarea reacţiei inflamatoare, în locul administrării suspensiei are loc ameliorarea microcirculaţiei, proliferarea ţesutului conjunctiv, intensificarea proceselor de resorbţie. Ca rezultat are loc un proces antiinflamator, reparativ şi regenerativ intens, cicatrizarea plăgii se produce mai rapid, provocând diminuarea edemului şi prevenind dezvoltarea lui datorită ameliorării microcirculaţiei în ţesuturile plăgii. Mononuclear cell culture contains biologically active substances that contribute to the activation of immune reactions, to the intensification of protein synthesis and fermentative processes, as well as to the inhibition of the inflammatory reaction, in place of the suspension administration there is an improvement in microcirculation, proliferation of connective tissue, intensification of resorption processes. As a result, an intense anti-inflammatory, reparative and regenerative process takes place, wound healing occurs faster, causing a decrease in edema and preventing its development due to the improvement of microcirculation in the wound tissues.

Metoda se realizează în modul următor: de la pacient, preliminar cu 24...48 ore, se prelevă 20...40 ml de sânge, la care se adaugă 20...100 UI de heparină la 1 ml de sânge. Din sânge se separă celulele mononucleare pe un gradient de densitate, se spală cu ser fiziologic steril, se includ în mediul de cultură (mediu Eagle, TC 199, RPMI 1640, etc.) şi se amplasează în termostat la temperatura de 37°C pentru 24...48 ore. După perioada de incubare cultura celulară se separă de mediul de cultură şi se amestecă, obţinându-se o suspensie în 5...15 ml de ser fiziologic steril. De asemenea se pregăteşte cheagul de fibrină, şi anume se prelevă 30...40 ml de sânge şi se centrifughează timp de 8...12 min la 3200 rot./min, apoi se scurge cheagul de fibrină bogat în trombocite. Plaga se curăţă de mase necrotice, de fibrină până la apariţia sângerării din marginile ei, se prelucrează cu soluţii antiseptice, apoi suspensia de celule, care conţine 3×107/ml de celule în cantitate de 5...15 ml, în dependenţă de suprafaţa plăgii, se administrează în jurul plăgii, la 1 cm de la marginea ei, în stratul subcutanat al membrului afectat, procedura se repetă peste fiecare 4 zile, iar seria de tratament constituie 8...10 proceduri. The method is performed as follows: 20...40 ml of blood is taken from the patient, 24...48 hours in advance, to which 20...100 IU of heparin per 1 ml of blood is added. Mononuclear cells are separated from the blood on a density gradient, washed with sterile saline, included in the culture medium (Eagle medium, TC 199, RPMI 1640, etc.) and placed in a thermostat at a temperature of 37°C for 24...48 hours. After the incubation period, the cell culture is separated from the culture medium and mixed, obtaining a suspension in 5...15 ml of sterile saline. Also, the fibrin clot is prepared, namely, 30...40 ml of blood is taken and centrifuged for 8...12 min at 3200 rpm, then the fibrin clot rich in platelets is drained. The wound is cleaned of necrotic masses, fibrin until bleeding appears from its edges, it is processed with antiseptic solutions, then the cell suspension, which contains 3×107/ml of cells in an amount of 5...15 ml, depending on the wound surface, is administered around the wound, 1 cm from its edge, in the subcutaneous layer of the affected limb, the procedure is repeated every 4 days, and the treatment series consists of 8...10 procedures.

Metoda revendicată a fost utilizată pentru tratamentul a 10 pacienţi. Rezultatele au fost satisfăcătoare. The claimed method was used for the treatment of 10 patients. The results were satisfactory.

Exemplul 1 Example 1

Pacientul K., 66 ani, internat în secţia chirurgie septică cu o plagă supurată de 6x9 cm în regiunea 1/3 inferioare a gambei drepte, care s-a dezvoltat timp de 2 săptămâni de la o rană în regiunea menţionată. Pacientul suferă de diabet zaharat de 18 ani. După investigarea clinică şi paraclinică a pacientului s-a utilizat metoda revendicată, şi anume la a doua zi de spitalizare s-au preluat 30 ml de sânge, la care s-au adăugat 600 UI de heparină. S-a separat cultura de celule mononucleare. S-a utilizat mediul de cultură RPMI 1640, care a fost amplasat în termostat la temperatura de 37°C pentru 48 ore. Apoi s-a separat cultura de mediul de cultură şi s-a amestecat cu 10 ml de ser fiziologic steril. De asemenea, cu 2 ore până la efectuarea metodei, s-a pregătit cheagul de fibrină pentru a fi aplicat pe suprafaţa plăgii. S-a efectuat prelucrarea plăgii. S-au administrat în jurul plăgii în stratul subcutanat 10 ml de suspensie de celule. Pe suprafaţa plăgii s-a aplicat cheagul de fibrină bogat în trombocite. Procedura a fost repetată peste fiecare 4 zile, seria de tratament a constituit 9 proceduri. Plaga s-a cicatrizat per primum, fără semne de inflamaţie. Pacientul a fost externat în stare satisfăcătoare. Patient K., 66 years old, admitted to the septic surgery department with a 6x9 cm suppurating wound in the lower 1/3 of the right leg, which developed over 2 weeks from a wound in the mentioned region. The patient has been suffering from diabetes mellitus for 18 years. After clinical and paraclinical investigation of the patient, the claimed method was used, namely on the second day of hospitalization, 30 ml of blood were taken, to which 600 IU of heparin were added. The mononuclear cell culture was separated. RPMI 1640 culture medium was used, which was placed in a thermostat at a temperature of 37°C for 48 hours. Then the culture was separated from the culture medium and mixed with 10 ml of sterile physiological serum. Also, 2 hours before the method was performed, the fibrin clot was prepared to be applied to the wound surface. The wound was processed. 10 ml of cell suspension was administered around the wound in the subcutaneous layer. The platelet-rich fibrin clot was applied to the wound surface. The procedure was repeated every 4 days, the treatment series consisted of 9 procedures. The wound healed per primum, without signs of inflammation. The patient was discharged in satisfactory condition.

Exemplul 2 Example 2

Pacienta N., 53 ani, internată în secţia chirurgie septică cu o plagă supurată de 4x6 cm în regiunea plantei drepte, care s-a dezvoltat timp de 5 luni de la o rană tăiată în regiunea menţionată. După investigarea clinică şi paraclinică a pacientei s-a utilizat metoda revendicată, şi anume la a doua zi de spitalizare s-au preluat 30 ml de sânge, la care s-au adăugat 600 UI de heparină. S-a separat cultura de celule mononucleare. S-a utilizat mediul de cultură RPMI 1640, care a fost amplasat în termostat la temperatura de 37°C pentru 48 ore. Apoi s-a separat cultura de mediul de cultură şi s-a amestecat cu 15 ml de ser fiziologic steril. De asemenea, cu 2 ore până la efectuarea metodei, s-a pregătit cheagul de fibrină pentru a fi aplicat de suprafaţa plăgii. S-a efectuat prelucrarea plăgii. S-au administrat în jurul plăgii în stratul subcutanat 10 ml de suspensie de celule. Pe suprafaţa plăgii s-a aplicat cheagul de fibrină bogat în trombocite. Procedura a fost repetată peste fiecare 4 zile, seria de tratament a constituit 9 proceduri. Plaga s-a cicatrizat per primum, fără semne de inflamaţie. Pacientul a fost externat în stare satisfăcătoare. Patient N., 53 years old, admitted to the septic surgery department with a 4x6 cm suppurating wound in the right plantar region, which developed over 5 months from a cut wound in the mentioned region. After clinical and paraclinical investigation of the patient, the claimed method was used, namely on the second day of hospitalization, 30 ml of blood were taken, to which 600 IU of heparin were added. The mononuclear cell culture was separated. RPMI 1640 culture medium was used, which was placed in a thermostat at a temperature of 37°C for 48 hours. Then the culture was separated from the culture medium and mixed with 15 ml of sterile physiological serum. Also, 2 hours before performing the method, the fibrin clot was prepared to be applied to the wound surface. The wound was processed. 10 ml of cell suspension were administered around the wound in the subcutaneous layer. Platelet-rich fibrin clot was applied to the wound surface. The procedure was repeated every 4 days, the treatment series consisted of 9 procedures. The wound healed per primum, without signs of inflammation. The patient was discharged in satisfactory condition.

1. RU 2359624 C1 2009.06.27 1. RU 2359624 C1 2009.06.27

2. RU 2322247 C1 2008.04.20 2. RU 2322247 C1 2008.04.20

3. RU 2184551 C2 2002.07.10 3. RU 2184551 C2 2002.07.10

4. RU 2144831 C1 2000.01.27 4. RU 2144831 C1 2000.01.27

5. RU 2228204 C2 2004.05.10 5. RU 2228204 C2 2004.05.10

6. MD 696 Y 2013.11.30 6. MD 696 Y 2013.11.30

Claims (1)

Metodă de tratament al ulcerelor trofice, care constă în aceea că cu 24...48 ore înainte de tratament se separă o suspensie de celule mononucleare din sângele pacientului, care conţine 3×107/ml de celule, de asemenea de la pacient cu 2...3 ore înainte de prima procedură se prelevă 30...40 ml de sânge, care se centrifughează timp de 8...12 min cu 3000...3500 rot./min cu obţinerea unui cheag fibrinic bogat în trombocite, suspensia se administrează subcutanat în jurul plăgii, la o distanţă de 1 cm de la marginile ei, în cantitate de 5...15 ml, după care pe plagă se aplică cheagul fibrinic, procedura se repetă peste fiecare 4 zile, iar seria de tratament este de 8...10 proceduri.A method of treating trophic ulcers, which consists in separating a suspension of mononuclear cells from the patient's blood 24...48 hours before treatment, containing 3×107/ml of cells. Also, 30...40 ml of blood is taken from the patient 2...3 hours before the first procedure, which is centrifuged for 8...12 min at 3000...3500 rpm to obtain a fibrin clot rich in platelets. The suspension is administered subcutaneously around the wound, at a distance of 1 cm from its edges, in an amount of 5...15 ml, after which the fibrin clot is applied to the wound. The procedure is repeated every 4 days, and the treatment series is 8...10 procedures.
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MD2229F1 (en) * 2003-04-18 2003-08-31 Maria Danu Method of treatment of the shank trophic ulcers on the background of chronic venous insufficiency of the lower extremities
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MD4197B1 (en) * 2012-10-22 2013-02-28 Maria Danu Method for treatment of trophic ulcers
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MD4197B1 (en) * 2012-10-22 2013-02-28 Maria Danu Method for treatment of trophic ulcers
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