Leeches have been used in Oriental medicine for the treatment of different diseases since ancient times. In Kazakhstan leeches were used for “improvement and cleaning of blood”. In the 1940s and 1950s leech therapy was mainly used for heart failure and vascular diseases. In the 70-80s leeches were seldom used because there was a shortage of leeches and lack of experienced personnel to apply the therapy. The revival of interest in leech therapy was caused due to the unsatisfactory results of conventional treatment of many cardiovascular diseases, and to new findings about the leech salivary components and its influence on the human organism.

The Department of Traditional Medicine of Almaty Advanced Training Institute for Physicians together with the first Kazakh Hirudotherapy Center organized a group of tutors for conducting advanced courses on hirudoreflexotherapy. Advanced courses were intended for physicians, who are familiar with acupuncture, because the site of leech application determines the efficiency of leech therapy. The educational program includes 60 hours of lectures and practical courses. Lectures are given on leech anatomy and biology, hemostasis, general characteristic of medical effects of leech saliva and general principles of hirudotherapy and usage of hirudotherapy in different fields of medicine. Classes are held at the hirudotherapy center. Each student is able to apply leeches under supervision and learn the principles of taking care of leeches. Since 1998 79 physicians have been trained in the use of hirudotherapy. It is necessary to complete the advanced course of hirudotherapy in order to obtain a license for the method. Kazakhstan has rich natural sources for leeches, which receive a Certificate of Quality and are included in the Kazakh Drug Directory.

The evolution of Larva Therapy (LT) or Maggot Debridement Therapy (MDT) has been influenced by a number historical, professional, social, economic and communicative factors. Historically, LT only became established when laboratories were set up, mostly in America, in the earlier part of the last century, supplying larvae of specific fly species for use in clinical wounds. With a younger age spectrum, in that pre-antibiotic era, chronic osteomyelitis was relatively common, and large numbers of such patients were treated at that time, mostly by ex-military surgeons.

Over the past fifteen years LT has been re-established, first in the USA, then in Israel, the UK and Europe, with a number of dedicated fly culture laboratories now providing larvae for clinical use. The greatest development has been over the past four years. At least ten thousand patient treatments have been undertaken to date, dealing mostly with chronic ulcers in the elderly. Reports on clinical outcomes will be reviewed.

Laboratory research has been directed at the larval biology, with particular study of the digestive exo-enzymes. By its very nature, it has been possible to introduce LT into the clinical arena in parallel with laboratory research. This gives the advantage of ongoing cross feeding of information to and from the laboratory and the patient. The time is now opportune for multi-centre clinical trials. These could be international. However, again by its very nature, LT involves the interaction of two highly complex multi-cellular organisms ­ the human patient and the maggot ­ and the challenge of mounting effective multi-centre trails at present remains unsolved.

The future holds great prospect for developments in this field. Some particular lines of development will be outlined, with LT becoming an integral part of modern wound care, as much in the West as in the Tropics.

Larva Therapy (LT), the use of the larvae of the fly species Lucilia sericata for the debridement of open ulcers and wounds, is now well stablished in a number of countries in Europe, and in Israel. Much is now known of the behavior of larvae in human wounds, and their efficacy in debridement, and in the removal of bacteria. However, little is understood of the physiological and pathological interactions that occur during larval feeding in the wound, particularly in chronic wounds, with their diversity and complexity.

The chronic wound is studied in terms of its numerous gradients, for instance of pH and oxygen tension, and rates, for instance of cell multiplication and destruction, contrasting those areas which are healing, with those which are deteriorating. Larval feeding behavior is then described. Larvae use their mouth hooks to anchor themselves onto the food source, but also to scavenge small particles. Salivary exo-enzymes are forcefully expelled into the wound, and partially digested material, which includes the bacteria released by this activity, is swallowed. Feeding is directed primarily at necrotic or partially decomposed material within the ulcer, but healing tissue and surrounding skin are exposed to the larval secretions. The effects of this intervention, in terms of host cellular behavior, and other secondary effects such as host immunological response, are considered. Similarly, positive and negative effects of the wound environment on the larvae and their behavior are also addressed.

Maggots are enclosed between two 0.5-mm thin layers of polyvinylalcohol-hydro-sponge, which are glued together over a small cube of a spacer material. The thin walls of the porous dressing are permeable enough for wound fluids and liquefied necrotic tissue components to guarantee the nourishment of the maggots. At the same time, the maggots` secretions pass through the Biobag to stimulate wound healing and control infection. Wound dressing with the Biobag is fast and very easily performed. The maggots can positioned directly on the area needing treatment. Pain is reduced as there is no mechanical irritation of the wound edges by the maggots` physical activity. Maggots cannot escape from the Biobag, which is essential for hospital hygiene. Patients` acceptance of maggots seems to be higher as they are not visible. During the 3-4 days of application the dressing might be removed for wound inspection at any time and replaced thereafter. Indication and technical details are demonstrated.

Hirudotherapy (HT) - the medicinal use of leeches (Hirudo medicinalis) - promotes local and systematic effects due to its anti-inflammatory, bacteriostatic, anti-coagulant, and analgesic properties, by bloodletting tissue decongestion. The literature contains isolated references to the dermatological aspects of H (Park et al., 1976; Ross, 1983; Smirnoff, 1994) and the majority of the results were obtained mostly from clinical observations. The aim of this report is to illustrate the efficacy of HT for different skin disorders in patients from the department of HT, Ust-Katchka Health Resort. The results of HT on 19 patients (10 males and 9 females, 54-73–years old) with chronic venous leg ulcers (CVLU) are presented. The medical history, routine biochemical, hematological and blood coagulation indices, planimetry, photography, and laser Doppler were recorded for all patients before and after treatment. The results of the HT were compared with results obtained using other treatment methods. In cases of CVLU aspirative HT was used. Five to seven leeches were applied to the affected limb, parallel to the inflamed vein, 1 cm from the border of the ulcer. The time of hemoextractiovaried considerably. Thresults after only one treatment of HT were encouraging. Almost all patients reported decrease in pain, edema and fatigue. In leech-treated patients improvement was assessed by the recovery of skin blood circulation and color, and by the reduction of the ulcer size by 32% in average. Complete healing was observed after 2-3 courses of HT in 37% of the ulcers treated. There were no secondary infections, systemic bleeding or anemia during HT. In two cases a skin irritation, itching and scarring of the wound was observed. One patient had an allergic skin eruption, which was successfully treated with oral antihistaminics. The mechanism of action of HT in chronic venous ulcers can be explained by decongestion in the tissue, injection of antihemostatic, vasodilatatory and antiphlogistic substances of the leech saliva into the wound, thus, restoring the micro- and macrocirculation, and optimizing regeneration. HT has provided and efficient treatment of sclerodermia (S). Smirnoff (1994) treated 26 cases of S (patching, lineal, diffuse, Bushke) with leeches with success in 58% of these cases. We used non-aspirative hirudoreflexotherapy for the treatment of local S in 4 females (median age 56 years), by applying the leeches to the points of acupuncture and around the focus of the lesion. The result of 3 courses of leeches was impressive in all patients. Two patients had a complete reduction of the skin lesion. In two patients there was more than 70% regression of the skin lesion. The improvement was visible after the first course of treatment when the focus and perifocal zone began to fade. There was a significant reduction of the thermogradient up to 0.6± 0.2, p< 0.05). There were no visible side effects or complications related to leeches. HT can be used in case of furuncles and carbuncles in the infiltrative phase (as monotherapy) and in abscess formation (together with surgery). We found that when leeches were used, infiltrates dissolved two days earlier, the exudation ceased more rapidly and healing time was shortened as compared to controls. We also recommend HT for the cosmetic correction of postoperative cicatrices. We investigated the effectiveness of HT in 9 cases (3 males and 6 females, aged 49-62) with Lichen ruber planus and had some positive results (decrease of edema, perifocal inflammation, and number of papules and increase of healing rate of ulcers in 66% of cases. The results of polarography (7^th day of the treatment) demonstrated significant increase of initial and maximal oxygen saturation, the rate of increase of oxygen saturation in the affected zones. H is thus may be considered to be an effective therapy for some skin disorders and diseases.

The anti-inflammatory, antiedematic and healing properties of leeching have been demonstrated clinically. The mechanism of action of leeches is not clear and is of scientific interest. In the first part of this study we examined the curative effect of aspirative and non-aspirative types of leeching on an electric wound model in 12 male Wistar rats (180-200 g). Five wounded rats, which had not been treated, were used as controls. Histological studies demonstrated the positive effect of leeching on the course of wound healing process: 1. Local necrotic and inflammatory reactions were decreased; 2. The period of inflammation was reduced; 3. Epithelialization and granulation were accelerated; 4. Eosinophils appeared sooner in the focus of lesion; 5. Increased and well-balanced activity of macrophages was promoted in the focus of lesion; 6. Synthesis of new blood and lymphatic vessels was enhanced; 7. More delicate, compact, vessel-free cicatrices were formed. Histological evaluation revealed that aspirative and non-aspirative types of leeching can influence the course, speed and outcome of wound healing . In the second part of the research we evaluated the anti-inflammatory effect and in particular the anti-exudative effect of leeching using the model of agar-induced inflammation (edema in the feet of rats, induced by the injection of 0,1 ml 1% agar under the plantar aponeurosis of 12 male Wistar rats). The results of oncometry indicated significant reduction of inflammation and edema after leech therapy. Three hours after administration of agar, a 30.0± 4.1% reduction was observed, and after 6 hours there was a 17.0± 2.3% reduction. Twenty-four hours later, when the inflammation was still present but less severe, leeching promoted the reduction of edema by 16.0± 3.0%. Thus, these results demonstrate the presence of an anti-exudative effect of leeching, which is more noticeable at the earliest stages of the inflammation.

According to K.Smorl, osteochondrosis is the clinical form of abnormal changes in intervertebral disks. The clinical picture of the disease is characterized by intensive and frequently occurring pains. The pain is especially acute if it is localized in the lumbosacral area. The muscles of the back are tense, and painful when pressed. The slightest movement, physical coughing and laughter can intensify the pain. The disease can be caused by physical strain, vibrations, sharp movements, changes in the intervertebral disks, trauma and other factors. The lumbar area is characterized by the presence of short muscles. Excessive strain of these muscles results in pain in the loins. The muscles can be tense for months, even for years, and hence the pain.

The intervertebral disks under the influence of body weight, strong movements and lifting heavy objects, results in degeneration of the vertebrae, especially after the age of thirty. The resilient core of a disk begins to be squeezed and its thickness decreases. Under these conditions and when bending or while lifting heavy objects, the resilient, jelly-like core breaks through a fibrous ring, which results in the formation of a hernia. The treatment of muscular blockade consists in releasing the spasm of the back muscles by relaxation, the treatment of edemas and physical rehabilitation. Today, reflexotherapy (acupuncture, electric puncture, and laser puncture) massages and physiotherapy are also used. These methods require highly qualified experts and are not always effective.

Apitherapy is the use of bee products such as venom, honey and propolis. The venom has a local and systemic effect in the nervous and vascular systems. The systemic effect is due to apamine and MSD (peptide 401), which increase the capillary permeability and have an anti-inflammatory action, which is stronger than hydrocortisone.

For the treatment of radiculitis with apitherapy a single application is normally sufficient. According to V.J. Prichodko (1972) the efficacy of the treatment with bees for all forms of lumbosacral radiculitis is 85,5 %. A course of treatment consists of 12 sessions and 150-250 bee stings. According to N.A. Bobrova et al. (1983) the efficacy of an injection of a bee venom extract (Apicaine) for the treatment of lumbosacral radiculitis is 59,4 %. Here, a course of treatment consists of 15-20 injections.

The extraction of the bee venom and the subsequent desiccation results in a loss of all aromatic substances and about 25 % of the greasy acids, and hence a decrease in its medicinal qualities. Therefore, in the "Vympel" Institute live bees are used for the treatment of lumbosacral radiculitis. In addition massages with honey, and treatment with creams of propolis and apilak are used. The treatment consists of 3-7 sessions and 2-25 bees are used for each session. Bees are kept in the institute throughout the year.

Case report: A 50-years old male patient who was a colonel in the army suffered from chronic lumbosacral radiculitis for the last 25 years. He was treated with traditional medicine, e.g., massages, physiotherapy, acupuncture, unguents,grindings and different physicexercises, but without success. The pains were stronger during hot weather. Due to the physical impairment he was dismissed from the army. In May 1999 a course of apitherapy with 4 sessions was initiated using 10 bee-stings. The pains disappeared, the movements became free, and there was no deterioration during hot weather.

Maggot or larva therapy (LT) has a fascinating past, which involves many great names from the history of surgery. Its zenith was in the 1930’s, when hundreds of patients were treated. Several major papers were published, and the documented results were very encouraging, especially when the only alternatives to LT were surgical debridement and topical antiseptics. Larvae were commercially available in the USA, though many LT users bred their own maggots. The advent of antibiotics and vastly improved hospital technology from the l940’s onwards led to the rabid “extinction” of LT. Consequently, most doctors toady are either entirely ignorant of LT, or see it as a historical dead end, with no further relevance. This presentation reviews our current experience, over four years, with LT in a district hospital in the UK. Forty orthopaedic patients have been treated for a wide variety of septic conditions, with variable outcomes. Most patients were aged between 60 and 80 years. Nearly one thrid had diabetes; some others were complicated by peripheral vascular disease, neuropathy, or presence of an implant. Only four upper limbs were treated. The general conclusion is that with proper case selection, healing or improvement of localized orthopaedic sepsis will result in over 80% of cases treated by LT. Particular cost benefits may follow larval debridement of amputation stumps. In four cases, a more proximal amputation was avoided. We believe these cases alone would offer ample justification for use of LT, if its use, or cost, is challenged. Aesthetic acceptance of the treatment by patients and nurses has been almost total. The response of the microbiologists has been neutral, which is disappointing, since LT offers a unique form of “germ killing” without antibiotics. There is no doubt that LT has now re-established itself as a valid option in current orthopaedic infection management, which merits wider acceptance, especially as those patients most likely to benefit are almost by definition disadvantaged. They include the elderly, severe diabetics, amputees and those with chronic sepsis. Its potential in developing countries is also obvious, although largery unrealized.

The number of patients with facial furuncles and carbuncules who have been cured in the clinic of maxillo-facial surgery has increased 3.7 times during the last 4 years. There was also an increase in the number of patients with “malignant” forms, which lead to cavernous sinus thrombosis, metastatic pneumonia and mediastinitis. The treatment is based mainly on the dissection of the purulent center, necrectomy and crossing and suturing the facial vein, especially when there is suspicion of a thrombosis. However, conventional drugs such as antibiotics often inadequately cure the furuncles and carbuncles. In these cases more traditional methods such as leech therapy is indicated. Leeches were used because their saliva has anti-inflammatory, thrombolitic, immunomodulatory, anesthetic and regeneratory effects. During 1998-99 leech therapy was applied mainly in complicated cases, e.g., thrombosis of the angular vein. Four patients who suffered from cavernous sinus thrombosis or purulent meningitis were treated successfully by leech therapy. This method made it possible to reduce the period of hospitalization, to eliminate venous thrombosis, inflammatory infiltration and intoxication. The practice shows us that leech therapy is an effective method of cure.

Since 1996 our team treated 159 wounds in 97 patients (54 males and 43 females), aged 35-95 (median 67), suffering from intractable wounds. The underlying diseases were diabetes (42), venous stasis (26), paraplegia (6), dementia (6), hemiplegia (3), vascular disease (3), PVD (3), Birger’s disease (1), lymphostasis (1) and thalassemia (1), polycytemia (1), basal-cell carcinoma (1) and unknown (4). The wounds had been present for 1-240 months (median:5) before MT was applied. Most of the wounds (138) were sloughy and necrotic and were located on the foot or the thigh, while the 20 pressure sores were located on the lower back and perianal region. One patient with basal-cell carcinoma had wounds all over his body but only the wound on the finger of the left hand was treated by maggot therapy. Fifty-five patients were hospitalized when they received treatment and 42 patient were ambulatory. Patients were treated in several Departments of the Hadassah Hospital (Dermatology, Hematology, Vascular Surgery and Internal Medicine, Diabetes and Day Care Units, Intensive Care, Orthopedics, Geriatrics and Plastic Surgery) in the Cardiac Surgery Unit of Sharei Zedek Hospital, in four geriatric hospitals (Neve Horim, Veyizrah Yitzhak, Kupat Holim Klalit and Hod Yerushalayim), one outpatient clinic (Kiriat Menachem) in Jerusalem and in the Geriatric Department of the Tel Hashomer Hospital in Tel Aviv. Maggot therapy was selected because all the other surgical and non-surgical treatment methods such as hydrogel dressings, antibiotics, hyperbaric oxygen and disinfectants, had failed to cure the wounds.

Sterile maggots (24-48 hrs old) of the green bottle fly, Lucilia sericata were administered to the wound 2-5 times weekly and replaced every 1-3 days. Depending on the size of the wound the number of treatments varied between 1 and 48 (median: 4), and lasted for a period of 1-210 days (median: 6).

Complete debridement was achieved in 75.3% of the wounds, significant debridement in 19.6%, partial debridement in 3.1% and in only 2.0% of the cases the wound remained unchanged. In at least eighteen cases leg amputation was prevented as a result of maggot therapy. As the therapy progressed, new layers of healthy tissue were formed over the wounds, while the offensive odor emanating from the necrotic tissue and the intense pain accompanying the wound decreased significantly. The majority of the patients did not complain about any major discomfort during the treatment. Twenty-four patients with superficial, painful wounds complained of increased pain during the treatment with maggots and were treated with analgesics.

It was observed that maggots are capable of entering any part of the wound wherever necrotic tissue exists and clean minute areas without harming healthy tissue in a manner resembling micro-surgery; a task which is very difficult to attain by the conventional surgery. After MDT, patients were sent either for a skin transplant or their wounds were treated with hydrocolloidal sheets or disinfectants.

It is known that maggots are capable in feeding on the necrotic tissue, and significantly reducing the number of bacteria inside their body as well as in the wound through their excretions, which contain antibacterial substances. GFP-producing E. coli were used to investigate the fate of these bacteria in the alimetract of sterile grown maggots. The presence obacteria in the different parts of the intestinal tract was studied using a laser scanning confocal microscope. A computer program was applied to analyze the intensity of the fluorescence and to quantify the number of bacteria. The crop and the midgut were the most heavily infected areas of the intestine. A significant decrease in the amount of bacteria toward the end of the midgut was observed. The number of bacteria decreased even more significantly in the anterior part of the hindgut and practically no bacteria were seen in the posterior end, near the anus. Bacterial examination of the gut sections on LB agar showed that there were viable E. coli in 66.7% of the crops, 52.8% of the midguts, 55.6% of the anterior hindguts and 17.8% of posterior hindguts.

In order to study the mechanisms by which the excreta of maggots of the green bottle fly, Lucilia sericata stimulate the granulation of the debrided wound and the immune system of the treated patient, we evaluated their potential to secrete cytokines in vitro. The liver-agar medium on which maggots were left for 24 hrs, was washed with PBS to extract the excretion products of maggots. As a control extracts of liver-agar without maggots and PBS were used. The presence of cytokines in 8 different lots of maggot secretions was invetigated

In an attempt to evaluate the cytokine trend production we studied IFN? for TH1 and IL-10 for TH2. As a general immune system activation marker sIL-2R was also analysed. All tests were assessed by a specific ELISA technique.High amounts of IL-10 (7-27 pg/ml) and IFN? (20-35 pg/ml) were produced by 8 and 5, respectively extracts tested, as compared to the negative controls (<3.4 and <6.8 pg/ml, respectively), while sIL-2R activation was not observed in any of the extracts.Although no clear trend towards TH1 or TH2 was observed, we conclude that maggot extracts do initiate specific cytokine production, which may explain their beneficial effect in the quick granulation of the debrided tissue.

The secretions of the maggots contain ammonia, urea and allantoin, which are known to also stimulate granulation. In addition maggot secretions contain calcium carbonate, which changes the wound pH from acid to neutral (7-8), and which also has a stimulatory effect to granulation.

The age-dependent changes in facial tissue are among the most important reasons for treatment by plastic surgery. In the arsenal of plastic surgery and cosmetology there are different methods for treating edemas, and resorption of hematomas including electrophoresis, phonophoresis, and anti-inflammatory drugs such as Lydasum, Ronidasum, heparin and Troxevasin Unguentum and gel. However, these remedies are not always effective and therefore, a study with “Hirudon” gel for epicutaneous application for patients during the postoperative period was initiated. The basic component of the gel is the saliva of medicinal leech, which contains biologically active substances with antithrombotic, thrombolytic, hypotensive, regenerating, antiartherogenous, antihypoxic, immunostimulating and analgesic actions.

"Hirudon" gel is prepared from leeches, which are maintained in the bio-factories of the "International Centre of the Medical Leech". The leeches are standardized according to their age, weight and period of starvation. "Hirudon" gel is a complex drug containing 0,5 % of leech saliva (based on Ultrez-10, England) and is manufactured by the "Biokon" Corporation JV (Donetsk, Ukraine). The drug contain ingredients, which are permitted by the EEC.

The experimental and clinical evaluation of the gel was carried out in the Institute of Plastic Surgery and Cosmetology of the Ministry of Public Health of the Russian Federation. Its toxicity was studied on white male rats (n=16), and its irritating and sensitizing action on white rats (n=16). Morphological studies were carried out on the skin and liver, and the anti-inflammatory action on rats by the Trinus method. Clinical trials were conducted on 20 patients (38-65 years-old) during the postoperative period, who were treated for dermal abrasions, removal of skin excesses from face and neck, and removal of excess tissue from upper and lower limbs and body. Long-term epicutaneous application (21 applications) of the "Hirudon" gel did not cause any generalized toxic effects in the animals. A comparative study showed that there were no differences between test and control animals with regards to body weight, state of internal organs and skin. No irritating and sensitizing effects of the gel were observed. Morphological studies of the skin and liver revealed no pathological changes. It was found that "Hirudon" gel has anti-inflammatory properties and decreases the inflammation by 19.6% after1 hour, by 23.0% after 2 hours and by 29.6% after 3 hours.

"Hirudon" gel was applied epicutaneously 2-3 times daily in the area of a posttraumatic edema of the periorbital area, such as postoperative scars of lid and hematomas of the face. All patients were examined and supervised by a doctor or cosmetologist. The efficacy of "Hirudon" gel was evaluated visually and with the help of photographs. A good anti-inflammatory effect and a better resorption of hematomas were observed after the application of the "Hirudon" gel for 5-7 days. The application of the "Heparin" Unguentum caused a resorption of hematomas only after longer periods (10-14 days). Application of the "Hirudon" gel (2-3 times daily) for 7 - 14 days decreased the postoperative edema in 50% of the patients within 2-3 days. It was also effective in the resorption of postoperative hematomas, especially in the periorbital area, as compared to patients who were treated with drugs such as Unguentum Heparini. Thus, "Hirudon" gel is indicated for epicutaneous application during the postoperative period for the resorption of hematomas and treatment of edemas.

There is currently a search for natural products, which improve man's psychosomatic status.. It is important to have non-toxic preparations, which are capable to cause relaxation. That is, either to take the necessary measures to prevent stress or to remove its consequences. The leech Hirudo medicinalis was selected for this purpose, which apart from its effective pharmacological actions (e. g., antithrombotic and thrombolytic), improves the mood, appetite, sleep and work capacity. Because the medical leech is not convenient both for the doctor and for the patient, we produced a series of cosmetic preparations capable of improving the psychosomatic status of a person.

In our work we used the medicinal leech Hirudo medicinalis which was maintained under artificial conditions developed at the Biokon (Ukraine). Eight to nine month old leeches, which starved for 4-5 months were used. After cryotreatment, homogenization, gel-filtration (Sephadex 6-75) and lyophilization, a preparation (NIK) was obtained. NIK contains hirudin (18,02,5 ATUmg^-1), blood plasma kallikrein inhibitor (14024 U.mg^-1), prostacyclin-like substances (88865 gmg^-1) and trypsin inhibitor (243 ATUmg).

Studies were conducted with NIK on the contractility of smootand striated muscle in vitro, on the state of muscle tone, and on the normal mental status of volunteer. It was shown that the preparation has a dose dependent effect on the state of a muscle tone and also on mental state of a person. Low doses and long lasting contact with receptors (isolated muscle tissue or skin receptors) cause a relaxation of the muscle and has a calming effect on the patient, e. g., normalization of sleep patterns and psychic tension. Higher doses however, have the opposite effect.

One group of white mice (n=36) were treated with LD₅₀=7.5 gkg^-1, LD₁₀₀ =9.5 gkg^-1 body weight by oral application of a water suspension of the preparation. At 2.5 gkg^-1 a slight decrease in obesity, with no damage to the liver cells was observed. At a dose of 7.5 gkg^-1 small drops of lipids were seen in hepatocytes. Epicutaneous applications of a water suspension of NIK preparation on guinea-pigs (180-200 g, n=24) during 21 days (at 1/30 and 1/15 of the LD₅₀, 20 mgkg^-1 and 40 mgkg^-1 body weight, respectively) no toxic, irritating or sensitizing effects on the animal organism was detected.

Two tablet formulations for bath, "Tonus" and "Relax" ("Biorica"), with standardized quantities of the active ingredients were developed. The toxicity of the tablets was studied on rats (200-220 g, n=25) by bathing them for 21 days in water treated with "Tonus" and "Relax" preparations. No toxic, irritating or sensitizing effect on animals was found. At the autopsy of the animals no pathological changes of internal organs (kidneys, lungs, spleen, heart) were detected.

An additional study was carried on healthy people (n=50, 30-65 years old). The volunteers took a bath every second day for 20 days and no irritating and sensitizing effects of the preparations were observed. These remedies soften the skin, improve microcirculation and help small wounds to heal.

NIK is a remedy prepared from biologically active substances extracted from the leech Hirudo medicinalis. Clinical trials were carried out in the Department of Neurology, Internal Medicine, Faculty of Fundamental Medicine of the State University (Kharkov, Ukraine).

Thirty patients (10 males and 20 females, 35-45-years-old) were divided in three groups. The patients did not receive any treatment one month prior to beginning of the study. The patients in the first group received a bath daily for 4 days (38 C?; 15 minutes). The patients of the second and third group were treated as above, however, the water was treated either with "Relax" or "Tonus" tablets.

The level of anxiety of the patient as an indication of his/her personality was measured by the Spielberg technique. A patient was considered suitable for the study, if his level of anxiety was moderate (31-45 points). The vegetative and hormonal tonus of the patient was measured three times with the help of a computer program, i.e., the "Heart-rate-variability" was measured before, immediately after and one hour after having a bath. The differences between the background and hourly measurements of heart rate indexes (HR), the variability of the R-R intervals, the magnitude of the hormonal (Pvlf), sympathetic (Plf) and parasympathetic (Phf) changes were evaluated.

There were no significant differences in the decrease of the HR or K-K interval variability in patients using a regular bath or such with "Relax" or "Tonus". The hormonal changes were 8% with a regular bath and 5-7% with “Tonus” bath. However, a significant change was observed with “Relax” bath. Baths with or without "Tonus" and "Relax" tablets equally diminished the activity of the sympathetic nervous system. A clear effect of the drugs was obtained in the activity of the parasympathetic nervous system. The regular bath diminished it by 2 %, the “Tonus” by 5% and the "Relax" by 22 %. Patients having a bath with the "Relax" tablets for four days had a decrease in level of anxiety to 25-31 points. No such decrease was observed with "Tonus" tablets.

The following series of trials on "Relax" tablets were conducted in the Neurosis Clinic "Gravieres" (France, Bordeaux). The clinic is specializes in the treatment and rehabilitation of people with moderate mental disorders, however, people with marked depressions or neurosis are also treated. Normally the patients are treated with psychopharmaca. Ten patients (21-45 years old) suffering from neuroses (40% of them alcoholics) were included in the study. The patients were treated in water baths (34-36 C?; 20 minutes) once every second day, during two weeks, usually 1 hour before the sleep, and were observed round the clock.

This study has shown that the drugs have a vasodilatory effect. The patient fell asleep easily, they were more relaxed, were sleeping without interruption wake up earlier with a feeling of having a good rest. A reduction of the medication was observed in 65 % of the cases. It should be stressed that the drugs significantly increased the working capacity of the individuals for a long period. It is possible that the drug also increases sexual potency. As a result of these studies new cosmetic preparations with anti-stress properties such as “Aura” and “Madame Latrille” were developed.

At the Department of Dermatology in the University Hospital of Basel, we started using larvae of Protomorphia terraenovae. The maggots were provided from external sources. Breeding and sterility were performed as published. Obviously the procedure used here was not adequate to achieve sterility of the larvae. Five out of 24 patients, who were treated with approximately 100 cycles of maggots for leg ulcers, developed sepsis, which could only be attributed to this therapy. The first patient developed a Candida sepsis on the fourth day of the second treatment. We could find no other focus of infection than his ulcer. The second patient developed sepsis on the fourth day of the treatment. Providentia stuarti was isolated from the hemoculture. The pathogen could not be isolated from a smear of the ulcer at admission. However, it was isolated after maggot therapy from a smear of the ulcer, before treatment with antibiotics. A third patient developed polymicrobial sepsis with Streptococcus group C and P. stuarti. Both pathogens were not present in the initial smear of the ulcer. In the fourth case four, a 74-year-old man developed sepsis with P. stuarti on day three and seven. The pathogen was isolated from the larvae and from the ulcer smear. Case five was a 75-year-old woman with an ulcer of mixed vascular type complicated by osteomyelitis on the left lower leg. On the first day of the second application of maggots, the patient developed sepsis with P. stuarti. The initial flora colonizing the ulcer was Staphylococcus aureus. After the first cycle of maggot therapy P. stuarti, Morganella morganii, enterococci, enterobacteriaceae and candida could be isolated from the ulcer. Genotyping the microorganisms of case two and five with pulsed field gel electrophoresis (PFGE) revealed the same type of microorganisms isolated from blood, larvae and ulcer after therapy. Where tested, the ulcer flora changed to become identical with the flora cultured from the larvae. Obviously, the procedure used in the laboratory, which supplied the maggots, was not sufficient to achieve sterility of the larvae. Bleeding and pain during the treatment were frequent symptoms. The antimicrobial properties of maggot secretion as postulated, were apparently not effective, at least not on the flora contaminating the larvae. Despite promising properties maggot therapy can have serioadverse events and should therefore only be used under strictly controlled conditions.

In 1999, 430 applications of maggots took place in 101 patients in our OPD and 563 applications in 114 patients in the hospital`s wards. This gives a total of 215 patients with 993 treatments which means that on average approx. 4 maggot applications were performed daily. Most of the patients had diabetes, diabetes in combination with arterial occlusive disease, stasis ulcers, arterial occlusive disease and chronic osteomyelitis. Indications, results and peculiarities of the treatment are discussed.

The importance of opportunistic microorganisms in the development of sexually transmitted diseases is steadily increasing. Non-pathogenic microorganisms such as Ureaplasma, Mycoplasma, Gardnerella vaginalis, Staphylococcus aureus and other representatives of the normal flora of the urogenital area, in association with more pathogenic microorganisms such as Chlamydia, Cytomegalovirus and Trichomonas, create powerful combinations, which are resistant to modern antibacterial substances.

It is suggested that the increase of microbial pathogenicity is due to local immunodeficiency in the pelvis area, resulting tiredness by overuse of hormone contraceptives.

Local immune rehabilitation can be achieved by using leeches in the area of lymph nodes, i.e., pubic area, the external aperture of inguinal channel in women and the inguinal area below Poparti ligament and medial to femoral vascular-nervous bundle in men. In addition thymus-dependent drugs and micro doses of antibiotics are injected in the area. This method cures Chlamydia, Cytomegalovirus and Mycoplasma infections in patients with chronic salpingo-oophoritis and urethroprostatitis., as it was demonstrated with the DNA test conducted within7-10 days.

The essence of this method is that thymus-derived drugs in combination with lymphotropic substances from leeches reach the thymus-dependent zones of lymphoglandulae. This results in a cascade of immunostimulatory reactions.

In conclusion it can be said that in the case of opportunistic infections a change of etiopathogenic factors can be observed, i.e., the reason for a strong inflammatory reaction is the local immunodeficiency, which allows opportunistic microorganisms to implant themselves and become more pathogenic.

One of the major components of treatment of malignant neoplasms of the pharynx and oral cavity is the radiation therapy, which is carried out usually prior to plastic surgery.

Radiation therapy frequently causes damages to healthy tissue in the treated area. The complications of the radiation can not always be easily treated. The complications are due to damage to the vessels, which is also the origin of thrombosis, disorders in vascular permeability and microcirculation. In addition to blood vessel damage, the lymphatic vessels suffer from obstruction of lymph drainage. Thus, the basic reason for complications during healing of postoperative defects of the oral cavity floor and pharynx (partial or total necrosis of the grafts) is due to disorders of microcirculation. Taking into consideration that additional trauma to tissues, which have already suffered from radiation injury, is not desirable, we have applied a gel, "Hirudon", which was prepared from the leech Hirudo medicinalis. "Hirudon" gel was applied to treat radiation injuries in 24 patients suffering from larynx cancer, with transition to the wall of pharynx and the floor of the oral cavity (I group). A control group including 10 patients with similar pathology received traditional medication (II group). All patients have undergone radical operations - extirpation of larynx with removal of the side wall of pharynx, removal of affected areas in the oral cavity, as well as pre- and postoperative teletherapy. During the postoperative period, all patients showed defects in the distal segment of the oral cavity floor or side wall of the pharynx. The majority of patients (26) developed a suberythematous dermatitis during the radiation. In 8 patients an erythematous dermatitis developed. A thin layer of "Hirudon" gel was applied on the affected are of the skin of patients from the 1st group, 3-4 times daily. The treatment was initiated immediately after the appearance of radiation damage and continued until healing. For the 2^nd group of patients Troxevasin and Heparin Unguentum were used. In addition, all patients were treated with anti-inflammatory, detoxifying, desensitizing and stimulating remedies. The following parameters were studied: blood coagulation; the dynamic of radiation damages on the skin and mucous membranes, according to the level of oxygen in tissues and the oxygen tension in tissues as it was measured by interstitial polarography.

A decrease by 4.5-6% in the level of oxygen in tissues exposed to radiation was observed. The low oxygen content can be explained by a vascularization disorder of these tissues as a result of radiation, decreased resistance of the organism and sometimes by steady immunodepression in given patients. During treatment with "Hirudon" gel the oxygen level in tissues increased by 10-11 %. In the control group and increase of 6.5-7.5 % was observed. The gradual healing of radiation damages occurred in parallel to the increase of oxygen quantity and the improvement in trophicity of the irradiated tissues. In patients with a suberythematous dermatitis the inflammation was controlled within 7-10 days after radiation. In patients with erythematous dermatitis the healing from radiation damage on the skin and mucous membranes occurred within 10-14 days after radiation. In the control group similar healing was seen in 4-6 weeks. Dermatitis was not observed in patients treated with "Hirudon" gel and the treatment by radiation was not stopped in any patient. No necrosis could be observed in patients who received skin graft.

The physicians noted that a gel is a more convenient way of treatment of radiation damages as compared to ointments, as the gel is absorbed quickly without leaving an oily layer on the treated surface.

Destabilase, a glu-lys isopeptidase, was first found in the saliva of Hirudo medicinalis in 1986. The enzyme’s activity is due to the hydrolysis of isopeptide links formed during accumulation of fibrin in aging cells and is responsible for the formation of eye cataracts.

The medicinal leech Hirudo medicinalis, which is maintained in the biofactories of the "International Centre of a Medical Leech" (Russia & Ukraine) was used in this study. The leeches were starved for 4 months. Destabilase is a large lipo-protein complex (MW 12.3 kDa) with high aggregation ability. As a result of destabilase aggregation the micelles change their spatial orientation according to the solvent.

The destabilase binds hirudin and blood plasma kallikrein inhibitor forming a water soluble liposome, which acts as the destabilase complex (DC), and consists destabilase, prostacycline analog, hirudin and kallikrein inhibitor). The DC has a MW of 25 kDa and has the shape of a monomeric liposome. In organic solvents only the prostacyclin analog of the destabilase is active.

The special structure of DC provides not only a stabilization of the components, but also facilitates their transport through the cell memb(trans-membrane transport). This was shown in experimental animals, which were treated with DC intravenously and per os.

Due to its liposomal nature DC demonstrates a thrombolytic activity: it passes quick through the cell membrane; attaches to the segment of a vascular wall and to the parietal thrombus at the expense of the destabilase lipid components; assists in the slow lysis of a fibrineous clot formed by the destabilase isopeptide activity; and blocks thrombogenesis at the expense of thrombin blockade, of blood plasma kallikrein, and aggregation and adhesion of thrombocytes. Thus, the natural liposome DC provides both preventive anti-thrombolytic and thrombolytic activities.

The resorption of microthrombi in the microcirculatory layer of the epidermis provides normal oxygen supply and nutrition, which significantly slows down the skin aging.

One can assume that the selective hydrolysis of isopeptide bonds by destabilase also protects of cells from aging, because these bonds are the predecessors of the outer desquamation layer of the epidermis.

The liposomal nature of this complex plays an important role in the transport and penetration of biologically active components of the destabilase complex through the epithelium.

Thus, the ability of destabilase to aggregate into micelles and to bind hirudin and kallikrein inhibitors is related to the properties of the destabilase complex and its liposomal strucuture, which is used as an active ingredient of the "Magia" regenerating cosmetic cream.

The choice of an adequate method for the treatment of carcinoma of the vulva depends on the features of the tumor process. Treatment by radiation is the method of choice. Damages to the skin and mucous membrane caused by gamma therapy of the vulva carcinoma are serious and require a search of new effective drugs with local effect.

"Hirudo" gel, which is prepared from biologically active substances (BAS) of the medicinal leech, was used for the prevention and treatment of radiation damage in 17 patients having carcinoma of the vulva. The leech saliva contains immunostimulatory, regenerating, antithrombotic and thrombolytic ingredients.

Gamma therapy was applied in 2 stages. “Hirudo" gel was applied before treatment on the areas of irradiation. The gel was applied a second time during the same night. The patients were also treated with anti-inflammatory drugs as well as with detoxifying and desensitizing medications.

In one case the application of the gel was discontinued due to intolerance. All patients had a small edema on the skin and of the mucous membrane of the vulval labia, at the beginning of the radiation. In 14 cases (82,3 %) there was an erythema and in 12 cases (85,7 %) there was no deterioration. In only 2 cases a wet dermatitis were noted.

The treatment with "Hirudo" gel was continued after the 1st stage of the radiation for 2 weeks, also during the 2nd stage of irradiation. The inflammatory reactions of the skin and mucous membrane were completely cured in all patients.

The present study showed that "Hirudo" gel has anti-inflammatory properties and prevents the development of heavy radiation damage to the skin and mucous membrane.

Recurrent external otitis is one of the most widespread ear diseases in workers of the coal- mining industry, resulting in loss of 10-21 workdays per year. Taking into account the etiology and pathogenesis of the recurrent external otitis, the preference should be given to drugs which have not only antibacterial but also anti-inflammatory, desensitizing and analgesic actions with a minimum of side effects. The drug of choice having the above-mentioned properties is "Hirudon" gel. This contains only natural ingredients, the basic active component being a complex of biologically active substances extracted from the medicinal leech Hirudo medicinalis. "Hirudon" gel decreases blood coagulation, ensures thrombolysis, is anti-inflammatory, improves microcirculation, and increases skin resistance.

We studied the clinical efficacy of "Hirudon" gel on 50 patients with recurrent external otitis (Group I). The gel was applied as follows: during the acute phase of the diseases the gel was applied twice daily in the ear canal and left for 2.5-3.5 hours. The treatment continued for 3-5 days, or until complete recovery. Thereafter, the gel was applied in the ear canal once daily for 30-45 minutes during a month. Concomitantly, 50 patients were treated with conventional drugs (Group II), i. e., antibiotics, analgesics and drugs for allergies, UHF, Linimentum according to Vishnevski, and Rivanol solution.

The efficiency of treatment in each group and the side effects of the drugs were compared according to the indices of patient’s general state (complaints, fever etc.), dynamics of local changes (infiltration and hyperemia of the ear canal), laboratory results of blood and urine analysis, oxygen tension in tissues as measured by a polarographic method, and bacteriological studies of the ear canal skin. The studies were carried out at the beginning of the treatment, on days 3, 5 and 7 of the treatment, and also one-month after the end of the treatment. The patients were also surveyed 6 and 12 months after the end of the treatment.

Improvement was observed in all 50 patients within 2-3 days of treatment with “Hirudon” gel, while in the control group decrease in inflammation began only on days 3-5. In the group of people using the “Hirudon" gel and six months after the end of the treatment no relapses were observed. After 12 months in 2 cases (4 %) a relapse of the external otitis was noted, which was probably due to damage to the skin of the ear canal during ear cleaning. In the control group 17 relapses (34 %) were observed. Three patients had 4 relapses and 6 patients three relapses during a year.

Important parameters, which were examined during the clinical trial included the measurement of oxygen supply to the tissue, the course of oxidizing processes and state of a capillary blood circulation. The oxygen tension in tissues was linearly related to the density of oxygen which was detected with the help of a polarograph (Polarographic analyzator PA-2) The clinical norm of the oxygen density in tissues is 30.0+1.2%. The average value for patients with an external otitis of the ear canal prior to the beginning of the treatment, was 18.6± 0,9%. The use of "Hirudon" gel increased the oxygen density in tissue during the first days of treatment by 24.8± 1.1%, which correlated with the decrease of pain and infiltrations in the ear canal. Patients treated with conventional methods had an increase of oxygen density in their tissue of 24%. The oxygen density in tissues of the ear canal in patients treated with "Hirudon" gel returned to normal 3-5 days after of treatment, while in the control group the patients the density normalized after 4-7 days. The oxygen tension in the skin of patients using the "Hirudon" gel was stable during the entire period of observation and remained at a level of 28.,9± 1.8%.

In 20 patients (12 females and 8 males; 18-78-year old) with chronic leg ulcers of mixed origin, maggot therapy (bios) was used for debridement. The effect of a single application for 3 – 5 days was evaluated by a clinical wound score and contact-free spectroscopy. Debridement was rapid and selective. The wound secretion was temporary increased. We observed a significant improvement of the wound score with a decrease from 12.2±1.9 to 4.0±2.0 (p<0.0). The treatment was well tolerated in all but one patient suffering from pyoderma gangraenosum who needed analgesic treatment. The remittance spectra disclosed an improvement of tissue oxygenation as shown by the characteristic oxygen doublet peak (548nm and 575nm) and an improvement of the edema as shown by reduced remittance between 730 to 1400 nm. Biosurgery is an effective and rapid treatment for chronic wounds with better selectivity and fewer side effects than mechanical debridement.