Method of treating malignant lung neoplasms in experiment

FIELD: medicine.

SUBSTANCE: invention relates to medicine, oncology, and can be used for treatment of malignant lung neoplasms in experiment. For this purpose performed is extracorporal irradiation of autoblood with red incoherent light λ=0.67 mcm dose W=3.06 J/cm2 in continuous mode with exposition duration to three minutes. After that 15-20 minutes later into said blood added is cyclophosphane in dose 40 mg/kg and the mixture is incubated for 40 minutes at T=37°C. After that it is re-infused into subclavian vein of the animal.

EFFECT: claimed method allows to reduce terms of anti-tumour treatment, increase efficiency of cyclophosphane action, reduce its toxic manifestations, increase animals' life term, increase non-specific anti-tumour resistance, activate anti stressor mechanisms.

2 tbl

 

The invention relates to medicine, namely to Oncology, and can be used in the experiment in the complex treatment of malignant tumors, transplantable tumors in the lungs by applying autohemotherapy with extracorporeal irradiation of blood red.

Biological fluids, as a complex multicomponent systems and having the properties of living matter, react structural alteration of a substance, even at very low external physical impact. The presence in the blood formed elements (erythrocytes, leukocytes, platelets, etc.) significantly increases the susceptibility and sensitivity of liquid media of the organism to the external influence of various physical factors, including light radiation.

It is known that electromagnetic waves in the optical range, including the red spectrum can manifest mechanisms aimed at the stimulation of natural killer cells tumor is able to activate various systems antitumor protection, to block proliferation and induce apoptosis of tumor cells (Sheiko E.A., DR. Zlatnik, Sakura GI Monochromatic radiation in the red spectrum as a factor stimulating the natural mechanisms of tumor cell death in vitro // Laser medicine, 2008. - vol.12, No. 1. - P.15-18). It was shown that the COI is whether the radiation is monochromatic red light with λ=0.67 microns, filed in continuous mode with a dose of exposure W=0,05 j/cm2for stimulation of lymphocytes from lung cancer patients, and investigated their influence on the culture of tumor cells C. In the cytotoxic test revealed an increase in the percentage of dead tumor target cells, in particular, by apoptosis, under the action of monochromatic LED on lymphocytes, as well as increased functional activity of T-lymphocytes and natural killer cells involved in the implementation of anti-tumor activity. However, in this work it was studied the effect of incoherent red light radiation in the conditions of the whole organism with transplantable tumor and chemotherapy. So the question remains relevant to developing methods to optimize the effectiveness of the combined anticancer treatment effects of light, not causing stimulation of the growth of the tumor itself.

There is a method of transcutaneous irradiation of blood (Sheiko E.A., salanova A.I., Zhukov, GV and other Patent No. 22811795 from 20.06.2006), taken as a prototype. However, this method has some significant limitations and drawbacks. The method was developed for transplantable under the back skin tumors Plissa. When implementing this method is used the impact of led red light with λ=0.67 microns with a maximum radiated power of 48 mW and Maxim is enoy the power flux-density (PPM)=7.5 ml W/cm 2in continuous mode percutaneously in the femoral vein. The duration of the procedure up to three minutes. Just spent 8 treatments every morning in the same hours. The disadvantages of the method include weakly pronounced antitumor efficacy.

The aim of the invention is to achieve an anti-tumor effect by increasing the effectiveness of anticancer drugs, weakening their damaging effect, prevent the development of severe intoxication.

This goal is achieved by the fact that the model ectopiceski growing tumor in the lungs of rats on the third week after inoculation was carried out antitumor effects. In all animals, blood was collected from the subclavian vein into tubes in quantities of 0.3 ml, was added the same amount of pulizia, the blood was irradiated with led light (DNS), then after 15-20 min was added cyclophosphamide at a dose of 40 mg/kg and incubated the mixture for 40 min at T=37°C, then repusively in the subclavian vein. Antitumor therapy was performed twice with an interval of seven days. DNS exposure was carried out with the help of led red light with λ=0.67 microns, obtained from the machine "Spectrum LC", dose W=3,06 j/cm2in continuous mode.

The invention is a Method of treatment of malignant neoplasms of the lung in the experiment is new, the AK not known as such a method for improving the treatment of lung tumors in the experiment with drugs, put on autologous blood, in combination with in vitro exposure LED red range of the spectrum.

Comparative analysis of the claimed solution with the prototype shows that the claimed method differs from the known fact that in the experiment on animals with lung tumor carried out on the third week after inoculation antitumor effects by sampling from the subclavian vein blood in a test tube with an equal amount of pulizia in the amount of 0.3 ml and its radiation in vitro DNS with λ=0.67 microns, obtained from the machine "Spectrum LC", dose W=3,06 j/cm2in continuous mode, then after 15-20 min in the blood has added cyclophosphamide at a dose of 40 mg/kg and incubated the mixture for 40 min at T=37°C, then repusively animal in the subclavian vein. Antitumor therapy was performed twice with an interval of seven days.

Thus, the claimed method meets the criteria of the invention of "Novelty."

The invention is a Method of treatment of malignant neoplasms of the lung in the experiment is industrially applicable, as it can be used in healthcare, scientific research institutes, experimental and clinical research opportunities DNS in complex anticancer treatment.

The invention "Method of treatment of malignant novobrdo the deposits of light in the experiment is as follows:

To obtain the led radiation (DNS) red light was used for the laser-led physiotherapeutic apparatus "Range-LC", the purpose of which is the implementation and selective laser led photogrammetrie (FHT). The impact of DNS was performed using the led red light with λ=0.67 microns, dose W=3,06 j/cm2in continuous mode, in animals with ectopic growth of transplantable tumors sarcoma 45 in the lungs at the third week after inoculation, extracorporeal blood obtained from the subclavian vein in tubes with an equal number of pulizia in the amount of 0.3 ml, then after 15-20 min in the blood has added cyclophosphamide at a dose of 40 mg/kg and incubated the mixture for 40 min at T=37°C, then repusively animal in the subclavian vein. Antitumor therapy was performed twice with an interval of seven days. Experiments carried out on outbred albino male rats weighing 250-300 g

The first group of animals blood in vitro were irradiated DNS, 15-20 minutes later incubated at T=37°C for 40 minutes, then was repusively in the subclavian vein. The second group of animals blood in vitro were irradiated DNS, then after 15-20 min was added cyclophosphamide at a dose of 40 mg/kg and incubated for 40 min at T=37°C, then repusively in the subclavian vein. Antitumor therapy was performed twice with an interval of seven days. DNS per the op perate were carried out using the led red light with λ=0.67 microns, received from the machine "Spectrum LC", dose W=3,06 j/cm2. Radiation was applied in a continuous mode. The third group served as a control. On the sixth week after inoculation, animals were scored by decapetala, the animals are left to track the time of their lives.

The results of histological studies have shown that microscopic examination of lung tissue of the control group was detected total lung tumor. In the Central part of the lung was observed diffuse growth of the tumor with foci of necrosis, on the periphery of the lung was determined by a separate small tumor foci. Tumor invasion was observed in mealeasy partitions, the main respiratory structures, including the bronchioles. Tumor cells were presented fusiform, oval and round shape with different shapes of pathological mitoses. In some areas showed hyperplasia of the connective tissue, forming the multi-layer of fibroblasts and histiocytes.

In animals after exposure to DNS during slaughter macroscopically in the lungs were isolated field hemorrhage was determined signs of perifocal inflammation, a major tumor sites were not found. Microscopic analysis of lung tissue was determined individual modified tumor cells, losing faith is tobrasol form. In tumor cells were observed degenerative changes, manifested in the presence of large vacuoles in the cytoplasm, nuclei - hypochromia or randomly generated chromatin network in the nucleus of some cells was determined karyorhexis, karyolysis and other necrotic changes.

Under a combination of DNS and the introduction of cytostatic agents on autologous blood lungs of rats did not contain large and small nodules. There were signs of interstitial and intraalveolar edema. Microscopic analysis of the tumor cells was not determined. However, in lung tissue was attended by a large number of cells and detached polymorphic bare nuclei, which are located in areas of proliferation of young connective tissue.

Table 1 presents some of the indicators in animals from the control and experimental groups obtained during the slaughter of animals. The thymus weight was significantly higher in control group than in the experimental groups, while the weight of the spleen was significantly higher in the second group than in other groups. The odds ratio of the weight of the thymus and adrenal glands, indirectly reflecting the level of balance of the two major subsystems of the organism, the immune and endocrine, was not significantly different from each other. Bleeding in the gastrointestinal tract were zafiksirovannyyo in control. Leukocyte index of intoxication (LII) Calif-Khalifa can be seen as one of the integral indicators of the state of immunity of the body containing the tumor. LEAH represents the ratio of the percentage of the sum of the cells of the neutrophilic series to the sum of the percentage of remaining cells (Ostrovsky VK, Kishina L.A., Plaksina NV and others, 2005). Normal indicators LII $ 1 to $ 3 (Ostrovsky VK, Alimov P.P., Mashchenko A.V. and others, 2003). Index of intoxication in the first and third control group had low values to units, but in the first group, after exposure to the Institute values LEAH exceeded the control by a factor of 1.2. Index of intoxication second group was close to the upper limit of normal and exceeded LEAH first group of 15 times, and the third group 17 times (respectively). Thus, the values of LEAH in the first and third group confirmed the presence in animals of different severity of inflammatory and destructive processes, and the reduction of antitumor immunity, while in the second group on indicators LEAH were high, suggesting a high functional capacity of the neutrophilic series. These data agree well with the figures of lysosomal test for the determination of cationic proteins (KB) in granulocytes is Ravi. The values of KB are equally high in the first and second group and exceed the value of KB control 4.5 times. It is known that the enhancement of nonspecific antitumor resistance is accompanied by the mobilization of the cellular link natural antitumor resistance of the body, manifested in the increase in the number and quality of neutrophils with higher level content indicators KB (Sheiko E.A., 2005), therefore we can conclude that with the help of DNS in the red spectrum, you can have a similar impact on the level of nonspecific antitumor resistance to increase.

Table 1
Some of the indicators obtained during the slaughter of the animals from the control and experimental groups (M±M)
Indexgroup 1 n=6group 2 n=6group 3 n=5
The presence of tumor in the lungsNot foundNot found
The thymus weight per 100 g of animal mass, mg60,2±2,1347,1±3,4 376,8±2,31,2
The adrenal mass in 100 g of animal mg14,6±2,714,9±2,316,5±1,3
The ratio of the mass of the thymus to the mass of the adrenal gland, cu4,1±2,43,2±2,24,7±2,4
The spleen weight per 100 g weight of the animal, mg548,9±212,3911,6±241,3691,1±202
Bleeding in the stomach and intestinesPartiallyNot found
LEAH $ 0,21±0,423,15±0,11,30,18±0,022
Indicators of cationic proteins of neutrophils in the blood1,74±0,131,59±0,130,38±0,051,2
Note. 1 - significantly compared to group 1 (P<0,05), 2 - significantly compared to group 2 (P<0,05), 2 - reliably on Rel is the solution to group 3 (P< 0,05)

Table 2 shows the timing of life left after the experience of the animals. As can be seen from table 2, the animals of the second group lived in 1.6 times, and the first group 1.4 times longer than control (sootvetstvenno).

Table 2
The lifespan of animals (M±m)
Indexgroup 1 n=7group 2 n=7group 3 n=6
The life span of rats, d101±3,53110±1,5370±31,2
Attitude control, %144,3157,1-
Note. 1 - significantly compared to group 1 (P<0,05), 2 - significantly compared to group 2 (P<0,05), 3 - significantly compared to group 3.

Thus, on the basis of the obtained experimental data we can conclude that monochromatic incoherent red light in the studied regime can affect immunocompetent blood cells, help the mother to improve the antitumor efficacy of cyclophosphamide, put on autologous blood, to increase the duration of the lifetime of these animals to increase their own nonspecific antitumor resistance of the organism.

Technical and economic efficiency "method of treatment of malignant neoplasms of the lung in the experiment is to reduce the terms of cancer treatment, effectiveness of the actions of cyclophosphamide, reducing its toxic effects, the increase in terms of animal life, a more pronounced increase in nonspecific antitumor resistance, activation of anti-stress mechanisms.

The method of treatment of malignant neoplasms of the lung in the experiment, including chemotherapeutic treatment and light exposure, characterized in that conduct in vitro irradiation of autologous blood red incoherent light λ=0.67 microns dose W=3,06 j/cm2in continuous mode, the exposure time up to three minutes, then after 15-20 min in the blood add cyclophosphamide at a dose of 40 mg/kg and incubated the mixture for 40 min at T=37°C, and then is re-infused animal in the subclavian vein.



 

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