Method of preparation of the suspension of xenolithic pork spleen for use in the treatment of patients with pathological changes in immune status

 

(57) Abstract:

The invention relates to medicine and can be applied to immune and immunomodulation. Method of preparation of the suspension of xenolithic includes perfusion transverse slices of pork spleen sterile saline solution, centrifugation, removal of the Packed fluid and the dilution of sediment saline solution in a ratio of 1:10. Technical result: suspension xenolithic pork spleen, cooked in this way, when introduced into the patient's body acts as humoral and cellular immunity of the person and reduces the frequency of adverse and allergic reactions in patients on the introduction of xenophobia. 3 Il.

The invention relates to medicine.

In medical practice the different ways of getting xenophobia pork spleen by perfusion of whole body sterile solutions for inclusion of xenophobia in the traditional scheme of treatment of patients with compromised immune status to immune and immunomodulation. A prototype of the selected method proposed by S. D. Nikonov: "Application of the perfusion solution xenoculture in the treatment of patients with extensive Novosibirsk, 1992.). When this methodology is taken at the slaughterhouse slaughterhouse pig spleen in sterile conditions is transported to a medical facility where the catheterization of the splenic artery minor artery and vein tied, then washed the blood and clots from the vascular bed of the spleen by injection with a syringe of saline solution with heparin in an artery of the spleen. As soon as the blood begins to flow through the splenic vein, the last fix and kanyoro. Then proceed to the perfusion of the spleen. For this system for drip infusion is attached to the bottle with saline solution, which is installed in a tripod at a height of 120 cm above the level of the tray with the spleen, and then connected to the blood catheter. During perfusion flowing from the splenic vein perfusion solution collected in sterile vials. For the removal of cells passengers use the techniques settling, or centrifugation. This supernatant under conditions of asepsis and antisepsis is pumped into a sterile container. For cooking xenophobia can be used and other solutions, for example, reopoliglyukin. Technical support and the way the sentence is ykov from staff when catheterization and kanalirovanii vessels.

The technique of obtaining xenophobia allows you to prepare a mixture of soluble biologically active substances of the spleen, however, cellular elements, represented mainly by lymphocytes which play a role in the implementation of immunocorrective and immunomodulating effect, thus artificially removed or destroyed, lyse.

There are other methods: Burdina, Century Manuilov B. M."the Role of factors secreted by the spleen, in the activation of phagocytes." Transplantation and artificial organs, M. M., 1986, S. 48-52. Slobod A. L., Bekchintaev I.e. "A modified technique using xenogenic spleen with purulent-septic diseases. Bulletin of the surgery. I. I. Grekova, 1991, No. 2, S. 96-100. Derived, A. "the Use of extracorporeal xenoplanets and perfusion solution of xenoculture in complex treatment of patients with peritonitis." M3 Grew. Federation. The research Institute of Transplantology and arts. the bodies of the candles. the honey. Sciences. M. 1993. C. 24. Nikonov, S. D. "the Effectiveness of perfusate xenoculture in the treatment of purulent-septic diseases in pulmonary surgery." The Novosibirsk. the honey. in-t, Kida. the honey. of Sciences, Novosibirsk. 1992. C. 22. Sitnikov C. A., Stagnina S. N., Tikhonov centuries, Deryabin, A., "Experimental oblecenie purulent-septic complications in surgery. "Nizhny Novgorod honey. journal 1991. N 2. C. 3-6.

The inventive method of preparation has the task of obtaining a suspension of living xenolithic porcine spleen in high concentration 15 105(ml) a solution of biologically active substances spleen by reducing the amount in the solution of tissue debris and cellular debris, which is achieved by centrifugation of xenophobia and save the bottom portions containing these cellular elements, with tissue debris and cellular debris (like a lighter) will be in the supernatant. Received a suspension of xenolithic when introducing it into the body of the patient will affect both humoral and cellular immunity of the person. Reduced the amount of tissue debris and cellular debris in the solution will reduce the number of adverse and allergic reactions of the patient to the introduction of suspended Xenocrates.

In Fig. 1 shows a system for otemanu crushed spleen (schematically shown assembled system to launder the spleen, showing the main components, the fluid flow in the system during operation;

in Fig.2 is a system for obtaining xenophobia ( schematically shown a system for receiving Xeno and perfusion of the spleen);

in Fig. 3 - remove the supernatant liquid from the vial containing the centrifuged xenophontos ( schematically indicated the correct location of the devices for removing the supernatant liquid from the vial xenophontos after centrifugation).

The proposed method is as follows.

The first step is to fence pork spleen, performed at the factory in the slaughterhouse to the lines. The fence is held by a special team under conditions of sterility. Extracted spleen in a sterile container (thermos) is transported to a medical facility, where it will be prepared suspension xenolithic porcine spleen. The transport duration should not exceed one and a half hours after casting. Preparation of spleen is carried out in a specially designed space (clean dressing, procedural, Department of detoxification, etc.,) in sterile conditions. Sterile tray spleen crushed scissors or a scalpel. The thickness of the pieces should not exceed 1 cm, weight - 2-3 g, the spleen is cut transversely to the longitudinal axis. Crushed spleen is placed in sterilized glass jars tsya sterile solution (for this you can use isotonic NaCl solution, latosol, disol), shake vigorously, the solution is poured.

This procedure is repeated twice to remove blood and tissue detritus. Then cooked the spleen is filled with a solution of the antibiotic with the aim of eliminating trapped in the container when cooking and the fence spleen microorganisms. You can use metronidazole (100 ml per 1 jar) or cephalosporins (average dose of 1 g per 1 Bank). The exposure time of the spleen with antibiotics from 4 to 24 hours, depending on what time you will need suspension xenolithic porcine spleen. At the time of exposure to antibiotic spleen is placed in the refrigerator storage temperature 0 - -4oC.

Suspension xenolithic pork spleen is prepared immediately before its introduction to the patient. This requires a roller pump 1, a slit-like nozzle 2, one dropper (preferably domestic production)3, the system for blood transfusion 4, 30-cm rubber tube 5 with an inner diameter of 0.6-0.8 cm, sterile solutions (physiological NaCl, 4 bottles of 400 ml) 6. The initial stage of preparation of the suspension of xenolithic pork spleen is getting xenopho, going system for hemosorption, where the capacity of the sorbent replaced the bottle with crushed spleen (see Fig. 1) and is laundering column of sterile saline. Pure saline when this system is connected to the blood collection and roller pump is forced through the column at a speed of 60 - 80 ml/min Volume of the leaching solution is on average 1200 ml. then to 400-ml bottle with sterile physiological saline, is connected to the system for blood transfusion ("fence"), connects the drip, which returns the liquid which has passed through a column of crushed spleen ("return"). Thus the roller pump will be "chasing" the saline solution in a closed loop (see Fig. 2). Xenophobic for the subsequent preparation of the suspension of xenolithic pork spleen is obtained by circulating the saline solution with a speed of 40 - 60 ml/min in a closed loop for 15-20 minutes At the end of this time the saline solution in the vial becomes light yellow color becomes slightly cloudy. The concentration of cells in it an average of 3.5 to 5.0 105in ml, the cell viability 70 - 79%. Cells that are in the solution of biologically active weselna presents lymphocytes - about 95% of all cells) is the so - called xenophobic.

The next stage bottle with xenophontos centrifuged in a centrifuge at a speed of 1200 revolutions per minute for 10 minutes. Then the vial is removed from a glass centrifuge, is opened and the supernatant is aspirated by a thin wall with a sterile glass pipette (8) attached to the vacuum-suction (see Fig. 3). On bottom of the vial should be about 3 milliliters of sedimentary fluid. Add 20-30 ml of sterile saline, the contents of the vial gently shaken, stirred. The obtained liquid volume presents dissolved biologically active substances spleen (cytokines are soluble factors of the immune cells, interleukins 1 and 2, interferon) and suspended in them xenolithic porcine spleen (mainly lymphocytes about 95% of all cells). The concentration of cells and their viability in the percentage of dead cells determined immediately after preparation of the suspension and is according to our data 15 105and 79% and 85% respectively. Obtained by this method of suspension xenolithic pork spleen with the introduction of it in the human body effect on humoral immunity is deystviy immunocompetent cells of the porcine and human organisms - cognate interaction) implementing fully its immunomodulating and immunostimulating effect, which allows its use in medicine.

Method of preparation of the suspension of xenolithic pork spleen for use in the treatment of patients with pathological changes in immune status in which perfusion of porcine spleen sterile saline solution, characterized in that prepared by perfusion of transverse slices of fresh pork spleen sterile solutions xenophobic centrifuged, the supernatant removed, the residue diluted with sterile saline solution in a ratio of 1 : 10.

 

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