| Clinical glioma is the most common tumor of the brain, and its high proportion of brain tumor (44.69),about 77% of the malignant glioma patients died within 1 year after diagnosis. For glioma treatment, the majority of scholars believe that the present should be used to surgery, chemotherapy,radiation therapy and biological composition of the combined therapy. Although glioma treatment has been achieved certain effects, but the recurrence rate is still relatively high mortality, poor prognosis is still very difficult neurosurgical problems. The reasons for these: 1. Showed invasive growth, it is difficult to find obvious borders in conventional surgery, there is a high recurrence.2.The majority of glioma cells on the two radiotherapy, chemotherapy-sensitivity is not high;3.Poor specificity of immune therapy. Clearly, the treatment of glioma need to continue efforts to explore and develop a new way.Cryotherapy is the use of partially ultra-low-temperature or low-temperature which cause cells mechanical damage, or metabolism injury to achieve the metabolic lesions removed a method of treatment. Frozen brain surgery in 1961 is a milestone, the first time Cooper and Lee frozen and stereotactic techniques for thalamotomy for the treatment of Parkinson's disease. 1962 for the basal ganglia surgery, and after resection of the brain used for benign and malignant tumors. 1982 Feasdale report that using stereotactic technology frozen pituitary tumor resection to control and improve the Acromegaly. Cryotherapy have brought the hope for tumors refractory, but because of traditional treatment methods are frozen with liquid nitrogen, not because of better control of its cooling rate, it is difficult to protect normal tissue and prone to complications such factors limiting the therapy applications. With the continuous development of refrigeration technology and innovation, the United States Endocare company successfully developed a new type of ultra-low-temperature helium intervention cold argon minimally invasive targeted ablation therapy (Endocare CryocareTMSurgical System)which overcome the above drawbacks. In 1998 Endocare CryocareTMSurgicaI System got through the United States FDA approval by the IEC, EMC CE certification and the European Union, will soon be used in hospitals around the world. It is the invention of refrigeration technology for the development of the latest achievements, not only inherit and develop the ultra-low-temperature treatment of the basic and clinical research results, but also introduced the new concept of refrigeration and new technologies. The system of combining the aerospace, bio-sensor, a computer-based monitoring and appropriate treatment targeting a number of technical, super low-temperature refrigeration and intervention targeting organic combination of hyperthermia. Comparing with the traditional method of comparison frozen in liquid nitrogen, argon helium refrigeration have unique advantages: (1) Injury in patients is limit, no requisition of surgery, less bleeding (2) Good success rate and low complication rate, (3) Normal organ tissue cells without toxicity, the patient recovered quickly (4) Minor surgical damage, and can be reused repeatedly;(5) Effective, easy to operate, easy for patients to accept, (6) Being implemented individually, but also implemented with chemotherapy, radiotherapy or Surgical Therapy, (7) Being used by other therapy or no treatment in patients with treatment failure. The emergence of the system bring a breakthrough of the development for ultra-low-temperature technology which will become a new and effective method, in the treatment of tumors.Through rapid freezing by high-pressure argon gas and thawing by high-pressure helium gas, Endocare Cryocare?Surgical System can destroy the tumor cells for purposes. The mechanisms include (1) -5°C, extracellular fluid first to be frozen for ice crystals to extracellular solute concentration, and water in cells educing and cells dehydration, high intracellular solute concentration, resulting in electrolyte concentration , pH, and dynamic change of the enzyme system, and their metabolic system damage; (2) When the temperature to below -15℃, the formation of ice crystals in the cells, stromal cells integration into internal and external blocks, damaged cells and Subcellular structure of macromolecules This is the main frozen injury mechanism, (3) high-pressure helium in the rapid wanning, in the -20℃- 40℃, the expansion of the intracellular ice crystals thus undermin the cell membrane structure to break it down and induce the death of the injury (4) After refrigeration, capillary endothelium cell damage,then thrombokinesis induce thrombokinesis which is the main reason of the cellular necrosis.(5) experiment liver cancer has confirmed, the frozen remnants of necrotic tumor cells can form specific antigen, and induce the production of specific tissues and organs and species-specific antibodies, that is, ultra-low-temperature-induced immune response, enhancing the body's immune function of the tumor.In clinical after frozen,the judgement of the frozen tissue necrosis scope is mainly dependent on imaging of the information provided. In the clinical treatment ,some doctors discovered that treatment is to be ineffective, prone to residual or recurrent in the edge of the ice hockey, which is one reason why hockey edge in the course of treatment no more than marginal tumor target. This is because, according to our medical imaging of the information provided to the clear conclusion that hockey formed within the scope of the death of tumor cells completely, ice hockey will be a marginal part of the residual tumor; the brain tissue, because of its function of the very fine , a little too much damage, it may bring about permanent irreversible loss of function, or even death. After the cryoablation, After refrigeration, the research about the frozen zone and the marginal organizations pathological changes to clear whether refrigeration scope of ice balls is agree with the scope of the rhagiocrine cell have a very important clinical significance. There are many factors for freezing and warming, thus resulting histopathological changes also affected by many factors. For a complete process Cryocare cryosurgery, the following five parameters are essential for: freezing temperature, the rate frozen, chilled, rewarming rate and the number of freeze-thaw cycles. Frozen different time and temperature, different frozen and rewarming rate, the different cycles of freezing temperature, the different organs histopathological changes have some differences. Cryocare frozen in the course of treatment, with different pathological changes in the relationship between the treatment and control parameters need regular in-depth experimental study. There were related research reports about Endocare CryocareTMSurgical System used in the liver, lung and prostate and other organs. In the treatment of different parameters, whether brain tissue compare with liver and other organizations in the organization changes going to be different. In this regard, there are little corresponding reports about Endocare CryocareTM Surgical used in the brain. Therefore, this study is intended to normal dogs for the study of the brain, with different diameter superconducting knife Cryocare frozen different time, optical microscopy and electron microscopy of histopathological changes were observed, clear cell the size of the area of necrosis, for the clinical treatment of brain tumor treatment parameters for reference. As follows:Chapter 1 Research of Endocare Cryocare?Surgical System for the treatment of canine brain frozen superconducting knife diameter of the variables and time-relatedObjective: To investigate the targeted therapy system Cryocare (Cryocare knife) in the superconducting knife (that is frozen probe) diameter and the time variable, frozen canine brain necrosis of the histopathological and ultrastructural changes.Methods: Randomly divided into three gpoups :the normal control group superconducting knife Group 2 mm in diameter and 3 mm group. 3% pentobarbital satisfied by 1 ml / kg by intraperitoneal injection of anesthesia animals,cut head hair, from prone position, local skin disinfectant iodine activity in the brain midline right next to a 1.5 cm, were cut before the beginning of the word Gap Skin exposed skull, a circular bone window, open dura mate encephal, to 2 mm, 3mm frozen probe in diameter perpendicular to the surface of the brain into the right frontal lobe of about 1.5 cm, respectively, using high-pressure argon quick frozen for 3 min, 5min, and then High-pressure helium rewarming 1 min; repeat. 48 h after reperfusion, infuse and get brain tissue, observe histopathological changes in the light and electron microscopy measuring diameter of the area of necrosis frozen.Results:1. Animals not died. In cryosurgery process, the local ice hockey surface in the brain are round, with the surrounding normal brain tissue boundaries clear. When dissecting, the boundaries between the necrotic area and the surrounding normal tissue are clear.2. Optical microscopy: clearly distinguish between normal and frozen zones frozen zone sector directly transform to normal zone, no fiber scar formation between them. Frozen uniformity of the central area was coagulation necrosis. Externa the central Region field is a inflammatory response, there are a number of cell debris and a lot of red, leukocyte infiltration, there is a hemorrhage—edema zone next to the zone of inflammatory reaction, a large number of the red cells in microvessels, platelets and set thrombosis, and that focal hemorrhage. Electron microscopic observation: the frozen area, marginal zone? All neuronal cell death and found no survival of the cell structure, no microvascular endothelial cells. This shows that some of the marginal apoptosis of cells Structure: nuclear pyknosis, nuclear fragmentation and melting of the chromatin condensed into coarse granular, some side-chromatin, the nuclear membrane swelling Obviously, part of the nuclear membrane rupture; intracellular organelles (within endoplasmic reticulum, mitochondria) swelling, and some broken disintegration; microvascular endothelial cells in the marginal damage, blood stagnation and microvascular thrombosis.3. the average diameter of necrosis in 3mm group is more than of 2 mm group, there were significant differences, p <0.05; both group 2 mm, 3 mm group, formed by frozen 5 min of the necrotic area are more than 3 min, there were significant differences, p <0.05 .Conclusion: take 2 freeze-thaw cycles with argon treatment of helium refrigeration system on canine brain tissue, frozen 3 min, 5min, causing frozen and eventually all of the marginal cell necrosis, cell death is on the verge of frozen edge. Diameter of 2 mm, 3mm Cryocare superconducting average diameter of a knife were 21.0 mm and 30.0 mm of spherical frozen necrotic area.
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