本文選題：VX2腫瘤細胞　切入點：動物模型　出處：《承德醫(yī)學院》2017年碩士論文

【摘要】：目的:通過實驗設立兔VX2骨轉(zhuǎn)移瘤動物模型,觀察確定適合穿刺裝置穿刺活檢的時間,明確適宜的穿刺時間后,通過PCNA免疫組化技術(shù),驗證自制改良穿刺裝置的效果。方法:實驗所應用的動物為人工養(yǎng)殖的家兔,第一部分所用家兔數(shù)量為25只,第二部分35只,實驗所用的各種設備來自于承德醫(yī)學院附屬醫(yī)院實驗室,所用的腫瘤細胞以及試劑采購于試劑公司。1、第一部分為制造一種適合本實驗的模型,觀察腫瘤模型隨著時間的推移而進展的各期變化,大體明確腫瘤生長需要的時間;采購VX2腫瘤細胞,植入家兔后側(cè)肢體的脛骨近端,通過將腫瘤細胞懸液置入家兔脛骨骨孔內(nèi)的方法建立模型;隨機抽取5只為對照組,其余20只以雙側(cè)后肢為實驗肢體,左側(cè)肢體采用腫瘤組織懸液植入法建立兔VX2骨轉(zhuǎn)移瘤模型(左側(cè)組),右側(cè)肢體植入生理鹽水(右側(cè)組),分別于種植后每周通過影像學及病理學檢查觀察腫瘤生長情況,進而明確VX2腫瘤進展各期大體時間。2、第二部分為驗證自制骨腫瘤穿刺裝置效果的實驗,在已成熟的制造家兔VX2脛骨骨腫瘤模型以及明確髓腔內(nèi)瘤體成型但尚未突破骨皮質(zhì)這一階段的時間基礎上,通過PCNA免疫組化技術(shù)來驗證該裝置的效果;選購35只實驗兔,按上述方法制造家兔VX2脛骨骨腫瘤模型,雙側(cè)后肢均植入腫瘤組織,隨機選出5只,作為對照組,其余30只分為左側(cè)組及右側(cè)組,左側(cè)組應用自制改良穿刺針于皮質(zhì)骨上打孔后封堵,右側(cè)組應用傳統(tǒng)的穿刺針進行打孔(此時穿刺針于皮質(zhì)上打孔的位置應與造模時植入腫瘤組織的孔相對,避免造模時形成的孔道影響實驗效果),兩組穿刺術(shù)完成后仔細縫合,同樣層數(shù)無菌敷料包扎(3層),術(shù)后于實驗后次日觀察兩組實驗動物手術(shù)部位滲出情況,進而明確兩組實驗動物術(shù)后手術(shù)部位出血情況有無差異;將兩組實驗動物于穿刺術(shù)完成后第10天處死,獲取穿刺針遺留孔周圍的軟組織,通過PCNA免疫組化技術(shù)查看穿刺針遺留的孔周圍有無腫瘤細胞擴散,對比兩種裝置的效果。結(jié)果:1、第一部分制造成功家兔VX2脛骨骨腫瘤模型,通過觀察影像學及病理學檢查,確定植入腫瘤組織后第2周末,即第14天可見髓腔內(nèi)成型瘤體,并且未侵犯皮質(zhì),左側(cè)組植入腫瘤細胞后1~2周腫瘤組織于髓內(nèi)生長,第3周開始腫瘤組織髓內(nèi)生長并侵犯部分骨皮質(zhì),第4周末腫瘤組織突破骨皮質(zhì)侵犯周圍軟組織。2、穿刺術(shù)實施后的次日見30只實驗動物中,左側(cè)組手術(shù)部位敷料可見血性滲出者1只,余下者敷料均干燥無滲出;右側(cè)組手術(shù)部位有25只可見大量血性滲出,2只少量血性滲出,3只敷料干燥無滲出;第10天處死所有實驗動物,獲取穿刺針所形成的孔道周圍軟組織行PCNA免疫組化技術(shù)處理,左側(cè)組即應用自制改良穿刺針組穿刺孔道周圍軟組織無腫瘤細胞侵及,而應用傳統(tǒng)穿刺針組穿刺孔周圍可見腫瘤細胞侵及。結(jié)論:成功地建立了一種兔VX2骨腫瘤模型,此模型可很好的應用于骨腫瘤方面各類研究的動物實驗中,利用自制的穿刺裝置實施穿刺術(shù),降低了穿刺術(shù)實施后各項并發(fā)癥的發(fā)生率,證明自制改良穿刺裝置可以有效地應用于腫瘤組織的穿刺活檢,是一種新型的有效的裝置和技術(shù)。這個裝置的特點為組合式的各部器械,通過穿刺針穿過骨皮質(zhì)后,利用裝置外側(cè)的套管來阻斷骨皮質(zhì)與骨髓內(nèi)的腫瘤細胞,同時使髓腔與外界形成通道,在我們獲取需要的足夠組織之后,通過套管將裝置的關鍵物即封閉栓置入穿刺孔內(nèi),進而避免了骨髓腔內(nèi)的腫瘤細胞通過穿刺孔擴散,并且此方法創(chuàng)傷較傳統(tǒng)穿刺針無增大,但對于穿刺后各項并發(fā)癥的預防卻有了顯著地提高。該裝置可在病理檢查未明確之前,盡量減少腫瘤組織的擴散,同時可減少穿刺部位的出血等并發(fā)癥發(fā)生,降低血腫形成的幾率,對于臨床工作有很大的意義。
[Abstract]:Objective: to set up animal model of rabbit VX2 bone metastases, observe the biopsy puncture device to determine the appropriate time, clear suitable time of puncture, by PCNA immunohistochemical technique, test the effect of modified puncture device. Methods: the application of the animal for artificial breeding of rabbits, the first part of the number of rabbits for the 25, second part 35, all kinds of equipment used in the experiment from the laboratory of Affiliated Hospital of Chengde Medical College, the tumor cells and reagent to the reagent company.1, the first part is to make a suitable experimental model, to observe the tumor model with the passage of time and the progress of each stage of change, generally clear tumor growth needs the time of purchase; VX2 tumor cells implanted in the posterior limb of the proximal tibia, the tumor cell suspension liquid into rabbit tibia bone hole is built with the model; The machine selected 5 rats as the control group, the remaining 20 only to the hind limbs of experimental limb, left limb tumor metastasis by established rabbit model of VX2 bone suspension implantation (left group), the right limb was filled with saline (right), respectively, after planting in a week by imaging and pathological examination to observe the growth situation the tumor, and then clear the approximate time of tumor progression of VX2.2, the second part is to verify the self-made bone tumor puncture device experiment in time based manufacturing VX2 has mature rabbit tibial bone tumor model and clear intramedullary tumor forming but not breaking the cortical bone in this stage, by PCNA immunohistochemical technique to verify the efficacy of the device; choose 35 rabbits, rabbit VX2 bone tumor model made by the above method, the hind limbs were implanted into the tumor tissues, 5 rats were randomly chosen as the control group, the remaining 30 were divided into the left The left and right group group group using self-made puncture needle punched in cortical bone after the closure, the needle right by applying the traditional drilling (the position of the puncture needle from the cortical perforating and when modeling implanted tumor tissue relative to avoid the formation of holes, when modeling the effects of pore effect), puncture in two groups after careful suture, the same number of sterile dressing (3), after the operation on the day after the experimental observation of the two groups of experimental animal surgical site exudation, and then clear the surgery site bleeding in two groups of experimental animal postoperative difference; two groups of experimental animal to puncture tenth days after death, get the needle from the soft tissue around the hole, by PCNA immunohistochemical techniques around the needle hole is left to see without the spread of tumor cells, comparing the two kinds of device. Results: 1, the first part of the manufacture of successful VX2 rabbit tibia Bone tumor model by observing the imaging and pathological examination, determine the tumor tissue second weeks after implantation, fourteenth days intramedullary tumor forming visible, and not involving the cortex, the left group at 1~2 weeks after implantation of tumor cells in tumor tissue growth in intramedullary tumor tissue third weeks intramedullary bone growth and invasion at the end of the fourth breakthrough cortex, tumor invasion of cortical bone soft tissue around.2, puncture time is after the implementation of the 30 experimental animal, dressing group left surgical site visible blood exudation in 1 cases, the rest of them were dressing dry without exudation; right surgical site 25 showed a large amount of blood exudation, 2 is only a small amount of blood 3 dry dressing exudation exudation; tenth days all the experimental animal, get the puncture needle formed around the channels of soft tissue by PCNA immunohistochemical technique, the left group application of modified puncture needle puncture group No way of soft tissue around the tumor cells invasion, and the application of traditional needle puncture holes around the tumor cells invasion. Conclusion: a rabbit VX2 bone tumor model was successfully established, and this model can be well applied in animal experiment of bone tumor research, the implementation of self puncture puncture device system and reduce the complications of puncture after the implementation of the incidence of biopsy proved that modified puncture device can be effectively applied to the tumor tissue, is a new and effective device and technology. The utility model is a combination of the device through the puncture needle through the cortical bone, bone block bone marrow tumor cells in the cortex and the outer sleeve device, the medullary cavity and the outside channel formed after we get enough tissue to, through the casing will be key device that closed bolt placement wear Puncture, and avoid the bone marrow tumor cells through the puncture hole diffusion, and this method is better than the traditional wound puncture needle does not increase, but for the prevention of complication has been significantly improved. The device can be before pathological examination is not clear, try to reduce the spread of tumor tissue, and can reduce hemorrhage the occurrence of complications such as puncture, reduce the risk of hematoma formation, has a great significance for clinical work.

【學位授予單位】：承德醫(yī)學院
【學位級別】：碩士
【學位授予年份】：2017
【分類號】：R-332;R738

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