本文關(guān)鍵詞： 法醫(yī)病理學(xué) 電擊死 骨骼肌 MuRF1　出處：《重慶醫(yī)科大學(xué)》2013年碩士論文　論文類型：學(xué)位論文

【摘要】：目的：電擊死和他殺偽裝電擊是法醫(yī)學(xué)實(shí)踐中常見的案件類型之一,但實(shí)際檢案過程中,法醫(yī)工作者對(duì)其診斷仍沒有統(tǒng)一的標(biāo)準(zhǔn),尤其是對(duì)無電流斑情況的死因推斷或特殊情況的水中電擊。許多學(xué)者從電擊死后各個(gè)組織器官的形態(tài)學(xué)及分子水平進(jìn)行了研究。當(dāng)電擊案件中無電流斑形成時(shí),組織學(xué)改變不明顯時(shí),電擊死后根據(jù)骨骼肌的變化推斷早期死亡時(shí)間鮮有報(bào)告。我們?cè)O(shè)計(jì)了以下實(shí)驗(yàn)檢測(cè)通電肢體骨骼肌MuRF1的表達(dá)情況,以期鑒別生前電擊死和死后電擊、生前電擊死和死后電擊早期死亡時(shí)間的推斷、電損傷對(duì)不同骨骼肌類型的影響。 方法：60只健康的Wistar大鼠,分成電擊死組、死后電擊組、空白對(duì)照組,每組20只。電擊死組大鼠用220v交流電的兩極分別連接右前肢與左后肢,通電90s致死,于電擊死后0h、1h、3h、6h取材；死后電擊組大鼠采用頸椎脫臼法處死后,分別于0h、1h、3h、6h電擊90s取材；空白對(duì)照組大鼠直接頸椎脫臼法處死,不進(jìn)行電擊,于死后0h、1h、3h、6h取材。取左后肢腓腸肌、脛骨前肌,進(jìn)行HE染色、免疫組化染色和實(shí)時(shí)熒光定量PCR處理。 結(jié)果：1、HE染色結(jié)果 電擊死組：可見骨骼肌細(xì)胞呈空泡樣改變、灶狀變性、壞死；部分細(xì)胞核極化、部份骨骼肌細(xì)胞斷裂、波浪樣改變,部分骨骼肌橫紋不清；胞漿凝聚、均質(zhì)化、嗜酸性變；胞核固縮、深染、扭曲,部分核伸長；骨骼肌間細(xì)小血管擴(kuò)張,紅細(xì)胞聚集,血管內(nèi)皮細(xì)胞腫脹、核深染。隨著死亡時(shí)間的延長,細(xì)胞溶解、壞死加重,可見嗜酸性粒細(xì)胞聚集。 死后電擊組：可見骨骼肌細(xì)胞灶狀壞死；骨骼肌細(xì)胞溶解、斷裂骨骼肌細(xì)胞濁腫顯著；胞核扭曲、固縮、深染,部分核伸長；骨骼肌細(xì)胞內(nèi)肌絲間隙增寬明顯；嗜酸性粒細(xì)胞浸潤。 空白對(duì)照組：可見骨骼肌細(xì)胞變性、壞死；胞漿凝聚、均質(zhì)化、嗜酸性；胞核固縮、深染；血管內(nèi)皮細(xì)胞腫脹；骨骼肌間細(xì)小血管擴(kuò)張,紅細(xì)胞聚集。 2、免疫組化染色結(jié)果 電擊死組通電肢體即刻腓腸肌胞漿可見棕黃色顆粒聚集,隨著時(shí)間的推移棕黃色顆粒逐漸減弱；死后電擊組通電肢體即刻腓腸肌可見胞漿棕黃色顆粒聚集,平均密度比電擊死組低；電擊死組通電肢體即刻脛骨前肌可見胞漿棕黃色顆粒聚集,1h平均密度達(dá)峰值,隨后降低；死后電擊組通電肢體即刻脛骨前肌可見胞漿棕黃色顆粒,與電擊死組脛骨前肌的變化趨勢(shì)相同,平均密度比電擊死組低；電擊死組、死后電擊組、空白對(duì)照組通電肢體脛骨前肌胞漿棕黃色顆粒平均密度大于腓腸肌。 3、RT-PCR定量結(jié)果 電擊死組大鼠左后肢腓腸肌MuRF1-mRNA表達(dá)量于死后即刻升高,1h、3h下降,與空白對(duì)照組相比有顯著差異,6h低于空白對(duì)照組無顯著意義；電擊死組大鼠左后肢脛骨前肌MuRF1-mRNA表達(dá)量于死后即刻升高,1h達(dá)峰值,3h開始降低,與空白對(duì)照組相比均有顯著差異；死后電擊組左后肢腓腸肌、脛骨前肌較空白對(duì)照組高,但升高不如電擊死組明顯；電擊死組、死后電擊組左后肢脛骨前肌MuRF1-mRNA含量均較腓腸肌高。 結(jié)論：1、MuRF1可為電擊死后早期死亡時(shí)間的推斷提供一定的參考依據(jù)。 2、MuRF1可作為生前電擊死與他殺死后偽裝電擊鑒別的輔助指標(biāo)之一。 3、脛骨前肌、腓腸肌對(duì)電擊反應(yīng)不同,脛骨前肌更易受電損傷影響。
[Abstract]:Objective: electrocution and homicide disguise shock is one of the most common types of cases in forensic medicine, but the case inspection process, forensic workers still no unified standard for the diagnosis, especially the shock for non current marks of autopsy or special conditions of water. Many scholars from morphological and molecular level of various tissues and organs electrocution was studied. When the shock cases without current spot formation, the histologic change is not obvious, after the early death of electrocution according to the changes of the skeletal muscles to infer the time little report. We designed the following experiments to detect expression of electrified limb skeletal muscle MuRF1, in order to distinguish antemortem death and death after the shock, before and after the death of electrocution shock early estimation of the death time, electrical injury effects of different skeletal muscle types.
Methods: 60 healthy Wistar rats were divided into electrocution group, after the death of shock group, blank control group, 20 rats in each group. Electrocution group rats right forelimb and left hind limb connected respectively with 220V AC power poles, 90s death, in shock after the death of 0h, 1H, 3h, 6h materials; after the death of electric shock rats were killed by cervical dislocation, respectively in 0h, 1H, 3h, 90s based 6h shock; blank control group rats were sacrificed by cervical dislocation, not to shock, after the death of 0h, 1H, 3h, 6h respectively. Take the left hindlimb gastrocnemius and tibialis anterior muscle. HE staining, immunohistochemical staining and real-time fluorescence quantitative PCR.
Results: 1, HE staining results
Electrocution group: vacuolar changes were seen in skeletal muscle cells, focal degeneration and necrosis; nuclear polarization, part of skeletal muscle cells were broken, wave like change, some skeletal muscle stripes is unclear; condensation of cytoplasm, homogenization, eosinophilic change; nuclear pyknosis, anachromasis, twist, nuclear elongation; small blood vessels expand, erythrocyte aggregation, endothelial cell swelling, nuclear stained. Along with the extension of the time of death, cell lysis, necrosis increased, visible accumulation of eosinophils.
Postmortem shock group: skeletal muscle cells showed focal necrosis, skeletal muscle cells dissolved and broken, skeletal muscle cells were turbid and swollen, nuclei were twisted, compressed, deeply stained, and some nuclei elongated.
Blank control group: skeletal muscle cell degeneration and necrosis, cytoplasmic condensation, homogenization, eosinophilia, nuclear condensation, deep staining, vascular endothelial cell swelling, skeletal muscle microvascular dilatation, red blood cell aggregation.
2, immunohistochemical staining results
Electrocution group power cell gastrocnemius limb immediately visible Brown particle slurry, with the passage of time Tan particles gradually weakened; after the death of the group of electrical shock rats' gastrocnemius muscle's cytoplasmic Brown particle, the average density was lower than the electrocution; electrocution group electrified limb immediately anterior tibial muscle visible cytoplasmic Brown particle, the average density of 1H reached the peak, then decreased; the group of electrical shock after death the body immediately the tibialis anterior muscle's cytoplasmic brown granules, and the same trend of electrocution group of the tibialis anterior muscle group is lower than the average density of electrocution; electrocution group, after the death of shock group, blank the control group average density of electrified limb anterior tibial muscle cytoplasm brown granules than gastrocnemius.
3, RT-PCR quantitative results
Electrocution group rats left hindlimb gastrocnemius MuRF1-mRNA expression increased, immediately after the death of 1H, 3h decreased, with significant difference compared with the control group, no significant 6h lower than the blank control group; electrocution group rats left hindlimb tibialis anterior muscle MuRF1-mRNA expression immediately after the death of the high rise, reached the peak at 1H. 3h, began to decrease, compared with the blank control group had significant difference; after the death of shock group left hindlimb gastrocnemius and tibialis anterior muscle than the control group, but higher than electrocution group significantly; electrocution group, shock group after the death of the left hind limb of tibialis anterior muscle MuRF1-mRNA content were compared with gastrocnemius.
Conclusion: 1, MuRF1 can provide some reference for the early death time of electric shock after death.
2, MuRF1 can be used as one of the auxiliary indicators for the identification of pre shock and kills of camouflage electric shock.
3, the anterior tibial muscle and the gastrocnemius are different to the electric shock, and the anterior tibial muscle is more susceptible to electrical damage.

【學(xué)位授予單位】：重慶醫(yī)科大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2013
【分類號(hào)】：D919.4

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