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  本文選題：年齡推斷　切入點：軟組織　出處：《汕頭大學(xué)》2008年碩士論文　論文類型：學(xué)位論文

【摘要】： 背景與目的 目前,推斷年齡主要是依據(jù)人體骨骼、細胞、生物大分子,以及組織中各類物質(zhì)的物理和化學(xué)特性等隨年齡增長的時序性變化。根據(jù)骨骼的多項指標聯(lián)合觀察,能較準確地推斷出年齡,是國內(nèi)外推斷年齡的主要手段,為破案偵察和法庭訴訟提供了準確可靠的證據(jù)。但對于軟組織檢材,如皮膚、血痕、毛發(fā)等,尚無公認的客觀準確的推斷年齡方法。近年來,隨著分子生物學(xué)技術(shù)的發(fā)展和普及,有報道檢測軟組織檢材的端粒酶等,為推斷年齡提供了一種新思路。但受諸多因素的影響,其推斷年齡的誤差較大,目前還處于研究階段。本實驗基于生物體各組織形態(tài)結(jié)構(gòu)隨年齡變化的自然規(guī)律,應(yīng)用生物力學(xué)的理論和技術(shù),探討不同年齡、性別組大鼠軟組織的生物力學(xué)性狀與年齡的相關(guān)性,旨在為用軟組織推斷年齡提供一種新的數(shù)值量化的技術(shù)思路。 材料與方法 SD大鼠96只,雌雄各8組,共16組,分別為新生兒組(≤1w)、嬰幼兒組(1～4W)、青少年組(1～2個月)、青年組(2～3個月)、中年早期組(3～7個月)、中年中期組(7～11個月)、中年晚期組(11～15個月)、老齡組(≥18月齡),每組6只。 于每個分組點頸椎脫臼處死大鼠,立即縱向切取腹壁正中皮膚、腹白線右側(cè)肌肉,小腸上段、結(jié)腸下端和胸主動脈壁全段各約5cm,小心剝離各組織周圍的結(jié)締組織,備制約1:4的長方形軟組織生物力學(xué)試件。同時,于相應(yīng)部位提取組織切片檢材及提取腦、心、肝、腎等全身臟器,放入10%的中性甲醛溶液內(nèi)固定約48小時。 步驟:將軟組織試樣固定于萬能材料拉力機特制夾具上,室溫下進行4～5次預(yù)調(diào),得到較理想的狀態(tài)后,進行軸向一維拉伸,直至軟組織被拉斷。每次測試過程不超過3分鐘,期間用生理鹽水噴灑軟組織以保持試件原有水分。電腦軟件自動采集檢測過程中的試樣數(shù)據(jù),主要測算指標有:最大應(yīng)力、斷裂時應(yīng)變、斷裂力、最大力變形、彈性模量等。應(yīng)用SPSS13.0和Excel統(tǒng)計軟件進行數(shù)據(jù)的分析處理。 組織切片檢材常規(guī)脫水、石蠟包埋、切片、HE、VG染色,光鏡下觀察各組軟組織與腦、心、肺、肝、脾、腎等內(nèi)臟組織的形態(tài)學(xué)特征。同時,采用顯微圖象處理技術(shù)進行膠原纖維、彈性纖維、肌纖維、腸絨毛高度等的觀察測量。 結(jié)果 1軟組織生物力學(xué)性狀 皮膚和肌肉組織的應(yīng)力隨年齡增長而呈先增后減的開口向下“拋物線”型變化,雌鼠的最大應(yīng)力值在36W齡左右,皮膚、肌肉最大應(yīng)力值分別約為6.00±1.02 Mpa,1.19±0.17Mpa,雄鼠應(yīng)力則于20W齡左右達到最大值,皮膚、肌肉最大應(yīng)力值分別約為6.93±1.13 Mpa,1.36±0.20Mpa,之后隨年齡的增長逐漸呈逐漸下降趨勢。52W齡以后,雌雄組的應(yīng)力差別不明顯。腸組織的應(yīng)力性別差異不太明顯,10W齡左右達到最大值,之后逐漸下降,而52W齡左右應(yīng)力又略有增加。血管應(yīng)力未觀察到與年齡和性別相關(guān)的明顯變化。雌雄鼠皮膚、肌肉、小腸的應(yīng)變10W左右最大,10～36W處于相對穩(wěn)定期,52W后開始逐漸呈下降趨勢,相同年齡組雌雄大鼠應(yīng)變不存在顯著差別。 2 HE與VG染色的組織形態(tài)學(xué)觀察 各年齡段大鼠腹部皮膚全層膠原纖維為主,彈性纖維較少。3W齡的皮膚膠原纖維與彈性纖維最細、最少,膠原纖維束的直徑約13.54um。隨年齡的增長,彈性纖維與膠原纖維逐漸增粗、排列致密。雌鼠36W左右纖維束最粗大,直徑約31.06 um,雄鼠20W左右膠原纖維束最粗大,直徑約35.12 um。之后,隨年齡的增加,皮膚膠原纖維變細、排布較稀疏紊亂。彈性纖維也隨年齡增長變細、變短,72W齡,膠原纖維束的直徑約20.67um。 大鼠腹壁肌肉橫斷面直徑隨年齡增長而增加,1W時肌纖維最細,與成年鼠差異最大。雌鼠約36W、而雄鼠約20W時肌纖維最粗壯、橫斷面最大,36W齡雌鼠與20W齡雄鼠肌纖維橫斷分別為11.57um、15.69um,以后隨年齡增長呈遞減趨勢,各年齡段比較P＜0.05。 大鼠從6W齡開始小腸絨毛高度、肌層厚度逐漸增加,肌纖維排列逐漸致密。10W齡左右絨毛高度達高峰、肌層最厚、肌纖維排列最致密,以后開始下降,52W齡又略有增加,至72W后小腸絨毛稀疏、短粗,腺體數(shù)量減少,腸壁肌層菲薄。雌雄鼠結(jié)腸下段除了腸絨毛外,其余變化與小腸基本上一致。 胸主動脈20W齡左右彈性膜折疊最豐富。52W后彈性膜折疊的數(shù)量與幅度逐漸減小。隨著年齡的增加,血管內(nèi)膜與管壁厚度增加,管腔擴張,管壁厚度與管腔直徑比值隨年齡的增加而增加。 大鼠內(nèi)臟組織也存在增齡過程中的變化,如老年大鼠腦回變窄、腦溝加寬,神經(jīng)元數(shù)量減少,神經(jīng)膠質(zhì)細胞數(shù)量和體積增加。老年大鼠心內(nèi)膜增厚、膠原纖維增多。老年大鼠肝細胞核體積增大,形態(tài)大小不一,肝細胞排列疏松,肝小葉中央靜脈周圍肝細胞排列較紊亂,細胞脂肪空泡增加。 結(jié)論 1、不同年齡和性別大鼠的皮膚、肌肉應(yīng)力隨年齡增長呈“拋物線”型變化規(guī)律,兩者呈二元回歸關(guān)系,雄鼠比雌鼠早一個年齡段達到最大應(yīng)力值;腸應(yīng)力呈“M”型變化規(guī)律。 2、皮膚膠原和彈力纖維形態(tài)結(jié)構(gòu)隨年齡的變化、肌纖維直徑隨年齡的變化,與組織生物力學(xué)變化之間存在物質(zhì)-功能的相關(guān)性,鑒于軟組織的生物力學(xué)特性呈隨年齡增長的“拋物線”型變化規(guī)律及二元回歸關(guān)系,應(yīng)配合組織形態(tài)學(xué)變化推斷年齡。
[Abstract]:Background and purpose
At present, age estimation is mainly based on human bone cells, biological macromolecules, sequential changes and all kinds of material in the physical and chemical characteristic with age. According to the observation of several indexes of skeleton, can accurately infer the age, is the main means to estimate the age at home and abroad, to provide accurate and reliable the evidence for the case of reconnaissance and court proceedings. But for the soft tissue samples, such as skin, blood, hair, objective and accurate method to estimate the age is not recognized. In recent years, with the development of molecular biology and the popularization, has reported the detection of soft tissue samples of telomerase, provides a new way of thinking to estimate the age. But under the influence of many factors, the age estimation error is large, is still in the research stage. The experimental nature of the organizations of the organism structure changes with age based on the application of biological The theory and technology of mechanics are used to explore the correlation between biomechanical properties of soft tissue and age in different age and gender groups, so as to provide a new quantitative method for inferring age from soft tissue.
Materials and methods
96 SD rats, male and female each 8 group, 16 group, respectively (1W), group of newborn infant group (1 ~ 4W), youth group (1~2 months) and young group (2~3 months), early middle age group (3~7 months), middle age group (7~11 month Interim), the late middle age group (11~15 months), aged group (aged 18 month old), 6 rats in each group.
In each group of rats were killed by cervical dislocation point, immediately cut the longitudinal midline abdominal skin on the right side of the linea alba muscle, small intestine, colon and lower thoracic aortic wall intact by about 5cm, be careful around the tissue dissection of connective tissue, by restricting rectangular soft tissue mechanics 1:4 specimens. At the same time, the corresponding parts extraction of tissue samples and extraction of brain, heart, liver, kidney and other organs, neutral fixation into 10% Formaldehyde Solution for about 48 hours.
Steps: soft tissue specimens were fixed in a universal material testing machine special fixture, 4~5 preset at room temperature, get the ideal state, the axial tensile, until soft tissue off. Each test process should not exceed 3 minutes, during spraying with saline soft tissue specimens in order to maintain the original water. The sample data in the detection process of automatic acquisition of computer software, the main indexes are: maximum stress, fracture strain, fracture strength, maximum deformation, elastic modulus and so on. The application of SPSS13.0 and Excel statistical software for analysis of the data processing.
Tissue samples of conventional dehydration, paraffin embedding, slicing, HE, VG staining, observe the soft tissue under light microscope and the brain, heart, lung, liver, spleen, kidney and other viscera tissue morphological characteristics. At the same time, by using collagen fiber, microscopic image processing technology of elastic fibers, muscle fiber, observation and measurement villus height and so on.
Result
1 biomechanical properties of soft tissue
The skin and muscle tissue stress is opening down "parabola" firstly increased and then decreased with the increase of age, the maximum stress value at about 36W in the skin, muscle of female rats, the maximum stress value of approximately 6 + 1.02 Mpa, 1.19 + 0.17Mpa, male rats stress in 20W age as to the maximum value of the skin, muscle, the maximum stress value of about 6.93 + 1.13 Mpa, 1.36 + 0.20Mpa, with age gradually decreased after.52W age group, male and female stress difference is not obvious. Intestinal tissue stress gender difference was not significant, reached the maximum at about 10W the value, then gradually decreased, and at about 52W stress and increased slightly. Vessel stress is not observed with age and sex related changes in male and female rats. The skin, muscle strain, intestinal 10W about 10 ~ 36W maximum, in a relatively stable period, 52W began to gradually decreased, the same age There was no significant difference in strain between the male and female rats.
Histomorphological observation of 2 HE and VG staining
Abdominal skin of rats of all ages full-thickness skin collagen fibers. The collagen fiber and elastic fiber elastic fiber less.3W age of the most fine, at least, collagen fiber bundle diameter of 13.54um. with age, elastic fibers and collagen fibers became thicker, dense. Female rats around 36W fiber bundle the thick diameter about 31.06 um male rats, collagen fibers around 20W thick, after about 35.12 um. in diameter. With the increase of age, the skin collagen fibers became thinner and more sparse arrangement disorder. Elastic fibers with age became thinner and shorter, 72W age, collagen fiber bundle diameter of about 20.67um.
Rat abdominal wall muscle cross-sectional diameter increased with age, 1W muscle fiber fine, the biggest difference with adult rats. Female rats and male rats is about 36W, about 20W of the muscle fiber thick, maximum, 36W old female rats and male rats aged 20W and muscle fiber cross-section were 11.57um, 15.69um, to after the trend of decrease with the increase of age, the age of P is less than 0.05.
The rats began to villus height from 6W age, muscle layer thickness increases gradually, muscle fibers arranged in dense villus height at about.10W reached the peak, the most thick muscle layer, muscle fibers arranged in the most dense, then began to decline, 52W age and slightly increased to 72W after intestinal villi sparse, short, reduce the number of intestinal glands. Thin wall muscle layer. Male and female rats colon segment except the intestinal villi, the change and small intestine is basically the same.
The size and size of elastic membrane were reduced after 20W age. The number and extent of elastic membrane folding decreased gradually. With the increase of age, the thickness of vascular intima and wall increased, lumen dilated, and the ratio of.52W to wall diameter increased with age.
There are also changes in visceral tissues of rats during the aging process, such as the brain of aged rats narrowed sulci widened, reduce the number of neurons, glial cell number and volume increased. Heart intimal thickening in aged rats, increased collagen fibers in aged rat liver nuclei. The increase in size, shape size, arrangement of hepatic cells osteoporosis, lobular liver cells around the central vein disordered cells, fat vacuoles increased.
conclusion
1, the skin and muscle stress of different ages and sexes showed a parabolic type change rule with age. Both of them showed a two element regression relationship. Male rats reached the maximum stress value in one age group, and the intestinal stress showed a "M" rule.
2, the skin collagen and elastic fiber morphology changes with age, muscle fiber diameter changes with age, correlation function between material and tissue biomechanical changes, in view of the biomechanical properties of soft tissue was increased with age "parabola" variation and two regression relationship, with morphological changes of age estimation.

【學(xué)位授予單位】：汕頭大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2008
【分類號】：D919

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