大鼠肾脏缺血再灌注损伤与重组成骨蛋白-1的关系

目的探讨鼠重组成骨蛋白1(rOP1)在大鼠肾脏缺血再灌注损伤过程中对氧自由基、细胞凋亡的影响及其对肾脏的保护作用。方法观察静脉应用rOP1250μg/kg体重,大鼠肾脏缺血60min,再灌注24h后,血浆超氧化物歧化酶(SOD)、脂质过氧化物丙二醇(MDA)、内皮素1(ET1)的变化,及其对肾脏细胞凋亡的影响。结果rOP1组和缺血再灌注组血浆SOD分别是(108±20)和(86±11)KNU/L,MDA分别是(8.90±0.49)和(11.70±0.81)μmol/L,ET1分别是(130±39)和(179±45)ng/L,rOP1组SOD水平显著性高于缺血再灌注组(P<0.05),而MDA和ET1水平显著性低于缺血再灌注组(P<0.05)。两组肾脏细胞凋亡指数分别是(5.72±3.08)%和(8.85±3.52)%,rOP1组显著性低于缺血再灌注组(P<0.01)。上述两组肾脏的的Miller’s评分分别是(2.0±0.3)和(3.4±0.5),rOP1组显著低于缺血再灌注组(P<0.01)。结论rOP1减轻氧自由基对肾脏缺血再灌注损伤,降低肾脏细胞凋亡指数,对肾脏缺血再灌注损伤具有保护作用。     

洛沙坦对缺血再灌注大鼠胰腺氧自由基、细胞凋亡和Bcl-2表达的影响

目的： 探讨洛沙坦（losartan）对大鼠胰腺缺血再灌注（I/R）损伤的保护作用及机制。方法：SD大鼠72只随机分为假手术组、I/R组和Losartan组，每组24只。采用钳闭大鼠腹腔干、肠系膜上动脉15,30,60min，再灌注6h，制成胰腺I／R损伤模型。losartan组给予losartan(40mg/kg)灌胃预处理；假手术组和I/R给予等容积的无菌蒸馏水。3组均于术后6ｈ断颈处死动物。用TUNEL法检测I/R区胰腺细胞凋亡、免疫组化法检测Bcl-2蛋白的表达, 并观察胰腺组织病理改变。结果：losartan可逆转胰腺组织炎症细胞浸润、腺泡萎缩等异常改变。缺血15,30min时段，losartan组胰腺细胞凋亡率为（6.5±2.9）％和（10.5±4.3）％显著低于I/R组的（10.2±3.2）％和（18.4±3.1）％（P<0.05）；丙二醛水平为（17.9±2.1）nmol/g（湿重）和（25.2±3.3）nmol/g（湿重）显著低于I/R组的（20.1±1.2）nmol/g（湿重）和（34.9±2.6）nmol/g（湿重）（P<0.05）；Bcl-2阳性细胞为（11.3±2.2）％和（16.2±2.7）％显著高于I/R组的（6.1±1.7）％和（10.3±2.1）％（P<0.05）。结论： losartan可减轻I/R对大鼠胰腺病理改变、抑制细胞凋亡。     

表皮生长因子对急性胰腺炎大鼠胰腺的保护作用

目的探讨表皮生长因子(EGF)对急性胰腺炎(AP)大鼠胰腺保护作用的机理。方法SD大鼠72只,随机均分为3组:对照组、AP组及AP-EGF组。AP-EGF组动物在诱导致AP模型后皮下注射EGF0.1μg/g体重,其余2组动物皮下注射相同体积的生理盐水。各组动物分别于术后6、12及24h剖杀取材(每个时相8只动物),经心脏取血4ml,其中2ml采用全自动生化分析仪检测血清淀粉酶,其余用于检测血浆中丙二醛(MDA)含量;以干纱布吸沾腹水,腹水量为浸满腹水的纱布重量与干纱布重量之差;各时相剖腹作大体观察,切取相同部位的胰腺组织5g检测其MDA含量,并观测胰腺组织病理学改变和细胞凋亡。结果AP-EGF组动物胰腺病理损伤较AP组轻;术后12及24h,AP-EGF组动物腹水量显著低于AP组〔(4.53±1.29)g vs(6.58±1.47)g、(7.64±1.85)g vs(11.96±2.13)g,P<0.05,P<0.01〕,AP-EGF组动物血清淀粉酶也显著低于AP组〔(142.0±8.3)U/L vs(187.9±10.4)U/L、(194.3±10.4)U/L vs(253.3±8.6)U/L,P<0.05,P<0.01〕;术后24h,AP-EGF组动物血浆中MDA含量为(2.34±0.23)μmol/L,胰腺组织中MDA含量为(5.21±1.46)μmol/g,均显著低于AP组的(3.15±0.38)μmol/L和(7.68±1.63)μmol/L,P<0.05,P<0.01;术后各时相,AP-EGF组动物胰腺细胞凋亡指数显著高于AP组〔(16.22±3.53)%vs(7.35±1.04)%、(11.67±2.40)%vs(4.81±0.86)%、(6.38±1.42)%vs(1.97±0.21)%,P<0.01〕。结论EGF能减轻AP时大鼠胰腺的病理损伤,迅速恢复血清淀粉酶正常活性,降低AP大鼠血浆及胰腺组织中MDA含量,通过诱导AP时腺体细胞的凋亡发挥对胰腺的保护作用。     

胰腺缺血-再灌注损伤细胞凋亡及其相关因素的研究

我们对胰腺缺血 再灌注损伤细胞凋亡及其相关因素如肿瘤坏死因子(ＴＮＦ) α、组织Ｃａ2 等在其过程中的作用进行研究 ,现将结果报道如下。一、材料与方法成年Ｗｉｓｔａｒ大鼠 60只 ,体重 (2 0 0±2 5 ) ｇ ,雌雄不拘。随机分为对照组 (Ａ组 ,10只 )、缺血 再灌注损伤组 (Ｂ组 ,3 0只 )、异搏定保护组 (Ｃ组 ,10只 )和ＴＮＦ α拮抗组 (Ｄ组 ,10只 )。戊巴比妥钠 45ｍｇ/ｋｇ体重腹腔注射麻醉 ,Ａ组直接取胰腺组织标本 ,心脏穿刺取血 2ｍｌ ;Ｂ组分别以血管夹阻断腹腔干和肠系膜上动脉 15、3 0、60ｍｉｎ(各 10只 )后再灌注 6ｈ ,取标…     

氯化钆抑制枯否细胞对大鼠肝脏缺血再灌注损伤的影响

目的探讨抑制枯否细胞对大鼠肝脏缺血再灌注损伤的影响。方法制作部分肝脏缺血再灌注大鼠模型80只,实验组注射氯化钆,对照组注射生理盐水,检测两组大鼠缺血前、再灌注后5min、1和6h血压、心率的变化,血清转氨酶(AST)、肿瘤坏死因子α(TNFα)和白细胞介素1(IL1)的水平及肝组织超微结构的改变。结果实验组再灌注6h血清TNFα和IL1为(0.475±0.069)μg/L和(0.221±0.056)μg/L,显著低于对照组的(0.831±0.167)μg/L和(0.335±0.127)μg/L(P<0.05),两组血压、心率和AST变化的差异也有统计学意义(P<0.05),实验组大鼠肝脏超微结构的损伤程度轻于对照组。结论抑制枯否细胞活化可减轻肝脏缺血再灌注损伤,枯否细胞在肝脏缺血再灌注损伤中的作用很重要。     

中性粒细胞明胶酶相关脂质运载蛋白对大鼠肾脏缺血再灌注损伤肾小管上皮细胞的保护作用

目的 探讨中性粒细胞明胶酶相关脂质运载蛋白(NGAL)对大鼠缺血再灌注损伤肾脏肾小管上皮细胞凋亡的保护作用及机制.方法 建立大鼠肾脏缺血再灌注模型,雄性SD大鼠随机分为对照组、缺血再灌注模型组、NGAL组 ;HE染色观察3组大鼠肾组织病理变化 ;TUNEL法检测肾小管上皮细胞凋亡 ;实时定量PCR、Western印迹法检测凋亡蛋白fas、bcl-2的表达变化.结果 与缺血再灌注模型组比较,NGAL组肾小管上皮细胞凋亡数量显著减少[(8.6±3.4)/HP比(20.8±3.7)/HP,P＜0.05] ;NGAL组肾组织fas mRNA(2.34±0.51比6.84±2.34,P＜0.05)、fas蛋白(0.65±0.05比0.95±0.08,P＜0.05)表达显著下调,bcl-2蛋白(0.33±0.05比0.24±0.03,P＜0.05)表达显著上调,但bcl-2 mRNA表达无明显改变.结论 NGAL对大鼠缺血再灌注损伤肾小管上皮细胞有保护作用,其作用可能与减少细胞凋亡、改变凋亡蛋白的表达有关.     

血红素氧化酶-1在脊髓缺血再灌注脊髓损伤后的表达

目的　探讨血红素氧化酶 1(HO 1)在大鼠脊髓缺血再灌注损伤中的表达。方法　制备脊髓缺血再灌注损伤模型 ,于再灌注后 4、8、16、2 4h及 2、5d收集脊髓标本。以假手术组大鼠脊髓为对照 ,采用免疫组织化学和原位杂交方法分别检测脊髓组织中HO 1的表达。以原位末端脱氧核糖核苷酸转移酶介导dUTP标记法测定细胞的凋亡。结果　对照组未发现HO 1表达 ,亦未见细胞凋亡。实验组在再灌注 4h左右HO 1开始表达上调 ,2d达峰值 (35 6± 2 96 ) ,与假手术组相比 ,差异有显著性 (P <0 0 1)。在伤后 4h见凋亡细胞 ,16h达高峰 (2 9 1± 0 4 4 )。HO 1与凋亡细胞的相关系数为r=- 0 731,P =0 0 0 5 ,具有统计学意义。结论　脊髓缺血再灌注损伤诱导HO 1表达上调 ;同时引起大量神经细胞凋亡 ,表明再灌注中脊髓存在明显、持续性的损伤。HO 1在脊髓缺血再灌注损伤中表达显著增加 ,有一定的保护作用。     

人白介素-10腺病毒转染对大鼠肝缺血再灌注损伤的保护作用

目的探讨静脉注射携带hIL-10基因腺病毒载体对大鼠肝常温缺血再灌注损伤的保护作用。方法经SD大鼠的尾静脉注射Ad-hIL10-EGFP 1×109PFU／ml。72 h后建立大鼠常温半肝缺血再灌注损伤模型(缺血60min,再灌注240min)。取血进行肝功能生化检测;HE染色观察肝组织形态变化;分别采用酶联免疫吸附法和免疫组化方法检测大鼠血清内及肝组织内hIL-10表达,用荧光显微镜观察肝脏冰冻切片中EGFP表达;Tunel法检测大鼠肝脏内细胞凋亡情况。结果治疗组大鼠肝功能较两对照组明显改善,肝细胞凋亡数量明显减少,(P<0．01);治疗组大鼠肝组织中可见EGFP表达及hIL-10染色;血清中hIL-10的表达量为(815．74±284．76)ng/ml。结论hIL-10腺病毒基因治疗可以减轻大鼠肝缺血再灌注损伤。     

缺血预处理对大鼠心肌细胞凋亡的影响及其机制

目的 观察心肌细胞凋亡及其机制在大鼠心肌缺血再灌注损伤和缺血预处理中的作用.方法 将40只大鼠随机分成4组:对照组(A组)、60min缺血再灌注组(B组)、120min缺血再灌注组(C组)、缺血预适应组(D组);应用TUNEL法检测心肌组织的凋亡;心脏病理改变用光学和电子显微镜观察;检测肿瘤坏死因子(TNF)-α、bcl-2和bax因子.结果 D组心肌细胞凋亡指数(11.36±4.23)%明显优于B组(18.14±7.69)%和C组(15.59±6.31)%,D组TNF-α、bel-2和bax的表达也明显优于B、C二组.结论 缺血预处理可减轻大鼠心肌缺血再灌注后心肌细胞的凋亡;TNF-α、bcl-2和bax表达的抑制与缺血预处理保护机制有关.     

细胞凋亡在大鼠体外肝缺血再灌注损伤中的作用

目的探讨体外切肝不同术式对肝脏缺血再灌注损伤中肝细胞凋亡的影响。方法利用SD大鼠建立完全离体、半离体切肝动物模型,术后4、24h处死大鼠采取标本,通过病理、电镜观察肝脏形态学改变,免疫组织化学检测肝组织的核增殖抗原(PCNA)表达,原位末端标记法(TUNEL)检测细胞凋亡。结果完全离体组肝脏损害最严重,细胞凋亡阳性指数最高,再灌注后4h和24h分别为24.4±1.9,32.6±2.7,而半离体组为17.0±2.6,26.4±1.1,两组间差异有统计学意义(P<0.05);完全离体组再灌注4h和24h后PCNA的表达分别为66.6±2.5,77.2±1.5,明显高于半离体组的57.4±1.5,68.6±2.5(P<0.05)。结论肝窦内皮细胞凋亡及之后伴随的肝细胞凋亡是肝缺血再灌注后肝脏损伤的方式之一,在肝细胞凋亡的同时也存在肝细胞的增生,半离体体外肝手术对肝脏损伤相对轻、暴露良好,因此临床上应优先选择。     

Hazards and complications of endoscopic transurethral ureterolithotripsy and lithoextraction and means of prevention

Complications related to ureterolithotomy and ultrasonic ureterolithotripsy performed under the control of visual endoscope were analyzed in 86 ureterolithiasis patients, methods of their prevention discussed. All the aforementioned complications were distributed into three groups: inapplicability of surgery due to anatomic and functional defects of lower and upper urinary tracts, intraoperative, and postoperative complications. The commonest ones were ureteral abruption and perforation, acute pyelonephritis, temporary vesicoureteral reflux. Their control measures were considered as relative methods of treatment: immediate surgical intervention in case of ureteral abruption, renal catheterization in patients with insignificant ureteral perforation or acute pyelonephritis. Adequate ureteroscopy, careful consideration of pro- and contraindications, catheterization of renal pelvis and urinary bladder performed within 2-3 days after the surgery and adequate antibacterial therapy are the most decisive steps in the control of aforementioned complications.     

牙体牙弓颌骨阻力中心及其临床意义

牙体、牙弓及颌骨的阻力中心在正畸矫治力系统中具有重要的意义,也是正畸学领域争论较多的一个问题。Dermaut等研究表明,当力作用于物体阻力中心时,物体将发生平动,否则将发生平动和转动的复合运动。目前,国内外多数学者认为牙体、牙弓及颌骨存在阻力中心,但其位置存在争议。本文就牙体、牙弓及颌骨的阻力中心及其临床意义作一综述。     

Characterisation and differentiation of chondroblastomas and chondromyxoidfibromas - presence and expression of collagen types I and II

AIM: Chondroblastomas and chondromyxoidfiibromas are rare benign skeletal neoplasms with reported overlapping histology. Aim of this study was to analyse the biochemical composition of the matrix of these tumour entities in order to further characterise the cellular phenotypes of these neoplasms using typical cell biological marker genes. METHODS: The matrix compositions of chondroblastomas and chondromyxoidfibromas were analyzed by HE-histology, histochemistry, and immunolocalization techniques. Cellular gene expression patterns were detected by mRNA in situ hybridization. RESULTS: Chondroblastomas are rich in collagen type I and show foci of an osteoid-like matrix, whereas collagen type II as a typical marker of chondrocytic differentiation was not detected in any of the specimens. Chondromyxoidfiibromas had foci of chondroid appearance with chondroblastic cellular differentiation characterised by collagen type II expression. CONCLUSION: These results characterise chondroblastomas and chondromyxoidfiibromas as skeletal neoplasms that have a different biology and which can be distinguished by matrix protein expression products: collagen type II, the typical marker of chondroblast differentiation, could only be detected in chondromyxoidfibromas, but not in chondroblastomas. Thus, both neoplasms are clearly different on the cell biological level.     

Histochemical and contractile properties of striated muscles of urethra and levator ani of dogs and sheep

AIMS: To understand their possible importance in long- and short-term control of continence, some properties of the striated muscles of the urethra and pelvic floor (levator ani) of dogs and sheep were investigated, especially fiber types and contractile characteristics. MATERIALS AND METHODS: Striated muscles of urethra and levator ani of 29 male and 6 female dogs and 11 male and 6 female sheep were removed and cut into strips. Some strips were frozen and stained for ATPase at pH 9.4 and 4.3 for fiber typing; others were set up in an organ bath to study contractile responses to nerve stimulation. RESULTS: All muscles contained both type I (slow) and type II fibers, ranging from 97% type II in female greyhound urethra to 60% in female sheep levator ani. For each muscle, there were fewer type II muscles in sheep than in dog. The diameters of the urethral fibers were about 60% of the levator ani in dogs and 34% in sheep. Contraction of the urethral muscle was faster than for levator ani and declined to about 80% of the peak, 500 msec after the beginning of stimulation at 20 Hz. The levator ani contraction rose to a steady level as long as stimulation continued. CONCLUSIONS: Both the levator ani and urethral striated muscles contain slow and fast fiber types. The levator ani muscles are capable of sustained contraction with rapid onset which will produce long-term closure of the urethra. The circular urethral muscle contraction was faster but less well maintained.     

Incorporation and release of85Sr and14C-proline-hydroxyproline in mineral and collagen of bone and incisor teeth of growing rats

The extent to which exchange and reutilization processes of mineral tracers affect skeletal mineral accretion and resorption measurements was evaluated by comparing the rates of appearance and disappearance of⁸⁵Sr and¹⁴C-proline-hydroxyproline in bones and teeth in growing rats for 12 days following simultaneous parenteral injection of these tracers. Expressions for the relative rates of collagen synthesis and breakdown, which unlike mineral metabolism are considered not to be complicated by exchange phenomena, were based on¹⁴C-proline conversion to¹⁴C-hydroxyproline; the specific activity of the latter was determined. Both the mineral and the collagen specific activities reflected the rates and patterns of growth of the samples assayed; rapid growth and a short interval of time between formation and resorption of tissue in themetaphyseal bone which contains the cartilagineous growth plate, slow growth and an interval of time between formation and resorption of tissue indiaphyseal bone and incisor teeth which is longer than the 12 days of the experiment. However, in metaphyseal bone the specific activity collagen/mineral ratio dropped by one half during the 4–12 day interval in contrast to diaphyseal bone and incisor teeth in which no change in this ratio was observed during this period of time. The data indicate that collagen in the metaphyseal growth zone is removed by resorption before it has become fully mineralized, and that exchange is a relatively unimportant factor in the long term kinetics of bone mineral.

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Zusammenfassung Das Ausmaß, bis zu welchem Austausch- und Wiederverwendungsprozesse der mineralen Tracer die Messungen des mineralen Skelett-Auf- und Abbaues beeinflussen können, wurde ausgewertet; zu diesem Zweck wurde die Geschwindigkeit des Auftretens und Verschwindens von⁸⁵Sr und von¹⁴C-Prolin-Hydroxyprolin in Knochen und Zähnen von wachsenden Ratten während der 12 auf die simultane parenterale Injektion dieser Tracer folgenden Tage verglichen.Der Ausdruck für die relative Geschwindigkeit des Kollagen-Auf- und Abbaues, bei welchem im Gegensatz zum Mineralmetabolismus kein Mitwirken des Austauschphänomens vermutet wird, basiert auf der Umwandlung von¹⁴C-Prolin zu¹⁴C-Hydroxyprolin; die spezifische Aktivität des letzteren wurde bestimmt.Aus der spezifischen Aktivität des Minerals sowie jener des Kollagens konnten die Geschwindigkeit und die Art des Wachstums der untersuchten Proben ersehen werden, d.h.schnelles Wachstum und ein kurzes Zeitintervall zwischen Bildung und Resorption des Gewebes imKnochen der Metaphyse, die auch die knorpelige Wachstumsplatte enthält, und andererseitslangsames Wachstum und längeres Zeitintervall (länger als die 12 Tage des Experimentes) zwischen Bildung und Resorption des Gewebes imKnochen der Diaphyse und in den Schneidezähnen. Immerhin fiel die spezifische Aktivität des Kollagen/Mineral-Anteils im Knochen der Metaphyse während dem 4–12tägigen Zeitintervall auf die Hälfte, im Gegensatz zum Knochen der Diaphyse und der Schneidezähne, bei welchen während dieser Zeitspanne kein Unterschied in diesem Verhältnis beobachtet wurde.Diese Ergebnisse zeigen, daß Kollagen in der Wachstumszone der Metaphyse durch Resorption verschwindet, bevor es ganz mineralisiert ist, und daß der Austausch ein relativ unwichtiger Faktor in der Kinetik auf lange Sicht des Knochenminerals ist.