瘦素对成骨细胞增殖及CollagenⅠ和Cbfa1 mRNA表达的影响

目的研究瘦素在体外对原代培养乳鼠颅盖骨成骨细胞增殖及对CollagenⅠ(Ⅰ型胶原蛋白)和Cbfa1 mRNA表达的影响。方法培养乳鼠成骨细胞,以乳鼠成骨细胞为体外实验模型,采用MTT法检测不同浓度瘦素(0、40、80和160ng/ml)作用于成骨细胞后的增殖情况;通过RT-PCR方法检测不同浓度瘦素对CollagenⅠ和Cbfa1 mRNA的表达的影响。结果瘦素作用于成骨细胞后,可促进其增值,OD值显著增加(P0.01);瘦素增加CollagenⅠ和Cbfa1 mRNA的表达,呈剂量效应关系。结论瘦素促进成骨细胞增殖,同时通过调节CollagenⅠ和Cbfa1的表达,促进成骨细胞分化,进而促进骨形成。     

rhIGF-1对成骨细胞增殖、BMP-2及Cbfa1基因表达的影响

目的通过重组人胰岛素样生长因子(rhIGF-1)对体外成骨细胞增殖、骨形态蛋白-2(BMP-2)及核心结合因子1(Cbfa1)基因表达的影响,探讨rhIGF-1对骨代谢影响的可能机制。方法不同浓度的rhIGF-1(0、10、50、100ng/ml)刺激体外培养的大鼠成骨细胞,采用噻唑蓝(MTT)法检测细胞增殖能力,用半定量RT-PCR法检测成骨细胞BMP-2、Cbfa1基因的表达。结果rhIGF-1可明显促进成骨细胞增殖(P0.05),并促进成骨细胞BMP-2、Cbfa1基因的表达(P0.01),具有浓度依赖性。结论rhIGF-1可促进成骨细胞的增殖、分化及成熟,可能是通过增强BMP-2、Cbfa1基因的表达来实现的。     

痩素对小鼠成骨细胞MC3T3-E1增殖及OPG/RANKL mRNA表达的影响

目的初步探讨瘦素对小鼠成骨细胞MC3T3-E1细胞增殖及骨保护蛋白、核因子κB激活受体配体mRNA表达的影响。方法以小鼠成骨细胞MC3T3-E1为体外实验模型,用MTT法检测瘦素作用MC3T3-E1细胞后的增殖情况,RT-PCR方法检测OPG、RANKL mRNA表达。结果瘦素（10、20、40、80和160ng.mL-1）作用于MC3T3-E1细胞24h后,可促进其增殖,OD值显著增加（P〈0.01）,增加骨保护蛋白mRNA表达,同时降低核因子κB激活受体配体mRNA表达,且呈浓度依赖性。结论瘦素促进MC3T3-E1细胞增殖,同时通过调节骨保护蛋白mRNA和核因子κB激活受体配体mRNA的表达,从而促进骨形成,抑制骨吸收。     

17-α雌二醇对体外培养成骨细胞影响的实验研究

目的 观察17—α雌二醇对体外培养成骨细胞增殖和成骨功能的影响。方法 用植块培养法从乳鼠中分离出成骨细胞，倒置显微镜观察其形态，碱性磷酸酶(ALP)染色来鉴定细胞。然后用不同浓度的17—α雌二醇作用于成骨细胞，使用四甲基偶氮唑蓝(MTT)比色分析、ALP含量测定、细胞钙含量测定来检测其对成骨细胞增殖和成骨功能的影响。结果 17—α雌二醇以剂量方式促进各项指标的表达，促进成骨细胞的增殖和骨形成。结论 17—α雌二醇具有促进成骨细胞的增殖和骨形成作用。值得作进一步的研究。     

胰岛素样生长因子-1对甲状旁腺激素介导成骨细胞增殖及成骨活性的影响

目的：通过构建乳鼠颅盖骨成骨细胞分离培养与鉴定方法,研究胰岛素样生长因子－1（Insulin－like growth factor－1,IGF－1）及IGF－1联合重组人甲状旁腺激素1－34（recombinant human parathyroid hormone 1－34,rhPTH1－34）对成骨细胞增殖及I型胶原蛋白mRNA表达的影响。方法培养乳鼠成骨细胞,并观察其形态及功能,以原代培养乳鼠成骨细胞为实验模型,分空白组、PTH组、IGF－1组及不同浓度IGF－1作用的PTH介导的成骨细胞组（0、10、50、100 ng／L）。通过不同剂量的胰岛素样生长因子－1（0、10、50、100 ng／L）联合10－9 mol／L重组人甲状旁腺素刺激体外培养的成骨细胞,用噻唑蓝（ MTT）法检测细胞的增殖能力,RT－PCR法检测成骨细胞I型胶原蛋白mRNA的表达。结果 PTH和IGF－1均可促进成骨细胞增殖;IGF－1联合PTH可以促进成骨细胞增殖,且具有剂量依赖性。 PTH联合IGF－1使成骨细胞增殖能力增强、I型胶原蛋白mRNA表达增强。 IGF－1可以促进成骨细胞I型胶原蛋白mRNA的表达,且具有剂量依赖性。 IGF－1可以促进成骨细胞I型胶原蛋白mRNA的表达,且具有剂量依赖性。结论 PTH与IGF－1均可刺激成骨细胞增殖和分化,IGF－1可促进PTH对成骨细胞的分化、增殖。两者合用其作用增强,有协同促进作用。     

盐酸二甲双胍对人成骨细胞增殖及Collagen I 、 LRP5 mRNA表达的影响

目的观察不同浓度二甲双胍对体外培养人成骨细胞增殖、Collagen I及LRP5mRNA表达的影响,探讨二甲双胍对成 骨细胞作用的可能机制。方法不同浓度二甲双胍(0、25、50、100、200 ^mo^L)刺激体外培养人成骨细胞72 h,采用CCK~8比 色法检测细胞增殖情况,用荧光定量RT~PCR法检测Collagen I、LRP5mRNA表达。结果显示二甲双胍可促进其增殖（P < 0.05),并促进成骨细胞CollagenI及LRP5mRNA表达（P <0.05),在200 pmoVL时增殖、表达最明显,呈剂量效应关系。结 论二甲双胍在一定浓度范围内可促进人成骨细胞增殖,可能通过Wnt/LRP5信号通路增强Collagen I及LRP5mRNA的表达 来调控。     

铁饱和乳铁蛋白对大鼠成骨细胞增殖与分化的影响

目的 探讨铁饱和乳铁蛋白对体外培养的大鼠成骨细胞增殖和分化的影响.方法 用酶消化法分离大鼠颅盖骨成骨细胞进行原代培养;乳铁蛋白螯合Fe~(2+)获得铁饱和乳铁蛋白;将其稀释到不同浓度作用于成骨细胞,四唑盐比色法(MTT)测定细胞增殖;碱性磷酸酶法(ALP)测定细胞碱性磷酸酶活性.结果 随着时间的增加,各浓度组均可促进成骨细胞增殖及增强碱性磷酸酶活性.其中100 μg/mL浓度组在72 h时细胞增殖数目最大、碱性磷酸酶活性最高,较对照组有统计学意义(P＜0.01).结论 铁饱和乳铁蛋白能促进大鼠成骨细胞的增殖和分化.     

蓇密钙片与马鹿角多肽对成骨细胞增殖分化的研究

目的观察蓇密钙片及马鹿角多肽对鼠胚成骨细胞MC3T3-E1的增殖分化作用及细胞内OPG、RANKL mRNA的表达。方法采用MEM(10%FBS)培养液培养细胞,细胞计数法描绘成骨细胞生长曲线;MTT法检测蓇密钙片(0、50、100、500、1000μg/ml)、马鹿角多肽(0、5、50、500μg/ml)和钙尔奇D对照组对鼠胚成骨细胞MC3T3-E1增殖的作用;蓇密钙片(0、50、100、500、1000μg/ml)、马鹿角多肽(0、5、50、500μg/ml)和钙尔奇D对照组干预鼠胚成骨细胞MC3T3-E1 72 h后,比色法检测细胞内碱性磷酸酶(alkaline phosphatase,ALP)的含量,RT-PCR法检测骨保护蛋白(osteoprotegerin,OPG)、核因子КB激活受体配体(receptor activator of NK-КB ligand,RANKL)mRNA的表达。结果蓇密钙片及马鹿角多肽作用于MC3T3-E1成骨细胞72 h后,可促进其增殖,OD值增加(P0.05),增加ALP的含量,促进OPG mRNA表达,同时抑制RANKL mRNA表达。结论蓇密钙片及马鹿角多肽促进MC3T3-E1成骨细胞增殖分化,可能与促进成骨细胞OPG mRNA表达、抑制RANKL mRNA表达有关,从而促进骨形成,抑制骨吸收。     

依普拉芬对新生大鼠颅盖骨成骨细胞BMP-2和TGF-β1 mRNA表达的影响

目的研究依普拉芬对体外培养的原代大鼠成骨细胞BMP-2(Bone Morphogenic Protein-2,骨形态发生蛋白-2)和TGF-β1(Transforming Growth Factor-β1,转化生长因子-β1)mRNA表达的影响,探讨其预防绝经后骨质疏松的机制。方法体外分离培养新生大鼠颅盖骨成骨细胞,取第二代细胞,将不同浓度的依普拉芬加入培养液,72h后用RT-PCR技术检测各组细胞BMP-2和TGF-β1mRNA表达情况。结果与阴性对照组比较,各浓度组的依普拉芬均可使成骨细胞BMP-2和TGF-β1mRNA表达量显著增加(P<0.01),其中,依普拉芬浓度在10-8~10-5mol/L之间作用最为显著。结论10-8~10-5mol/L浓度范围内的依普拉芬均可促进成骨细胞BMP-2和TGF-β1mRNA的表达,从而间接促进其增殖和分化,增加骨形成。     

甲状旁腺激素联合辛伐他汀对成骨细胞分化及OPG和RANKL mRNA表达的影响

目的联合甲状旁腺激素(rhPTH1-34)和辛伐他汀(SIM)在体外对乳鼠颅盖骨成骨细胞分化及骨保护素(OPG)和核因子κB受体活化因子配体(RANKL)基因表达的影响。方法以乳鼠成骨细胞为体外试验模型,rhPTH1-34(10-9mol/L)联合不同浓度SIM(10-8、10-7、10-6mol/L)作用于体外培养的乳鼠成骨细胞,采用对硝基苯磷酸盐(PNPP)法测定碱性磷酸酶(ALP)活性;RT-PCR法测定OPG和RANKL基因的表达。结果 rhPTH1-34和SIM单独给药均可促进成骨细胞ALP活性及OPG基因、降低RANKL基因表达(P0.05);两者联合后与SIM单独作用组比较,ALP活性明显增加,并能协同促进OPG、降低RANKL基因表达(P0.05)。结论 rhPTH1-34和SIM联合应用对成骨细胞分化和代谢有协同作用。     

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.