芳香化酶、雌激素、雌激素受体α和β在文昌鱼消化管上皮和肠神经元的免疫定位

用免疫组织化学技术和鼠抗人胎盘芳香化酶抗体、兔抗人雌二醇抗体及雌激素受体ER-α与ER-β多克隆抗体对文昌鱼消化管进行免疫组织化学定位.研究结果显示,芳香化酶、雌激素和雌激素受体α和β的免疫活性蛋白在肝盲囊、中肠前部和后部的上皮细胞及肠神经元表达,后肠显免疫阴性反应,表明文昌鱼肠道像哺乳类大鼠胃壁细胞和肠上皮细胞一样,能够合成雌激素并具有内分泌作用.提供了确凿的形态学证据,并对阐明雌激素在调节文昌鱼肠道消化功能及其作用机制以及雌激素功能的演化有十分重要的理论和学术意义.     

循环荷载下黄土地区桩基础抗拔承载力计算

为研究竖向循环荷载对桩基抗拔承载力的影响,基于现场试验分析了循环荷载下循环次数、循环荷载比对循环位移比的影响规律,总结了循环作用下的抗拔系数与循环位移比的关系,建立了基于循环位移比的桩基础抗拔承载力计算模型。研究结果表明：相对于单一拉拔荷载,循环荷载下桩基础的承载能力衰减更快,两种情况下桩基的承载力计算方法不可等同;循环荷载比和循环次数是影响桩基承载能力的重要因素,循环拉拔作用下循环次数大于2或循环拔压作用下循环拔压荷载比大于5∶4时,循环荷载对桩基承载力有较大的削弱作用;循环位移比对桩基承载能力的衰减存在放大作用,当循环位移比达到0.8时,桩基承载能力的衰减率达到50%,因此实际工程中应合理控制循环位移比的大小;提出了黄土地基中循环荷载作用下桩基承载力计算模型,通过案例表明,新提出模型计算的抗拔承载力与实测值误差较小,验证了模型的准确性和可靠性。研究结果为循环荷载作用下桩基础抗拔承载力的计算提供了宝贵的借鉴。     

雌激素防治阿尔茨海默病的作用机制

目的:研究雌激素防治阿尔茨海默病(AD)的作用机制.方法:对国内外近年文献进行复习和综述.结果:雌激素能对抗β淀粉蛋白的沉积和毒性作用、促进胆碱能神经元的存活和突触可塑性、抗氧化和维持神经元的钙稳态、调节载脂蛋白E的表达、减轻炎症反应、增加脑血流量和脑的能量代谢,从而影响AD的发生、发展.结论:雌激素可通过多条途径影响AD的发生、发展,雌激素替代治疗对于防治AD具有重要作用.     

激素和抗菌素结合治疗奶牛子宫内膜炎的疗效

根据奶牛产后不同患病时期其生殖内分泌的机能状态,应用激素治疗途径人为调理下丘脑—垂体—性腺轴系与子宫的关系,增强病畜自我防卫能力,并与抗菌素相结合治疗73头子宫内膜炎奶牛。其中三合激素和土霉素结合治疗20头,治愈率为70％;雌激素、催产素和四环素结合治疗15头,治愈率为80％;雌激素和青、链霉素结合治疗21头,治愈率为76.19％;前列腺素和氯霉素结合治疗17头,治愈率为82.35％。     

干湿循环下花岗岩残积土抗剪强度及边坡稳定性分析

为研究干湿循环对花岗岩残积土抗剪强度特性和抗剪强度参数的影响,分析花岗岩残积土边坡的稳定性,改进了常规的单元试验方法.以整个花岗岩残积土边坡为试验对象进行干湿循环模拟,在每次循环后直接在边坡坡面上切取花岗岩残积土试样进行直剪试验.在此基础上,利用有限元软件ABAQUS对一实际边坡进行了干湿循环的数值模拟,在每次循环模拟前对上一次模拟结果进行分析,将等效塑性应变较大区域的土体强度进行适当折减,分析每次循环后花岗岩残积土边坡的应力应变、稳定性以及塑性变形.试验结果表明:花岗岩残积土的抗剪强度随干湿循环次数、循环幅度的增加而减小,其中黏聚力值衰减明显,内摩擦角的变化相对较小,在3～5次干湿循环后抗剪强度趋于稳定;通过数据分析得到干湿循环的黏聚力公式,进一步得到了干湿循环条件下花岗岩残积土的抗剪强度公式;干湿循环作用导致边坡稳定性降低,安全系数减小,且第一次干湿循环后安全系数的降幅最大.     

木脂素类植物雌激素对ER_α及ER_β结合亲和力的分子动力学研究

本研究采用分子动力学模拟方法探究了4种木脂素类植物雌激素与雌激素受体ER_α及ER_β之间的结合亲和力。结果表明:四种木脂素类植物雌激素对ER_β的结合亲和力高于对ER_α的结合亲和力;罗汉松脂酚和开环异落叶松脂酚对ER_α及ER_β的结合亲和力高于肠内脂和肠二醇对ER的结合亲和力;范德华能、静电能和非极性溶剂化能自由能更有利于配体与受体的结合。     

干湿循环条件下陶瓷粉再生混凝土抗硫酸盐侵蚀性能及可靠性分析

为了研究陶瓷粉再生混凝土抗硫酸盐干湿循环损伤的能力,设计了陶瓷粉、再生粗骨料取代率不同的6组配合比,检测各组试件质量损失和相对动弹模量,评判出最优配比,在Wiener退化过程的基础上进行可靠度建模并预测试件的剩余寿命.结果表明:在硫酸盐干湿循环过程中,动弹性模量和质量呈现先增加后下降的趋势.陶瓷粉代替部分再生细骨料能有效提高再生混凝土抗硫酸盐侵蚀能力;再生粗骨料的掺入导致试件性能大幅度下降且影响更为显著.当陶瓷粉、再生粗骨料的替代率为15%、0%时,陶瓷粉再生混凝土抗硫酸盐侵蚀能力最强,且试件经受178次硫酸盐干湿循环后完全失效.     

旱区陡坡黄土的低压力抗剪强度特性试验研究

黄土强度指标的准确确定是地质体稳定性分析的基础依据。以延安地区某边坡上更新统黄土为研究对象,利用改进的直剪仪,开展不同初始含水率、密度、干湿循环条件下原状与重塑黄土的低压力抗剪强度试验,研究原状与重塑黄土的低压力抗剪强度及抗剪强度指标随初始含水率、密度及干湿循环次数的变化规律,建立黄土的低压力抗剪强度指标与初始含水率、密度及干湿循环次数之间的函数关系。结果表明,黄土的低压力抗剪强度与初始含水率的关系可采用指数函数描述,与土体密度和干湿循环次数的变化关系可采用线性函数描述;原状与重塑黄土的黏聚力、内摩擦角与初始含水率之间呈负相关,且原状黄土抗剪强度指标高于重塑黄土;黄土的抗剪强度指标随土体密度增加而显著提高,随干湿循环次数增加呈明显减小趋势。     

雌激素对心肌Nrf2及GST和GCL的影响

雌激素的抗氧化作用与核转录相关因子(Nrf2)信号分子及其Nrf2/ARE信号路径有密切关系.Nrf2/ARE在调节机体抗氧化酶和Ⅱ相解毒酶的表达过程中起着非常重要的作用.利用原代培养心肌细胞缺氧/复氧模型,在雌激素预孵化条件下,研究了雌激素对谷胱甘肽-S-转移酶(GST)和谷氨酸半胱氨酸连接酶(GCL)表达及Nrf2核转移的影响,结果表明:雌激素能够明显增加GST和GCL的表达,促进Nrf2核转移.雌激素通过促进Nrf2入核及GST和GCL的表达发挥抗氧化作用.     

循环预剪作用对饱和粉土抗液化强度和孔压发展的影响

利用DDS-70微机控制电磁式振动三轴仪,在均等固结条件下,对黄河口饱和粉土进行了不同预剪应力和预剪振次的循环三轴试验,探讨了循环预剪作用对饱和粉土的抗液化强度和孔隙水压力发展规律的影响.试验结果表明:低水平的循环预剪应力不会使饱和粉土在循环预剪阶段破坏;随着循环预剪应力和循环预剪振次的增加,饱和粉土的抗液化强度得到提高,但当循环预剪振次达到某一值时,抗液化强度趋于稳定.对振动孔隙水压力比-振次比进行归一化后发现:无循环预剪作用时饱和粉土孔压发展模式为双曲线,有循环预剪作用时为反正弦曲线,反正弦曲线拟合参数较小,约为0.2～0.4.     

利用漂浮组织块获得高纯度兔成骨细胞的方法

建立了一种能够获得高纯度成骨细胞的简便可靠的培养方法,为成骨细胞的相关研究提供稳定的种子细胞来源。在常规培养方法的基础上进行改良,建立了组织块漂浮培养法,从新生兔颅骨分离、培养成骨细胞。观察细胞的生长状态及形态,绘制生长曲线并计算细胞倍增时间,并进行碱性磷酸酶染色和矿化能力的检测。该方法分离培养的细胞显示典型的成骨细胞生物学特性,获得的成骨细胞纯度高、性状稳定、增殖能力强。说明组织块漂浮培养法是一种可行的培养方法,能够为成骨细胞的相关研究提供稳定可靠的种子细胞来源。     

Effects of genistein, daidzein and glycitein on the osteogenic and adipogenic differentiation of bone marrow stromal cells and on the adipogenic trans-differentiation of osteoblasts

The 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT), alkaline phosphatase (ALP) activity and oil red O assays were used to examine the effects of genistein, daidzein and glycitein on the osteogenic and adipogenic differentiation of primary mouse bone marrow stromal cells (MSCs) and the adipogenic trans-differentiation of primary mouse osteoblasts. The results indicated that daidzein, genistein and glycitein at concentrations from 1×10^-8 mol/L to 1×10^-5 mol/L promoted the proliferation of MSCs and osteoblasts; genistein, daidzein and glycitein promoted osteogenic differentiation and inhibited adipogenic differentiation of MSCs, and inhibited adipocytic transdifferentiation of osteoblasts at appropriate concentrations as 17β-estradiol. It suggests that genistein, daidzein and glycitein regulate a dual differentiational process of MSCs into the osteogenic and adipogenic lineages, and trans-differentiational process of primary osteoblasts into the adipocyte lineages, causing a lineage shift toward osteoblast. Protective effects of them on bone may be mediated by a reversal of adipogenesis which may promote the proliferation, differentiation and mineralization of osteoblasts, and make adipocytes secrete less cytokines which may promote osteoclast formation and activation. In addition, the results also indicated that genistein, daidzein and glycitein may be helpful in preventing the development of steroid induced osteonecrosis.     

The Effect of Cyclic Stretching on Matrix Production, Mineralization and Differentiation of Osteoblasts

A four-point bending apparatus is used to investigate the effects of stretching on collagen synthesis, mineralization and differentiation of osteoblasts. Cells are stretched at 1500με for 24 hours. The responses of osteoblasts to mechanical signal of physiological stretching are evaluatedfrom three aspects: collagen production, extracellular inorganic calcium secretion and ALP activity. The results show that osteoblasts decrease the collagen synthesis, calcium secretion and ALP activity compared to the control cells (65.82 %, 73.51%, 48.10 96 respectively ), confirming that cyclic stretching at 1500με inhabits the ohvsiological activity of osteoblasts.     

Physiological responses of osteoblasts to cyclic stretching and the change of intracellular calcium concentration

The development of bone tissues is regulated by mechanical stimulation.Cyclic stretching was applied to the osteoblasts that were delivered from rat calvarie,The results showed that stretching at 500με increased the cell proliferation while loading at 1000με and 1500με inhabited cell growth ,Loading also increased the adhesive force between cells and substrate as well as spreading areas of osteobalsts.Furthermore,the fluorescence probe Fluo-3/AM was used to investigate the effect of stretching stimulation on the intracellular calcium concentration of osteoblasts.The intracellular calcium concentration of osteoblasts that were stretched at 500 με for 5 min was 92.9% higher than the control ,After being treated with the panax ontoginseng saponins,the streteched osteoblasts still expressed 28.6% higher intracellular calcium concentration than that of the control ,which proved that both the influx of extracellualr calcium and the release of intracellular calcium store were involved in the increase of intracellular calcium concentration when osteoblasts responded to the cyclic stretching And the influx of extracellular calcium through transmembrance channel played a main role.     

羟基聚磷酸钙钠骨细胞界面形态

研究一种新的骨替代材料－羟基聚磷酸钙钠骨细胞界面形态,评价其生物相容性。方法：采用体外细胞培养技术,选用ＳＤ乳鼠颅顶骨细胞,从光镜和扫描电镜两个层面观察细胞和材料之间的相互关系。结果随着培养时间的延长,细胞在材料表面的密度增加,未见细胞死亡。     

成骨细胞诱导骨髓基质细胞体外成骨的初步研究

目的:探讨在不使用细胞因子或化学药物的情况下,成骨细胞(Osteoblast,OB)与骨髓基质细胞(Bone Marrow Stromal Cells,BMSCs)混合培养时,成骨细胞提供的成骨微环境能否在体外诱导BMSCs向成骨细胞分化,并复合支架形成成熟的骨组织.研究成骨细胞诱导BMSCs有效成骨的最小比值(指成骨细胞与骨髓基质干细胞数量的比值).方法:SY别培养SD乳鼠的成骨细胞与SD大鼠的BMSCs,将成骨细胞和BMSCs以1:9、2:8、3:7、1:0的不同比例进行混合培养,通过测定第3、6、9天培养液上清中的碱性磷酸酶(ALP)的含量,研究成骨细胞促BMSCs有效成骨的最小比值.将两种细胞以该最小浓度比混匀接种于涂附Ⅰ型胶原壳聚糖材料支架上(直径9 mm,高3mm)作为混合培养组,相同终浓度的单纯成骨细胞和单纯BMSCs分别接种于相同支架作为阳性对照及阴性对照.另设置低比值成骨细胞对照组(仅含有共培养组中相同的成骨细胞数,但不含有共培养组中的BMSCs).全部标本均于体外培养8周后取材,通过大体观察、组织学及免疫组织化学等相关检测对新生骨进行评价.结果:成骨细胞和BMSCs以3:7的比例进行混合培养时已可实现有效成骨.3:7比例的混合培养组及阳性对照组(成骨细胞组)体外培养8周后大体观察和苏木素-伊红染色(HE)、ALP染色基本相同,均表达骨特异性细胞外基质Ⅰ型胶原,形成了较成熟的骨组织.阴性对照组(单纯BMSCs组)和低比值成骨细胞组,原细胞支架复合物变小、变形.低比值成骨细胞组在局部形成了少量的骨组织,阴性对照组(单纯BMSCs组)未能发现骨样组织形成.结论:在不使用细胞因子或化学药物的情况下,成骨细胞提供的成骨微环境能够在体外诱导BMSCs向成骨细胞分化并形成成熟的骨组织.混合细胞中成骨细胞与BMSCs的比例为3:7时是有效成骨的最小比值.     

Alternative splicing and expression of the insulin-like growth factor （IGF-1） gene in osteoblasts under mechanical stretch

Insulin-like growth factor 1 (IGF-1) promotes osteoblasts differentiation and bone formation, and its expression is induced by mechanical stretch, thus IGF-1 has been considered an effector molecule that links mechanical stimulation and local tissue responses. In this study, a mechanical stretching device was designed to apply physiological level static or cyclic stretching stimulation to osteoblasts. Different isoforms of IGF-1 mRNA were amplified by RT-PCR from the cells using respective primers and these amplified products were sequenced. An iso-form of IGF-1 splicing product was found to be selectively produced by osteoblasts under stretching stimulation. This IGF-1 isoform had identical sequence with the mechano growth factor (MGF) which was originally identified in muscle cells. Regulations of the expression of the liver-type IGF (L.IGF-1) and MGF in osteoblasts under stretch stimulation were further studied using semi-quantitative RT-PCR. Stretch stimulation was found to promot the expression of IGF-1 (L.IGF-1 and MGF), and for both isoforms expression was more effectively stimulated by cyclic stretch than static stretch. MGF was detected only in osteoblasts subjected to mechanical stretch, suggesting MGF was a stretch sensitive growth factor. Expression of MGF peaked earlier than that of L.IGF-1, which was similar to their regulation in muscle and suggested similar roles of MGF and L.IGF-1 in bone as in muscle cells. The functions of MGF and LIGF-1 in osteoblasts shall be established by further experimental studies.     

成骨细胞在纳米材料表面上粘附特性

在利用静电自组装技术制备的具备不同纳米形貌的S iO2纳米粒子薄膜及空白试片上,进行了SD大鼠成骨细胞的粘附特性考评实验,并对比了成骨细胞分别在纳米粒子基片和空白试片上的粘附数、细胞活性及形态的差异.实验结果表明:利用静电自组装技术获得的纳米结构表面对细胞的粘附特性确实存在良性影响;在所采用的3个纳米尺度下,细胞粘附数随纳米尺度的增加而增加,M TT检测结果表明细胞活性也随之增加.     

Behavior of MC3T3-E1 Osteoblast Cultured on Chitosan Modified with Polyvinylpyrrolidone

The physical and chemical properties of four kinds of modified chitosan materials made by blending chitosan with polyvinylpyrrolidone （PVP） were investigated. All four of these modified chitosan materials were hydrophilic with water contact angles ranging from 59°to 69°. Fourier transform-infrared spectra of the modified materials showed a new band at 1288 cm^-1, implying formation of a surface physical interpenetrating network structure. Enzyme linked immunosorbent assay results indicated that much less fibronectin was adsorbed on the modified materials than on only chitosan. The viability of MC3T3-E1 osteoblasts cultured on the materials was assessed by 3-（4,5-dimethyl-2-thiazolyl）-2,5-diphenyl- 2H-tetrazolium bromide assay. The results show that adding PVP10000 into the chitosan promotes adhesion of MC3T3-E1 osteoblasts on the modified materials, but has no effect on cell growth and proliferation; while adding PVP40000 reduces cell adhesion, growth, and proliferation. The results suggest that the increased hydrophilicity of the material surface does not always improve its biocompatibility, which will influence the selection and design of biomaterials.     

大鼠骨髓基质干细胞分离培养及向成骨、成脂诱导分化的研究

目的利用细胞原代及传代培养技术将大鼠骨髓基质干细胞诱导分化为成骨细胞和脂肪细胞,为其进一步应用奠定基础.方法全骨髓法分离大鼠骨髓基质干细胞,传代后分别在成骨、成脂诱导条件下继续培养,采用碱性磷酸酶染色、茜素红染色及油红"O"染色观察其成骨及成脂分化结果.结果第2代大鼠骨髓基质干细胞成骨诱导9 d后碱性磷酸酶染色呈阳性细胞,连续诱导14 d后可见矿化结节形成,成脂诱导14 d后可见脂肪细胞形成.结论随着诱导条件的不同,大鼠骨髓基质干细胞在体外可定向分化为成骨细胞和脂肪细胞.