Combination of bone tissue engineering and BMP-2 gene transfection promotes bone healing in osteoporotic rats

OBJECTIVE: The aim of this study was to develop a feasible approach to promote bone healing in osteoporotic rats using autogenous bone tissue-engineering and gene transfection of human bone morphogenetic protein 2 (hBMP-2). METHODS: Bone marrow stromal cells (BMSCs) from the left tibia of osteoporotic rats were transfected with the hBMP-2 gene in vitro which was confirmed by immunohistochemistry, in situ hybridization and Western blotting. Autogenous transfected or untransfected BMSCs were seeded on macroporous coral hydroxyapatite (CHA) scaffolds. Each cell-scaffold construct was implanted into a defect site which was created in the ramus of the mandible of osteoporotic rats. Four or eight weeks after implantation in situ hybridization was performed in BMSCs transfected with hBMP-2, X-ray examinations, histological and histomorphological analyses were used to evaluate the effect of tissue-engineered bone on osseous defect repair. RESULTS: Newly formed bone was observed at the margin of the defect 4 weeks after implantation with BMSCs transfected with BMP-2. Mature bone was observed 8 weeks after treatment. In the control group there was considerably less new bone and some adipose tissue was observed at the defect margins 8 weeks after implantation. CONCLUSIONS: Autogenous cells transfected with hBMP-2 promote bone formation in osteoporotic rats. BMSC-mediated BMP-2 gene therapy used in conjunction with bone tissue engineering may be used to successfully treat bone defects in osteoporotic rats. This method provides a powerful tool for bone regeneration and other tissue engineering.     

High-resolution nonlinear optical imaging of live cells by second harmonic generation

By adapting a laser scanning microscope with a titanium sapphire femtosecond pulsed laser and transmission optics, we are able to produce live cell images based on the nonlinear optical phenomenon of second harmonic generation (SHG). Second harmonic imaging (SHIM) is an ideal method for probing membranes of living cells because it offers the high resolution of nonlinear optical microscopy with the potential for near-total avoidance of photobleaching and phototoxicity. The technique has been implemented on three cell lines labeled with membrane-staining dyes that have large nonlinear optical coefficients. The images can be obtained within physiologically relevant time scales. Both achiral and chiral dyes were used to compare image formation for the case of single- and double-leaflet staining, and it was found that chirality plays a significant role in the mechanism of contrast generation. It is also shown that SHIM is highly sensitive to membrane potential, with a depolarization of 25 mV resulting in an approximately twofold loss of signal intensity.     

外源基因表达造成的群体感应细菌生存压力的建模分析

外源基因的表达及其对细菌种群的影响对于群体感应系统和合成生物学产业的研究具有重要意义。然而,人们对于表达外源蛋白的细菌本身的行为模式仍然知之甚少。为了研究菌落生长和外源基因表达的过程究竟受到哪些因素的影响,文中测量了受Lux类受体调控的外源基因在N-酰基高丝氨酸内酯 (N-acyl homoserine lactone,N-AHL) 信号分子诱导下的表达,并模拟了其对细菌种群动态的影响。文中建立了一个假设性的数学模型,对信号分子诱导表达下细菌种群生长受影响的现象进行了分析。先前的研究通常将细菌种群生长受群体感应系统影响的现象归咎于合成群体感应信号分子的消耗与N-AHL信号分子的毒性,文中提供了对于这种生存压力的另一种可能的解释。     

3D-QSAR studies of checkpoint kinase 1 inhibitors based on molecular docking and CoMFA

Three-dimensional quantitative structure–activity relationship (3D-QSAR) studies were performed on a series of substituted 1,4-dihydroindeno[1,2-c]pyrazoles inhibitors, using molecular docking and comparative molecular field analysis (CoMFA). The docking results from GOLD 3.0.1 provide a reliable conformational alignment scheme for the 3D-QSAR model. Based on the docking conformations and alignments, highly predictive CoMFA model was built with cross-validated q ² value of 0.534 and non-cross-validated partial least-squares analysis with the optimum components of six showed a conventional r ² value of 0.911. The predictive ability of this model was validated by the testing set with a conventional r ² value of 0.812. Based on the docking and CoMFA, we have identified some key features of the 1,4-dihydroindeno[1,2-c]pyrazoles derivatives that are responsible for checkpoint kinase 1 inhibitory activity. The analyses may be used to design more potent 1,4-dihydroindeno[1,2-c]pyrazoles derivatives and predict their activity prior to synthesis.     

海洋芽孢杆菌（Bacillus marinus）B-9987菌株抑制病原真菌机理的研究

摘要：【目的】：探讨海洋芽孢杆菌（Bacillus marinus）B-9987菌株的代谢产物BMME-1,对植物病原真菌茄链格孢菌的抑菌作用机理。【方法】分别使用分光光法、气相色谱-质谱GC-MS联用技术、红外光谱法等,检测了BMME-1处理病原真菌后,菌体渗透性、细胞壁及细胞膜成份的变化。【结果】BMME-1对茄链格孢菌的抑菌中浓度（MIC50）为6.2 mg/L,最小杀菌浓度（MFC）为50 mg/L,在MIC50浓度或高于此浓度处理靶标菌,将导致菌体蛋白质、核酸等大分子物质的外流;处理菌株葡聚糖结     

桃ERF转录因子家族生物信息学分析及 芽萌发相关基因筛选

以中油四号油桃(Prunus persica var. nectarina)为研究对象, 利用MEGA 6.0、MEME、GSDS和DNAMAN 6.0等软件对桃ERF家族数据进行生物信息学分析, 鉴定得到102个ERF转录因子家族基因, 并通过构建系统进化树将这102个基因分为10个子家族(I-X)。基因结构分析表明, 有81个基因不含内含子, 20个基因含有1个内含子, 有1个基因与其它成员差异较大, 含有5个内含子。保守元件分析表明, ERF家族包含20个保守元件, 其中Motif 1、Motif 2和Motif 4都属于AP2/ERF结构域, 同一个保守元件主要出现在同一个子家族中, 并且大部分保守元件的功能未知。VIII子家族基因的荧光定量PCR分析表明, 在桃叶芽处于不同的发育状态时, PpeERF068的表达量存在较大差异, 光照培养箱中培养的桃芽在萌发过程中各时期表达量变化趋势进一步表明该基因可能与叶芽萌发有关, 将其命名为PpeEBB1。该研究为进一步揭示PpeEBB1的分子机制奠定了基础, 并为桃树的栽培管理和熟期调控了提供理论指导。     

与链霉菌发育分化有关的启动子——PTH4的调控研究

依赖于天蓝色链霉菌分化关键基因——whiG的发育调控启动子(P_TH4)所控制的下游基因被克隆了,用双脱氧链终止法进行了双链测序.结果表明在1597bp的DNA片段中含有一个完整的开读框架(ORF).在计算机的蛋白文库比较中未找到与该基因产物同源的已知蛋白,可能是一个新的蛋白产物.用基因破坏的策略初步研究了该基因的生物学功能,发现该基因的破坏影响了放线紫红素的产生,即与天蓝色链霉菌放线紫红素的生物合成有关.这进一步证明启动子——P_TH4在链霉菌分化中的多级调控作用.     

链霉菌分化调控启动子——PTH4和PTH270的体内转录研究

天蓝色链霉菌分化调控启动子P_TH4和P_TH270被分别亚克隆到链霉菌启动子探针载体pIJ4083后,构建的重组质粒被命名为pIJ4470和pIJ4471.当pIJ4470和pIJ4471转化天蓝色链霉菌的白色分化阻断突变株(C85,C70,C71,C17和C119)后,通过pIJ4083上儿茶酚加氧酶报告基因的表达可知启动子的活性.从构建的重组菌株中进行了总RNA的提取.用同位素标记P_TH4和P_TH270的5’-端制备成了探针,以不同来源的RNA为模板与制备的探针分别进行了DNA-RNA杂交.S1 mapping的结果表明,来自C85/pIJ4470,C85/pIJ4471,C70/pIJ4470,C70/pIJ4471及C17/pIJ4470,C17/pIJ4471菌株的RNA杂交后都给出了较强的阳性杂交信号,而来自C71/pIJ4470,C71/pIJ4471菌株的RNA杂交未出现阳性信号,来自C119/pIJ4470与C119/pIJ4471菌株的RNA杂交后有弱的信号.上述结果表明启动子P_TH4和P_TH270的转录依赖于链霉菌分化关键基因whiG,部分依赖于分化基因whiH,而不依赖于分化基因whiA,whiB及whiI.     

Identification of the cardiac and circulating form of atriopeptin in rabbit

Analysis of peptides purified from high and low molecular weight fractions of rabbit atrial extracts indicates that the sequence of the first 30 residues of rabbit atriopeptigen exhibits 80% homology with the rat peptide, and that the low molecular weight rabbit peptide (28 residues) is identical to rat atriopeptin 28 (AP 28). The effects of infused 1-deaminoarginine8-vasopressin (dAVP) and phenylephrine, volume expansion, and water immersion on AP release into the circulation of the rabbit was studied. Neither dAVP, nor water immersion elevated right atrial pressure (RAP) or plasma AP levels in the anesthetized rabbits. Phenylephrine induced a sustained increase in systemic blood pressure and right atrial pressure which was accompanied by elevated plasma AP immunoreactivity which appeared to be identical to rat AP-28 on HPLC. There is obviously a preferential conservation of the AP sequence, since the C-terminal peptide is exactly the same in rabbit, rat and mouse and differs from human, dog, cow and pig only by the single substitution of an isoleucine for a methionine residue.     

北京地区红隼的迁徙路线和活动区域

红隼(Falco tinnunculus)被列为国家二级重点保护野生动物,是能同时适应农村和城市环境的小型猛禽,对维持城市生态系统稳定具有重要意义。2022年4月至7月,为在北京救助的7只红隼佩戴了卫星追踪器,追踪其活动轨迹,依据追踪的动物活动位点数据,采用净平方位移-时间曲线依次对各红隼的迁徙模式进行了判别,深入分析了迁徙红隼的迁徙时间、距离和路线等,并采用核心密度法分别计算了迁徙及留居型红隼95%及50%活动区面积。研究结果表明,在北京地区红隼的迁徙模式为部分迁徙,追踪的7只红隼个体(N01 ~ N07)中,4只为留鸟,1只为迁徙鸟,2只居留类型无法准确判断。N01为迁徙红隼,其度夏地和越冬地分别在内蒙古锡林郭勒盟和河北廊坊,此红隼秋季迁徙速度明显高于春季,其春季迁徙距离551 km,历时25 d,平均迁徙速度为22 km/d,而秋季迁徙距离412 km,历时2 d,平均迁徙速度为203 km/d,河北承德滦平县是其春季迁徙的重要中途停歇地。不同红隼个体间95%及50%活动区面积均存在较大差异,迁徙红隼N01 95%、50%活动区面积在度夏区分别为93.10 km²、17.50 km²,在越冬区分别为7.03 km²、0.99 km²;留居型红隼95%、50%活动区面积均值分别为1 165.34 km²、178.71 km²(n = 4),其中最大95%、50%活动区面积分别为4 320.26 km²(N02)、648.22 km²(N02),最小95%、50%活动区面积分别为2.80 km²(N03)、0.29 km²(N03)。本研究揭示了北京地区红隼的迁徙模式、迁徙路线、重要停歇地及活动区状况,为红隼的针对性保护和管理提供了科学依据。