Role of kisspeptin/GPR54 signaling pathways in prompting formation of precocious puberty

AIM: To explore the role of kisspeptin/GPR54 signaling pathway in the pathogenesis of precocious puberty based on female precocious puberty rat model induced by the single dose of danazol. METHODS: Female SD rats aged 3 d were randomly divided into normal group, vehicle group and model group. On the 5th day, the model rats were given a single subcutaneous injection of danazol with ethanol and ethylene glycol mixture. All rats were executed on 15 d, 25 d, 30 d, 35 d and 40 d, and the samples were collected to observe the sexual organ development. The levels of E₂, FSH and LH in peripheral blood were measured by ELISA. The Kiss-1 mRNA expression of Kiss-1, GPR54 and GnRH in hypothalamus was detected by real-time PCR. The kisspeptin expression in rat hypothalamus was observed by immunofluorescence. RESULTS: The time for puberty onset and sexual maturation in the model rats was significantly earlier than that in normal group and vehicle group. On days 25 and 30, the levels of peripheral sex hormones and uterine coefficients in model group were significantly higher than those in normal group and vehicle group. On days 25 and 30, the ovarian morphological development in the model rats was significantly earlier than that in normal group. On day 25, the mRNA expression of hypothalamic Kiss-1 and GnRH, and kisspeptin expression of hypothalamic arcuate nucleus in the model rats significantly increased compared with normal group and vehicle group. On day 30, kisspeptin expression of hypothalamic arcuate nucleus in the model rats decreased compared with normal group and vehicle group. On day 35, the mRNA expression of Kiss-1 and GnRH in the model rats decreased compared with normal group and vehicle group. The mRNA expression of GPR54 had no obvious difference among all groups. CONCLUSION: The Kiss-1 mRNA and kisspeptin expression in the model rats with precocious puberty is significantly increased in the hypothalamus during onset of puberty, suggesting that kisspeptin is an initiating factor for precocious puberty. Kisspeptin/GPR54 signaling pathway may play an important role in the occurrence of precocious puberty.     

Development of cell model of polymer/liquid crystal and effect of their elasticity on adhesion of rBM-MSCs

AIM: To develop the cell model of polymer/liquid crystal and to study the effect of their elasticity on the adhesion of rat bone marrow mesenchymal stem cells (rBM-MSCs). METHODS: Using the method of solvent evaporation induced phase separation, the cell model of polymer/liquid crystal was constructed. The surface morphology and phase separation structure were determined by polarized optical microscopy (POM), scanning electron microscopy (SEM) and small angle X-ray scattering (SAXS). rBM-MSCs were separated and expanded by adherent culture. The surface markers of rBM-MSCs were detected by flow cytometry. The cells were induced to osteogenic differentiation and adipogenic differentiation for 2 weeks. After 3 passages, the cells were divided into 4 groups, including total PU control group, 10% membrane group, 30% membrane group and 50% membrane group. The cells were then incubated with rhodamine phalloidin for cytoskeleton staining and were observed under the confocal laser scanning microscope after cultured for 24 h. RESULTS: The cell model of polymer/liquid crystal was constructed successfully using the method of solvent evaporation induced phase separation. Flow cytometry results showed that the rBM-MSCs positively expressed CD29, CD44 and CD90, and negatively expressed CD34 and CD45. After stained with alizarin red S and oil red O, the calcium nodule and lipid droplets in rBM-MSCs were observed obviously. The cytoskeleton staining result indicated that the area in total PU control group, 10% membrane group and 30% membrane group were greater, and the actin microfilaments were also clearer than that in 50% membrane group. CONCLUSION: The cell model with suitable content of liquid crystal made a contribution to the rBM-MSCs' adhesion, but too much liquid crystal inhibits cell adhesion.     

金银花和红银花挥发性成分的顶空固相微萃取#br# 气质联用检测与比较

 采用顶空固相微萃取气质联用（SPME–GC–MS）的方法检测红银花与金银花新鲜花蕾和 干花蕾的挥发性成分。4 种花蕾样品共鉴定出67 种挥发性物质,其中新鲜红银花与金银花分别检测出48 种和46 种,经烘干处理的干红银花与金银花分别检测出34 种和37 种。[3.2.1.0（1,5）]三环辛烷、β– 芳樟醇、5–甲基–2–己醇和以及十六烷是4 种样品共有的成分。其中红银花挥发性成分主要以烃类、 醇酮类为主,β–芳樟醇含量显著高于金银花;样品在由新鲜到烘干这一过程中,烃类和酯类挥发性物质 含量降低,醛类和醇酮类挥发性物质显著增加。     

不同菜豆品种4种抗营养因子水平差异分析

菜豆中植物凝集素、皂苷、胰蛋白酶抑制剂和植酸等抗营养因子是引起菜豆食物中毒的内源物。以56份食荚菜豆品种为材料,测定了鲜荚中4种抗营养因子含量或活性。结果表明,供试菜豆品种群体4种抗营养因子水平均存在极显著差异。植物凝集素含量(均值为1.743 mg·g~(-1))和胰蛋白酶抑制剂活性(均值为1.680 mg·g~(-1))变异系数均在100%以上,品种频率分布曲线主峰明显,但在极高和极低区域均有品种间断分布,出现了品种水平的数量等级差异;皂苷含量(均值为3.730 mg·g~(-1))和植酸含量(均值为3.102 mg·g~(-1))变异系数均低于41%,品种频率分布曲线呈近正态分布,主峰明显,双尾有低频率连续分布。相关分析表明,植物凝集素含量与胰蛋白酶抑制剂活性呈极显著正相关。聚类分析将供试56份菜豆品种划分为3个品种群,近80%品种聚于抗营养因子中等水平品种群,近12%品种居于较低水平。由于菜豆植物凝集素是食用致毒性的主要内源物,同时其和胰蛋白酶抑制剂等均与菜豆田间抗病虫性密切相关,预示着筛选出极低水平植物凝集素的菜豆品种是可行的,但同时可能会降低其田间抗病虫性。     

Effect of rhynchophylline on TGF-β1/Smad pathway for processing ventricular remodeling in spontaneously hypertensive rats

AIM: To investigate the effect of rhynchophylline (Rhy) on blood pressure, cardiac hypertrophy and myocardial fibrosis in spontaneously hypertensive rats (SHR). METHODS: Spontaneously hypertensive rats were randomly divided into model group, high dose (10 mg·kg^-1·d^-1) and low dose (2.5 mg·kg^-1·d^-1) group of rhynchophylline, captopril group (17.5 mg·kg^-1·d^-1). Wistar-Kyoto rats were used as normal control. Respectively, systolic blood pressure was measured by tail cuff every 2 weeks. After 10 weeks, heart weight index and left ventricular weight index were calculated. The myocardial hydroxyproline and plasma angiotensin Ⅱ were detected. Moreover, basic myocardial histopathological changes and myocardial collagen fibres were observed by HE staining and Masson staining, respectively. The protein expression of TGF-β₁ and Smad3 in the myocardium was measured by the methods of immunohistochemistry and Western blot. RESULTS: Compared with SHR model group, Rhy significantly reduced blood pressure (P<0.05), the levels of HYP in the myocardium (P<0.05) and the levels of AngⅡ in the plasma (P<0.01). The pathological damages of the myocardial tissues and collagen deposition were attenuated. The protein expression of TGF-β₁ and Smad3 was significantly reduced by the treatment with Rhy (P<0.01). CONCLUSION: Rhynchophylline reduces blood pressure and adjusts to improve ventricular remodeling of SHR. The mechanism may be involved in the TGF-β₁/Smad pathway and reducing AngⅡ content.     

Expression of Jagged2/Notch3 signaling molecules in pulmonary hypertensive rats induced by monocrotaline

AIM: To study the expression of Jagged2/Notch3 signaling molecules in pulmonary vascular wall of pulmonary hypertensive rats induced by monocrotaline. METHODS: SD rats were randomly divided into normal control group (C group,n=15), solvent control group (S group,n=15) and monocrotaline model groups (M group,n=15). The model of pulmonary hypertension was established by a single subcutaneous injection of monocrotaline (50 mg/kg). The rats in S group were given a single subcutaneous injection of the same dose of solvent. After 4 weeks, the pulmonary vascular remodeling was assessed by HE staining, and the mean pulmonary artery pressure (mPAP) and right ventricular systolic pressure (RVSP) were determined by right heart catheterization. The expression of Jagged2/Notch3/Hes5 molecules in the pulmonary vascular wall was detected by immunohistochemical method and real-time PCR. RESULTS: Compared with S group and C group, the percentage of medial wall thickness of smaller arteries in model group increased significantly (P<0.01). The levels of mPAP and RVSP in M group were significantly higher than those in S group and C groups (P<0.01). The results of real-time PCR showed that the expression of Jagged2, Notch3 and Hes5 was significantly increased in M group compared with S group and C group. The data from immunohistochemical detection indicated that Jagged2 mainly expressed in the intima of small lung artery, Notch3 and Hes5 mainly expressed in the medial smooth muscle cells. Compared with S group and C group, the expression of Jagged2 and Notch3 was significantly increased in the lung small arteries of M group. CONCLUSION: The activation of Jagged2/Notch3 signaling pathway might play an important role in the formation of pulmonary hypertension.     

Cre/loxP定位重组系统在植物雄不育和杂种优势中的利用研究

Cre—loxP是来源于噬菌体P1的一种位点特异性重组系统。目前，它已广泛应用于基因功能鉴定，动植物及微生物基因组的修饰以及基因的表达和调控。雄性不育系在杂种优势的利用中具有重要意义。本实验是以Cre—loxP系统调控细胞毒素基因barnase在植物雄性器官发育的特异时期在特异组织表达，从而达到控制植物育性的目的。1．在本实验中，通过PCR方法克隆了解淀粉芽孢杆菌Bacillus amyloliquefaciens的barnase基因，及其抑制因子barstar的DNA序列，同时，构建成功双元表达载体及基因枪转化载体，并已获得经分子险测呈阳性的小麦植株。2．osg6B是来源于水稻花药绒毡层的特异性的启动子，它在小孢子发育的四分体时期至单核花粉期在花药绒毡层中特异启动基因的表达，它具有较为严紧的时空特异性。以osg6B为启动子，构建了一组受控于Cre／loxP位点特异性重组系统的适用于基因枪法转化小麦的载体系统。其中，转不育基因(og6B—barnase)小麦植株生长正常，但开花后，花粉量明显减少，自交不能得到正常种子。进行花粉活力检测发现，转基因小麦大部分花粉表现为败育。花粉离体萌发试验表明，转不育基因小麦的花粉离体萌发率极低，几近败育。说明以osg6B驱动barnase在小麦花药中的表达可以导致雄性不育。3．以osg6B为启动子，构建了一组受控于Cre／loxP的用于控制双子叶植物育性的双元表达载体系统。以转cre基因的烟草作为受体，以含阻断序列的质粒农杆菌进行二次转化。二次转化株营养生长正常，但开花期花器官表现异常。具体表现为花瓣异常开裂或不开裂，花丝变短或粘连，花药提早萎蔫或异常开裂。推测这种现象的出现与启动子的组织特异性或barnase的渗露表达有关。4．二次转化株花粉染色因不同株系出现全部败育、部分不育和多数可育的情况。花粉离体萌发实验也得到了相似的结果。这说明通过筛选，利用Cre／loxP得到完全不育的植物株系是完全可行的。5．获得了9个barnase的突变体，初步推测第十四位氨基酸及第八十位氨基酸与核糖核酸酶活性有关。同时，发现barnase的第316位核苷酸序列为易突变位点，自然发生的突变多插入一个C，从而产生终止位点的改变。这或许会为今后barnase的克隆与利用提供一些参考。     

Similarities and differences between Amaranthus species and cultivars and estimation of outcrossing rate on the basis of electrophoretic separations of urea-soluble seed proteins

Wheat-rye translocation line (2BS/2RL) has been developed for resistance to biotype L of Hessian fly and agronomically useful traits. AFLP analysis using 64 primer pairs was conducted in order to identify 2RL-specific polymorphisms between “Coker 797” (non-2RL), near-isogenic line (NIL) carrying 2RL, and “Hamlet”. Nine primer combinations identified twelve reproducible polymorphic fragments in the NIL carrying 2RL. These twelve fragments were cloned and sequenced with an aim towards converting AFLP markers into sequence tagged sites (STS). A comparison of the 12 sequences with non-redundant accessions in the NCBI database using the BLAST search option indicated that one fragment of approximately 200 bp in length (amplified using primer combination E+AAC / M+CTA) was highly homologous with the rye-specific repetitive sequence R173-1 and Wis-2-lA, a retrotransposon-like element in wheat. Two STS primers (SJ07 and SJ09) out of twelve STS primer sets enabled the detection of polymorphisms between Coker 797 and NIL carrying 2RL. In order to verify whether the polymorphism detected by primers SJ07 and SJ09 was in fact the result of the presence of 2RL, additional plant material was examined. Amplified products of about 260 bp fragment with the SJ07 primer set were generated in rye cvs.“Chilbohomil” and “Jochunhomil”, triticale experimental line Suwon 15, and wheat experimental line K-14 (1AL/1RS & 2BS/2RL), as well as NIL carrying 2RL and Hamlet, but not in Coker 797 (non-2RL), “Keumgangmil” (non-translocation wheat), KS92WGRC17 (PI592729 /;/ 6BS/6BL-6RL), KS92WGRC19 (P1592731 /;/ 4BS/4BL-6RL), “TAM200” (1AL/1RS), and “Siouxland” (1BL/1RS). Our data suggest that primer set SJ07 amplifies a “2RL-specific” fragment of diagnostic value. This revised version was published online in August 2006 with corrections to the Cover Date.     

The Effect of Chromosome 1B/1R Translocation on the Yield Potential of Certain Spring Wheats (Triticum aestivum L.)

Nearly 50 percent of the 1988 advanced breeding lines of the CIMMYT bread wheat breeding program possess the 1B/1R homozygous translocation. Hence, a trial was conducted to estimate the effect of 1B/1R chromosome translocation on the yield potential of some of our high-yielding spring wheats, where non-limiting levels of fertility, moisture, preventive pest and disease programs were used. In conclusing the 1B/1R lines seemed to have increased their above-ground biomass yield, number of spikes per meter², 1000-grain weight and test weight. They also exhibited a slight advantage over the 1B homozygous lines on grain yield. The observed difference, however, was non-significant, as was the plant height difference observed among the two groups. Varietal comparisons indicated that the 1B/1R group headed later than the 1B group.     

STUDY ON SELF -SCANNING CIRCUIT OF LOW POWER DISSIPATION MOS IMAGE SENSOR

The paper studies a self- scanned circuit used in MOS image sensor with the demand of developing of high -order.low power disspation MOS image sensor and puts forward a kind of self -scanned circuit .which uses a three tramister dynamic nocomparing circuit whith a changing capacitor bootstrap circuit as MOS image sensor. The circuit adopted a silicon gote P -MOS technique, which is a high -speed low-dissipation dynamic no-coparing circuit. It is a practical unit circuit in the high -order array and solves the problem that the power dissipation of array rises with the increase of the array bite.