玉米大斑病菌Velvet基因家族生物信息学分析

采用生物信息学方法对玉米大斑病菌Velvet基因家族成员进行序列鉴定,并从理化性质、保守结构域、二级及三级结构以及功能域等方面进行预测和分析。结果表明,该家族各成员无信号肽结构。玉米大斑病菌Velvet家族基因与其他植物病原真菌Velvet家族基因具有较高同源性。该家族成员具有典型的Velvet超家族保守结构域。二级结构分析结果显示,该家族各成员蛋白质的结构元件均主要由无规则卷曲构成。家族成员可能通过可逆磷酸化在植物病原真菌的次生代谢中发挥作用。上述结果为进一步研究Velvet基因家族在植物病原真菌次生代谢中的作用打下了基础。

Combination of Supercritical Fluid Extraction with Ultrasonic Extraction for Obtaining Sex Hormones and IGF-1 from Antler Velvet

Supercritical CO2 (SC-CO2) extraction technology and ultrasonic technology were used to extract two active sex hormones, estradiol and progesterone, and insulin-like growth factor-1 (IGF-1) from antler velvet. The effects of SC-CO2 extraction condition on the extraction yield and content of sex hormones, the ultrasonic extrac-tion condition on the content of IGF-1 and the SC-CO2 extraction condition on the activity remaining of IGF-1 were studied. The optimal conditions were obtained. The experimental results showed that, in presence of 75% ethanol as the co-solvent, the mean yield and content of estradiol and progesterone were 87.67 pg·g-1 and 1224.10 pg·g-1, 12.38 ng·g-1 and 354.06 ng·g-1, respectively, with extraction pressure of 30 MPa, temperature of 35°C, extraction time of 30 min and CO2 consumption of 15 L·g-1 at the flow rate of 2.0 L·min-1. The highest content of IGF-1 was 7425.75 ng·g-1 antler velvet residue, when the pH10 ammonia-ammonium chloride buffer solution was used as the solvent, the ratio of solvent to sample was 20/1 (volume/mass), the extraction temperature was 0-35°C, and the ex-traction time was 4×15 min. Under these conditions, 93.68% activity remaining of IGF-1 in the residue was ob-tained, while little IGF-1 activity exists in traditional residue. The experimental results indicate that the technology of SC-CO2 with co-solvent is of advantage for getting high content sexual hormones and keeping high activity of IGF-1 in the residue, which can not be achieved by traditional extraction methods.

Determination of Sex Hormones in Antler Velvet by High Performance Liquid Chromatography Tandem Mass Spectrometry

Eighteen sex hormones in antler velvet were determined by high performance liquid chromatography tandem mass spectrometry.The solid phase extraction was applied to eliminating the matrix effect.The experimental conditions were examined and optimized.Under the optimal conditions,the proposed method provides the good linearities and determination limits（0.2―1.0 μg/kg） of the analytes investigated.The recoveries ranging from 72.3% to 149.5% were obtained for the target analytes at two concentration levels.This method was applied to the determination of eighteen sex hormones in different kinds of antler velvet samples and the obtained results are satisfactory.The results indicate that the proposed method is suitable for the determination of sex hormones in antler velvet samples.

ACE-I inhibitory peptide fractions from enzymatic hydrolysates of velvet bean (<i>Mucuna pruriens</i>)

The hydrolysis of velvet bean (Mucuna pruriens) protein in the presence of Alcalase?-Flavourzyme? and Pepsin-Pancreatin was investigated. The results showed that Alcalase?-Flavourzyme? (29.08%) sequential system catalyzed the hydrolysis most efficiently that Pepsin-Pancreatin (24.78%). In addition, the higher ACE-I inhibitory activity was achieved with the sequential system Alcalase?-Flavourzyme? (33.13%). Furthermore, the concentration of peptides employing an ultrafiltration (UF) system or their purification by gel filtration chromatography showed that the oligomeric peptides with lower molecular weight registered the highest ACE-I inhibitory activity. It has been demonstrated that Mucuna pruriens protein hydrolysates could serve as a source of peptides with ACE inhibitory activity and this activity can be attributed mainly to the mixture of short peptides in the hydrolysate.

Antibacterial Activities of <i>Psidium guajava</i>(Guava) and <i>Velvet tamarin</i>(Icheku) Local Chewing Sticks on <i>Streptococcus mutans</i>Isolated from Human Mouth

Globally dental diseases are mainly caused by Streptococcus mutans, it is one of the leading causative agents of dental caries worldwide, because of its resistance to conventional antibacterial agents, alternative therapies are used to control resistance of oral pathogens. This research was done to determine the antibacterial activities of Psidium guajava (guava) and Velvet tamarin (Icheku) chewing sticks on Streptococcus mutans isolated from the oral cavity. The study was conducted in Owerri Imo State Nigeria during November-December period. Phytochemical analysis of the plant extracts was done using appropriate techniques. The procedure used for antimicrobial susceptibility test was disk diffusion method. Serial dilutions of Psidium guajava (guava) and Velvet tamarind (Icheku) extracts were prepared, Muller-Hinton media was used to put together the extract of serial dilutions of Psidium guajava (guava) and Velvet tamarin (Icheku) and a microbiological procedure were used for visually determining the minimum inhibitory concentration as well as minimum bactericidal concentration. Phytochemical evaluation of the plants’ extracts revealed that it contains saponins, tannins, alkaloid, steriods, glycosides and phenol. The results obtained from the antibacterial susceptibility testing of the extracts against Streptococcus mutans showed that the zones of inhibition recorded ranged from 18 mm to 27 mm. Ethanol (Soxhlet) extract of Icheku twig showed no zone of inhibition on the isolated organism. The ethanol (soxhlet) extract of the individual Psidium guajava (guava) and Velvet tamarin (Icheku) has a better antibacterial effect when compared to their aqueous extracts and combined forms. Psidium guajava (Guava) and Velvet tamarin (Icheku) twigs are made up of composite that is active against S. mutans and can be used in oral hygiene. There is a need for further investigation on the plant extracts as the rural poor make use of it because it is cheap, readily available and the rich also use it once they are in the village mostly in Eastern Nigeria. Similarly chewing sticks has been reported to be practiced by 90 of rural population in Nigeria.

Therapeutic effects of velvet antler polypeptides on hepatic fibrosis in rats

OBJECTIVE To explore the therapeutic effects and underlying mechanisms of velvet antler polypeptides(VVAPs)in CCl4-induced experimental hepatic fibrosis in rats.METHODS Anti-hepatic fibrosis properties of VAPs were tested by Subcutaneous injection(SC)into male Wistar rats of CCl4- induced experimental hepatic fibrosis.After SC injections for 45 consecutive days at doses of 5mg·kg-1(low dose,VAPsL),10mg·kg-1(mid-dose,VAPsM)and 20mg·kg-1(high-dose,VAPsH),the rats were sacrificed and the various indicators were evaluated and tested.Observed hepatic cells degeneration and necrosis,inflammatory infiltration and levels of serum enzymes to assess treatment of VAPs;The expression levels of superoxide dismutase(SOD),glutathione peroxidase(GSH-Px),MDA,and hydroxyproline(HYP)in liver tissue were analyzed;RT-PCR analysis was carried out to detect the expression levels of matrix metalloproteinases2(MMP-2)and tissue inhibitor of metalloproteinases 1(TIMP-1)in liver tissue.RESULTS VAPs has obvious anti-hepatic fibrosis effects.Hepatocyte swelling,fatty degeneration was significantly reduced,reducing infiltration of inflammatory cells.Release of alanine aminotransferase(ALT)and aspartate aminotransferase(AST)decreased significantly,reduction of hyaluronic acid(HA)and laminin(LN)obviously,at the same time,the content of total protein and albumin increased significantly in serum.Activity of SOD and GSH-Px was significantly raised and the content of MDA and HYP was reduced significantly in liver tissue.Expression levels of MMP-2and TIMP-1 mRNA in liver were decreased significantly.These improvements were more significant in high-dos and mid-dose groups(P<0.05 or P<0.01 vs model group).CONCLUSION These findings suggest VAPs can significant treat the hepatic fibrosis,which may be due to protect liver cells and improve liver functions by hydroxyl radical scavenging activity and great effect of antioxidation,and decrease the gene expression of MMP-2,improving exist-environment of liver cells and decreasing the gene expression of TIMP-1,prompting degradation of extracellular matrix.

Velvet＇s multi-pulsed emission and multi-pulsed electron beams

The velvet electron emission characteristics and beams＇ brightness are investigated with a multi-pulsed mode. The results indicate that in the multi-pulsed mode the velvet emission is not uniform and the periphery emission is much stronger than that from the centre. The periphery emission contributes much more to the formation of the cathode plasma than the centre emission, which leads to diode impendence breakdown. The relationship between the cathode plasma expansion and the initial emittance of the cathode is deduced to describe the characteristics of the multi-pulsed vacuum diode. The emittance of the multi-pulsed beams is measured to be less than 1000mm·mrad. The brightness of the electron beams is better than 1× 10^8A/（m·rad）2.

Female Body Discourse in Tipping the Velvet

Being known as the lesbian-themed novel,Tipping the Velvet has always been analyzed with queer theory and Judith But?ler’s sexual performance.Female body discourse is a new perspective to analyze it with the combination of the text analysis and feminine writing put forward by Helene Cixous.Narrating the female body in a novel is a way to deconstruct the patriarchal society and to awaken women to fight against it.It is still of significance in 21st society where gender inequality and discrimination exist everywhere.

禾谷炭疽菌Velvet蛋白CgrVosA的生物学功能

【背景】禾谷炭疽菌(Colletotrichum graminicola)是侵染玉米的主要病原菌之一,每年造成严重的经济损失,直接威胁全球粮食安全。Velvet蛋白是真菌特有的一种保守性功能蛋白,它们在调控真菌生长及发育过程中发挥着重要作用。【目的】从禾谷炭疽菌中鉴定Velvet蛋白CgrVosA,并分析其生物学功能。【方法】利用同源重组的方法构建禾谷炭疽菌CgrvosA敲除突变株和互补菌株,通过测定其营养生长、胁迫因子响应、分生孢子产量及萌发、致病性等明确其功能。【结果】CgrvosA编码一个含有425个氨基酸的蛋白,有一个典型的Velvet蛋白结构域。与野生型菌株相比,敲除突变株ΔCgrvosA生长速度减慢,并且对NaCl和H_(2)O_(2)敏感;ΔCgrvosA卵圆形分生孢子的产量和萌发率显著降低,并且形态明显大于野生型菌株。【结论】禾谷炭疽菌Velvet蛋白CgrVosA参与调控菌株营养生长、对高渗和氧化应激的响应、卵圆形分生孢子的产生和萌发等过程。

Comparison of Structure and Biological Activity of Natural Polypeptide from Velvet Antlers of Cervus elaphus with Those of Synthesized Polypeptide

Biochemical techniques including ion-exchange chromatography, gel filtration chromatography and re- versed-phase high-performance liquid chromatography（RP-HPLC） were used to isolate and purify the natural poly- peptide from velvet antler（nVAP） of Cervus elaphus（C, elaphus）, which has a molecular weight of 3215.8 and the primary structure of VLSAADKSNVKAAWGKVGGNAPAFGAEALLRM. The homology of the protein sequence in nVAP with known protein sequence is less than 50%, suggesting that nVAP appears to be a new bioactive substance. At a level of 0.4--50 gg/mL, nVAP promotes mitosis in epidermal ceils, chondrocytes and NIH3T3 fibroblasts pri- marily cultured in a significant way. Given that a yield of high-purity nVAP isolated from C. elaphus is 0.001%, nVAP is artificially synthesized to prepare synthetic velvet antler polypeptide（sVAP） according to its primary struc- ture. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis（SDS-PAGE） of sVAP shows a single band, and its HPLC spectrum displays a single peak. The matrix-assisted laser desorption ionization time-of-flight mass spectro- metry（MALDI-TOF-MS） was used to identify sVAP to be of a molecular weight of 3200 and the consistency between primary structures of sVAP and nVAE Bioactivity test shows that at a dose of 5--40 μg/mL, sVAP promotes the pro- liferation of primarily cultured epidermal cells and NIH3T3 cell line. From the traditional Chinese medicine theory, velvet antler from Cervus nippon（C, nippon） and velvet antler from C. elaphus are considered as the same medicine, but differences between biochemical base and pharmacological effect of these two velvet antlers have been observed. We compared the total polypeptide mapping of the two velvet antlers, discovering that nVAP is active polypeptide and only exists in the velvet antler of C. elaphus, sVAP is similar to nVAP in physicochemical property and biological activity. These studies extend the possible utility of sVAP to be the promising compound to prepare velvet antler polypeptide of C. elaphus.

Velvet和LaeA蛋白调控药用真菌发育及活性成分合成相关的研究进展

药用真菌在中国传统医学中占有重要地位,但面临野生资源稀缺和药用活性成分含量低等问题。Velvet和LaeA蛋白在大多数真菌(低等壶菌类到高等担子菌类)中均存在,对真菌发育与活性成分合成调控具有重要作用,本文阐述了这两类蛋白的结构与功能。Velvet蛋白家族包括VeA(Velvet)、VelB(Velvet like B)、VosA(viability of spores A)和VelC(Velvet like C)4个成员,均具有真菌特异的Velvet结构域,在真菌有性生殖、无性生殖和次生代谢产物合成等方面发挥调控功能,其作用受到光、温度等环境因素影响。LaeA蛋白具有S腺苷甲硫氨酸(SAM)依赖的甲基转移酶结构域(MTD),通过光受体蛋白介导,调控及修饰Velvet蛋白家族,协同调控真菌的生长发育和次生代谢。因此,阐明药用真菌Velvet和LaeA蛋白作用机制对重要药用真菌资源培育及活性成分合成调控具有指导作用。

丝状真菌中的Velvet家族蛋白

丝状真菌是重要的微生物资源,能够产生大量的小分子活性化合物,如β-内酰胺类抗生素、夫西地酸、环孢霉素和他汀类药物等。这些小分子次级代谢产物的生物合成受全局性调控因子的调控,除丝状真菌中已发现的Lae A外,还存在一类具有Velvet结构域的特殊调控因子,它们在丝状真菌的生长发育和次级代谢调控中发挥着重要的作用。本文综述了丝状真菌Velvet家族蛋白(Ve A、Vel B、Vos A和Vel C)的结构特征及其对有性生殖、无性生殖和次级代谢产物合成等方面的调控作用,并探讨了可能的分子调控机制通路,为揭示次级代谢调控网络和激活沉默基因产生新型结构的化合物提供一定的参考价值。

Effects of velvet antler polypeptide on sexual behavior and testosterone synthesis in aging male mice

Twenty-four-month-old male C57BU6 mice with low serum testosterone levels were used as a late-onset hypogonadism （LOH） animal model for examining the effects of velvet antler polypeptide （VAP） on sexual function and testosterone synthesis. These mice received VAP for 5 consecutive weeks by daily gavage at doses of 100, 200, or 300 mg kg-1 body weight per day （n = 10 mice per dose）. Control animals （n = 10） received the same weight-based volume of vehicle. Sexual behavior and testosterone levels in serum and interstitial tissue of testis were measured after the last administration of VAP. Furthermore, to investigate the mechanisms of how VAP affects sexual behavior and testosterone synthesis in vivo, the expression of steroidogenic acute regulatory protein （STAR）, cytochrome P450 cholesterol side-chain cleavage enzyme （P450scc）, and 3β-hydroxysteroid dehydrogenase （3βHSD） in Leydig cells was also measured by immunofluorescence staining and quantitative real-time PCR. As a result, VAP produced a significant improvement in the sexual function of these aging male mice. Serum testosterone level and intratesticular testosterone （ITT） concentration also increased in the VAP-treated groups. The expression of STAR, P450scc, and 3β-HSD was also found to be enhanced in the VAP-treated groups compared with the control group. Our results suggested that VAP was effective in improving sexual function in aging male mice. The effect of velvet antler on sexual function was due to the increased expression of several rate-limiting enzymes of testosterone synthesis （STAR, P450scc, and 3β-HSD） and the following promotion of testosterone syothesis in vivo.

Biological effect of velvet antler polypeptides on neural stem cells from embryonic rat brain

Background Velvet antler polypeptides (VAPs), which are derived from the antler velvets, have been reported to maintain survival and promote growth and differentiation of neural cells and, especially the development of neural tissues This study was designed to explore the influence of VAPs on neural stem cells in vitro derived from embryonic rat brain Methods Neural stem cells derived from E12 14 rat brain were isolated, cultured, and expanded for 7 days until neural stem cell aggregations and neurospheres were generated The neurospheres were cultured under the condition of different concentration of VAPs followed by immunocytochemistry to detect the differentiation of neural stem cells Results VAPs could remarkablely promote differentiation of neural stem cells and most neural stem cells were induced to differentiate towards the direction of neurons under certain concentration of VAPs Conclusion Neural stem cells can be successfully induced into neurons by VAPs in vitro , which could provide a basis for regeneration of the nervous system

THE EVALUATION METHODS OF VELVET ANTLER AND THE DETERMINATION OF NUCLEOSIDES AND STEROID HORMONES IN DIFFERENT PARTS FROM SIKA DEER AND RED DEER

Velvet antler is an important and precious traditional Chinese animal medicine,and was used for strengthening the kidney,anti-fatigue,improving sexual function and so on.But the evaluation methods of velvet antler and the scientific evidence for the use of antler is insufficient.Materials and Vacuum freeze drier was used for dehydration of the fresh antler at-60.,6 pa for 24 h.

鹿茸多肽对顺铂所致卵巢早衰大鼠卵巢功能的保护作用及其机制

目的:探讨鹿茸多肽(VAP)对顺铂(DDP)致卵巢早衰模型大鼠的影响及机制。方法:取动情周期正常的雌性大鼠并将其随机分为对照组、模型组、VAP干预组,每组各10只。模型组和VAP干预组通过连续20 d腹腔注射DDP(4 mg·kg^(−1)·d^(−1))建立卵巢早衰大鼠模型,干预组同时连续20 d灌胃VAP(400 mg·kg^(−1)·d^(−1))。HE染色观察卵巢形态学表现,并对各级发育卵泡计数。ELISA检测血清雌二醇(E2)、促卵泡生成素(FSH)、抗穆勒管激素(AMH)水平。生化法检测卵巢组织中谷胱甘肽(GSH)和丙二醛(MDA)水平。探针标记法检测卵巢组织中活性氧(ROS)水平。普鲁士蓝染色观察卵巢组织中铁蓄积情况并检测亚铁离子(Fe^(2+))水平。免疫组化和Western blot检验卵巢组织中溶质载体家族7成员11(SCL7A11)、谷胱甘肽过氧化酶4(GPX4)和酰基辅酶A合成酶长链家族成员4(ACSL4)的表达。通过生育力监测观察各组大鼠生育能力。结果:与对照组相比,模型组卵巢萎缩,铁蓄积(P<0.01),各级发育卵泡和总卵泡减少(P<0.05),闭锁卵泡增多(P<0.01),产仔数减少(P<0.01),血清E2、AMH水平降低(P<0.01),FSH水平提高(P<0.01),卵巢组织中GSH水平、SCL7A11和GPX4蛋白表达均降低(P<0.05),Fe^(2+)、MDA和ROS水平及ACSL4蛋白表达均升高(P<0.01)。与模型组相比,VAP能改善DDP导致的上述指标改变(P<0.01)。结论:鹿茸多肽可改善顺铂致卵巢早衰大鼠的卵巢功能、维持正常的卵泡发育并提高其生育能力,机制可能与脂质过氧化反应和铁死亡有关。

20世纪50—60年代外销绒织物设计特征探微

20世纪50—60年代是中国丝绸新品种、新花样开发设计的高峰时期之一,外销绒织物设计在这个时期也获得了长足的进步。本文以上海丝绸进出口公司刊印的《新品种》系列丝绸样本为基本线索,围绕不同类型绒织物的设计观念、设计方法、工艺特征等展开探索和研究。着重对经起绒织物、纬起绒织物、素绒织物上的印花、雕绒及静电植绒织物的经典样本进行起绒工艺、纹样设计、艺术特征的分析,探讨此时期绒织物发展的历史价值和现实意义。

莱赛尔长丝/棉交织面料的原纤化染整工艺研究

设计了一款莱赛尔长丝与棉交织并适合原纤化的家纺面料,同时研究了此面料的两种原纤化染整工艺流程。染色前进行原纤化控制生产的面料颜色饱和度高、布面精致平整;染色后进行原纤化控制生产的面料颜色饱和度低、布面微皱有怀旧效果。且两种原纤化染整工艺生产的面料绒感细腻、表面光洁、手感蓬松、触感微凉。

鹿茸多肽对骨质疏松模型大鼠的改善作用及对SIRT1/FOXO1信号通路的影响

目的:探讨鹿茸多肽(VAP)对骨质疏松(OP)模型大鼠的保护作用,并阐明其可能的作用机制。方法:60只12周龄SD大鼠随机分为对照组、模型组、阳性药组(1 mg·kg^(-1)·d^(-1)阿仑膦酸钠灌胃)、低剂量(100 mg·kg^(-1)·d^(-1))VAP组、中剂量(200 mg·kg^(-1)·d^(-1))VAP组和高剂量(300 mg·kg^(-1)·d^(-1))VAP组,每组10只。除对照组外,其余各组大鼠肌肉注射地塞米松(2 mg·kg^(-1))复制OP大鼠模型,对照组大鼠肌肉注射等体积生理盐水,每周2次,连续11周。双能X射线骨密度仪检测各组大鼠股骨骨密度(BMD),酶联免疫吸附试验(ELISA)法检测各组大鼠血清中血钙(Ca^(2+))、血磷(P)、骨保护素(OPG)、碱性磷酸酶(ALP)和骨钙素(OCN)水平,生化法检测各组大鼠血清中丙二醛(MDA)水平和超氧化物歧化酶(SOD)活性,HE染色观察各组大鼠骨组织病理形态表现,Western blotting法检测各组大鼠骨组织中沉默信息调节因子1(SIRT1)、过氧化氢酶(CAT)、Runt相关转录因子2(RUNX2)和叉头框蛋白O1(FOXO1)蛋白表达水平。结果:与对照组比较,模型组大鼠股骨BMD明显降低(P<0.05);与模型组比较,阳性药组、中剂量VAP组和高剂量VAP组大鼠股骨BMD明显升高(P<0.05或P<0.01)。与对照组比较,模型组大鼠血清中Ca^(2+)、P和OPG水平及SOD活性明显降低(P<0.05),ALP、OCN和MDA水平明显升高(P<0.05);与模型组比较,低剂量VAP大鼠血清中OPG水平明显升高(P<0.05),阳性药组、中剂量VAP组和高剂量VAP组大鼠血清中Ca^(2+)、P和OPG水平及SOD活性明显升高(P<0.05或P<0.01),阳性药组和各剂量VAP组大鼠血清中ALP、OCN和MDA水平明显降低(P<0.05或P<0.01)。HE染色,与对照组比较,模型组大鼠骨组织中骨细胞数量减少且排列混乱,骨小梁纤细,出现大片断裂,髓腔扩大;与模型组比较,阳性药组、中剂量VAP组和高剂量VAP组大鼠骨组织中骨小梁粗壮,排列紧密。Western blotting法,与对照组比较,模型组大鼠骨组织中SIRT1、CAT、RUNX2和FOXO1蛋白表达水平明显降低(P<0.05);与模型组比较,阳性药组、中剂量VAP组和高剂量VAP组大鼠骨组织中SIRT1、CAT、RUNX2和FOXO1蛋白表达水平明显升高(P<0.05或P<0.01)。结论:VAP对OP模型大鼠具有保护作用,其作用机制可能与介导SIRT1/FOXO1信号通路抗氧化应激作用有关。

开埠城市文化遗产的艺术演变探析——以烟台绒绣为例

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