叶面喷施PP_(333)和6-BA对甜荞花芽分化及内源激素的影响

以甜荞品种‘北早生’和‘赤甜荞1号’为材料,利用多效唑(PP_(333))和6-苄基腺嘌呤(6-BA)在不同幼苗时期进行叶片喷施处理,并采用扫描电镜观察花芽分化过程,记录开花时间,调查开花和结实数,通过液质联用方法检测3种内源激素生长素(IAA)、赤霉素(GA_3)、脱落酸(ABA)及人工合成细胞分裂素(6-BA)含量,探究两种植物生长调节剂对甜荞的花芽分化及内源激素含量的影响,为甜荞花期调控提供依据。结果显示:(1)第二片真叶期喷施100mg·L~(-1)PP_(333)和150mg·L~(-1)6-BA能显著提高甜荞结实率和单株产量,两种处理的结实率分别较对照提高49.5%、39.4%,单株产量分别提高41.8%、23.0%。(2)观察甜荞花芽分化过程并将其划分5个时期,分别为生长锥分化前期、生长锥分化期、小花原基分化期、雌雄蕊分化期和雌雄蕊形成期,在第二片真叶期喷施PP_(333)和6-BA均加快了甜荞花芽分化进程,使其开花时间提前。(3)喷施PP_(333)在幼苗期和现蕾期降低了内源IAA、6-BA含量,增加GA_3、ABA含量,而在盛花期增加内源IAA、6-BA含量,降低ABA、GA_3含量;喷施6-BA在幼苗期和现蕾期增加了内源GA_3、IAA含量,在盛花期降低了内源GA_3、IAA含量,3个时期均降低了内源ABA含量,增加了6-BA含量。研究发现,第二片真叶期是调控甜荞小花发育的关键时期,在此时叶面喷施100mg·L~(-1)PP_(333)和150mg·L~(-1)6-BA的调控效果最佳,可加快甜荞花芽分化进程,使开花时间提前,增加单株结实粒数,提高甜荞品种的结实率和单株产量;PP_(333)主要是通过减少甜荞开花数,增强弱势小花活力来提高结实粒数,而6-BA主要是利用增加开花数,提高有效可孕小花数来增加结实粒数。     

Synthesis,in vitro stability,and antiproliferative effect of d‐cysteine modified GnRH‐doxorubicin conjugates

Overexpression of gonadotropin‐releasing hormone (GnRH) receptor in many tumors but not in normal tissues makes it possible to use GnRH analogs as targeting peptides for selective delivery of cytotoxic agents, which may help to enhance the uptake of anticancer drugs by cancer cells and reduce toxicity to normal cells. The GnRH analogs [d ‐Cys⁶, desGly¹⁰, Pro⁹‐NH₂]‐GnRH, [d ‐Cys⁶, desGly¹⁰, Pro⁹‐NHEt]‐GnRH, and [d ‐Cys⁶, α‐aza‐Gly¹⁰‐NH₂]‐GnRH were conjugated with doxorubicin (Dox), respectively, through N‐succinimidyl‐3‐maleimidopropionate as a linker to afford three new GnRH‐Dox conjugates. The metabolic stability of these conjugates in human serum was determined by RP‐HPLC. The antiproliferative activity of the conjugates was examined in GnRH receptor‐positive MCF‐7 human breast cancer cell line by MTT assay. The three GnRH‐Dox conjugates showed improved metabolic stability in human serum in comparison with AN‐152. The antiproliferative effect of conjugate II ([d ‐Cys⁶, desGly¹⁰, Pro⁹‐NHEt]‐GnRH‐Dox) on MCF‐7 cells was higher than that of conjugate I ([d ‐Cys⁶, desGly¹⁰, Pro⁹‐NH₂]‐GnRH‐Dox) and conjugate III ([d ‐Cys⁶, α‐aza‐Gly¹⁰‐NH₂]‐GnRH‐Dox), and the cytotoxicity of conjugate II against GnRH receptor‐negative 3T3 mouse embryo fibroblast cells was decreased in comparison with free Dox. GnRH receptor inhibition test suggested that the antiproliferative activity of conjugate II might be due to the cellular uptake mediated by the targeting binding of [d ‐Cys⁶‐des‐Gly¹⁰‐Pro⁹‐NHEt]‐GnRH to GnRH receptors. Our study indicates that targeting delivery of conjugate II mediated by [d ‐Cys⁶‐des‐Gly¹⁰‐Pro⁹‐NHEt]‐GnRH is a promising strategy for chemotherapy of tumors that overexpress GnRH receptors.     

高粱LEA基因家族的鉴定及表达分析

有研究表明,干旱、低温和盐等环境胁迫能够诱导LEA基因的表达。为了探索LEA基因家族在高粱响应外界刺激过程中起到的作用,本研究通过生物信息学的方法对LEA基因家族在高粱全基因组水平进行鉴定和分析,于高粱全基因组中共鉴定出35个基因家族成员,不均匀地分布于高粱8条染色体上,结合系统进化树和保守结构域分析结果,将高粱LEA基因家族成员分为7组。亲水性分析和结构无序性预测表明高粱LEA蛋白绝大多数为亲水性且结构无序。基因结构分析显示了各分组基因结构上的保守性。高粱LEA基因的启动子分析发现了一些与激素和非生物胁迫响应相关的顺式作用元件。对激素和干旱胁迫下高粱LEA基因的表达分析发现外界胁迫能够诱导部分高粱LEA基因的表达。     

Extracellular vesicles from thyroid cancer harbor a functional machinery involved in extracellular matrix remodeling

Extracellular vesicles (EVs) participate in cell-stroma crosstalk within the tumor microenvironment and fibroblasts (Fb) contribute to tumor promotion in thyroid cancer. However, the role of tumor-stroma derived EVs still needs to be deciphered. We hypothesized that the interaction of thyroid tumor cells with Fb would liberate EVs with a specific proteomic profile, which would have an impact on EV-functionality in thyroid tumor progression-related events. Tumor (TPC-1, 8505c) and non-tumor (NThyOri) thyroid cells were co-cultured with human Fb. EVs, obtained by ultracentrifugation of conditioned media, were characterized by nanoparticle tracking analysis and western blotting. EV-proteomic analysis was performed by mass-spectrometry, and metalloproteinases (MMPs) were studied by zymography. EV-exchange was evaluated using immunofluorescence, confocal microscopy and FACS. EVs expressed classical exosome markers, with EVs from thyroid tumor cell-Fb co-cultures showing a proteomic profile related to extracellular matrix (ECM) remodeling. Bidirectional crosstalk between Fb and TPC-1 cells produced significantly more EVs than their isolated cells, and potentiated EV-functionality. In line with this, Fb-TPC-1 derived EVs induced MMP2 activation in NThyOri supernatants, and MMP2 activity could be evidenced in Fb and TPC-1 contact-independent co-cultures. Besides, MMP2 interactors allowed us to discriminate between EVs from thyroid tumoral and non-tumoral milieus. Interestingly, Fb internalized more EVs from TPC-1 than from NThyOri producing cells. Fb and thyroid tumor cell crosstalk produces specialized EVs with an ECM remodeling proteomic profile, enabling activation of MMP2 and possibly facilitating ECM-degradation, which is potentially linked with thyroid tumor progression.     

外源保幼激素对异色瓢虫卵巢发育及生殖相关基因转录水平的影响

【目的】本研究旨在明确外源保幼激素(juvenile hormone, JH)对异色瓢虫Harmonia axyridis成虫卵巢发育以及生殖信号通路中关键基因转录水平的影响。【方法】以萝卜蚜Lipaphis erysimi饲喂的异色瓢虫雌成虫为空白对照组,以点滴同体积丙酮的雌成虫为溶剂对照组,对人工饲料饲喂的羽化后第2天的雌成虫点滴不同剂量(80, 120和160 ng/头)JHⅢ 1, 3, 5, 7和9 d后,解剖成虫卵巢,拍照并测量卵巢长度及其第一卵室的长度和宽度。利用qPCR分析在最适JHⅢ剂量(120 ng/头)处理后1, 5和9 d的异色瓢虫雌成虫生殖信号通路中关键基因JH受体methoprene-tolerant (Met)基因、krüppel homolog 1(Kr-h1)基因、卵黄原蛋白(vitellogenin, Vg)基因(Vg1和Vg2)和卵黄原蛋白受体(vitellogeninreceptor, VgR)基因表达水平。【结果】与溶剂对照组相比,点滴80和120 ng/头剂量JHⅢ可促进异色瓢虫成虫卵巢发育,其中120 ng/头剂量JHⅢ处理羽化后第2天雌成虫后7 d卵巢中出现大量卵黄沉积,与萝卜蚜饲喂的空白对照组卵巢发育状态一致,卵巢长度以及第一卵室的长度和宽度均显著高于溶剂对照组的;160 ng/头剂量JHⅢ处理则抑制卵巢发育。qPCR结果表明,120 ng/头剂量JHⅢ处理后5和9 d雌成虫中HaMet, HaKr-h1, HaVg1, HaVg2和HaVgR基因表达量较对照组(丙酮处理组)均增加,其中处理后5 dHaMet, HaVg1和HaVgR基因表达量分别为对照组的2.78, 13.14和8.28倍,处理后9 d分别为对照组的2.36,1.85和2.01倍,差异极显著。【结论】120 ng/头JHⅢ可促进异色瓢虫成虫卵巢发育,上调处理后5和9 d时雌成虫HaMet, HaKr-h1, HaVg和HaVgR基因表达量;推测这些关键基因在保幼激素调控异色瓢虫卵巢发育及卵黄形成过程发挥重要作用。     

Synthesis and structure–activity relationship of 16,17-modified gibberellin derivatives

Gibberellins (GAs) are a group of diterpenoid plant hormones that control plant growth and development at various stages. Biologically active GAs share the common structures of a 3β-hydroxy group, a carboxy group at C-6, and a γ-lactone between C-4 and C-10. Hydroxylation at C-2β is a major deactivation step in many plant species, and hydroxylation at C-13 has been shown to weaken the binding affinity of GAs to their receptor proteins. In rice, bioactive GA₄ has also been shown to be deactivated through 16α,17-epoxidation. Moreover, 16,17-dihydro-16α,17-dihydroxy GA₄ has been identified as an aglycon of its glucoside from rice. However, our knowledge on the biological activity of 16,17-epoxidized GAs is currently limited to 16,17-dihydro-16α,17-epoxy GA₄. Moreover, the bioactivity of 16,17-dihydro-16α,17-dihydroxy GA₄ remains unknown. Here, we synthesized 16,17-epoxidized or dihydroxylated GA derivatives and performed a structure–activity relationship study using rice seedlings. 16,17-Epoxidation of bioactive GA₁ and GA₄ reduced their activity to promote elongation of rice leaf sheaths. Moreover, 16,17-dihydroxylation significantly decreased the activities of 16,17-dihydro-16α,17-epoxy GAs. These results suggest that GAs are deactivated in a stepwise manner via 16,17-epoxidation and hydrolysis of these epoxy groups.     

雌激素与丹栀逍遥丸对更年期女性生殖内分泌系统及免疫功能的调节作用

目的:探究雌激素联合丹栀逍遥胶囊对更年期综合征妇女生殖内分泌系统及免疫功能的调节作用。方法:选取2014年5月至2014年9月我院收治的女性更年期综合症患者60例并将其随机分为两组,其中对照组给予雌激素替代等临床常规疗法;实验组在雌激素替代治疗基础上加用丹栀逍遥胶囊联合治疗。观察并比较两组患者生殖内分泌激素(E2、FSH、LH)水平、T淋巴细胞亚群(CD3+、CD4+、CD8+、CD4+/CD8+)比例以及IL-2活性的变化。结果:治疗后,两组血清E2、CD3+、CD4+、CD4+/CD8+水平及IL-2活性均较治疗前显著升高,而FSH、LH水平均明显低于治疗前,且实验组治疗后E2、CD3+、CD4+、CD4+/CD8+水平及IL-2活性显著高于对照组,FSH、LH水平均明显低于对照组,差异均有统计学意义(P0.05)。结论:雌激素联合丹栀逍遥胶囊能更加有效调节女性更年期综合征患者生殖内分泌激素水平,提高免疫功能,值得临床推广。     

女性甲状腺癌的病理与预后分析

目的:探讨女性甲状腺癌的病理特点以及预后影响因素。方法:收集2005年1月至2009年8月,齐齐哈尔和平医院收治的女性甲状腺癌患者148例,回顾分析其临床病理特点以及预后影响因素。结果:病理检查显示131例(88.5%)甲状腺乳头状癌(PTC),6例(4.1%)为滤泡状癌(FTC),3例(2.0%)为髓样癌(MTC),6例(4.1%)为未分化癌(ATC);病理分期Ⅰ-Ⅱ期占45.3%,Ⅲ-Ⅳ期占54.7%;27例(18.24%)为周围组织侵犯,9例(6.1%)为远处转移,10例(6.76%)对侧甲状腺转移,56例(37.84%)颈部淋巴结转移;1年、3年、5年生存率依次为97.3%、93.2%、83.8%;年龄、病理床分期、病理类型、淋巴结转移、远处转移以及周围组织侵犯均是影响临床预后的重要因素(P0.05)。结论:女性甲状腺癌存在病理分期晚、病理分型差、淋巴结转移、远处转移及局部侵犯率高等不良预后因素,早期影像学检查对临床治疗具有指导意义。     

糖尿病大鼠ghrelin和nesfatin-1表达动力学及其调控

目的:探讨STZ诱导糖尿病大鼠ghrelin和nesfatin-1动力学及分泌调节变化。方法:STZ诱导糖尿病大鼠模型;采用葡糖糖脱氢酶分析法测量血浆葡萄糖水平;免疫放射分析检测血浆ghrelin、nesfatin-1、胰岛素、胰岛素样生长因子1(IGF-1)、生长激素(GH)含量;采用real-time PCR检测ghrelin m RNA水平变化;免疫组化观察ghrelin和nesfatin-1免疫活性细胞数量。结果:糖尿病大鼠体重显著降低(t=23.16,P<0.01),血糖水平显著升高(t=22.55,P<0.01),血浆胰岛素和IGF-1水平显著降低(t=6.50,t=24.13,P<0.01),但GH水平显著升高(t=3.30,P<0.05)。糖尿病大鼠血浆总ghrelin(t=7.03,P<0.01)和活性ghrelin(t=3.33,P<0.05)水平均显著升高,血浆nesfatin-1水平则显著降低(t=6.24,P<0.01);糖尿病大鼠血浆总ghrelin与GH(r=0.81,P<0.01)和IGF-1水平(r=-0.58,P<0.01)呈显著相关性;与对照组大鼠相比,糖尿病大鼠胃总ghrelin(t=16.86,P<0.01)和活性ghrelin(t=3.30,P<0.05)水平均显著降低;而胃nesfatin-1(t=7.93,P<0.01)水平则显著升高。胃总ghrelin水平与血浆IGF-1水平呈明显相关性(r=0.65,P<0.01);与对照组大鼠相比,糖尿病大鼠胃ghrelin m RNA表达水平显著升高(t=16.8,P<0.01),胃底ghrelin免疫活性细胞数量显著减少(t=3.98,P<0.01);实验中给予大鼠自由饮食,糖尿病大鼠血浆总ghrelin水平显著增加(t=7.53,P<0.01),nesfatin-1水平显著降低(t=5.46,P<0.01)。糖尿病大鼠注射胰岛素后,可使增加的ghrelin水平(t=1.76,P=0.11)和降低的nesfatin-1水平接近正常(t=1.96,P=0.06);且胰岛素可显著反转糖尿病大鼠胃总ghrelin(t=8.54,P<0.01)和nesfatin-1水平(t=2.42,P<0.05);以及注射胰岛素后,糖尿病大鼠胃底ghrelin细胞显著增加,nesfatin-1细胞明显减少(t=3.21,t=2.59,P<0.05)。结论:Ghrelin或nesfatin-1参与糖尿病大鼠能量平衡调控。