6个品种樱花在北京地区的适应性研究

为丰富北京地区樱花种植品种,选择大渔樱(Cerasus×kanzakura ‘Tairyo-zakura’)、修善寺寒樱(Cerasus×kanzakura ‘Rubescens’)、阳春(C. spachiana ‘Yoshun’)、天之川(C. serrulata ‘Erecta’)、兰兰(C. serrulata ‘Ranran’)、红华(C. serrulata ‘Kouka’)等6个品种樱花进行实验性种植。在田间观测的基础上,测定低温胁迫下根系脱氢酶活性、枝条脯氨酸含量,高温胁迫下叶片的电导率、可溶性蛋白含量。结果表明,6个品种中,随着温度下降,6个品种樱花根系脱氢酶还原量、枝条脯氨酸含量均呈现先上升后下降的趋势。大渔樱、修善寺寒樱、兰兰在-10 ℃时脱氢酶活性与脯氨酸含量达到最大值,阳春、天之川、红华在-15 ℃脱氢酶活性与脯氨酸含量达到最大值。6个品种的叶片相对电导率,均随着温度的上升不断升高。而叶片可溶性蛋白含量变化趋势,品种间差异较大。阳春、天之川、红华在北京地区适应性良好,大渔樱、修善寺寒樱、兰兰可在小环境较好的区域栽植。     

顶空固相微萃取-气相色谱-质谱联用法分析香花秋海棠及其变种花香气成分

利用顶空固相微萃取结合气相色谱-质谱联用技术对盛花期的香花秋海棠(Begonia handelii)雄花及其变种铺地秋海棠(B. handelii var. prostrata)雌花和红毛香花秋海棠(B. handelii var. rubropilosa)雄花的挥发性香气成分进行分析。结果表明,从铺地秋海棠花中鉴定出香气成分32 种,其中醇类物质含量最高,占总成分50.10,其次分别为碳氢类、醛类和酸类物质,分别占总成分25.39、13.87、4.34;从香花秋海棠花中鉴定出香气成分为21种,醛类物质含量最高,占总成分57.12,其次是醇类、碳氢类、杂环类物质,分别占总成分17.37、15.31、4.88;从红毛香花秋海棠花中鉴定出香气成分44 种,醇类物质含量最高,占总成分38.22,其次是碳氢类、醛类和醚类物质,分别占总成分34.36、16.83、6.91。铺地秋海棠和红毛香花秋海棠香气成分较接近,富含具有清香、甘甜气息的芳樟醇氧化物等物质;香花秋海棠花中醛类物质含量最高,具有淡焦甜香气。     

miR-92a-3p和miR-25-3p海绵上调Kiss1并影响雌性小鼠的青春期启动及动情周期

下丘脑Kiss1神经元产生的神经肽kisspeptin通过影响促性腺激素释放激素的分泌,参与青春期的启动、生殖系统的成熟以及排卵等过程的神经内分泌调节。Kiss1基因的表达受到包括多种反式调控因子及表观遗传的调控。预测与前期研究表明,miR-92a-3p、miR-25-3p的种子序列能够与Kiss1的3′-UTR直接结合,抑制Kiss1的表达。为进一步研究miR-92a-3p、miR-25-3p在Kiss1表达调控中的作用,本研究分别构建了对miR-92a-3p、miR-25-3p具有抑制作用的特异性吸附海绵载体：sponge-miR-92a和sponge-miR-25,以实现miRNA的功能缺失。流式细胞术和双荧光素酶报告基因系统分别证实,这2个海绵载体均能够非常有效地吸附外源性或内源性靶miRNA。sponge-miR-92a和sponge-miR-25载体被进一步包装成慢病毒LV-sponge-miR-92a和LV-sponge-miR-25。实时荧光定量PCR结果显示,经LV-sponge-miR-92a和LV-sponge-miR-25感染的下丘脑原代神经元细胞中,Kiss1表达水平均显著上调（P<0.05）;将LV-sponge-miR-92a注射到下丘脑后,雌性小鼠阴门开启时间明显提前（P<0.05）;下丘脑注射LV-sponge-miR-92a和LV-sponge-miR-25扰乱了雌性小鼠的正常动情周期。综上所述,成功构建了能够有效吸附miR-92a-3p、miR-25-3p的海绵载体,证明它们在解除miR-92a-3p、miR-25-3p对Kiss1的抑制中的作用,下丘脑注射海绵对雌性小鼠的阴门开启时间和动情周期均产生一定程度的影响,提示miR-92a-3p、miR-25-3p可能在青春期的启动和生殖成熟过程中发挥重要的作用。     

被动游泳运动对小鼠抑郁样行为及其肠道菌群组成的影响

被动游泳运动可诱发小鼠抑郁样行为,游泳环境的改变已成为抑郁样行为严重程度影响因素之一。观察不同水质、水温及持续时间对被动游泳小鼠抑郁样行为的影响,并初步探讨肠道菌群组成与抑郁样行为的关系。通过不同条件下的被动游泳运动建立抑郁样行为小鼠模型。采用糖水偏好实验及强迫游泳实验评价其行为学变化;采用16S rDNA高通量测序技术及实时荧光定量PCR技术对小鼠肠道菌群进行分子生态学分析。被动游泳16周后,各模型组小鼠体质量及糖水偏爱度均较正常对照组降低,而不动时间则有所延长。其中,室温海水游泳15 min小鼠体质量及糖水偏爱度降低程度最大,不动时间最长,与正常对照组比较,均具有显著性差异（P<0.05）。各模型组小鼠肠道菌群Chao指数、Shannon指数及PCoA分析均较正常对照组具有显著性差异（P<0.05）,其从门水平到属水平的丰度也发生不同程度改变。其中,室温海水游泳15 min小鼠肠道菌群组成变化程度最大,并发生拟杆菌属、普氏菌属等多个菌属的富集以及乳杆菌属丰度的减少,实时荧光定量PCR实验也得到了较为一致的结果（P<0.05）。以上结果表明,被动游泳运动可导致小鼠抑郁样行为的发生,其肠道菌群组成也发生明显改变;同时,菌群组成的改变会随着抑郁样行为的严重程度而有所变化。     

野野村氏菌FIM02-765的分类鉴定及Simaomicin α的抗肿瘤活性

明确海洋沉积物来源的放线菌FIM02-765的分类地位,揭示其代谢产物Simaomicinα的抗肿瘤活性。通过形态学、生理生化特征、细胞糖分组成分析以及16S rRNA序列分析,对海洋放线菌FIM02-765进行菌种鉴定;通过大孔吸附树脂柱层析和Sephadex LH-20凝胶柱层析从该菌的发酵产物中分离得到稠环氧杂蒽酮类化合物Simaomicinα;通过MTT法对化合物Simaomicinα的体外抗肿瘤细胞活性进行了研究。结果表明,菌株FIM02-765属于野野村氏菌(Nonomuraea sp.),同时该菌产生的稠环氧杂蒽酮类化合物Simaomicinα具有较强体外抑制多种肿瘤细胞增殖的活性。研究表明1株经鉴定的海洋野野村氏菌FIM02-765产生稠环氧杂蒽酮类化合物Simaomicinα对肿瘤细胞具有较强体外抑制活性,为深入开展该菌的遗传育种以及Simaomicinα的生物合成研究提供参考。     

bFGF Promotes the Proliferation of Rat Peripheral Blood Mesenchymal Stem Cells Through β-Catenin Signaling

bFGF (basic fibroblast growth factor, bFGF) could promote the proliferation of bone marrow and cord blood mesenchymal stem cells. However, the effect of bFGF on the proliferation of peripheral blood mesenchymal stem cells (PBMSCs) needs further research. This study aimed to investigate the role of bFGF on the culture and expansion of PBMSCs in vitro. Firstly, arterial blood was collected from rats abdominal aorta. After mononuclear cells (MNCs) were separated with Ficoll separation fluid, MNCs were cultured in the DMEM medium without bFGF (served as control group) or with bFGF (10, 20 ng/mL, served as 10 or 20 ng/mL bFGF group). PBMSCs were obtained by adherent culture method. The third passage of PBMSCs was detected for the MSC surface markers and the effect of bFGF on the cell cycle of PBMSCs using flow cytometry. The effects of bFGF on colony formation, cell growth, and the expressions of cyclin D1, cyclin E, p21 and β-catenin were evaluated. PBMSCs showed no difference in morphology among the three groups. PBMSC clonies appeared 14 days after cultivation. Compared with the control group, the cell growth confluence of PBMSCs was obviously increased by 40% and 80% in groups treated with 10 ng/mL bFGF or 20 ng/mL bFGF respectively after culture of 21 days (all P<0.05). Compared with the group treated with 10 ng/mL bFGF, the confluence of PBMSCs in 20 ng/mL group was further increased by 28% (P<0.05). Cells of the third passage were positively stained for CD29 and CD90, while were negative for CD45. These results were consistent with the phenotypic characteristics of MSCs. Compared with the control group, the colony number of PBMSCs in the 10 ng/mL and 20 ng/mL bFGF groups was increased by 51% (P<0.05) and 92% (P<0.05), respectively. Compared with the 10 ng/mL group, the colony number of PBMSCs was further increased in 20 ng/mL group by 14% (P<0.05). The growth curve of PBMSCs showed that after 7 days of culture, the number of PBMSCs in 10 ng/mL bFGF group and 20 ng/mL bFGF group was increased by 41% (P<0.05) and 61% (P<0.05), respectively. Moreover, the cell number had a statistically significant difference between these two groups (P<0.05). Results from flow cytometry cell cycle showed that the numbers of PBMSCs in the G1 phase of experimental groups were significantly decreased as the concentration of bFGF increased when compared with the control group (P<0.05), whereas the number of PBMSCs in the S phase was significantly increased (P<0.05). Immunofluorescence experiments showed that, compared with the control group, bFGF significantly promoted the nuclear translocation and expression of β-catenin in PBMSCs. Compared with the 10 ng/mL group, the PBMSCs in 20 ng/mL bFGF group showed stronger nuclear translocation and expression of β-catenin. Western blot experiments showed that the levels of β-catenin and its target proteins cyclinD1 and cyclinE were significantly increased (all P<0.05), whereas expression of p21 was significantly decreased in PBMSCs in the bFGF groups in a concentration dependent pattern when compared with control group (P<0.05). The study firstly confirms that bFGF promotes the proliferation of PBMSCs by regulating the β-catenin signaling pathway, which may facilitate the aquisition of larger number of PBMSCs for stem cell engineering in vitro.     

福建省福州市马尾区药用植物资源研究

依托第四次全国中药资源普查工作,摸清马尾区药用植物资源种类、分布、重点中药材品种以及药材栽培等情况。根据第四次全国中药资源普查要求,通过实地调查、走访以及栽培基地调查等方法对马尾区药用植物资源进行调查,查阅相关资料确定野生药用植物的药用部位、生活型等。马尾区野生药用植物种类共计506种,隶属于132科360属,含2～5种的寡种科占绝对优势地位;草本为主要生活型,药用部位集中在根类和全草类;有福建省重点药材50种、特色药材22种。由于长期受人为干扰,马尾区原生植被破坏严重,药用植物种类较为贫乏。因此,可考虑林下种植中药改善土壤环境,实现中药资源可持续发展。     

湘西“蒿菜粑粑”原料植物鼠麴草总黄酮提取物体外抗氧化活性研究

为了解湘西特色食品“蒿菜粑粑”原料植物鼠麴草(Gnaphalium affine)总黄酮提取物体外抗氧化能力,采用DPPH、ABTS自由基清除实验,还原力实验和抑制β-胡萝卜素褪色实验等方法,测定鼠麴草总黄酮抗氧化活性。结果显示,鼠麴草总黄酮母液中总黄酮浓度为7.01 mg·mL–1mg/mL;总黄酮提取物对DPPH、ABTS自由基有较好的清除能力,其半数抑制浓度(IC50)分别为16.30 mg·L–1、30.16 mg·L–1,将胡萝卜素相对吸光度降为50%的时间延长至67.49 min,在还原能力、延缓胡萝卜素褪色和抑制脂质过氧化上也有较好效果。鼠麴草总黄酮提取物具有良好的体外抗氧化活性,可作为优质食用植物资源进一步开发与推广。     

Exosomal miR-21-5p derived from cisplatin-resistant SKOV3 ovarian cancer cells promotes glycolysis and inhibits chemosensitivity of its progenitor SKOV3 cells by targeting PDHA1

Ovarian cancer (OC) is a common reason for gynecologic cancer death. Standard treatments of OC consist of surgery and chemotherapy. However, chemoresistance should be considered. Exosomal miR-21-5p has been shown to regulate the chemosensitivity of cancer cells through regulating pyruvate dehydrogenase E1 subunit alpha 1 (PDHA1). However, the role of miR-21-5p/PDHA1 in OC is unclear. The levels of miR-21-5p and PDHA1 in clinical samples and cells were investigated. Exosomes derived from SKOV3/cisplatin (SKOV3/DDP) cells (DDP-Exos) were isolated and used to treat SKOV3 cells to test DDP-Exos effects on SKOV3 cells. Extracellular acidification rate and oxygen consumption rate were tested with a Seahorse analyzer. Cell apoptosis was analyzed by a flow cytometer. PDHA1 was overexpressed and miR-21-5p was silenced in SKOV3 cells to study the underlying mechanism of miR-21-5p in OC. Quantitative real-time PCR and immunoblots were applied to measure gene expression at mRNA and protein levels. The levels of PDHA1 in DDP-resistant SKOV3 or tumor tissues were significantly decreased while the levels of miR-21-5p were remarkably upregulated. miR-21-5p in DDP-Exos was sharply increased compared to that of Exos. Data also indicated that DDP-Exos treatment suppressed the sensitivity of SKOV3 cells to DDP and promoted cell viability and glycolysis of SKOV3 cells through inhibiting PDHA1 by exosomal miR-21-5p. miR-21-5p derived from DDP-resistant SKOV3 OC cells promotes glycolysis and inhibits chemosensitivity of its progenitor SKOV3 cells by targeting PDHA1. Our data highlights the important role of miR-21-5p/PDHA1 axis in OC and sheds light on new therapeutic development.