Selection for High Oridonin Yield in the Chinese Medicinal Plant Isodon (Lamiaceae) Using a Combined Phylogenetics and Population Genetics Approach

Oridonin is a diterpenoid with anti-cancer activity that occurs in the Chinese medicinal plant Isodon rubescens and some related species. While the bioactivity of oridonin has been well studied, the extent of natural variation in the production of this compound is poorly known. This study characterizes natural variation in oridonin production in order to guide selection of populations of Isodon with highest oridonin yield. Different populations of I. rubescens and related species were collected in China, and their offspring were grown in a greenhouse. Samples were examined for oridonin content, genotyped using 11 microsatellites, and representatives were sequenced for three phylogenetic markers (ITS, rps16, trnL-trnF). Oridonin production was mapped on a molecular phylogeny of the genus Isodon using samples from each population as well as previously published Genbank sequences. Oridonin has been reported in 12 out of 74 species of Isodon examined for diterpenoids, and the phylogeny indicates that oridonin production has arisen at least three times in the genus. Oridonin production was surprisingly consistent between wild-collected parents and greenhouse-grown offspring, despite evidence of gene flow between oridonin-producing and non-producing populations of Isodon. Additionally, microsatellite genetic distance between individuals was significantly correlated with chemical distance in both parents and offspring. Neither heritability nor correlation with genetic distance were significant when the comparison was restricted to only populations of I. rubescens, but this result should be corroborated using additional samples. Based on these results, future screening of Isodon populations for oridonin yield should initially prioritize a broad survey of all species known to produce oridonin, rather than focusing on multiple populations of one species, such as I. rubescens. Of the samples examined here, I. rubescens or I. japonicus from Henan province would provide the best source of oridonin.     

CD138免疫磁珠细胞分选的FISH技术在多发性骨髓瘤诊断中的应用

为了分析CD138免疫磁珠细胞分选的染色体荧光原位杂交(FISH)技术在提高多发性骨髓瘤(MM)细胞遗传学异常检测敏感性的作用。本研究选取我院收治的30例确诊MM的患者为研究对象,分离骨髓单个核细胞,应用探针组合,同时采用2种方法进行细胞遗传学检测:实验组采用CD138免疫磁珠分选浆细胞后行荧光原位杂交技术(MACS-FISH)检测;对照组直接荧光原位杂交技术(D-FISH)检测。结果:30例MM患者,实验组采用CD138 MACS-FISH检出率为83.3%,对照组D-FISH法细胞遗传异常检出率为46.7%,两组差异具有统计学意义(p<0.05)。研究结果表明:分析不同类型的细胞遗传异常,MACS-FISH法1q21检出率为46.7%,RB1检出率为50.0%,Ig H检出率为70.0%,P53检出率为20.0%;D-FISH法检出率分别为23.3%,30.0%、36.7%、10.0%。通过细胞核型分析,30例MM患者中,发现5例患者为异常核型,仅为16.7%,其中1例患者为单一结构异常,复杂异常核型患者为4例。我们的研究结论表明:进行CD138免疫磁珠分选浆细胞的FISH技术在多发性骨髓瘤诊断应用中可显著提高细胞遗传学异常检测敏感性,具有临床推广应用的价值。     

广西弄岗黑叶猴栖息地选择与利用

2005年9月至2006年8月对广西弄岗国家自然保护区内一群黑叶猴进行行为观察,采用瞬时扫描法观察并收集猴群的栖息地利用数据,分析黑叶猴对喀斯特石山栖息地的选择与利用规律。结果表明,黑叶猴对不同山体部位的利用有显著性差异(χ~2=43.063,df=4,P0.001)。黑叶猴对崖壁的利用频率最高(占总记录的36.67%±9.44%),其次是山坡(32.30%±9.57%),对山脚(14.15%±5.01%)、山顶(11.24%±8.42%)和平地(5.63%±2.92%)的利用较少。分析发现,黑叶猴对山体部位的利用没有显著的季节性差异(山顶:Z=-0.160,P=0.837;崖壁:Z=-0.320,P=0.749;山坡:Z=-0.480,P=0.631;山脚:Z=-1.601,P=0.109;平地:Z=0,P=1)。黑叶猴将崖壁作为主要的休息场所,山坡和山脚为主要的移动和觅食场所。黑叶猴对栖息地的利用受食物可获得性的影响。当食物中花的可获得性降低时,猴群增加对山脚的利用;当嫩叶可获得性降低时,猴群增加在崖壁移动的频率;当果实的可获得性升高时,猴群增加在平地觅食的频率。食物组成与黑叶猴栖息地利用也有关系。总体来看,黑叶猴对山顶的利用频率与花的觅食比例呈显著正相关;对山脚的利用频率与果实+种子的觅食比例呈显著正相关。猴群在山顶休息的频率随花和成熟叶的觅食比例的上升而上升;在山脚休息的频率随果实+种子的觅食比例的上升而上升。猴群在山坡觅食的频率随果实+种子的觅食比例的下降而上升;在山脚觅食的频率随嫩叶的觅食比例的下降而上升。另外,平均最低温度与猴群在平地觅食的频率呈负相关关系。分析表明食物的分布和数量对黑叶猴栖息地利用有重要影响,黑叶猴对栖息地的选择是在觅食利益与风险之间进行权衡的结果。     

Development and performance evaluation of TaqMan real-time fluorescence quantitative methylation specific PCR for detecting methylation level of PER2

Molecular Biology Reports - PER2 gene methylation is closely related to the occurrence and progress of some cancers, but there is no method to quantitatively detect PER2 methylation in conventional...     

广西弄岗黑叶猴的家域和日漫游距离

我们对广西弄岗国家级自然保护区内的一群黑叶猴的家域和漫游行为进行了为期12个月的研究,以探讨该物种利用喀斯特石山的规律.结果表明,黑叶猴的家域总面积为28.75 hm2,平均月家域面积为8.35±4.24 hm2.猴群平均日漫游距离为661.77±130.20 m.黑叶猴对家域面积的利用表现出雨季明显小于旱季的季节性变...     

Longli is not a Hybrid of Longan and Lychee as Revealed by Genome Size Analysis and Trichome Morphology

Lychee, longan, longli, and rambutan are closely related, commercially important fruit trees in the Sapindaceae family. Longli fruits are morphologically similar to both lychee and longan, displaying a yellow-brown pericarp like longan, and small, sharp protuberances like lychee. These similarities have led to the hypothesis that longli is the result of intergeneric hybridization between longan and lychee. Scanning electron microscopy and flow cytometry were used to examine trichome morphology and genome size, respectively, to test this hypothesis. Longan, lychee, longli (D. confinis), and ‘Malesianus’ (D. longan sub spp malesianus) had morphologically distinct trichomes. The genome sizes for lychee (554 Mb), longan (444 Mb), ‘Malesianus’ (404 Mb), and rambutan (339 Mb) are distinctive and in a narrow range. ‘Malesianus’ has a genome 9% smaller than that of longan and 27% smaller than that of lychee. It is likely a species that evolved independently in northern Borneo island, and could be classified as a species, Dimocarpus malesianus, not a sub-species of longan as presently stated. Flow cytometry revealed a 50% variation in genome sizes among longli varieties, with genome sizes ranging from 450 to 678 Mb, beyond the range between longan and lychee. The genome size variation and distinct leaf hair morphology suggest that longli is not an intergeneric hybrid, and it is likely a separate genus evolved independently. The tested cultivars with distinctive genome sizes within D. confinis could be classified as separate species.     

夏枯草PvGGPPS基因的克隆和诱导表达分析

为探究夏枯草中GGPPS基因的生物学特性及功能,该文在夏枯草转录组测序的基础上设计特异性引物,采用逆转录PCR技术获得夏枯草中GGPPS基因的全长核苷酸序列,并进行生物信息学分析;采用qPCR法分析PvGGPPS基因在不同外源性物质诱导下在夏枯草果穗中的表达量以及该基因在夏枯草不同组织中的表达量。结果表明:PvGGPPS基因开放阅读框1 092 bp,编码363个氨基酸,理论分子量为38 815.68 D,等电点为5.69。PvGGPPS蛋白具有异戊烯基焦磷酸合酶家族的特征结构域。系统进化树表明PvGGPPS蛋白与丹参、毛喉鞘蕊花GGPPS蛋白具有较高的亲缘关系。qPCR分析表明,PvGGPPS基因在叶中表达量高于果穗及茎。对果穗施加7种外源性物质处理24 h后,GA3处理组该基因表达量升高。PvGGPPS基因在夏枯草不同组织中表达量差异较大,且受外源物质诱导表达。该研究结果为进一步研究PvGGPPS基因对夏枯草萜类成分合成途径中的功能及表达调控奠定基础。     

七鳃鳗Lja-SHP2分子鉴定、重组表达及免疫学研究

高等脊椎动物的蛋白酪氨酸磷酸酶SHP2(SH2 domain-containing protein-tyrosine phosphatase-2)由ptpn11基因编码,催化酪氨酸残基去磷酸化,与其他能催化酪氨酸磷酸化的蛋白酪氨酸激酶共同调节机体内多种信号通路的信号传导。以往研究表明,SHP2在高等脊椎动物T细胞和B细胞的激活与信号转导过程中起着重要作用。为了研究无颌类脊椎动物日本七鳃鳗(Lampetra japonica)中与SHP2同源的分子——Lja-SHP2在免疫应答反应中的作用,本研究通过PCR扩增获取其Lja-SHP2开放阅读框序列,并构建到原核表达载体pET-32a中,成功在大肠杆菌中实现重组蛋白表达并制备了其兔源多克隆抗体。用混合菌免疫刺激日本七鳃鳗后,通过实时荧光定量PCR和免疫印迹方法检测了Lja-SHP2在日本七鳃鳗免疫相关组织中mRNA和蛋白水平表达谱。结果显示,混合菌免疫刺激后,Lja-SHP2 mRNA和蛋白表达在外周血白细胞和髓样小体中无显著变化,而在鳃组织中显著性上调(P<0.05),说明Lja-SHP2在混合菌刺激后主要参与了鳃组织的免疫应答反应。为了进一步探究Lja-SHP2与淋巴细胞亚群免疫应答反应的相关性,本研究分别使用B细胞有丝分裂原脂多糖(lipopolysaccharide,LPS)和T细胞的有丝分裂原植物凝集素(phytohemagglutinin,PHA)免疫刺激日本七鳃鳗。经LPS免疫刺激后,与对照组相比,白细胞中Lja-SHP2蛋白表达显著上调,鳃组织和髓样小体没有显著性差异表达;但经PHA免疫刺激后,与对照组相比,白细胞、鳃组织和髓样小体3种组织中Lja-SHP2均有上调,尤其在白细胞中上调最为显著,大约是对照组的2.5倍,说明Lja-SHP2参与了日本七鳃鳗由PHA介导的免疫应答反应。由于PHA能刺激日本七鳃鳗鳃组织中VLRA+淋巴细胞的活化,这表明Lja-SHP2可能参与了PHA介导的VLRA+淋巴细胞亚群的免疫应答反应。上述研究结果为进一步探索Lja-SHP2在七鳃鳗免疫应答过程中的功能奠定了基础,也为揭示SHP2分子家族的系统发生及探索高等脊椎动物适应性免疫系统的早期发生及其进化历程提供一定的线索。     

Aidi injection,a traditional Chinese biomedical preparation for gynecologic tumors: a systematic review and PRISMA-compliant meta-analysis

Aidi injection (ADI), a traditional Chinese biomedical preparation, is a promising adjuvant therapy for gynecologic tumors (GTs), including cervical cancer (CC), endometrial cancer (EC), and ovarian cancer (OC). Although studies have reported positively on ADI therapy, its exact effects and safety in GT patients remain controversial. Therefore, a wide-ranging systematic search of electronic databases was performed for this meta-analysis. Data from 38 trials including 3309 GT patients were analyzed. The results indicated that the combination of conventional treatment and ADI markedly improved the patients’ overall response rate (P<0.00001), disease control rate (P<0.00001), and quality of life (P<0.05) compared with conventional treatment alone. Furthermore, patient immunity was enhanced with combined treatment, as indicated by significantly increased percentages of CD3⁺ (P=0.005) and CD4⁺ (P<0.00001) and increased CD4⁺/CD8⁺ ratio (P=0.001). Most of the adverse events caused by radiochemotherapy such as gastrointestinal issues, leukopenia, thrombocytopenia, and hepatotoxicity, (P<0.05 for all) were significantly alleviated when ADI was used in the GT patients. However, other adverse events such as nephrotoxicity, diarrhea, alopecia, and neurotoxicity did not significantly differ between the two groups. Overall, these results suggest that the combination of conventional and ADI treatment is more effective than conventional treatment alone.     

云杉腐烂病菌Cytospora piceae全基因组测序及比较基因组分析

【目的】壳囊孢属(Cytospora)真菌引起的林木腐烂病和枝枯病,是一类重要的、分布广泛的枝干病害,可引起重大经济损失和生态破坏。通过全基因组测序和比较基因组学分析,探究不同腐烂病菌的全基因组特征,分析其与寄主选择和致病性相关的基因或基因家族的差异性,将有助于进一步揭示腐烂病菌与寄主互作的分子机制,为腐烂病的有效防治提供基础资料。【方法】采用PacBio测序技术对云杉腐烂病菌Cytosporapiceae进行了全基因组测序和组装,并通过比较基因组学方法,从基因组水平探究引起腐烂病的4种腐烂病菌的基因组的差异,分析其共有的和特有的与致病相关的基因家族。【结果】C. piceae基因组大小为39.25 Mb,GC含量为51.79%。基于单拷贝直系同源基因构建的系统发育树显示,C. piceae与Cytospora chrysosperma的进化关系相近,Valsamali和Valsapyri则更相近。比较基因组学分析表明,4种腐烂病菌均具有重复诱导的点突变(RIP)活性,其中,C. piceae的RIP活性最强。与其他3种腐烂病菌相似,与木质素降解相关的AA3和AA7家族在C. piceae中显著扩张,但木质素降解关键酶AA5家族均缺失;C. piceae和C. chrysosperma基因组中果胶降解关键酶GH28和CE8家族基因的数量与V. mali和V. pyri相近。4种腐烂病菌都含有较多数量的MFS(major facilitator superfamily)超家族转运蛋白和较少的ABC(ATP-binding cassette transporter)超家族转运蛋白,但C. piceae含有更多DHA2、PDR和MDR类转运蛋白。4种腐烂病菌的分泌蛋白的GO分类分子功能主要集中在水解酶活性,其中V. mali含有最多数量的该类别基因;而生物学过程则集中在碳水化合物代谢过程、果胶分解过程和氧化还原过程。在次生代谢核心基因中,C.piceae的PKS基因明显少于V.mali和V.pyri;在C.piceae含有的4个特异性次生代谢基因中,3个为NRPS基因。【结论】4种腐烂病菌含有的碳水化合物活性酶的种类和数量相似,且都具有较强的果胶降解能力。不同腐烂病菌的膜转运蛋白中多药转运体的选择性扩增,以及次生代谢核心基因中NRPS类基因的特异性存在和缺失,表明它们作为重要的致病因子很可能在腐烂病菌寄主选择中发挥了重要的作用。