萱草花粉中微管蛋白生物化学性质

微管（microtubule）是细胞骨架的重要成份,参与囊泡运输、信息传递等多种生命活动。我们从萱草花粉中纯化了植物微管蛋白,对其生物化学及生物物理学部分性质研究表明,纯化的微管蛋白经超离心法测定沉降系数为6．2S,SDS-PAGE分析α,β微管蛋白分子量为56kD、58kD,凝胶扫描分析纯度为93．7％。等电聚焦电泳测定等电点为pI=5．35。光谱学性质研究结果表明,最大紫外吸收峰为280．8nm,荧光光谱研究表明最大激发波长为282nm;此时的最大发射峰为338nm,圆二色光谱分析二级结构表明小螺旋占27．24％,β-折叠占24．48％,无规卷曲为48．28％,呈典型球蛋白特征。     

A meta-analysis of the association of glutathione S-transferase P1 gene polymorphism with the susceptibility of breast cancer

Glutathione S-transferase P1 (GSTP1) is one of the important mutant sites for the cancer risk at present. The conclusions of the published reports on the relationship between GSTP1 A/G gene polymorphism and the risk of breast cancer are still debated. This meta-analysis was performed to evaluate the association between GSTP1 and the risk of breast cancer. The association reports were identified from PubMed and Cochrane Library, and eligible studies were included and synthesized using meta-analysis method. 35 investigations were included into this meta-analysis for the association of GSTP1 A/G gene polymorphism and breast cancer susceptibility, consisting of 40,347 subjects (18,665 patients with breast cancer and 21,682 controls). The association between GSTP1 A/G gene polymorphism and breast cancer risk was not found for overall population, Caucasians and Africans. Interestingly, the GSTP1 A/G gene polymorphism was associated with the susceptibility of breast cancer in Asians (G allele: OR = 1.10, 95 % CI: 1.04–1.17, P = 0.001; GG genotype: OR = 1.36, 95 % CI: 1.14–1.62, P = 0.0008; AA genotype: OR = 0.92, 95 % CI: 0.85–0.98, P = 0.02). Furthermore, the GSTP1 A/G gene polymorphism was associated with the susceptibility of breast cancer for the analysis of the controls from hospital. In conclusion, GSTP1 A/G gene polymorphism is associated with the breast cancer susceptibility in Asians. However, more studies on the relationship between GSTP1 A/G gene polymorphism and the risk of breast cancer should be performed in further.     

Synthesis and antimicrobial activity of polyhalobenzonitrile quinazolin-4(3H)-one derivatives

A novel series of polyhalobenzonitrile quinazolin-4(3H)-one derivatives were synthesized and characterized by NMR, IR, MS, and HRMS spectra. All of the newly prepared compounds were screened for antimicrobial activities against four strains of bacteria (Gram-positive bacterial: Staphylococcus aureus and Bacillus cereus; Gram-negative bacterial: Escherichia coli and Pseudomonas aeruginosa) and one strain of fungi (Candida albicans). Among the synthesized compounds, 5-(dimethylamino)-8-(2,4,5-trichloro-isophthalonitrile) quinazolin-4(3H)-one (7k) exhibited significant activity towards Gram-positive bacterial, Gram-negative bacterial, and the fungi strains. The MIC (0.8–3.3 μg/mL) and MBC (2.6–7.8 μg/mL) for this compound were close to those of nofloxacin, chlorothalonil, and fluconazole, making it the most potent antimicrobial agents in the series.     

植物EXO70基因家族的研究进展

胞吐是存在于所有真核生物的一种极其重要的细胞活动,直接参与了激素和神经信号的分泌、细胞生长、细胞极性的建立,细胞分裂和细胞壁的形成等多项生理过程。在胞吐过程中,高尔基后转运膜泡与靶膜的识别是由进化上高度保守的胞泌复合体（exocyst）介导的。该复合体由8个蛋白亚基构成,其中EXO70是组成胞泌复合体功能的关键亚基,可与小G蛋白和膜脂互作,参与复合体在靶膜组装。目前,对植物胞泌复合体功能的了解非常有限,已有证据显示其广泛参与了细胞生长,细胞壁形成、细胞分裂等多种生物学过程。与酵母和动物相比,植物胞泌复合体的一个显著特征是：EXO70在高等植物基因组中存在多个同源基因,其具体生物学功能尚不清楚。本文综述胞泌复合体的研究进展,重点讨论植物EXO70的多基因家族,推测不同的EXO70可能参与了组织细胞或运载底物特异的膜泡转运过程。     

人嗅黏膜间充质干细胞来源外泌体的分离鉴定及生物学特性研究

外泌体是指释放到细胞外微环境中的直径约50～130 nm的纳米级的膜性囊泡。嗅黏膜间充质干细胞（olfactory mucosa mesenchymal stem cells,OM-MSCs）作为一类新发现的间充质干细胞,在许多疾病中均具有治疗作用,且其内在机制与其旁分泌的外泌体密切相关,但OM-MSCs外泌体的分离、鉴定及生物学特性的研究尚未见报道。本研究采用超速离心法提取OM-MSCs培养液中的外泌体,应用流式细胞术及免疫荧光进行细胞鉴定后,分别用透射电子显微镜、纳米粒径分析及Western印迹对外泌体形态、颗粒大小和表面的特异性分子标志进行分析鉴定。采用CCK8增殖实验,Western印迹和划痕实验,分析其对人脑微血管内皮细胞增殖和迁移的影响。电镜、Western 印迹和纳米粒径分析的结果显示：OM-MSCs来源外泌体形态多为圆形,直径约为40～150 nm;表达外泌体标记物CD63,CD81;CCK-8法检测显示：不同浓度的OM-MSCs源外泌体可提高人脑微血管内皮细胞的增殖活性,且其增殖促进作用具有浓度依赖性（P<0.05）。Western 印迹检测结果显示：相比空白对照组,OM-MSCs源外泌体可显著提高内皮细胞的增殖细胞核抗原蛋白质水平表达（P<0.01）,细胞划痕实验结果显示,OM-MSCs源外泌体可增强内皮细胞的迁移能力,且高于对照组（P<0.01）。本研究表明：通过超速离心法可以分离纯化获得OM-MSCs源外泌体,且该外泌体具有促进人脑微血管内皮细胞迁移和增殖的作用。     

Strategies for targeting energy metabolism in Kirsten rat sarcoma viral oncogene homolog -mutant colorectal cancer

Alterations in cellular energy metabolism play critical roles in colorectal cancer (CRC). These alterations, which correlate to KRAS mutations, have been identified as energy metabolism signatures. This review summarizes the relationship between colorectal tumors associated with mutated KRAS and energy metabolism, especially for the deregulated energy metabolism that affects tumor cell proliferation, invasion, and migration. Furthermore, this review will concentrate on the role of metabolic genes, factors and signaling pathways, which are coupled with the primary energy source connected with the KRAS mutation that induces metabolic alterations. Strategies for targeting energy metabolism in mutated KRAS CRC are also introduced. In conclusion, deregulated energy metabolism has a close relationship with KRAS mutations in colorectal tumors. Therefore, selective inhibitors, agents against metabolic targets or KRAS signaling, may be clinically useful for colorectal tumor treatment through a patient-personalized approach.     

(NH4)2SO4对瓜尔豆幼苗在不同光照下耐冷性的影响

通过温室穴盘育苗试验,研究叶面喷施(NH4)2SO4对低温胁迫12 h后不同光通量密度(PFD)条件下,瓜尔豆幼苗在常温培养过程中幼苗生存及叶片SOD活性、POD活性和MDA含量的影响。结果表明:低温处理一夜后,在温室自然光照(强光)下进行常温恢复时,瓜尔豆叶片中SOD和POD活性始终显著低于对照,MDA含量显著高于对照,幼苗死亡率显著高于对照;在90μmol.m-2.s-1弱光下恢复2~4 d后,叶片中保护酶活性显著高于对照,MDA含量保持较低水平,幼苗均正常生长。叶面喷施(NH4)2SO4能显著提高低温处理后的植株叶片抗氧化酶类活性,减少叶片中MDA的产生,降低幼苗死亡率,0.2%(NH4)2SO4处理效果最好,其次为0.4%(NH4)2SO4。短期低温后,使植物处在弱光环境下能减少活性氧的发生,为植物抗氧化系统达到新的平衡提供有利条件,从而减轻甚至消除低温伤害。     

旋覆代赭汤对食管癌细胞干性的影响*

目的: 探究旋覆代赭汤对食管癌细胞干性的影响。方法: 将BALB/c裸鼠随机分为对照组与实验组,每组5只,分别连续给予生理盐水和旋覆代赭汤(9.89 g/kg)灌胃处理,在给予灌胃第8日时皮下接种5×10⁶细胞数量的人源食管癌ECA-109细胞,每周测量肿瘤大小,4周后处死小鼠。收取肿瘤组织和小鼠血清,采用RT-qPCR、Western blot和免疫组化方法检测肿瘤细胞干性相关转录因子NANOG、OCT4、SOX2的表达。分别用含10%胎牛血清和上述两组小鼠的血清处理48 h食管癌ECA-109细胞,每组设置三个复孔,用RT-qPCR和Western blot方法检测NANOG、OCT4、SOX2的mRNA和蛋白表达水平,以及AKT及其磷酸化(p-AKT)的蛋白表达水平;用流式细胞术检测肿瘤细胞中ALDH酶活性;用成球实验检测肿瘤细胞的成球数量。结果: 与对照组相比,旋覆代赭汤显著抑制食管癌小鼠肿瘤的生长和大小(P＜0.01或P＜0.05);旋覆代赭汤药物血清显著降低食管癌细胞的中NANOG、OCT4和SOX2 mRNA和蛋白的表达水平、ALDH酶活性和成球数量,以及AKT和AKT的磷酸化(p-AKT)水平(P＜0.01或P＜0.05)。结论: 旋覆代赭汤具有抑制食管癌细胞干性的作用,其可能是治疗食管癌的潜在有效药物,为食管癌治疗探寻新的有效药物提供理论依据。     

环境条件对双叉犀金龟成虫存活及繁殖的影响

双叉犀金龟Allomyrina dichotoma是一种重要的森林资源昆虫,本文研究了环境温度（20℃、23℃、26℃、29℃、32℃）及菌糠含水量（35%、45%、55%、65%）对双叉犀金龟成虫寿命及子代数量的影响。结果表明,双叉犀金龟雌雄虫寿命均随温度的升高呈显著缩短,20℃~26℃下雌虫寿命在55~62 d之间,显著长于29℃~32℃处理;20℃~23℃下的雄虫寿命在33~48 d之间,显著长于26℃~32℃处理;环境温度及菌糠含水量均对双叉犀金龟成虫子代数量有显著影响,26℃时,4个菌糠含水量处理的平均子代数量为31.42 头/雌,显著高于其它温度处理,菌糠含水量55%~65%时,5个温度处理的平均子代数量为18.83~19.12 头/雌,显著高于其它菌糠含水量处理,其中以温度26℃、菌糠含水量55%处理的双叉犀金龟子代数量最高,达45.42 头/雌。各温度下双叉犀金龟繁殖子代数量均随时间的延长呈先上升后下降的趋势,各温度下产卵繁殖的高峰时间存在明显差异。在26℃时,双叉犀金龟各时间段子代数量均高于其它温度,在35~45 d时,达到产卵高峰期（12.33 头/雌）,综合各指标,建议利用菌糠饲养双叉犀金龟时,其成虫最佳环境条件为26℃,菌糠含水量55%。