植物细胞生长周期中内外源细胞激动素的变化

应用细胞同步培养技术、高压液相色谱分析（ＨＰＬＣ）和酶偶联免疫法（ＥＬＩＳＡ）分析了胡萝卜细胞在悬浮培养时生长分裂周期的变化并对内源激动素类物质（玉米素Ｚ－Ｚｅａｔｉｎ、核糖基玉米素ＺＲ－ｚｅａｔｉｎｒｉｂｏｓｉｄｅ、二氢玉米素ＤＨＺ－ｄｉｈｙｄｒｏｚｅａｔｉｎ、核糖二氢玉米素ＤＨＺＲ－ｄｉｈｙｄｒｏｚｅａｔｉｎｒｉｂｏｓｉｄｅ、异戊烯基核糖基腺嘌呤ＩＰ－ｉｓｏｐｅｎｔｅｎｙｌａｄｅｎｉｎｅ和核糖基异戊烯基腺苷ＩＰＲ－ｉｓｏｐｅｎｔｅｎｙｌａｄｅｎｉｎｅｒｉｂｏｓｉｄｅ）含量进行定量分析．从用荧光显微技术分析ＤＮＡ含量变化结果看出，附加有激动素０．１×１０－６·Ｌ－１的悬浮液中，胡萝卜细胞的分裂周期为８．５ｈ，比不加外源激素的处理短１ｈ．在无外源激素存在的情况下，细胞分裂期中内源细胞分裂素类物质除ＺＲ之外，其余的Ｚ、ＤＨＺ、ＤＨＺＲ、ＩＰ和ＩＰＲ的含量在分裂的第６．５ｈ均有一个峰值出现，ＺＲ在第４．５和８．５ｈ有峰值出现．外加细胞分裂素的处理中．Ｚ、ｉＰ和ｉＰＲ整个周期无明显含量变化，ＤＨＺ和ＤＨＺＲ在第６．５ｈ出现峰值．ＺＲ在第８．５ｈ时也出现峰值．说明附加细胞激动素可协调Ｚ．ｉＰ和ｉＰＲ的积累，加速利用?     

水稻花培获得大群体健壮绿苗的研究

通过使用混合激素诱发籼稻871015、871109花粉愈伤形成胚性细胞;对籼稻花粉胚性细胞团和粳稻绿芽丛进行分割培养,快速繁育大群体绿苗获得成功。籼稻两个组合的8块胚性细胞团,在两个月内经3—5次切割培养获得绿苗12690株;粳稻花培绿芽丛仅经一次分割培养后的净增绿苗率127.2%,大幅度地提高了水稻花药培养再生绿苗率。分割培养的再生苗,健壮的形成特征:苗粗、浓绿、根多、洁白。良好的生理素质:发根力强,平均每棵试管苗多增根1.8条,根长度增长168.4%;平均每棵再生苗鲜重和干重分别增加0.29克和0.175毫克。     

流式细胞术检测TcR Vα24/Vβ11双阳性NKT细胞及其在HCV感染者中的应用

目的研究丙肝病毒感染者外周血NKT细胞含量与HCV感染状态之间的相关性.方法应用流式细胞术检测TcR Vα24/Vβ11双阳性NKT细胞.结果抗-HCV阳性且HCVRNA也阳性的病例NKT细胞数量明显低于抗-HCV阳性而HCV RNA阴性的病例（P〈0.01）,明显低于正常对照组（P〈0.05）.结论由于NKT细胞数量减少或活化受阻,造成HCV的持续感染,不利于病毒的清除;另一方面,由于肝内有HCV的存在,NKT细胞大量向肝内趋化,导致外周血的数量相对减少.     

树木木质部细胞凋亡及心材形成机理研究进展

概述有关木纤维、管状分子与薄壁细胞等木质部细胞凋亡和心材形成方面的研究成果,探讨木质部细胞凋亡与心材形成之间的信号通路及调控机理,并基于当前研究现状,提出几点建议。     

余甘子精粉的体外抗氧化活性分析

The antioxidant activities in vitro of refined powder of Phyllanthus emblica L. were detected by methods based on chemical measurement and cell modeling. The results of anti-peroxidation ability and iron reduction ability trials showed that the refined powder of P. emblica L. had strong ability of anti-peroxidation, and it increased with the increasing of concentration with a clear dose-response relationship. When the concentration reached 800 μg·mL^-1, the ability of anti-peroxidation was almost equal to the positive control VC with the same concentration. Meanwhile, it also had iron reduction ability that accompanied by significant dose-response relationship. By in vitro cell culture method, H₂O₂ was used to induce SH-SY5Y cells damages to establish the cell model of oxidative damage. The author detected the effect of different concentrations of powder on cell survival rate for the model cells by MTT assay. The results indicated that refined powder could show better protective effect to oxidative damaged SH-SY5Y cells in the range of 600 1 200 μg·mL^-1. When the concentration reached 1 200 μg·mL^-1, the cell survival rate was the highest and refined powder exhibited the best antioxidant activity. Both methods proved the strong antioxidant activities of refined powder of P. emblica L. in vitro.     

Effect of CHOP expression knock-down on apoptosis of renal tubular epithelial cells

AIM:To study the effect of C/EBP homologous protein (CHOP) on the apoptosis of renal tubular epithelial HK2 cells. METHODS:The serum mRNA levels of CHOP in the patients with acute kidney injury and healthy controls were detected by qPCR. In vitro, renal tubular epithelial HK2 cells were divided into control group, negative group (transfected with negative control siRNA), si-CHOP group (transfected with CHOP siRNA), and induced by transforming growth factor-β1 (TGF-β1). The viability of the cells was measured by MTT assay, and the apoptotic rate was analyzed by flow cytometry. The protein levels of nuclear antigen Ki-67, proliferating cell nuclear antigen (PCNA), caspase-3 and cleaved caspase-3 were determined by Western blot. RESULTS:Compared with the healthy controls, the serum mRNA levels of CHOP in the patients with acute kidney injury were increased significantly (P<0.05). Transfection with CHOP siRNA significantly decreased the expression of CHOP in the renal tubular epithelial HK2 cells (P<0.05). Knock-down of CHOP expression by siRNA significantly increased the viability of renal tubular epithelial HK2 cells (P<0.05), decreased the apoptotic rate (P<0.05), increased the expression of Ki-67 and PCNA (P<0.05), and down-regulated the protein level of cleaved caspase-3 (P<0.05). CONCLUSION:The serum mRNA levels of CHOP were increased in the patients with acute kidney injury. Knock-down of CHOP expression inhibits the apoptosis of renal tubular epithelial cells by regulating the expression of proliferation-and apoptosis-related proteins.     

内生菌与雷公藤细胞悬浮共培养的生理生化特性

将2株雷公藤内生真菌Fusarium oxysporum NS33与Penicillium steckii NS6、2株内生细菌Enterobacter cloacae sub sp LG3与Serratia marcescens LY1及其组合分别与雷公藤细胞悬浮共培养,对不同培养体系内雷公藤细胞的生长及其生理生化特征进行研究。结果显示,在共培养前期,与对照相比,接种单一内生菌株提高了细胞的干重,其中菌株NS6的促生效果最明显;而在共培养后期,无论是单一内生菌还是混合内生菌均对细胞生长具有抑制作用。内生真菌和菌株组合处理的培养液p H值有明显的升高,而内生细菌LY1则明显降低了培养液的p H值,其具有产酸性。另外,当雷公藤细胞同混合菌株共培养时,培养基中总糖消耗量是最大的,而接种单独菌株时则对细胞可溶性蛋白含量具有一定的提高作用。接种内生菌会影响雷公藤细胞的POD、CAT及SOD活性,与对照相比,接种单独菌株会更加提升POD与CAT的活性,而细胞MDA含量则明显下降。     

猴头菌子实体提取物对人肺癌细胞SPCA-1的抑制作用

猴头菌(Hericium erinaceus)子实体的醇提物用不同极性有机溶剂分部提取,得到石油醚、氯仿、乙酸乙酯和正丁醇分部,其中仅石油醚分部能抑制SPCA-1细胞增殖和促进其释放ROS(reactive oxygen species),对石油醚分部的进一步研究发现石油醚分部可诱导SPCA-1细胞早期凋亡和降低G_0/G_1期细胞数量。     

Liraglutide ameliorates liver injury of type 2 diabetic mice via micro-RNA-33-AMPK pathway

AIM: To observe the effects of liraglutide on the level of microRNA-33 (miR-33) and the expression of AMP-activated protein kinase (AMPK) and apoptosis-related proteins in mice with type 2 diabetes mellitus (T2DM), and to explore its possible mechanism. METHODS: High-fat diet and intraperitoneal injection of streptozocin were used to establish the type 2 diabetic model in C57BL/6 mice. The mice were randomly divided into 4 groups (n=15):in control group, the normal mice were subcutaneously injected with equivalent volume of saline; in model group, the T2DM mice were subcutaneously injected with equivalent volume of saline; in low-and high-dose liraglutide treatment groups, the T2DM mice were subcutaneously injected with 100 and 200 μg·kg^-1·d^-1, respectively. After 4 weeks of administration, the levels of FBG, TG, TC, HDL-C, LDL-C, ALT and AST were determined. HE staining was used to observe the pathological changes of the liver tissues. The protein level of cleaved caspase-3 in the liver tissue was detected by the technique of immunofluorescence. The protein levels of p-AMPK/AMPK and apoptosis-related proteins were detected by Western blot. The expression of miR-33 in the liver tissues was detected by real-time PCR. RESULTS: Compared with model group, the contents of FBG, TG, TC, LDL-C, ALT and AST were decreased significantly, while the content of HDL-C was increased significantly in low-dose liraglutide group and high-dose liraglutide group (P<0.05). The protein levels of phosphorylated AMPK and Bcl-2 were up-regulated significantly, and the expression of cleaved caspase-3 was down-regulated significantly (P<0.05). The level of miR-33 was decreased significantly (P<0.01). CONCLUSION: Liraglutide alleviates liver injury in type 2 diabetic mice, and the mechanism may be associated with reducing the level of miR-33 and increasing the phosphorylation of AMPK in the liver tissues, thereby inhibiting hepatocyte apoptosis.     

细胞穿透肽介导的小黑麦小孢子遗传转化研究

本研究选用小黑麦小孢子作为供试材料,进行细胞穿透肽介导的植物单倍体细胞遗传转化可行性研究。提取单核靠边期的小黑麦小孢子进行悬浮培养,将细胞穿透肽与报告基因GFP的表达框复合物对小孢子进行转化。转化植株进行了PCR分子检测,并在荧光体视显微镜下观察外源GFP基因的表达情况。共获得21株候选转基因植株,其中染色体自然加倍植株为10株。T1代加倍植株的PCR检测结果显示,加倍的转基因株系中有3个株系的PCR检测结果呈阳性。通过荧光体视显微镜可在转基因植株的幼胚部位观测到GFP蛋白的荧光。以上结果表明外源GFP基因已经整合到小黑麦基因组上并且得到表达并稳定遗传。因此,由细胞穿透肽介导的植物单细胞转化是一种有效、可行的植物单细胞转基因方法。