乙酰水杨酸对采后玉露桃果实成熟衰老进程和乙烯生物合成的影响

20℃下用1.0mmol/L(pH3.5)的乙酰水杨酸(ASA)处理玉露桃Prunuspersica(L.)Batsch果实,分析其对果实成熟衰老相关因子的影响。结果表明,随着果实成熟衰老,LOX活性、超氧自由基()生成速率、ACC合成酶、ACC氧化酶活性和乙烯释放均呈峰形变化,果实硬度迅速下降,组织电导率持续上升;ASA处理显著抑制了峰前相应的LOX、ACC合成酶和ACC氧化酶活性变化以及乙烯释放,维持了较高的果实硬度和细胞膜的完整性,延缓了果实的成熟衰老。     

PG和LOX对采后番匣果实软化及细胞超微结构的影响

以番匣（Ｌｙｃｏｐｅｒｓｉｃｏｎ ｅｓｃｕｌｅｎｔｕｍ）品种‘丽春’为试验材料研究了果实采后多聚半乳糖醛酸酶（ＰＧ）和脂氧合酶（ＬＯＸ）的活性变化与果实硬度的关系,并观察了细胞超微结构的变化。结果表明,随着果实采后呼吸强度和乙烯释放量不断增加,ＰＧ和ＬＯＸ活性迅速增加,而果实硬度逐渐下降。呼吸强度和ＬＯＸ活性高峰出现在发白期（ＢＲ）,而乙烯释放量高峰延迟至转色期（ＴＵ）。ＰＧ活性随着果实的成熟逐渐增加,而ＬＯＸ活性在发白期之后保持较高水平。分别用外源ＬＯＸ和ＬＯＸ加底物处理番匣组织切片,发现组织细胞的衰老进程加速,进而导致细胞膜瓦解、胞壁纤维松弛以及细胞器空胞化。ＰＧ和ＬＯＸ都与果实的后熟软化有关,ＬＯＸ与衰老的启动有关。     

脱落酸在桃果实成熟过程中的作用

 以白凤桃为试材,分析研究了桃果实生长发育过程中脱落酸（ABA）的变化以及外源ABA和氟啶酮（Fluridone）处理对果实发育后期离体果实后熟衰老进程的影响,探讨了ABA与桃果实成熟的关系。结果表明：桃果实生长发育后期果实内源ABA的积累构成了果实成熟的启动信号,而ABA高峰的出现启动了果实的后熟衰老进程;外源ABA处理促进了采后果实的后熟衰老进程,而Fluridone处理抑制了果实内源ABA的生物合成和乙烯产生,延缓了桃果实的后熟软化;乙烯则可能作为ABA的一种辅助因子影响果实的后熟衰老进程。     

果实采后软化生理机制及调控技术研究进展

软化是果实采后成熟衰老的重要特征,包括一系列复杂有序的生理变化。该研究介绍了影响果实软化的主要因素,包括果实品质、细胞壁组成成分、细胞壁降解酶活性、乙烯浓度等变化,并总结抑制果实采后软化的物理、化学技术和方法,以期为掌握果实软化生理机制、调控果实软化提供参考依据。     

PG,ACC,乙烯对番茄果实成熟的影响

番茄果实在成熟过程中硬度下降迅速，ＰＧ活性急剧上升，表现出显著的负相关；用ＡＣＣ和乙烯利处理绿熟期果实，结果乙烯促进了正常番茄和ａｌｃ番茄果实成熟进程，其ＰＧ活性也升高，而外源乙烯和ＡＣＣ不影响ｎｏｒ基因突变体果实的成熟进程。ＡＣＣ和ＰＧ无直接关系。     

桃果实采后脂氧合酶活性与内源乙烯对果实软化衰老的影响

以“南京白凤”桃果实为试验材料，研究了常温、低温和GA3处理条件下果实采后LOX(脂氧合酶)活性变化和内源乙烯发生规律。获得了后熟期间LOX活性与乙烯发生量的跃变型变化趋势；在果实软化衰老过程中，LOX启动膜脂过氧化作用导致果实软化衰老，同时内源乙烯的大量合成加速了果实的软化衰老速度；低温和GA3处理通过降低LOX活性和抑制内源乙烯合成，并延迟其高峰期的出现而延缓果实衰老进度。     

乙烯调控非跃变型果实成熟衰老研究进展

 非跃变型果实成熟衰老过程产生的乙烯远低于跃变型果实,20世纪70年代以来,乙烯被认为在非跃变型果实成熟衰老中发挥的作用非常有限。然而,由于该类型包含许多重要经济价值的果实,如柑橘、草莓、葡萄、枇杷等,近年来乙烯参与非跃变型果实成熟衰老相关研究陆续有报道。大量结果明确表明,非跃变型果实诸多成熟相关过程也直接受乙烯调控,且非跃变型果实和跃变型果实品质变化的某些分子调控途径非常相似。本文中主要综述了乙烯调控非跃变型果实成熟衰老研究进展。     

温度和GA_3对桃果实采后乙烯合成及桃ETR1同源基因表达的影响

为明确桃果实采后成熟衰老期间乙烯的合成与桃ETR1同源基因之间的关系,进一步了解乙烯的转导和作用过程,设计合成了桃ETR1扩增引物RTetr1和RTetr2,用RT-PCR方法研究了温度和GA3处理对白凤桃(Amyg-daluspersicaL.)果实采后乙烯生成和桃ETR1同源基因表达的影响。结果表明,低温处理降低了果实乙烯释放量,低温+GA3处理不仅降低了果实乙烯释放量,还延缓乙烯释放高峰期。PCR分析表明,桃ETR1的cDNA合成受温度和GA3调节,随着乙烯合成能力的增强,ETR1同源基因的表达水平增加,在乙烯信号转导过程中呈正向表达模式。     

PG和LOX对采后番茄果实软化及细胞超微结构的影响

以番茄 (Lycopersiconesculentum)品种‘丽春’为试验材料研究了果实采后多聚半乳糖醛酸酶 (PG)和脂氧合酶 (LOX)的活性变化与果实硬度的关系 ,并观察了细胞超微结构的变化。结果表明 ,随着果实采后呼吸强度和乙烯释放量不断增加 ,PG和LOX活性迅速增加 ,而果实硬度逐渐下降。呼吸强度和LOX活性高峰出现在发白期 (BR) ,而乙烯释放量高峰延迟至转色期 (TU)。PG活性随着果实的成熟逐渐增加 ,而LOX活性在发白期之后保持较高水平。分别用外源LOX和LOX加底物处理番茄组织切片 ,发现组织细胞的衰老进程加速 ,进而导致细胞膜瓦解、胞壁纤维松弛以及细胞器空胞化。PG和LOX都与果实的后熟软化有关 ,LOX与衰老的启动有关     

BTH对南山甜桃采后生理和炭疽病的影响

以深圳南山甜桃为试材,研究了采前对树体进行BTH喷雾处理对采后果实炭疽病及成熟衰老的影响。结果表明,采前喷施BTH 50 mg.L-1可以有效防治采后果实炭疽病,减少腐烂;降低了采后甜桃果实呼吸速率和乙烯释放率,延缓了果实硬度的下降和果实的成熟衰老。     

猕猴桃衰老过程中PG,果胶质和细胞壁超微结构的变化

采后美味猕猴桃秦美果肉组织乙烯生成不断增加的同时，ＰＧ９多聚半乳糖醛酸酶）活性迅速提高，致使原果胶大量降解为可溶性果胶，细胞结构受损，胞壁纤维松弛、细胞器逐步空泡化，果肉由硬变软。ＰＧ活性和果肉硬度的变化呈明显的负相关。     

纤维素酶和多聚半乳糖醛酸酶与果实成熟

对果实成熟软化过程中组织超微结构变化以及Cx和PG对果实成熟软化的作用,Cx、PG在分子水平上的研究和尚待进一步研究的问题进行了系统综述:果实成熟软化与Cx、PG活性增强直接相关,PG在果实成熟软化过程中对果实着色也有着一定影响;编码Cx的基因已经在某些果实的cDNA文库中被鉴定;利用反义RNA技术对PG进行负调控是研究PG对果实成熟软化作用的主要手段。目前对Cx、PG在果实成熟过程中的作用研究主要集中在呼吸跃变型果实上,而在非呼吸跃变果实中的作用很可能有所不同。因此,研究Cx、PG在柑橘等非呼吸跃变果实成熟过程中的作用将是尚待研究的又一重要方向。     

子瓜果实贮存期细胞壁超显微结构及相关成分的变化

以大板黑瓜子子瓜为材料,西农8号西瓜为对照,测定和观察不同贮藏期果实硬度、果皮、果肉细胞壁超显微结构、中果皮果胶及其相关酶的变化,研究子瓜果实耐贮性机理。结果表明,贮藏前期子瓜果实硬度较大、中果皮原果胶含量呈上升趋势,25d后平缓下降,西农8号西瓜中果皮果胶酶(PE)、多聚半乳糖醛酸酶(PG)活性在贮藏期存在明显的高峰,子瓜相应酶的活性变化趋势平缓,果皮果肉组织相关细胞结构解离,破坏时期与程度早于、重于子瓜,至实验结束时,已基本丧失其原有的结构,而子瓜还维持一定程度的完整性。由此认为,子瓜贮存期原果胶含量较高,PE、PG活性变化平缓,细胞壁和细胞膜系统超显微结构相对稳定可能与耐贮性有关。     

Effect of Jinan injection on ultrastructure of lung cancer cell lines

AIM: To investigate the effects of Chinese medicine, Jinan injection, on ultrastructure and mitochondria in cultured lung cancer cell lines. METHODS: The cultured lung cancer cell lines PG and PAa were used and divided into 4 groups: control (C), cisplatin (DDP), Jinan (JA) and Jinan in combination with cisplatin (DJ), respectively. The changes of morphology and mitochondria membrane potential, intracellular Ca²⁺ and pH in every group were observed by inverted microscope and electronic microscope as well as by using flow cytometry, staining by rhodamine, Fluo-3 and BCECF, respectively. RESULTS: Degeneration cells showed chromatin condensation and peripheral congregation. In cytoplasm autophage lysosome increased and myelinoid body was seen easily. In mitochondria structure, where the space between the inner and outer membranes of these organelles expanded as the matrix was compressed. The electron-dense or swelled was observed as vacuole degeneration and its matrix showed electron-lucent. Compared to control, mitochondria membrane potential increased in every group after 24 h and 48 h treatment. DDP increased intracellular calcium ion in PG cells, however, in PAa cells, JA and DJ decreased it. Intracellular pH got lower at 24 h and higher at 48 h in PG and PAa cells. There were significance in every group vs control in PG and PAa by statistic t-test (P＜0.01). CONCLUSION: Apoptosis was induced in PG and PAa cell lines by Jinan injection and DJ. Mitochondria matrix displayed electron-dense, mitochondrial potential, intracellular calcium ion and pH showed an increasing trend. Mitochondria damages may play an important role in apoptosis induced by Jinan and DJ.     

苹果果实发育过程中淀粉代谢和淀粉粒超微结构研究

苹果果实发育过程中,伴随果糖、葡萄糖和蔗糖的积累,淀粉含量经历了从低到高,又从高到低的变化。发育中后期,果实淀粉与可溶性糖呈现互为消长的变化。淀粉粒超微结构的变化表明,幼果期果肉细胞代谢旺盛,可观察到发育初期的造粉质体和大量的线粒体;发育中期是果实淀粉的主要累积期,整个造粉质体内充满淀粉;成熟期的果肉细胞呈现降解特征,淀粉粒大部分被降解,但淀粉粒表膜依然保持完整。还研究了淀粉代谢及淀粉粒超微结构变化与果实发育的关系。     

‘徐香’与‘海沃德’猕猴桃冷藏期间组织结构与生理变化差异

以猕猴桃‘徐香’（较不耐贮藏）和‘海沃德’（耐贮藏）果实为试材,研究其在0 ℃贮藏期间生理及组织结构的变化差异,从而探究其与耐贮性的关系。结果表明：贮藏期间两品种果肉硬度均不断下将,‘海沃德’果肉硬度降至10 N的时间为150 d,而‘徐香’仅为95 d;‘徐香’的呼吸高峰和乙烯释放高峰的出现分别较‘海沃德’早20 d和10 d,且峰值分别高出16%和250%。贮藏期间‘徐香’果实的质量减少率也显著高于‘海沃德’。在整个贮藏期间,‘海沃德’果实的淀粉、原果胶、纤维素含量均高于‘徐香’,而淀粉酶（AM）、果胶代谢相关酶（多聚半乳糖醛酸酶PG、果胶甲酯酶PE、β–半乳糖苷酶β-Gal）和纤维素酶（Cx）的活性高峰值均低于‘徐香’。‘海沃德’的表皮毛极显著细短于‘徐香’;且角质层厚度及表皮细胞层数都大于‘徐香’;贮藏期间果肉细胞的形变程度‘海沃德’小于‘徐香’;‘海沃德’的细胞壁厚于‘徐香’,且贮藏期间中胶层分裂缓慢,线粒体较完整。总之,耐贮性好的猕猴桃品种角质层和表皮细胞厚,质量减少率较低,呼吸速率和乙烯释放速率慢,细胞壁酶活性低。     

青梅果实的采后成熟特性和肉质变化

研究了２０℃,相对湿度９３％条件下青梅果实的采后成熟特性和肉质变化,结果表明,青梅果实采后呼吸活性和乙烯生成量随着成熟作用的进展而加大,乙烯生成量极高,在呼吸类型上属于跃变型果实;果皮急速黄化,果肉急速软化,主要原因在于多聚半乳糖醛酸酶,果胶甲酯酶活性的增大,导致水溶性果胶含量上升和盐酸可溶性果胶含量的下降。     

柿果实采后软化中PG酶活性及其基因DkPG1的表达

为了进一步探索多聚半乳糖醛酸酶（PG）在柿（Diospyros kaki L.）采后软化中的分子调控机制,应用实时荧光定量PCR技术和DNS比色法检测常温下丙烯和1–甲基环丙烯（1-MCP）处理‘富平尖柿’果实中PG基因DkPG1的表达量和PG酶活性的变化。结果显示,随着柿果实成熟软化,DkPG1基因转录水平呈先上升后下降的趋势,其中丙烯处理4 d出现表达高峰,比对照提前12 d且峰值极显著高于对照,而1-MCP处理的果实中的表达高峰出现在处理后28 d,比对照推迟12 d峰值极显著低于对照。丙烯处理果实PG酶活性迅速增加,活性峰与DkPG1表达高峰出现在同一天,1-MCP处理的果实PG酶活性明显降低,活性峰出现在DkPG1表达高峰前4 d。此外,对不同处理下乙烯释放量和果胶组分变化分析的结果显示DkPG1基因的表达受乙烯调控,进而影响果胶组分代谢。     

Effects of berberine on the proliferation of human pulmonary carcinoma cells

AIM: To study the effect of berberine on the proliferation of human pulmonary carcinoma cells (PG cells). METHODS: The proliferation of PG cells was determined by using MTT assay. Cell cycle and apoptosis of PG cells were determined by using flow cytometry. Confocal scanning imaging system was used to assay the ROS-releasing level of PG cells. RESULTS: Berberine was shown to inhibit proliferation of PG cells directly and in a concentration-dependent manner (P<0.05，P<0.01). Berberine blocked the cell cycle of PG cells， and beberine at concentration of 40 mg/L induced the apoptosis of PG cells. After 6 or 12 h incubation, berberine began to induce ROS-releasing in PG cells at 10 mg/L. While incubated with berberine for 24 h, PG cells were induced to release ROS at lower concentration. CONCLUSION: Berberine has an inhibitory effect on the proliferation of PG cells, and these effect may be carried out by regulating the production of intracellular ROS and the process of cell cycle.     

Mechanisms underlying the protection of berberine against liver injury induced by lipopolysaccharide in mice

AIM: To investigate the protective effects of berberine against liver injury induced by lipopolysaccharide in mice and the mechanisms underlying its protective effect. METHODS: The male mice were divided randomly into control, berberine group, LPS group and berberine treatment group. Mice were administered intragastrically with distilled water (0.01 mL/g) or 5 g/L neutral sulfate berberine (0.01 mL/g) once a day for 5 days and injected intraperitoneally with normal saline or LPS （0.02 mL/g，28 mg/kg）at 1 h after gavage on day 5. Blood was collected for determining alanine aminotransferase (ALT) and aspartate aminotransferase (AST) activities, the content of tumor necrosis factors-α (TNF-α) at 10 h and 2 h after LPS or normal saline injection, respectively. Furthermore, the liver tissue was processed, and histological changes and ultrastructure in liver were observed with light and electron microscopy, malondialdehyde (MDA) content and superoxide dismutase (SOD) activity in liver were also detected. RESULTS: Both ALT and AST activities in serum in LPS group were higher than those in control and berberine treatment group. LPS increased the serum TNF-ɑ content at 2 h after injection, which was reversed by berberine pretreatment. The histological examination showed that LPS caused severe hepatic cell edema, degeneration, apoptosis and even necrosis, and ultrastructure observation demonstrated that LPS induced mitochondrial swelling, condensation and margination of chromatin, irregular nuclear envelope in hepatocytes. The above pathological changes produced by LPS were attenuated by berberine pretreatment. Moreover, MDA contents in liver tissue were higher in LPS group than control and berberine treatment group, but there were no significant difference in SOD activity between berberine treatment and LPS group. CONCLUSION: Berberine has a protective effect on LPS-induced liver injury in mice, the mechanisms may be related to its decreasing the production of TNF-α, inhibiting lipid peroxidation and protecting mitochondria.