Effects of salinity on the growth and lipid composition of Atlantic salmon (Salmo salar) and turbot (Scophthalmus maximus) cells in culture

The direct effects of osmotic pressure (salinity) on growth performance and lipid composition were investigated in fish cells in culture. Cell lines from a relatively stenohaline marine species, turbot (Scophthalmus maximus) (TF) and an anadromous species, Atlantic salmon (AS) were cultured in media supplemented with NaCl to produce osmotic pressures varying from 300 to 500 mOsm kg⁻¹. The growth rates of the two cell lines were affected in a similar manner by the salinity of the media with the rank order for both peak cell numbers and growth rates up to the day of peak cell number being 300 > 350 > 400 > 450 > 500 mOsm kg⁻¹. Cell death occurred in both cell lines in older cultures at all salinities with the greatest loss of viable cells in media of 300 and 350 kg⁻¹. However, there were quantitative and qualitative differences between the cell lines in their lipid metabolism in response to the salinity of the media. The lipid content expressed per cell showed a positive correlation between lipid per cell and salinity in TF cells, but this was less apparent in AS cells. The percentage of total polar lipid classes increased with increasing salinity in TF cells due mainly to graded increases in the percentages of choline phospholipids. In contrast, there were no significant differences in the proportions of polar and neutral lipid classes with salinity in AS cells. The only significant effect of salinity in AS cells was a decreased proportion of dimethylacetals in total lipid at the highest salinity. The same significant effect of salinity on dimethylacetal content of total lipid was observed in TF cells. However, in addition there was a graded decrease in the percentage of 18:2n-9 in TF cell total lipid with increasing salinity. This was accompanied by increased percentages of total n-3 and n-6 PUFA with higher proportions of both groups of PUFA at 450 and 500 compared with 300 mOsm kg⁻¹. The results show that environmental salinity, in the absence of hormonal or other physiological stimuli, has direct effects on the growth and lipid metabolism of fish cells and that these effects differ in cells from different fish species.     

Effect of transketolase-like protein 1 on proliferation of human nasopharyngeal carcinoma cells

AIM: To investigate the effect of transketolase-like protein 1 (TKTL1) on proliferation of human nasopharyngeal carcinoma cells in vitro. METHODS: The siRNA against TKTL1 mRNA was constructed and transfected into human nasopharyngeal carcinoma cells (CNE cell line). The activity of transketolase was detected before and after RNA interference.Real-time PCR was used to determine the mRNA expression of transketolase (TKT) gene family in the CNE cells.Flow cytometry and MTT test were used to detect the effect of anti-TKTL1 siRNA on cell proliferation and cell cycle in the CNE cells. RESULTS: The total transketolase activity was significantly decreased in the CNE cells transfected with siRNA TKTL1 construct compared with the cells transfected with control vector or untransfected CNE cells. No significant difference in the expression level of TKT and TKTL2 gene between the CNE cells transfected with siRNA TKTL1 construct and the cells transfected with control vector or untransfected CNE cells was observed (P>0.05). However, the expression level of TKTL1 gene was significantly downregulated in the CNE cells transfected with siRNA TKTL1 construct compared with the cells transfected with control vector.Cancer cells were arrested in G0/G1 phase, and cancer cell proliferation was significantly inhibited in the CNE cells transfected with siRNA TKTL1 construct. CONCLUSION: TKTL1 plays an important role in the total transketolase activity and cell proliferation of human nasopharyngeal carcinoma. TKTL1 may be considered as a potential target for novel anti-cancer therapy.     

鱼类解偶联蛋白（UCP）基因研究新进展

通过生理性解偶联调节氧化磷酸化效率可能是进化早期发展起来的一种普遍策略,对生命的存在至关重要. 解偶联蛋白（uncoupling protein,UCP）家族是位于线粒体内膜的转运蛋白,哺乳动物基因组中已发现5个成员,其中UCP1~UCP3紧密相关,而UCP4和UCP5（即BMCP1,brain mitochondrial carrier protein-1）与它们差异较大. UCP1~UCP4不仅在哺乳类、鸟类等恒温脊椎动物中普遍存在,而且在鱼类等变温动物中也被证实存在,但UCP5目前仅在哺乳类中被发现. 哺乳动物褐色脂肪组织的UCP1,介导呼吸链产生的质子梯度泄漏,使氧化产生的能量以热的形式散发,但鱼类等变温动物UCP1的功能尚待进一步研究确立. UCP2和UCP3在产热、控制活性氧生成、脂肪酸氧化以及肥胖、糖尿病的发生中均发挥重要作用,二者的转录调节较复杂. UCP4和UCP5与其他UCP关系较远,UCP4仅在脑中表达,而UCP5在脑中高表达,它们的功能还不十分明确,但研究表明很可能与UCP2和UCP3的功能相似.     

鲫肠道上皮细胞原代培养方法的研究

探讨了健康鲫(Carassius auratus)幼鱼肠道上皮细胞的原代培养方法。结果显示,用含有抗生素、胶原蛋白酶Ⅰ、EDTA和胶原蛋白酶Ⅳ组成的D-Hanks液消化无菌分离的鲫肠道可以获得大量细胞团及绒毛隐窝;用含有抗生素、胎牛血清(FBS)、表皮生长因子(EGF)、胰岛素的DMEM培养液进行培养,并根据肠道上皮细胞与成纤维细胞贴壁时间的差异进一步纯化,连续培养12d后可以得到纯化的肠道上皮细胞。     

杜仲叶提取物对人结直肠癌细胞HCT116及抗血管生成活性的影响

研究了杜仲叶提取物(EULE)对人结直肠癌细胞HCT116体外增殖和凋亡的影响以及其抗血管生成的活性。采用噻唑蓝(MTT)比色法测定了EULE及阳性对照药顺铂对HCT116和人脐静脉内皮细胞(HUVEC)体外增殖的影响;用流式细胞仪测定了EULE对HCT116细胞凋亡的影响。结果表明:EULE对HCT116细胞增殖具有一定的抑制作用,并表现出较好的量效关系,但抑制率稍低于顺铂;EULE对HCT116细胞的凋亡率的影响不显著;EULE对HUVEC细胞增殖的作用具有浓度依赖性,表现为低浓度促进增殖,高浓度抑制增殖。当EULE质量浓度分别为100、200、400和800 mg/L时,对HCT116细胞的抑制率分别为6.50%、10.52%、16.99%和56.49%,对HCT116细胞的凋亡率分别为0.00%、5.90%、7.00%和4.50%。EULE 800 mg/L时对HUVEC细胞的抑制率为41.59%,说明杜仲叶提取物只有达到一定浓度才具有抗新生血管生成的活性。     

Meiotic chromosome behavior in allotriploid cultivar of Lifium hybrids.

采用细胞遗传学方法,对Longiflorum×Asiatic系列异源三倍体百合‘Bonsoir’（2n=3x=36）小孢子母细胞减数分裂进行了研究和分析。结果显示,有64．31％的花粉具有活力,且有活力的花粉大小范围在986．33-3491．68μm2之间,并呈双峰分布。形成败育花粉的主要原因如下：高度不规则的染色体配对、落后染色体、染色体桥、不均等分离、微核等现象。另外,减数分裂中期Ⅱ纺锤体的异常定向导致了末期Ⅱ二分体和三分体的形成,产生未减数的配子,如融合纺锤体和三极纺锤体;但平行纺锤体和垂直纺锤体不参与未减数配子的形成。一些小孢子母细胞在胞质分裂过程中未形成细胞板,导致单分体的形成以及二分体和三分体的增加,提高了未减数配子的颛率。通讨对小袍子发毕讨稃中各种观象的观察．对异源=倍体百合存畜种申的府阳讲行讨论．     

施氮对优质小麦颖果腹部和背部胚乳细胞物质充实的影响

为直观揭示施氮对小麦籽粒灌浆的生理效应,以徐麦30（强筋小麦) 和扬麦13（弱筋小麦）为材料,采用树脂包埋样品进行半薄切片和Image Pro Plus图像分析软件从形态学角度研究了孕穗期施氮对小麦颖果腹部和背部胚乳细胞物质充实的影响。结果表明,施氮后颖果腹部和背部胚乳细胞蛋白体数目增加,花后11、15和19 d最显著,同时蛋白体的聚集方式发生改变,对照组（不施氮）胚乳细胞内以小蛋白体为主,呈散状分布,而施氮组胚乳细胞含有许多大蛋白体或蛋白聚积体,分布较为集中。徐麦30蛋白体发育对氮素的响应程度高于扬麦13。施氮后腹部胚乳细胞淀粉粒排列更为紧密,细胞充实率增加,且扬麦13的响应程度大于徐麦30。施氮也增加了徐麦30背部胚乳细胞充实率,而对扬麦13的影响表现相反。     

玉米子粒胚乳细胞增殖与库容充实的关系

采用生长分析法和细胞计数法研究了掖单13和石单3号两个玉米品种子粒灌浆与胚乳细胞增殖的关系,结果表明：同一品种粒重的差异是由灌浆速度决定的,而不同品种粒重的差异则是灌浆持续期的长短造成的。胚乳细胞数与灌浆速度和灌浆持续期均呈极显著相关,表明胚乳细胞数的不同是引起粒重不同的主要原因。同时对库容提出新的表达式。     

斜带石斑鱼肝细胞分离及原代培养方法的建立

本研究以斜带石斑鱼肝细胞为实验对象,在不同培养条件下进行原代培养,旨在探讨稳定可靠的斜带石斑鱼肝细胞分离及原代培养方法。采用组织块分离法和胰蛋白酶(含EDTA)消化法分离肝细胞,并通过密度梯度离心法分离纯化肝细胞,细胞悬液于DMEM/F-12、M199和L-15培养液中培养;细胞活力及数量采用血球计数板计数,并通过MTT法测定细胞增殖率;同时,测定不同时间培养上清液中乳酸脱氢酶(LDH)活性、白蛋白(ALB)和尿素氮(BUN)的含量,以分析肝细胞生长状态。结果表明,组织块方法不适于斜带石斑鱼肝细胞的培养,未见细胞从组织块中迁出,而胰蛋白酶消化法获得良好稳定的培养效果,细胞产量达到1.6×108个/g肝重,活细胞数达到95%;L-15培养基细胞生长明显优于DMEM/F-12和M199培养基;启动原代培养的48~72 h阶段肝细胞生长代谢旺盛,培养上清液中LDH活性显著降低,ALB和BUN含量显著升高。结果显示,0.25%的胰蛋白酶常温消化法适合斜带石斑鱼肝细胞的分离,斜带石斑鱼肝细胞原代培养的最适培养基为L-15培养基,肝细胞在启动原代培养的48~72 h生长代谢旺盛。     

喹乙醇诱导鲤肝细胞凋亡的研究

细胞凋亡(apoptosis)是受基因控制的细胞自主的有序死亡。凋亡细胞的形态特征主要表现为细胞核的染色质浓缩,边移,聚集在核膜下,进而核发生裂解,核碎片与细胞碎片有单层膜包裹形成凋亡小体(apoptosisbody)[1]。目前已发现某些物理因素(辐射、高温等)、细胞因子、病毒感染和