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.     

Salmonid fish viruses and cell interactions at early steps of the infective cycle

A flow cytometric virus-binding assay that directly visualizes the binding and entry of infectious pancreatic necrosis virus (IPNV), infectious haematopoietic necrosis virus (IHNV) and virus haemorrhagic septicaemia virus (VHSV) to several cell lines was established. The highest efficiency of binding was shown by the BF-2 cell line and this was used to study, at the attachment level, the interactions of these cells with salmonid fish viruses in coinfections, and to further determine if the earliest stage of the viral growth cycle could explain the previously described loss of infectivity of IHNV when IPNV is present. Our results demonstrated that IPNV binds to around 88% of cells either in single or dual infections, whereas IHNV attachment always decreased in the presence of any of the other viruses. VHSV binding was not affected by IPNV, but coinfection with IHNV reduced the percentage of virus-binding cells, which suggests competition for viral receptors or co-receptors. Internalization of the adsorbed IHNV was not decreased by coinfection with IPNV, so the hypothetical competence could be restricted to the binding step. Treatment of the cells with antiviral agents, such as amantadine or chloroquine, did not affect the binding of IPNV and VHSV, but reduced IHNV binding by more than 30%. Tributylamine affected viral binding of the three viruses to different degrees and inhibited IPNV or IHNV entry in a large percentage of cells treated for 30 min. Tributylamine also inhibited IHNV cytopathic effects in a dose-dependent manner, decreasing the virus yield by 4 log of the 50% endpoint titre, at 10 mm concentration. IPNV was also inhibited, but at a lower level. The results of this study support the hypothesis that IHNV, in contrast to VHSV or IPNV, is less efficient at completing its growth cycle in cells with a simultaneous infection with IPNV. It can be affected at several stages of viral infection and is more sensitive to the action of antiviral compounds.     

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。     

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．     

茶悬浮培养细胞玻璃化超低温保存研究

为保存具有稳定代谢能力的优良茶细胞系,本文对茶叶悬浮培养细胞玻璃化法超低温保存进行了初步研究。结果表明:预培养以4d最佳,60%PVS2冰浴装载以20min最好,100%PVS2冰浴脱水处理60min最优,40℃复温解冻细胞存活率最高,采用这些参数进行完整试验的细胞存活率达到76%,初步建立了茶悬浮培养细胞玻璃化超低温保存的冷冻程序。     

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。     

EGCG抑制Nicotine诱导肺腺癌H1299细胞JAK2/STAT3 mRNA的表达研究

为探索EGCG对Nicotine诱导肺癌细胞增殖的抑制作用,本研究通过MTT实验筛选出EGCG、Nicotine对肺腺癌细胞H1299的最佳作用浓度,利用实时荧光定量PCR技术检测EGCG、Nicotine对H1299细胞中Bax、Bcl-2、Jak2和Stat3基因mRNA相对表达量的变化。实验结果表明,EGCG对H1299细胞的半抑制浓度IC₅₀值约为32βμmol·L^-1（24βh）、15βμmol·L^-1（48βh）;1βμmol·L^-1的Nicotine对H1299细胞促增殖作用明显;以15βμmol·L^-1的EGCG预处理H1299细胞24βh可显著下调1βμmol·L^-1 Nicotine的促增殖作用（P<0.05）。1βμmol·L^-1的Nicotine处理H1299细胞可明显降低JAK2/STAT3信号通路中Bax基因mRNA的表达,增加Bcl-2、Jak2、Stat3基因的mRNA表达;15βμmol·L^-1 EGCG预处理H1299细胞可反向调控Nicotine诱导所致JAK2/STAT3信号通路中Jak2、Stat3和Bax、Bcl-2基因表达量的变化,结果具有显著性差异（P<0.05）。由此可知,EGCG对Nicotine诱导的肺腺癌H1299细胞增殖及JAK2/STAT3信号通路中促增殖基因mRNA的表达起抑制作用。     

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。     

斑节对虾体长与中肠腺细胞数相关性研究

通过现场测定养殖日龄为１６～１８ｄ的斑节对虾的体长、体重和全长,并应用石蜡切片技术对被测虾的中肠腺中部进行细胞计数,研究了斑节对虾体长与中肠腺细胞数的相关性。结果表明,对虾体长与中肠腺中部每个高倍视野的平均细胞数呈密切负相关（ｒ＝－０．９４７）,公式为：Ｃ＝３３０．３６２３２３ｅ－０．０８７３８４Ｌ。对虾体长与体重密切相关（ｒ＝０．９４８）,可用对数回归方程描述：ｌｇＷ＝－１．９２３５７８＋３．０８２８３２ｌｇＬ。对虾体长与全长密切相关（ｒ＝０．９９５）,其线性回归方程为：Ｙ＝－０．１８７１３７＋１．３１６７０５Ｘ。通过这些公式,中肠腺中部每个高倍视野的平均细胞数可由体长直接推算出或由体重和全长间接算出,使斑节对虾杆状病毒感染度的计算更加简单和准确。