Instability of anthocyanin accumulation inVitis vinifera L. var. Gamay Fréaux suspension cultures

The inherent instability of metabolite production in plant cell culture-based bioprocessing is a major problem hindering its commercialization. To understand the extent and causes of this instability, this study was aimed at understanding the variability of anthocyanin accumulation during long-term subcultures, as well as within subculture batches, inVitis vinifera cell cultures. Therefore, four cell line suspensions ofVitis vinifera L. var. Gamay Fréaux, A, B, C and D, originated from the same callus by cell-aggregate cloning, were established with starting anthocyanin contents of 2.73±0.15, 1.45±0.04, 0.77±0.024 and 0.27±0.04 CV (Color Value)/g-FCW (fresh cell weight), respectively. During weekly subculturing of 33 batches over 8 months, the anthocyanin biosynthetic capacity was gradually lost at various rates, for all four cell lines, regardless of the significant difference in the starting anthocyanin content. Contrary to this general trend, a significant fluctuation in the anthocyanin content was observed, but with an irregular cyclic pattern. The variabilities in the anthocyanin content between the subcultures for the 33 batches, as represented by the variation coefficient (VC), were 58, 57, 54, and 84% forV. vinifera cell lines A, B, C and D, respectively. Within one subculture, the VCs from 12 replicate flasks for each of 12 independent subcultures were averaged, and found to be 9.7%, ranging from 4 to 17%. High- and low-producing cell lines, VV05 and VV06, with 1.8-fold differences in their basal anthocyanin contents, exhibited different inducibilities tol-phenylalanine feeding, methyl jasmonate and light irradiation. The low-producing cell line showed greater potential in enhanced the anthocyanin production.     

Simultaneous saccharification and fermentation of acid-pretreated corncobs with a recombinant Saccharomyces cerevisiae expressing beta-glucosidase

To reduce the cellobiose inhibition of exoglucanase and endogulcanase and enhance cellulose hydrolysis during simultaneous saccharification and fermentation (SSF), a beta-glucosidase encoding gene named BGL1 was cloned from Saccharomycopsis fibuligera and integrated into the chromosomal rDNA region of the Saccharomyces cerevisiae industrial strain NAN-27 producing NAN-227. Compared with the parental strain, which had no detectable activity, the beta-glucosidase specific activity in NAN-227 was 1.02 IU/mg of protein. When cellobiose was used as the sole carbon source in a shake-flask, NAN-227 consumed 6.2g/L of cellobiose and produced 3.3g/L of ethanol in 48 h. The yield was 0.532 g/g. The parent strain only consumed 1.92 g/L of cellobiose and no ethanol was detected. During the SSF of acid-pretreated corncobs NAN-227 produced 20 g/L of ethanol at 72 h, which was similar to the parent strain when 20IU of beta-glucosidase/g of substrate was added.     

Efficacy of postoperative adjuvant transfusion of cytokine-induced killer cells combined with chemotherapy in patients with colorectal cancer

Purpose

To assess the activity and safety of postoperative adjuvant immunotherapy with transfusion of cytokine-induced killer (CIK) cells combined with chemotherapy in patients with colorectal cancer.

Methods

We retrospectively studied 96 consecutive patients with colorectal cancer who were treated with resection between January 2010 and December 2012 as well as adjuvant chemotherapy. Twenty-one of these patients accepted at least 1 cycle of CIK cell transfusion for immunotherapy (CIK group). Disease free survival (DFS), immune cells and treatment related side effects were assessed. The patients were followed up until May 2013.

Results

By the end of follow-up, 10 patients (10.42 %) had died. Eighteen patients (18.75 %) had withdrawn. All the patients in the CIK group are still alive, and only 1 patient had withdrawn. Patients in the CIK group had significantly longer DFS than those in the control group [HR = 0.28, 95 % CI (0.09, 0.91), p = 0.034]. The 2-year DFS rates of patients in the CIK group and the control group were 59.65 ± 24.80 % and 29.35 ± 6.39 %, respectively. The CD4⁺/CD8⁺ ratios were significantly lower during the period of chemotherapy than those before chemotherapy (p = 0.0038), while the ratios were significantly higher during the period of CIK cell transfusion than those before CIK therapy (p = 0.0484). There were no immediate adverse reactions to the CIK cell transfusions.

Conclusion

Adjuvant transfusion of CIK cells prolongs DFS in patients with colorectal cancer.     

二种淡水微囊藻rDNA16S-23S基因间隔区的序列测定与分析

本研究采用ＰＣＲ及序列测定的方法,对我国淡水铜绿微囊藻有毒株和另一低毒的种类惠氏微囊藻（Ｍ５７４）ｒＤＮＡ１６Ｓ－２３Ｓ基因间隔区进行了序列的测定和分析,结果表明：ｒＤＮＡ１６Ｓ－２３Ｓ基因间隔区可以作为一个精细且稳定的指标,用一微囊藻的分类和鉴定。并从分子水平提出了同囊藻与惠氏微囊藻在种系有较近缘的关系,本文首次对微落属ＭｉｃｒｏｃｙｓｔｉｓｒＤＮＡ基因间隔区全序列作了报道。为微囊藻属的鉴定及系     

甘蓝型油菜秦优10号杂交种纯度鉴定的SSR引物筛选

为了建立一套快速可靠的油菜杂交种纯度的鉴定方法,本文以秦优10号及其亲本、杂种ZZH2为试验材料,对前人开发的2对SSR引物(7号引物, 9号引物)进行了再次筛选.结果显示,7号引物不但能很好地区别出混入杂交种中的亲本,还能将杂交种子的同母异父组合种子从杂交种中分离出来,能够用于鉴别秦优10号杂交种子的真伪;9号引物能区分出杂交种和母本,但不能区分开杂交种和父本.同时,本试验利用人工制成的秦优10号杂交种标准样(纯度为100%)以及7份大田鉴定不同纯度梯度的杂交种子对7号引物鉴定结果的准确性进行了验证,鉴定结果与大田鉴定结果基本一致.本文结果将为鉴定秦优10号杂交种纯度提供更准确的技术资料.     

神经系统与内分泌系统的相互影响与协同作用

在人体内,神经系统和内分泌系统紧密联系,协调配合,相互作用。它们的基本功能都是信息传递,在此功能之上,两大系统几乎调控着机体全部的代谢活动。将以综述的形式,介绍神经内分泌系统的结构基础,神经系统与内分泌系统的相互作用,以及两大系统共同发挥作用的主要领域。     

分类多样性在大型底栖动物生态学方面的应用:以黄海底栖动物为例

根据文献资料,系统整理了黄海大型底栖动物物种名录,并得到了其平均分类差异指数和分类差异变异指数的理论平均值及95%置信区间漏斗图。结果表明,黄海共有大型底栖动物1,360种,分属于17门35纲91目368科842属。其平均分类差异指数的理论平均值为93.7,分类差异变异指数为213.6。将已知受到中度扰动的胶州湾部分站位真实值叠加到平均分类差异指数的95%置信区间漏斗图中,发现全部站位均显著低于95%置信区间,显示了平均分类差异指数在海洋污染监测方面的良好应用前景。     

抗菌肽的固相合成,分离纯化与构效关系的研究

应用手工固相ＤＣＣ／ＨＯＢｔ法合成ＣｅｃｒｏｐｉｎＢ、Ｓｈｉｖａｌ１、ＡＢＰ３、ＷＨＤ４种抗菌肽,各抗菌肽的Ｃ－端均为酰胺化,最终用ＨＰＬＣ分离纯化,这４种肽对大肠杆菌Ｋ１２Ｄ３１,野生型大肠杆菌,产气杆菌及４种植物病原菌青枯菌,临床致病理大肠杆菌,伤寒杆菌,硝酸盐杆菌均有明显的杀伤或抑制作用,对白血病细胞Ｋ５６０,肺癌细胞Ａ５４９也有杀伤作用,其中尤以ＷＨＤ杀伤作用最为明显,并用对其它３种肽不能