High-level secretory production of recombinant human platelet-derived growth factor-BB by <Emphasis Type="Italic">Saccharomyces cerevisiae</Emphasis> under the non-selective conditions

Recombinant human platelet-derived growth factor-BB (rhPDGF-BB) was produced by Saccharomyces cerevisiae. A two-stage cultivation strategy with mixture of glucose and galactose was developed to enhance rhPDGF-BB production, with its concentration being 32 mg/l fermentation broth under optimal conditions: corn steep powder as nitrogen source, 2 g/1 of glucose concentration at the beginning of induction phase, a pH of 5.0 and 6.5 for cell growth and rhPDGF-BB accumulation. The purification process consisted of yeast supernatant ultrafiltration followed by ion exchange chromatography and molecular sieve, and the recovery of rhPDGF-BB was estimated to be 20.28%. Biological activity of the purified rhPDGF-BB was 3.05 ng/ml. Published in Russain in Prikladnaya Biokhimiya i Mikrobiologiya, 2009, Vol. 45, No. 2, pp. 176–180. The article is published in the original.     

Interactions in family pairs and parental care in captive steppe lemming (Lagurus lagurus)

Partner interactions and parental activities in pair-bonded male and female steppe lemmings (L. lagurus) were estimated under laboratory conditions. Interactions between adults in family pairs were found to be neutral or peaceful with rare agonistic contacts. Compared to other rodent species living in family groups, the frequency and duration of partner grooming in adults were found to be low, indicating relatively weak pair bonding in steppe lemmings. At the same time, males spend much more time with pups in the nest than females, but they are inferior to the latter in grooming (licking) and nest material delivery. Parental behavior of L. lagurus is discussed in comparison with that of other species of the subfamily Microtinae.     

High-quality genomic DNA extraction from formalin-fixed and paraffin-embedded samples deparaffinized using mineral oil

Extracting DNA from formalin-fixed and paraffin-embedded (FFPE) tissue remains a challenge, despite numerous attempts to develop a more effective method. Polymerase chain reaction (PCR) success rates with DNA extracted using current methods remain low. We extracted DNA from 140 long-term archived FFPE samples using a simple but effective deparaffinization method, removing the wax with mineral oil, and a commercially available DNA extraction kit. DNA quality was subsequently tested in a genotyping experiment with 14 microsatellite markers. High-quality DNA was obtained with a mean PCR success rate of 97% (range: 88–100%) across markers. The results suggested that DNA extracted using this novel method is likely to be suitable for genetic studies involving DNA fragments <200 bp.     

Direct somatic embryogenesis and shoot organogenesis from leaf explants of Primulina tabacum

An efficient propagation system via somatic embryogenesis and shoot organogenesis and plant regeneration system for endangered species Primulina tabacum Hance was established. Thidiazuron (TDZ) was the key plant growth regulator for inducing somatic embryogenesis and kinetin (KIN) and 6-benzylaminopurine (BAP) were the key cytokinins for inducing shoot organogenesis from leaf explants. TDZ combined with BAP or KIN in the induction Murashige and Skoog medium induced both somatic embryos and adventitious shoots. Leaf explants with abaxial site in contact with the medium induced less somatic embryos or adventitious shoots compared to inversely placed leaf explants and the optimum pH was 6.5–7.0. Secondary somatic embryos or adventitious shoot could be induced from primary somatic embryos using TDZ and BAP. Shoots developed adventitious roots on rooting medium containing 0.5 μM indole-3-butyric acid and 0.2 % activated carbon. Over 90 % of plantlets survived following acclimatization and transfer to potting mixture (sand:Vermiculite:limestone; 1:2:1).     

Prediction model of DnBP degradation based on BP neural network in AAO system

A laboratory-scale anaerobic-anoxic-oxic (AAO) system was established to investigate the fate of DnBP. A removal kinetic model including sorption and biodegradation was formulated, and kinetic parameters were evaluated with batch experiments under anaerobic, anoxic, oxic conditions. However, it is highly complex and is difficult to confirm the kinetic parameters using conventional mathematical modeling. To correlate the experimental data with available models or some modified empirical equations, an artificial neural network model based on multilayered partial recurrent back propagation (BP) algorithm was applied for the biodegradation of DnBP from the water quality characteristic parameters. Compared to the kinetic model, the performance of the network for modeling DnBP is found to be more impressive. The results showed that the biggest relative error of BP network prediction model was 9.95%, while the kinetic model was 14.52%, which illustrates BP model predicting effluent DnBP more accurately than kinetic model forecasting.     

Production of xylooligosaccharides by xylanase from Pichia stipitis based on xylan preparation from triploid Populas tomentosa

Xylooligosaccharides (XOS) with DP 2-4 are important synbiotics used as food ingredients based on its prebiotic characteristics. In this work, the production of XOS from lignocellulosic material was performed by combined chemical-enzymatic methods. Xylan was prepared from triploid Populas tomentosa, and bioconverted into XOS by crude xylanase solution obtained from Pichia stipitis. The effects of reaction time, temperature, enzyme dosage, and pH value on the production of XOS were fully evaluated. Under the optimal condition (25 U g⁻¹ substrate, pH 5.4 and 50 °C), 36.8% of the xylan preparation was converted to XOS, equivalent to 3.95 mg/mL of the hydrolyzate. Xylobiose, xylotriose and xylotetrose were analyzed to be the main products of the enzymatic hydrolyzate, which together accounted for over 95% of the released oligosaccharides. Meanwhile, the effect of sonication pretreatment on the conversion efficiency of the xylan preparation was also investigated.     

肿瘤发生的细胞离子动态及其作为抗癌药标的综合效应

大量肿瘤细胞的有关研究表明,H+、Na+、Ca2+等细胞离子的动态对肿瘤发生起着关键作用.然而,单个离子作为代谢基础的作用是多方面的,其涉癌效果很不一致,有时还是相反的.因此,以控制单个离子动态作为抗癌药物的靶标,其效果并不理想.迄今对细胞离子的致癌作用的研究虽然是大量的,但均散见各章,缺少综合考虑.为此,本文简要介绍了上述离子的致癌作用,并探讨了以控制离子动态为药标,设计抗癌药物时应考虑的综合效应,供癌药物学研究者参考.     

A systematic investigation of leaf epidermis in <Emphasis Type="Italic">Camellia</Emphasis> using light microscopy

The characters of mature leaf epidermis of 58 species and 3 varieties belonging to 19 sections, representing all the 4 subgenera: Protocamellia, Camellia, Thea and Metacamellia were investigated under light microscope. The main conclusions are as follows: 1) The shape of the epidermal cells is elliptical, polygonal or irregular and the anticlinal walls are arched, straight or sinuolate, sinuous to sinuate. 2) The stomatal apparatus, consisting of a stoma (a pore plus a pair of guard cells) and 3–4 subsidiary cells, restricted to the abaxial surface of the leaves, were assigned to anisocytic, anisotricytic, isotricytic or tetracytic type. 3) Special structures in leaf epidermis, such as stomatal clusters, aborted stomata, secretory cells, cuticular intrusions and their taxonomic implications were also elucidated.     

Morphological observation of nuclear sub-localization of human angiogenin in HUVECs

Angiogenin, a potent angiogenic factor, was cloned and expressed by Escherichia coli and then purified with gel filtration chromatography. Approximately 90% pure angiogenin was obtained to generate a monoclonal antibody. Using western immunoblotting and ELISA, we confirmed that monoclonal antibody C46 secreted from hybridoma cells stably and specifically binds to angiogenin. The fused protein angiogenin-EGF was then expressed in HUVECs, and the subcellular localization of the recombinant protein was determined by confocal microscopy and TEM assay. Recombinant angiogenin was found to mainly concentrate in the pars granulosa of the nucleus, where the protein accumulates to form ribonucleoprotein particles.