Rapamycin attenuates BAFF‐extended proliferation and survival via disruption of mTORC1/2 signaling in normal and neoplastic B‐lymphoid cells

B cell activating factor from the TNF family (BAFF) stimulates B‐cell proliferation and survival, but excessive BAFF promotes the development of aggressive B cells leading to malignant and autoimmune diseases. Recently, we have reported that rapamycin, a macrocyclic lactone, attenuates human soluble BAFF (hsBAFF)‐stimulated B‐cell proliferation/survival by suppressing mTOR‐mediated PP2A‐Erk1/2 signaling pathway. Here, we show that the inhibitory effect of rapamycin on hsBAFF‐promoted B cell proliferation/survival is also related to blocking hsBAFF‐stimulated phosphorylation of Akt, S6K1, and 4E‐BP1, as well as expression of survivin in normal and B‐lymphoid (Raji and Daudi) cells. It appeared that both mTORC1 and mTORC2 were involved in the inhibitory activity of rapamycin, as silencing raptor or rictor enhanced rapamycin's suppression of hsBAFF‐induced survivin expression and proliferation/viability in B cells. Also, PP242, an mTORC1/2 kinase inhibitor, repressed survivin expression, and cell proliferation/viability more potently than rapamycin (mTORC1 inhibitor) in B cells in response to hsBAFF. Of interest, ectopic expression of constitutively active Akt (myr‐Akt) or constitutively active S6K1 (S6K1‐ca), or downregulation of 4E‐BP1 conferred resistance to rapamycin's attenuation of hsBAFF‐induced survivin expression and B‐cell proliferation/viability, whereas overexpression of dominant negative Akt (dn‐Akt) or constitutively hypophosphorylated 4E‐BP1 (4EBP1‐5A), or downregulation of S6K1, or co‐treatment with Akt inhibitor potentiated the inhibitory effects of rapamycin. The findings indicate that rapamycin attenuates excessive hsBAFF‐induced cell proliferation/survival via blocking mTORC1/2 signaling in normal and neoplastic B‐lymphoid cells. Our data underscore that rapamycin may be a potential agent for preventing excessive BAFF‐evoked aggressive B‐cell malignancies and autoimmune diseases.     

Fabrication and Characterization of Native and Oxidized Potato Starch Biodegradable Films

The need to replace conventional polymers due to environmental pollution caused by them has led to increased production of biodegradable polymers such as starch. Thus, the application possibilities of starch have increased. In this study, we produced and characterized biodegradable films derived from native and oxidized potato starch. The film-forming solution was prepared with different concentrations of extracted starch (native or oxidized) and a plasticizer (glycerol or sorbitol). Then, the mechanical, barrier, morphological, and structural properties of the films were characterized. The moisture content of the films varied from 15.35?±?1.31 to 21.78?±?0.49%. The elastic modulus of the films ranged from 219?±?14.97 to 2299?±?62.91 MPa. The film of oxidized starch plasticized with sorbitol in the lowest content was the most resistant and flexible; moreover, this film also presented lower water vapor permeability and low solubility in water. Fourier-transform infrared spectroscopic analysis of the biodegradable films indicated the presence of same functional groups as those of starch with bands in the same regions. The film thickness was lower for the films plasticized with glycerol whereas the color variation (Δ?) was lower for the ones plasticized with sorbitol. In case of both plasticizers, the increase in their content decreased the Δ? value. All the biodegradable films presented stability against water absorption owing to their low solubility in water. Morphological evaluation revealed the presence of partially gelatinized starch granules in the films. The roughness parameter (Rq) of the films varied from 3.39 to 10.9 nm, indicating that their surfaces are smooth. X-ray diffraction studies showed a B-type pattern for the starches, which is representative of tubers. Further, the films present higher relative crystallinity (RC) compared to the starches. The biodegradable starch films are uniform, transparent and with low solubility in water. The oxidation of starch and use of sorbitol as a plasticizer resulted in improved properties of the starch films, which is suitable for application.     

Depth of edge influence of the agricultural-forest landscape boundary,Southwestern China

The landscape boundary is an important component of a landscape and often plays an indispensable role in regulating ecological flows. The primary objective of this study was to estimate how far the edge effects on agricultural-forest landscape boundaries can penetrate into the forest and agricultural field. This will serve as a basis for understanding the interaction between forest and agricultural fields in the mountainous area of southwestern China and provide scientific basis for the practice of the policy of “returning agricultural field to forest.” Based on field investigations, three types of boundaries with six sampling transects were selected. We investigated the soil moisture, soil nutrients and vegetation diversity along the transect gradient and explored the depth of edge influence (DEI) with moving split-window techniques for analyzing the data. DEI for soil moisture changed with the seasons, ranging from 6 m in the pepper field to 2 m in the forest during the dry season, and from 12 m in the pepper field to 2 m in the forest after heavy rain. DEI based on soil organic matter ranged from 1.5 to 10 m in the forest field, while it was not detected based on other soil nutrient factors. DEI based on vegetation diversity varied from 4 to 26 m and from 10 to 31 m in the forest and agricultural fields, respectively. These results provide the scientific basis for the policy “returning agricultural field to forest”. Based on these field observations, reducing human disturbance and revegetating with natural shrubs and meadows could be more effective for vegetation conservation in terms of soil moisture and soil nutrient content in the arid valley of the Minjiang River.     

Tn7-mediated introduction of DNA into bacmid-cloned pseudorabies virus genome for rapid construction of recombinant viruses

lacZa-mini-attTn7 was inserted into the intergenic region between the gG and gD genes in a PRV bacterial artificial chromosome (BAC) by homologous recombination in E. coli. The resulting recombinant BAC (pBeckerZF1) was confirmed by PCR and sequencing. Green fluorescent protein (GFP) gene was then transposed into pBeckerZF1 by transposon Tn7 to generate pBeckerZF2. Recombinant viruses vBeckerZF1 and vBeckerZF2 were generated by transfection with the corresponding BAC pBeckerZF1 or pBeckerZF2. The titers and cytopathic effect (CPE) observed for by vBeckerZF1 and vBeckerZF2 was comparable to that of the parental virus vBecker3. vBeckerZF2 was serial passaged for five rounds in cell culture, and the mini-Tn7 insertion was stably maintained in viral genome. These results show that recombinant viruses can be rapidly and reliably created by Tn7-mediated transposition. This technology should accelerate greatly the pace at which recombinant PRV can be generated and, thus, facilitate the use of recombinant viruses for detailed mutagenic studies. Foundation item: Key technologies R&D program (2006BAD06A01) from the Ministry of Science and Technology of China.     

Comparison of three ELISA kits for the differentiation of foot-and-mouth disease virus-infected from vaccinated animals

A study was performed to validate 3 FMDV 3ABC-I-ELISA kits developed in China for the differentiation of FMDV infected and vaccinated animals. Sets of sera from naive and vaccinated cattle as well as from cattle that had been infected were tested for antibodies against nonstructural proteins (NSPs) of FMDV by commercial diagnosis kits, Ceditest® FMDV-NS (Ceditest® kit), UBI® FMDV NONSTRUCTURAL PROTEIN ELISA DIRECTION INSERT (UBI® kit) and a FMDV 3ABC-I-ELISA kit developed at the Lanzhou Veterinary Research Institute. The test parameters (sensitivity and specificity) of the three kits were determined, and the result obtained from FMD 3ABC-I-ELISA kit was compared with that obtained from two foreign kits. The results indicated that the coincidence rate between the FMDV 3ABC-I-ELISA and Ceditest® kits was 98.05%, and the coincidence rate between the FMDV 3ABC-I-ELISA and UBI® kits was 94.4%; the sensitivity of both Ceditest® and FMDV 3ABC-I-ELISA kit was 100%. However, the sensitivity of the UBI® kit was only 81.8%. With sera from naive or vaccinated non-infected animals, the specificity of all tests exceeded 90%.     

Developmental expression of the receptor for advanced glycation end-products (RAGE) and its response to hyperoxia in the neonatal rat lung

Background  

The receptor for advanced glycation end products (mRAGE) is associated with pathology in most tissues, while its soluble form (sRAGE) acts as a decoy receptor. The adult lung is unique in that it expresses high amounts of RAGE under normal conditions while other tissues express low amounts normally and up-regulate RAGE during pathologic processes. We sought to determine the regulation of the soluble and membrane isoforms of RAGE in the developing lung, and its expression under hyperoxic conditions in the neonatal lung.     

Properties,Microstructure and Heat Seal Ability of Bilayer Films Based on Fish Gelatin and Emulsified Gelatin Films

Bilayer films prepared by the lamination of fish gelatin film (GF) and its corresponding emulsified film (EF) with different thickness ratios (7:3, 5:5 and 3:7) were characterized. Bilayer films had the similar tensile strength (TS) to EF (p > 0.05) but showed lower elongation at break (EAB) (p < 0.05). All bilayer films showed the lower water vapor permeability (WVP) but higher oxygen permeability (OP) than GF. Bilayer films had varying ΔE* (total color different), where the highest value was observed in that laminated with higher thickness ratio of EF (p < 0.05). Lower light transmission and higher transparency value were obtained for bilayer films, compared to GF (p < 0.05). Based on scanning electron microscopic (SEM) cross-section micrographs, all bilayer films consisted of two layers. Differential scanning calorimetric (DSC) analysis revealed that the bilayer films had the higher glass transition temperature (T _g) than GF but lower than EF. All bilayer films were heat sealable, however their seal strength and seal efficiency were lower than those of GF (p < 0.05). Therefore, the thickness ratios of GF and EF had a marked effect on the mechanical and barrier properties as well as heat sealing ability of resulting bilayer films.     

Effect of organosolv pretreatment on mechanically pretreated biomass by use of concentrated ethanol as the solvent

In this study, we determined the effect of organosolv pretreatment on herbaceous biomasses corn stover and wheat straw, by using high-concentration ethanol as the solvent. A high-concentration of ethanol allows for the easy reuse and recycling of the solvent. First, we tested the effects of ethanol pretreatments at 60 and 99.5% (w/w) and found that highest solvent concentration resulted in low glucose digestibility. The maximum enzymatic glucose digestibility with 60% ethanol was 92.6% at 190°C for 120 min (using corn stover) and 86.9% at 190°C for 120 min (using wheat straw). In contrast, the digestion rates with 99.5% ethanol were 68.8 and 77.4% under the same conditions, respectively, indicating that there is a limit to the use of high-concentration ethanol as the solvent. To overcome this limitation, we applied a mechanical pretreatment step before the chemical pretreatment. Subsequently, glucose digestibility increased significantly to 93.1% with 99.5% ethanol as the solvent. Additionally the enzymatic digestibility of mechanically pretreated corn stover was higher than that of non-pretreated corn stover by about 40%. Taken together, these results confirm the efficacy of using high-concentration ethanol as a solvent for organosolv pretreatment when done in conjunction with mechanical pretreatment.     

黄心夜合不同季节鲜叶挥发性成分研究

利用气相色谱-质谱联用（GC-MS）技术,对黄心夜合（Michelia martinii Levl.）春、夏、秋、冬4季不同鲜叶挥发油成分及其含量进行了分析研究。结果显示,去掉峰值0.49以下的色谱峰从4个季节黄心夜合鲜叶样品中共鉴定出65种挥发油成分,其中,春、夏、秋、冬4个季节鲜叶分别检测出26种、29种、31种和32种成分,4个季节鲜叶样品共有成分为三环烯、α-蒎烯、莰烯、香桧烯、β-蒎烯等14种。黄心夜合4个季节鲜叶及干燥花蕾中的共有挥发性成分有α-蒎烯、莰烯、β-蒎烯、芳樟醇4种。黄心夜合鲜叶挥发油成分中的α-蒎烯、β-蒎烯、β-月桂烯、β-芳樟醇、芳樟醇、月桂烯含量高,且春季鲜叶挥发油得率最高,最适合挥发油提取。鲜叶具有保健功能,适于四季园林养生配置。本研究黄心夜合4个季节鲜叶挥发油成分差异及季节性变化,可为黄心夜合挥发油提取的季节选择及养生保健群落的营建提供科学依据和指导。