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Enteric virus concentration in large-volume water samples is crucial for detection and essential for assessing water safety. Certain dissolution and suspension components can affect the enrichment process. In this study, tangential flow ultrafiltration (TFUF) was used as an enrichment method for recovering enteric virus in water samples. Interestingly, the bacteriophage MS2 recovery in reclaimed water and the reclaimed water without particles were higher than that in ultrapure water. The simulated reclaimed water experiments showed that humic acid (HA) (92.16% ± 4.32%) and tryptophan (Try) (81.50 ± 7.71%) enhanced MS2 recovery, while the presence of kaolin (Kaolin) inhibited MS2 recovery with an efficiency of 63.13% ± 11.17%. Furthermore, Atomic force microscopy (AFM) revealed that the MS2-HA cluster and the MS2-Try cluster had larger roughness values on the membrane surface, making it difficult to be eluted, whereas MS2-Kaolin cluster had compact surfaces making it difficult to be eluted. Additionally, the MS2-HA cluster is bound to the membrane by single hydrogen bond with SO, whereas both the MS2-Try cluster and the MS2-Kaolin cluster are bound to the membrane by two hydrogen bonds, making eluting MS2 challenging. These findings have potential implications for validating standardized methods for virus enrichment in water samples., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

Two novel Gram-stain-negative, aerobic, non-motile and rod-shaped bacteria, designated as WL0004 T and XHP0148 T , were isolated from seawater samples collected from the coastal areas of Nantong and Lianyungang, PR China, respectively. Both strains were found to grow at 10-42 °C (optimum, 37 °C) and with 2.0-5.0 % (w/v) NaCl (optimum, 3.0 %). Strain WL0004 T grew at pH 6.0-9.0 (optimum, pH 7.0-8.0), while XHP0148 T grew at pH 6.0-10.0 (optimum, pH 7.0-8.0). The major cellular fatty acids (>10 %) of both strains included summed feature 8 (C 18 : 1  ω 6 c and/or C 18 : 1  ω 7 c ). In addition, strain WL0004 T contained 11-methyl C 18 : 1  ω 7 c and strain XHP0148 T contained C 12 : 0 3-OH. The respiratory quinone of both strains was ubiquinone-10. The G+C content of genomic DNA of strains WL0004 T and XHP0148 T were 62.5 and 63.0 mol%, respectively. Strains WL0004 T and XHP0148 T showed the highest 16S rRNA gene sequence similarity to Ruegeria pomeroyi DSS-3 T (99.4 and 99.0 %, respectively), and the 16S rRNA gene-based phylogenetic analysis indicated that the two strains were closely related to members of the genus Ruegeria . The average nucleotide identity and digital DNA-DNA hybridization values among the two strains and type strains of the genus Ruegeria were all below 95 and 70 %, respectively, and the phylogenetic tree reconstructed from the bac120 gene set indicated that the two strains are distinct from each other and the members of the genus Ruegeria . Based on this phenotypic and genotypic characterization, strains WL0004 T (=MCCC 1K07523 T =JCM 35565 T =GDMCC 1.3083 T ) and XHP0148 T (=MCCC 1K07543 T =JCM 35569 T =GDMCC 1.3089 T ) should be recognized as representing two novel species of the genus Ruegeria and the names Ruegeria marisflavi sp. nov. and Ruegeria aquimaris sp. nov. are proposed, respectively.

To explore the effect of different microbial strains on blueberry pomace with solid-state fermentation (SSF), three fungi strains and three lactic acid bacteria (LAB) strains were utilized to investigate with respect to polyphenol profiles, antioxidant capacities, and bioaccessibility. Different strains exhibited different capacities for metabolizing polyphenolic compounds in blueberry pomace. The contents of 10 phenolic acids and 6 flavonoids (except (+)-catechin) were increased in blueberry pomace fermented by Lactobacillus acidophilus (LA). A similar tendency was observed in blueberry pomace fermented by Aspergillus niger (AN) and Lactobacillus plantarum (LP), where the concentration of 8 phenolic acids and 5 flavonoids was enhanced, with the following exceptions: (+)-catechin, ferulic acid, vanillic acid, and quercitrin. Chlorogenic acid and quercetin were the maximum phenolic acids and flavonoids in blueberry pomace with SSF, upgraded at 22.96 and 20.16%, respectively. Contrary to the growth of phenolic acids and flavonoid compounds, all individual anthocyanins showed a decreased trend. Only in the blueberry pomace fermented by AN, all anthocyanidins exhibit a rising trend. After SSF, 2,2'-azino-bis (3-ethylbenzothiazoline-6-sulfonic acid) (ABTS), 2,2-diphenylpicrylhydrazyl (DPPH), and ferric reducing antioxidant power (FRAP) radical scavenging abilities were increased by up to 33.56, 59.89, and 87.82%, respectively. Moreover, the simulated gastrointestinal digestion system revealed that SSF improved the bioaccessibility of polyphenolic compounds. Compared with other strains, LA, LP, and AN showed better excellent capacities for metabolizing polyphenolic compounds, which led to a greater increase in antioxidant activity and bioaccessibility in fermented blueberry pomace., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2023 Tian, Li, Long, Liang, Chen, Huang and Ran.)

We investigated the effects of maize straw and its biochar application on soil organic carbon chemical composition, the abundance of carbon degradation genes ( cbh I) and the composition of cbh I gene community in a Moso bamboo forest, to provide the theoretical and scientific basis for enhancing carbon sequestration. We conducted a one-year field experiment in a subtropical Moso bamboo forest with three treatments: control (0 t C·hm -2 ), maize straw (5 t C·hm -2 ), and maize straw biochar (5 t C·hm -2 ). Soil samples were collected at the 3 rd and 12 th months after the treatment. Soil organic carbon chemical composition, the abundance and community composition of cbh I gene were determined by solid-state 13 C NMR, real-time fluorescence quantitative PCR, and high-throughput sequencing, respectively. The results showed that compared with the control, maize straw treatment significantly increased the content of O-alkyl C and decreased aromatic C content, while maize straw biochar treatment showed an opposite effect. Maize straw treatment significantly increased the abundance of cbh I gene and the relative abundance of Penicillium , Gaeumannomyces and Marasmius. However, maize straw biochar treatment reduced the abundance of this gene. The relative abundance of dominant cbh I in soils was positively correlated with the content of O-alkyl C and negatively correlated with the content of aromatic C. Results of redundancy analysis showed that maize straw treatment had a significant effect on the microbial community composition of cbh I gene by changing soil O-alkyl C content, while maize straw biochar affected the microbial community composition of cbh I gene by changing soil pH, organic carbon, and aromatic C content. Maize straw biochar treatment was more effective in increasing soil organic carbon stability and reducing microbial activity associated with carbon degradation in the subtropical Moso bamboo forest ecosystem compared with maize straw treatment. Therefore, the application of biochar has positive significance for maintaining soil carbon storage in subtropical forest ecosystems.

Three undescribed lignan glycosides, echiunines E-G (1-3), as well as eight known compounds (4-11) were isolated from Fritillaria verticillata Willd. Among them, compounds 1-3 were a series of lignan glycosides reported for the first time from genus Fritillaria. Their structures were elucidated by analyses of extensive spectroscopic data and comparison of the NMR data with those reported previously, the absolute configuration of compounds were further confirmed by calculated ECD method. The NO release inhibitory effects of compounds were evaluated in LPS-activated RAW264.7 macrophages. Compounds 7-8 showed inhibitory acitivities in a dose-dependent manner., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 Elsevier B.V. All rights reserved.)

β-cyclodextrin butenate was synthesized by using N, N'-Carbonyldiimidazole (CDI) activating reagent and 4-Dimethylaminopyridine (DMAP) as catalyst. The best preparation condition of β-CD butenate was described as below: reaction temperature was 25°C, concentration of 2-butenoic acid was 450 mmol/L, concentration of DMAP was 12.5 mmol/L and reaction time was 20 minutes and at this condition the yield of β-CD butenate was 0.83 mmol/g. According to the results of FT-IR spectrum, NMR spectroscopy and HPLC-QTof-mass spectrum of β-CD butenate, there were four types β-CD butenate synthesized, which were β-CD-2-butenoic acid monoester, β-CD-2-butenoic acid diester, β-CD-2-butenoic acid triester and β-CD-2-butenoic acid tetraester, respectively., Competing Interests: There are no conflicts of interest with other people or organizations within 3 years of beginning the work submitted that could inappropriately influence the work submitted., (© 2023 The Author(s). Published by Informa UK Limited, trading as Taylor & Francis Group.)

Methane (CH 4 ) and nitrous oxide (N 2 O) are two extremely important greenhouse gases in the atmosphere. Nitrogen fertilizer is an important factor affecting CH 4 and N 2 O emissions in rice fields. Rational application of nitrogen fertilizer can not only promote high yields of rice but also reduce greenhouse gas emissions. Existing studies have shown that nitrogen reduction and optimal application can effectively improve the nitrogen use efficiency of rice on the basis of ensuring the yield and reduce the loss of N 2 O caused by nitrification and denitrification of excessive nitrogen in soil. Fertilization times and fertilizer types have significant effects on CH 4 and N 2 O emissions in paddy fields. In this study, a field experiment was conducted for two consecutive years (2019-2020) to study the effects of fertilizer application on CH 4 and N 2 O emissions from rice fields by setting up four treatments consisting of no fertilizer (CK), customary fertilizer application by farmers (CF), twice fertilizer (TT), and 20% replacement of chemical fertilizer by organic fertilizer (OF) using static chamber-gas chromatography. Additionally, the effect of integrating rice yield and integrated global warming potential (GWP) on the greenhouse gas emission intensity (GHGI) per unit of rice yield was analyzed to explore fertilizer application for yield increase and emission reduction in a typical rice growing area in the middle and lower reaches of Yangtze River. The results showed that:① compared with those of CK, the fertilizer treatments reduced CH 4 emissions by 14.6%-25.1% and increased N 2 O emissions by 610%-1836% in both years; ② compared with those of CF, both the TT and OF treatments showed a trend of increasing CH 4 emissions and reducing N 2 O emissions. CH 4 emissions increased by 1.8% ( P >0.05) and 14.0% ( P <0.05), respectively. The annual average of N 2 O emissions decreased by 63.3% ( P <0.05) and 49.2% ( P <0.05) in both the TT and OF treatments, respectively. ③ Compared with that of CK, both fertilizer applications increased rice yield and reduced GHGI; compared with that of CF, the OF and TT treatments increased the average annual rice yield by 17.0% and 10.7%, respectively, and reduced GHGI by 6.8% and 13.7%, respectively. The OF treatment had a better yield increase than that of the TT treatment, and the TT treatment had a slightly better emission reduction than that of the OF treatment. In terms of combined yield and GHG emission reduction, both twice fertilizer (TT) and 20% replacement of chemical fertilizer by organic fertilizer (OF) could reduce the intensity of GHG emission per unit of rice yield and achieve yield increase and emission reduction while ensuring rice yield.

In July 2019-July 2020, we conducted a field trial to examine the effects of nitrogen addition (60 kg N·hm -2 ·a -1 ), biochar application (10 t·hm -2 ), and their combination on soil N 2 O emission and the relationship between soil N 2 O emission and environmental factors in a typical Moso bamboo ( Phyllostachys edulis ) plantation in Hangzhou City of Zhejiang Province. Soil N 2 O flux of Moso bamboo plantation was measured by the static chamber-gas chromatography technique. The results showed that nitrogen addition treatment increased the annual cumulative N 2 O emission by 14.6%, while biochar application and the combination treatment reduced it by 20.8% and 10.6%, respectively. Soil N 2 O flux rate was significantly correlated with soil temperature, NO 3 - -N concentration, urease and protease activities, and soil NH 4 + -N concentration across all treatments. In conclusion, under the background of nitrogen deposition, the application of biochar would have a significant reduction effect on soil N 2 O fluxes in Moso bamboo plantations.

Gardenia jasminoides Ellis is a traditional aromatic and medicinal plant in China. Here, the complete chloroplast genome of a wild-type gardenia adapted to island climate was assembled. The assembled genome was 155,247 bp in length, with four typical regions, i.e., a large single-copy (LSC) region (85,414 bp), a small single-copy (SSC) region (18,235 bp) and two inverted repeats (IRs) regions (25,799 bp each). In total, 138 genes were predicted, including 90 protein-coding genes, 40 tRNA genes and eight rRNA genes. The overall GC content of the chloroplast genome was 37.5%. The chloroplast genome would provide more information for the phylogeography and phylogeny study of G. jasminoides ., Competing Interests: No potential conflict of interest was reported by the author(s)., (© 2021 The Author(s). Published by Informa UK Limited, trading as Taylor & Francis Group.)

We conducted a pot experiment to investigate the effects of planting broadleaf tree species (i.e., Cinnamomum camphora , Schima superba , and Quercus glauca ) and Moso bamboo ( Phyllostachys edulis ) on soil carbon mineralization and microbial community structure. The rates of soil carbon mineralization were measured via alkali trapping method. The structural and functional diversity of soil bacterial and fungal communities were analyzed by terminal restriction fragment length polymorphism (T-RFLP) and real-time quantitative PCR techniques. The soil planted with Moso bamboo exhibited a significantly higher carbon mineralization rate and labile carbon content than those in the soils planted with broadleaf tree species. The underground biomass of Moso bamboo was higher than that of broadleaf tree species. The soil bacterial communities were more sensitive than fungal communities to the planting of different plant species . Moreover, soil fungal diversity of Moso bamboo was distinctly different from that of broadleaf tree species. Compared to the diversity of soil bacterial communities, the diversity of soil fungal communities was more closely related with soil pH, organic carbon content, and carbon mineralization. In comparison to the broadleaf tree species, the Moso bamboo planting could substantially increase soil organic carbon minera-lization, which was affected mainly by the soil fungal community structure.

Cerasus serrulata (Rosaceae) is an important flowering cherry resource which is valuable for developing new cultivars of flowering cherries. It is broadly distributed and possesses abundant variations. In this study, phylogeographic analysis was conducted to reveal the evolutionary history to better understand the genetic diversity and genetic structure of C. serrulata so as to provide more accurate molecular insights into better conservation and utilization of the germplasm resources. A total of 327 individuals from 18 wild populations were collected. Three chloroplast DNA (cpDNA) fragments (matK, trnD-E, and trnS-G) and the nuclear internal transcribed spacer (ITS) were utilized. The results showed a high genetic diversity at both species level and population level of C. serrulata . High genetic differentiation and the existence of the phylogeographic structure were detected. No significant expansion events were discovered. Two geographic lineages were inferred. One was confined to the Qinling Mountains and the Taihang Mountains. The other was from the Wuling Mountains to the Jiangnan Hilly Regions and then went northeast to the coast of Asia. In addition, some taxonomic treatments of the C. serrulata complex are discussed and reconsidered. Conservation and utilization strategies of wild C. serrulata germplasm resources were recommended., Competing Interests: None declared., (© 2020 The Authors. Ecology and Evolution published by John Wiley & Sons Ltd.)

We investigated the effects of tea plantation age on soil microbial community structure and diversity with surface and subsurface soil samples (0-20 and 20-40 cm) from tea plantation at different ages (0, 20, 25, 38 and 48 years). We analyzed soil bacterial and fungal communities by terminal restriction fragment length polymorphism (T-RFLP) and real-time quantitative PCR techniques. The results showed that soil physicochemical properties changed significantly after planting tea. The contents of soil organic carbon, available nitrogen, and available phosphorus increased at first, and gradually decreased with the increases of tea plantation age. The contents of organic carbon and total nitrogen in the topsoil were significantly higher than those in the subsoil. Moreover, soil bacterial community composition varied across tea plantation age. Bacterial diversity index decreased with increasing tea plantation age. The composition and diversity of soil fungal communities did not show a clear relationship with the tea plantation age. Overall, soil bacterial communities was more sensitive to the tea plantation age than fungal communities. With the increases of tea plantation age, the ratio of soil fungi to bacteria showed increasing trend. Soil microbial community in the tea plantation changed from a "bacterial type" with low fungi/bacteria ratio (F/B) to a "fungal type" with high F/B.

Supramolecular gels constructed from low-molecular-weight gelators via noncovalent interactions have received increasing attention. The rapid development of stimuli-responsive supramolecular gels with attractive properties is highly desirable to meet the ever-growing demand of materials science and chemistry. The inherent reversible and dynamic nature of noncovalent interactions in supramolecular gels endows the materials with sensing, processing, and actuating functions in response to specific environmental changes and offers them great potential in flexible biomaterials and intelligent devices. In particular, pillar[n]arenes with symmetrical pillar-shaped architectures have been recognized as an emerging class of synthetic macrocycles after crown ethers, cyclodextrins, calixarenes, and cucurbiturils, and proven to be excellent candidates for the fabrication of functional supramolecular gels due to their many advantages including facile synthesis, diverse functionalization, and appealing host-guest properties. This review provides a comprehensive overview of recent progress in supramolecular gels involving pillar[n]arenes and their derivatives as synthetic macrocyclic arenes, from the viewpoints of the synthetic approach, controllable assembly, stimuli-responsiveness, and functions. Perspectives of this burgeoning field of research are also given at the end.

A novel supramolecular organic framework gel (PQPA) has been constructed by a novel bilateral 8-hydroxyquinoline modified pillar[5]arene host (P5Q) and a bilateral bromohexyl functionalized pillar[5]arene guest (P5C6). Interestingly, the gel PQPA shows special selective and ultrasensitive sensing property for Fe 3+ . The limit of lowest detection (LOD) of gel PQPA for Fe 3+ is 0.102 nM. Meanwhile, by adding Fe 3+ into the gel PQPA, a no fluorescence Fe 3+ coordinated metallogel (PQPA-Fe) could be constructed. Due to the competitive coordination of F - , fluorescent "turn-on" ultrasensitive response of the Fe 3+ coordinated metallogel (PQPA-Fe) for F - is achieved. The limit of lowest detection of the Fe 3+ coordinated metallogel (PQPA-Fe) for F - is 9.79 nM. In addition, the xerogel of the supramolecular organic framework gel (PQPA) is capable for removing Fe 3+ in a lower concentration aqueous solution at an adsorption rate of 99.9%. More importantly, the supramolecular organic framework gel (PQPA) could be used to detect Fe 3+ in the Yellow River Water samples. The supramolecular organic framework gel (PQPA) and Fe 3+ coordinated metallogel (PQPA-Fe) could act as ultrasensitive reversible fluorescent chemosensor, test kits and fluorescent display materials for Fe 3+ and F - ., (Copyright © 2019 Elsevier B.V. All rights reserved.)

Elevated atmospheric CO 2 concentration, altered precipitation regime, increased nitrogen deposition, and land cover change have not only changed the physical and chemical properties of forest soils, but also affected plant growth and microbial activity, with concequences on soil carbon and nitrogen cycles, including soil CH 4 uptake. In this study, we summarized the important role of soil CH 4 uptake in forests under global change scenarios. The differences of responses as well as the underlying mechanisms of soil CH 4 uptake in forests to global change were reviewed. Elevated atmospheric CO 2 concentration inhibits soil CH 4 uptake. Reduced precipitation tends to promote soil CH 4 uptake. Increased nitrogen input inhibits soil CH 4 uptake in nitrogen-rich forests, but promotes or has no effects on soil CH 4 uptake in nitrogen-poor forests. Conversion of forests to grassland, farmland, or plantations would reduce soil CH 4 uptake, while afforestation increases soil CH 4 uptake. The future research should explore the long-term and multiple effects of global changes on forest soil CH 4 uptake. In addition, molecular biology methods should be developed to explore the microbial mechanism of soil CH 4 uptake.

Here, a novel pseudorotaxanes-type crosslinker of a supramolecular polymer network (WP5-PN) has been constructed from a host water-soluble pillar[5]arene (WP5) and a guest naphthalene dimethylamine derivative (PN) via a stepwise process involving multiple non-covalent interactions. The obtained supramolecular polymers were able to transform into a supramolecular polymer gel (WP5-PN-G) and show AIE properties in DMSO-H2O binary solution. Interestingly, due to the dynamic and reversible nature of non-covalent interactions, the resultant supramolecular polymer gels exhibited external stimuli-responsiveness to different parameters, such as temperature, acid-base, competitive guest and mechanical stress. Moreover, WP5-PN-G showed fluorescent response for Fe3+ and Cu2+, while its xerogel showed excellent recyclable separation properties for these metal ions with adsorption rates up to 98.07% and 95.38%, respectively. Moreover, by rational introduction of these metal ions into the WP5-PN-G, corresponding metal ion coordinated metallogels, such as WP5-PN-FeG and WP5-PN-CuG were obtained. These metallogels could selectively and sensitively sense F- and CN-, respectively. The detection limits of these metallogels for F- and CN- were about 1 × 10-8 M. The WP5-PN-G has potential applications in multi-analytes detection and separation as well as fluorescent display materials.

The β-cyclodextrin-acrylamide (CDM-AM) copolymer was prepared from acrylamide and β-CD maleate (CDM) using 60 Co γ-ray irradiation method. The optimized preparation conditions for the CDM-AM copolymer are as follows: CDM:AM mass ratio of 1:1; irradiation dose of 4 kGy; and using 20 mL of DMF water solution. The yield rate of CDM-AM was 75% in grams using these synthetic conditions. The effects of the CDM-AM copolymer on the solubility and fungicidal activity of natamycin (NM) and carbendazim (MBC) were investigated. The stability constant of NM·CDM-AM and MBC·CDM-AM complexes at 303 K were of 13,446.06 M -1 and 2595.3 M -1 , respectively. The complexes were characterized using phase solubility diagrams, NMR spectra and FT-IR spectra. The analysis of the biological activities of these two complexes indicated that they possessed enhancing fungicidal activities compared to NM and MBC alone.

Ultrasensitive response properties are an intriguing concern for stimuli-responsive materials. Herein, we report a novel method to achieve an ultrasensitive response by introducing the competition of cationπ and ππ interactions into a pillar[5]arene-based supramolecular organic framework (SOF-AMP). SOF-AMP was constructed with a novel bis-naphthalimide functionalized pillar[5]arene, which was able to form a stable supramolecular gel (SOF-AMP-G) in cyclohexanol. Interestingly, SOF-AMP-G shows an ultrasensitive response to Fe3+ through the competition of cationπ and ππ interactions. Meanwhile, the Fe3+ coordinated SOF (MSOF-Fe) shows an ultrasensitive response to H2PO4-. SOF-AMP-G displayed yellow fluorescence whereas, after the addition of 0.5 equiv. of Fe3+ to SOF-AMP-G, the yellow fluorescence was quenched. The detection limit of SOF-AMP-G for Fe3+ is 7.54 × 10-9 M. More interestingly, the Fe3+ coordinated SOF gel (MSOF-Fe-G) could sense H2PO4- with a fluorescence "turn-on". The detection limit of MSOF-Fe-G for H2PO4- is 4.21 × 10-9 M. Simultaneously, the Fe3+ and H2PO4- responsive thin films based on these SOF gels were prepared. Moreover, these SOF gels could be used as ultrasensitive ion sensors, fluorescent display materials and sensitive logic gates.

In order to investigate the effect of fertilization on soil fungal community of Phyllostachys violascens stand, we characterized the abundance and community structure of soil fungi using the methods of terminal restriction fragment length polymorphism (T-RFLP) and real-time quantitative PCR. The experimental treatments included blank control (CK), chemical fertilizer (CF), organic manure (M), mixture of chemical fertilizer and organic manure (CFM) and mixture of chemical fertilizer and organic manure plus mulching (CFMM). The results showed that fungal community structure and diversity were markedly affected by different fertilization regimes. In 0-20 cm soil layer, CF and CFMM treatments resulted in a significant reduction of Shannon and evenness indices. Soil fungi under CK, M, CFMM treatments in 0-20 cm soil layer and CK, CF, CFMM treatments in 20-40 cm soil layer formed independent community structures. Soil fungal abundance showed significantly positive correlation with the concentrations of soil total nitrogen, ammonium nitrogen and nitrate nitrogen. Organic matter, ammonium nitrogen, available phosphorus and available nitrogen concentrations were the main factors that influenced the variation of fungal community composition. The above findings indicated that 0-20 cm and 20-40 cm soil layers had different characteristics in response to different fertilization regimes under P. violascens stand. The soil fungal community in the 0-20 cm layer was mainly affected by soil organic matter, while that in the 20-40 cm layer was sensitive to the input of chemical fertilizer. The effect of fertilization on the diversity of fungal community mainly occurred in the 0-20 cm soil layer.

Modifications at either end, both ends, or in-between the ends of a Morpholino oligo provide functional groups for further conjugation. Amino groups are the most useful and efficient reactive entities for chemical bonding with other molecules. The combination of modifications at both ends, especially with double functionalization at the 3'-end, yields myriad opportunities for diverse applications. An orthogonally protected diamine for advanced 3'-end double modification on the solid phase synthesis support allows the convenient assembly of a vast variety of custom-designed molecules. A particular application is the assembly of a class of Vis-Vivo-Morpholino where at the 3'-end an optically visible fluorophore is installed at one side for fluorescent detection and an in vivo delivery moiety is attached at the other side for intracellular activity studies.

In order to elucidate the effects of intensive management on soil carbon pool, nitrogen pool, enzyme activities in Moso bamboo (Phyllostachys pubescens) plantations, we collected soil samples from the soil surface (0-20 cm) and subsurface (20-40 cm) layers in the adjacent Moso bamboo plantations with extensive and intensive managements in Sankou Township, Lin'an City, Zhejiang Province. We determined different forms of C, N and soil invertase, urease, catalase and acid phosphatase activities. The results showed that long-term intensive management of Moso bamboo plantations significantly decreased the content and storage of soil organic carbon (SOC), with the SOC storage in the soil surface and subsurface layers decreased by 13.2% and 18.0%, respectively. After 15 years' intensive management of Masoo bamboo plantations, the contents of soil water soluble carbon (WSOC), hot water soluble carbon (HWSOC), microbial carbon (MBC) and readily oxidizable carbon (ROC) were significantly decreased in the soil surface and subsurface layers. The soil N storage in the soil surface and subsurface layers in intensively managed Moso bamboo plantations increased by 50.8% and 36.6%, respectively. Intensive management significantly increased the contents of nitrate-N (NO 3 - -N) and ammonium-N (NH 4 + -N), but decreased the contents of water-soluble nitrogen (WSON) and microbial biomass nitrogen (MBN). After 15 years' intensive management of Masoo bamboo plantations, the soil invertase, urease, catalase and acid phosphatase activities in the soil surface layer were significantly decreased, the soil acid phosphatase activity in the soil subsurface layer were significantly decreased, and other enzyme activities in the soil subsurface layer did not change. In conclusion, long-term intensive management led to a significant decline of soil organic carbon storage, soil labile carbon and microbial activity in Moso bamboo plantations. Therefore, we should consider the use of organic fertilizer in the intensive mana-gement process for the sustainable management of Moso bamboo plantations in the future.

Converting natural forests to plantations significantly affects the characteristics of soil organic carbon (C) pools, due to the changes of vegetation cover and management practices. In this paper, to investigate the effects of conversion from evergreen broad-leaved forest (EBF) to Chinese chestnut plantation (CP) on soil organic C pool, the soils from surface (0-20 cm) and subsurface (20-40 cm) layers were sampled from the above two forests (the CP was converted from the EBF and had been intensively managed for 10 years) in Lin'an City, Zhejiang Province. The soil orga-nic C storage, labile organic C pools, and other basic soil properties were determined. The chemical composition of soil organic C was determined by nuclear magnetic resonance (NMR) technique. Results showed that the soil organic C storage, water soluble organic C (WSOC), hot water soluble organic C (HWSOC), microbial biomass C (MBC) and readily oxidizable C (ROC) concentrations in the surface layer decreased by 19.7%, 34.4%, 25.8%, 30.4% and 25.2%, respectively, after the conversion from EBF to CP. Such values for the subsurface layers decreased by 13.5%, 38.4%, 19.8%, 34.1% and 22.2%, respectively. The O-alkyl C content, aromatic C content and aromaticity of soil organic C in the surface layer decreased signi-ficantly by the land-use conversion, while alkyl C content, carbonyl C content and alkyl C to O-alkyl C (A/O-A) ratio increased signi-ficantly. The O-alkyl C content of soil organic C in the subsurface layer decreased significantly by the land-use conversion, alkyl C content and A/O-A ratio increased significantly, and aromatic C content, carbonyl C content and aromaticity of soil organic C were not changed. In conclusion, conve-rting EBF to CP and subsequent intensive management significantly decreased the soil organic C sto-rage and labile C pool contents and altered the chemical composition of soil organic C.

Objective: To evaluate clinical results of early total care (ETC) treatment for elderly patients with intertrochanteric femur fractures., Methods: Clinical data of 106 elderly patients with intertrochanteric fracture treated from January 2012 and February 2015 were retrospectively studied. According to whether receiving the early total care mode, the patients were divided into 2 groups, 34 cases were diagnosed and treated with early total care pattern (ETC group), including 14 males and 20 females with an average age of (74.88 ± 4.38) years old ranging from 70 to 86. According to Evans types, 4 cases were type I, 5 cases were type II, 13 cases were type III, 11 cases were type IV, 1 case was type V. Seventy-two patients were treated with conventional trauma method (conventional group), including 35 males and 37 females with an average age of (74.46 ± 3.63) years old ranging from 70 to 85. According to Evans type, 8 cases were type I ,13 cases were type II, 25 cases were type III, 25 cases were type IV, and 1 case was type V. All fractures were treated with proximal femoral nails anti-rotation (PFNA). Operative time, hospital stays, leaving bed time, complications, cases of death at 1 year after operation, postoperative Harris score at 12 months were observed and compared., Results: All patients were followed up, the time of ETC group ranged from 9 to 18 months with an average of 13.29 ± 1.51, and the time in conventional group ranged from 12 to 16 months with an average 12.93 ± 1.15, while there was no significant difference between two groups in time of following-up (t = 1.368, P = 0.174). There was no significant meaning in cases of death between ETC group (2 cases) and conventional group (8 cases). Three cases occurred complications in ETC group, and 20 cases in conventional group,there was obvious meaning between two groups (χ² = 0.739, P = 0.318). Operative time,hospital stays,leaving bed time in ETC group respectively was (2.03 ± 0.67) d, (15.41 ± 2.87) d and (3.62 ± 0.74) d; while in conventional group respectively was (4.17 ± 1.59) d, (20.11 ± 4.24) d and (5.35 ± 1.22) d; there were significant differences between two groups in operative time, hospital stays, leaving bed time. Postoperative Harris scores at 12 months in ETC group was (82.32 ± 4.56), and (79.24 ± 5.52) in conventional group, there was obvious meaning between two groups (t = 2.833, P = 0.006)., Conclusion: ETC pattern is a novel method for diagnosis and treatment of intertrochanteric femur fractures in elderly, it could shorten operative time, hospital stays, leaving bed time, decrease complications and promote recovery of function.

Objective: To establish a neonatal pig model of hemolytic jaundice., Methods: Twelve seven-day-old purebred Yorkshire pigs were randomly divided into an experimental group and a control group (n=6 each). Immunization of New Zealand white rabbits was used to prepare rabbit anti-porcine red blood cell antibodies, and rabbit anti-porcine red blood cell serum was separated. The neonatal pigs in the experimental group were given an intravenous injection of rabbit anti-porcine red blood cell serum (5 mL), and those in the control group were given an intravenous injection of normal saline (5 mL). Venous blood samples were collected every 6 hours for routine blood test and liver function evaluation., Results: The experimental group had a significantly higher serum bilirubin level than the control group at 18 hours after the injection of rabbit anti-porcine red blood cell serum (64±30 μmol/L vs 20±4 μmol/L; P<0.05). In the experimental group, the serum bilirubin level reached the peak at 48 hours (275±31 μmol/L), and decreased significantly at 96 hours after the injection (95±17 μmol/L), but all significantly higher than that in the control group (P<0.05). At 18 hours after the injection, the experimental group had a significantly lower red blood cell (RBC) count than the control group [(4.58±0.32)×10(12)/L vs (5.09±0.44)×10(12)/L; P<0.05]; at 24 hours, the experimental group showed further reductions in RBC count and hemoglobin level and had significantly lower RBC count and hemoglobin level than the control group [RBC: (4.21±0.24)×10(12)/L vs (5.11±0.39)×10(12)/L, P<0.05; hemoglobin: 87±3 g vs 97±6 g, P<0.05]. The differences in RBC count and hemoglobin level between the two groups were largest at 36-48 hours., Conclusions: The neonatal pig model of hemolytic jaundice simulates the pathological process of human hemolytic jaundice well and provides good biological and material bases for further investigation of neonatal hemolysis.

To investigate variation of soil fungal community in response to invasion of Phyllostachys edulis into native broadleaf forest, we characterized the community structure and the abundance of fungi in soil under bamboo (BB), mixture forest of bamboo and broadleaf (MF) and broadleaf forest (BL) using terminal restriction fragment length polymorphism (T-RFLP) and real-time quantitative PCR. The results showed that the most obvious difference in the soil fungal community structure was observed between the BB and BF stands, followed by that between the MF and BL. Shannon index and evenness index of soil fungi were higher in the MF than in the BB and BL. pH and NH4+-N content were the most important environmental gradients on the distribution of fungal community under BB, while NO3(-)-N content significantly affected the distribution of the fungal community under BL. The abundance of fungi in BL was significantly higher than that in BB and MF, and the fungi abundance showed a negative correlation with soil pH but a positive correlation with NO3(-)-N content. These results implied that heterotrophic nitrification driven by fungi could occur in soil of BL, and this process might be changed by the bamboo invasion.

Soil CO2 effluxes in natural broad-leaved forest and the conversed Chinese fir plantation in Linglong Mountains Scenic of Zhejiang Province were evaluated by using static closed chamber and gas chromatography method. The results showed that soil CO2 efflux showed consistent seasonal dynamics in natural broad-leaved forest and Chinese fir plantation, with the maximums observed in summer and autumn, the minimums in winter and spring. Soil CO2 effluxes were 20.0-111.3 and 4.1-118.6 mg C . m-2 . h-1 in natural broad-leaved forest and Chinese fir plantation, respectively. The cumulative soil CO2 emission of natural broad-leaved forest (16.46 t CO2 . hm-2 . a-1) was significantly higher than that of Chinese fir plantation (11.99 t CO2 . hm-2 . a-1). Soil moisture did not affect soil CO2 efflux. There was a significant relationship between soil CO2 efflux and soil temperature at 5 cm depth. There was no significant relationship between soil CO2 efflux of natural broad-leaved forest and water soluble organic carbon content, while water soluble organic carbon content affected significantly soil CO2 efflux in Chinese fir plantation. Converting the natural broad-leaved forest to Chinese fir plantation reduced soil CO2 efflux significantly but improved the sensitivity of soil respiration to environmental factors.

Dynamic changes (from April 2013 to March 2014) in soil respiration components were investigated by Li-8100 in the Moso bamboo plantation in Lin' an City, Zhejiang Province. Results showed that the average annual values for the soil total respiration rate, heterotrophic respiration rate, and autotrophic respiration rate in the Moso bamboo plantation were 2.93, 1.92 and 1.01 imol CO2 . m-2 . s-1, respectively. The soil respiration rate and its components exhibited strongly a seasonal dynamic pattern. The maximum appeared in July 2013, and the minimum appeared in January 2014. The annual cumulative CO2 emissions through soil respiration, heterotrophic respiration, and autotrophic respiration were 37.25, 24.61 and 12.64 t CO2 . hm-2 . a-1, respectively. The soil respiration and its components showed a close relation with soil temperature of 5 cm depth, and the corresponding Q10, values at 5 cm depth were 2.05, 1.95 and 2.34, respectively. Both the soil respiration and heterotrophic respiration were correlated to soil water soluble organic C (WSOC) content, but no significant relationship between autotrophic respiration and WSOC was observed. There were no significant relationships between soil respiration components and soil moisture content or microbial biomass C. The seasonal changes in soil respiration components in the Moso bamboo plantation were predominantly controlled by the soil temperature, and the soil WSOC content was an important environmental factor controlling total soil respiration and soil heterotrophic respiration.

The β-cyclodextrin-acrylamide (CDM-AM) copolymer was prepared from acrylamide and β-CD maleate (CDM) using K2S2O8 as initiator. The effects of the CDM-AM copolymer on the solubility and fungicidal activity of natamycin (NM) and carbendazim (MBC) were investigated. The stability constant of NM·CDM-AM and MBC·CDM-AM complexes at 303 K were of 10,725.45 M(-1) and 3000.89 M(-1), respectively. The complexes were characterized using phase solubility diagrams, NMR spectra and FT-IR spectra. The analysis of the biological activities of these two complexes indicated that they possessed enhancing fungicidal activities compared to NM and MBC alone., (Copyright © 2015 Elsevier Ltd. All rights reserved.)

β-CD maleate (CDM) and β-CD itaconate (CDI) were prepared by esterification of β-cyclodextrin (β-CD) with maleic acid and itaconic acid using phosphate as a catalyst in a semi-dry process. The esterification of β-CD was carried out using [Itaconic acid ] or [Maleic acid ] 4 mol/mol of CD; M/L ratio 1:0.6; temperature 110 °C; [4-methoxyphenol] 2.5% amount of acid; reaction time 3.5h. The esterification rates of CDM and CDI are 70.38% and 21.02%, respectively. We found that CDM and CDI were both monoesters. Here, we also established a new evaluation method for the rate of esterification., (Copyright © 2014 Elsevier Ltd. All rights reserved.)

CO2, N2O and CH4 are important greenhouse gases, and soils in forest ecosystems are their important sources. Carya cathayensis is a unique tree species with seeds used for high-grade dry fruit and oil production. Understory vegetation management plays an important role in soil greenhouse gases emission of Carya cathayensis stands. A one-year in situ experiment was conducted to study the effects of understory removal on soil CO2, N2O and CH4 emissions in C. cathayensis plantation by closed static chamber technique and gas chromatography method. Soil CO2 flux had a similar seasonal trend in the understory removal and preservation treatments, which was high in summer and autumn, and low in winter and spring. N2O emission occurred mainly in summer, while CH4 emission showed no seasonal trend. Understory removal significantly decreased soil CO, emission, increased N2O emission and CH4 uptake, but had no significant effect on soil water soluble organic carbon and microbial biomass carbon. The global warming potential of soil greenhouse gases emitted in the understory removal. treatment was 15.12 t CO2-e . hm-2 a-1, which was significantly lower than that in understory preservation treatment (17.04 t CO2-e . hm-2 . a-1).

Effect of biochar addition on soil CO2 efflux in a typical Chinese chestnut (Castanea mollissima) plantation in Lin'an, Zhejiang Province, China was investigated from July 2012 to July 2013 by the static closed chamber-GC technique. Soil temperature, soil moisture, WSOC and MBC concentrations were determined as well. Results showed that soil CO2 efflux exhibited a strong sea- sonal pattern. Compared with the control (without biochar application), the biochar treatment increased the soil CO2 efflux only in the first month since application, and then the effect diminished thereafter. There were no significant differences in the annual cumulative value of soil CO2 efflux between the biochar and control treatments. The annual mean value in soil MBC concentration (362 mg · kg(-1)) in the biochar treatment was higher than that (322 mg · kg(-1)) in the control. However, no significant difference in the soil WSOC concentration was found between the biochar and control treatments. Strong exponential relationships between soil temperature and soil CO2 efflux were observed regardless of the treatment and soil layer. The apparent temperature sensitivity (Q10) of soil CO2 efflux in the biochar treatment was higher than that in the control. Soil CO2 efflux was related to soil WSOC concentration but not with soil MBC or moisture content. To conclude, the application of bamboo leaf biochar did not affect the annual cumulative CO2 emission in the Chinese chestnut plantation but increased the Q10, and the CO2 efflux was predominantly controlled by the soil temperature and soil WSOC level.

A novel bacterial strain, CMG1240(T), was isolated in 1988 from mixed soil samples collected from the United States and South America in a selective enrichment medium with guar gum as the sole carbon source. This microbial isolate showed β-mannanolytic activity to hydrolyse the galactomannans present in guar gum. Strain CMG1240(T) was aerobic, Gram-stain-variable, non-motile, rod-shaped and endospore-forming. It was further examined based on a combination of phenotypic, physiological and genetic characterization. On the basis of 16S rRNA gene sequence similarity, cellular lipid profile and fatty acid composition, strain CMG1240(T) was shown to belong unequivocally to the genus Paenibacillus. Quinone analysis showed that MK-7 was the only menaquinone detected. The main cell-wall sugar was xylose with trace amounts of mannose and glucose. The major polar lipids were diphosphatidylglycerol, phosphatidylglycerol, phosphatidylethanolamine, and unknown glycolipids, phospholipids, phosphoglycolipids and other lipids. The peptidoglycan structure was A1γ (meso-diaminopimelic acid-direct). The major fatty acids were anteiso-C15 : 0 and C16 : 0. The DNA G+C content was 46 mol% as determined experimentally and by analysis of the genomic sequence. The 16S rRNA gene sequence of strain CMG1240(T) shared highest similarity with that of Paenibacillus fonticola ZL(T) (97.6 %) while all other tested Paenibacillus strains showed lower sequence similarities (≤95.3 %). The results of DNA-DNA hybridization and chemotaxonomic tests enabled the genotypic and phenotypic differentiation of strain CMG1240(T) from P. fonticola. Based on these results, strain CMG1240(T) ( = ATCC BAA-2594(T) = DSM 25539(T)) should be designated the type strain of a novel species within the genus Paenibacillus, for which the name Paenibacillus lentus sp. nov. is proposed.

Carbon (C) occluded in phytolith (PhytOC) is highly stable at millennium scale and its accumulation in soils can help increase long-term C sequestration. Here, we report that soil PhytOC storage significantly increased with increasing duration under intensive management (mulching and fertilization) in Lei bamboo (Phyllostachys praecox) plantations. The PhytOC storage in 0-40 cm soil layer in bamboo plantations increased by 217 Mg C ha(-1), 20 years after being converted from paddy fields. The PhytOC accumulated at 79 kg C ha(-1) yr(-1), a rate far exceeding the global mean long-term soil C accumulation rate of 24 kg C ha(-1) yr(-1) reported in the literature. Approximately 86% of the increased PhytOC came from the large amount of mulch applied. Our data clearly demonstrate the decadal scale management effect on PhytOC accumulation, suggesting that heavy mulching is a potential method for increasing long-term organic C storage in soils for mitigating global climate change.

In June 2011-June 2012, a fertilization experiment was conducted in a typical Castanea mollissima stand in Lin' an of Zhejiang Province, East China to study the effects of inorganic and organic fertilization on the soil CO2 flux and the relationships between the soil CO2 flux and environmental factors. Four treatments were installed, i. e., no fertilization (CK), inorganic fertilization (IF), organic fertilization (OF), half organic plus half inorganic fertilization (OIF). The soil CO2 emission rate was determined by the method of static closed chamber/GC technique, and the soil temperature, soil moisture content, and soil water-soluble organic carbon (WSOC) concentration were determined by routine methods. The soil CO2 emission exhibited a strong seasonal pattern, with the highest rate in July or August and the lowest rate in February. The annual accumulative soil CO2 emission in CK was 27.7 t CO2 x hm(-2) x a(-1), and that in treatments IF, OF, and OIF was 29.5%, 47.0%, and 50.7% higher than the CK, respectively. The soil WSOC concentration in treatment IF (105.1 mg kg(-1)) was significantly higher than that in CK (76.6 mg x kg(-1)), but was obviously lower than that in treatments OF (133.0 mg x kg(-1)) and OIF (121.2 mg x kg(-1)). The temperature sensitivity of respiration (Q10) in treatments CK, IF, OF, and OIF was 1.47, 1.75, 1.49, and 1.57, respectively. The soil CO2 emission rate had significant positive correlations with the soil temperature at the depth of 5 cm and the soil WSOC concentration, but no significant correlation with soil moisture content. The increase of the soil WSOC concentration caused by fertilization was probably one of the reasons for the increase of soil CO2 emission from the C. mollissima stand.

Objective: To study the effects of biological clock protein on circadian disorders in hypoxic-ischemic brain damage (HIBD) by examining levels of CLOCK and BMAL1 proteins in the pineal gland of neonatal rats., Methods: Seventy-two 7-day-old Sprague-Dawley (SD) rats were randomly divided into sham-operated and HIBD groups. HIBD model was prepared according to the modified Levine method. Western blot analysis was used to measure the levels of CLOCK and BMAL1 in the pineal gland at 0, 2, 12, 24, 36 and 48 hours after operation., Results: Both CLOCK and BMAL levels in the pineal gland increased significantly 48 hours after HIBD compared with the sham-operated group (P<0.05). There were no significant differences in levels of CLOCK and BMAL proteins between the two groups at 0, 2, 12, 24 and 36 hours after operation (P>0.05)., Conclusions: Levels of CLOCK and BMAL1 proteins in the pineal gland of rats increase significantly 48 hours after HIBD, suggesting that both CLOCK and BMAL1 may be involved the regulatory mechanism of circadian disorders in rats with HIBD.

Objective: To explore efficacy of modified funnel method for transpedicular bone grafting in treating thoracolumbar vertebras fracture., Methods: From May 2006 to November 2011, 35 patients (19 males and 16 females, ranged in age from 21 to 66 years with an average of 34.6 years) with thoracolumbar vertebras fracture were treated by posterior pedicle screw fixation, reduction and modified funnel method for transpedicular autogenous iliac bone grafting. Of the 35 cases, 9 cases were severe compression fracture and 26 cases were burst fracture. The anterior body height and Cobb's angle of injured vertebral were measured by X-ray; the effect of implantation bone and CT value were assessed by radiograph CT scan; Nerve function were evaluated according to Frankel's neurological function classification and back pain were evaluated by visual analogue scale (VAS)., Results: Thirty-two patients were followed up from 18 to 37 months with an average of 19.7 months. No nerve injuries aggravated, and 11 patients with partial nerve injuires preoperatively were improved at least 1 level recovery postoperatively; Breakage or loosen of screws were not found. Vertebral bone grafting filled well, bone fusion were got after 6 months' treatment and without cavity gap in grafting bone area. The anterior vertebral height was improved respectively from preoperative (50.17 +/- 8.26)% to postoperative (90.79 +/- 4.85)%, and (90.34 +/- 4.03)% at the final follow-up. The Cobb's angle improved from preoperative (28.7 +/- 6.24) degrees to postoperative (7.26 +/- 3.79) degrees, with (7.34 +/- 4.05) degrees at the final follow-up. CT value of injured vertebras at the final follow-up were significantly higher than adjacent vertebras'. The average VAS was 1.06., Conclusion: Posterior pedicle screw fixation, reduction and modified funnel method for transpedicular autogenous iliac bone grafting is a feasible and safe method for the treatment of thoracolumbar vertebras fracture. It can effectively prevent bone loss of injured vertebral height, progressive deformity of kyphosis, and keep spinal movement function at the maximum.

To investigate the effects of the conversion from native shrub forest (NF) to Chinese chestnut plantation (CP) on the soil carbon (C) and nitrogen (N) pools, soil samples were collected from the adjacent NF and CP in Anji County of Zhejiang Province, with their water-soluble organic C (WSOC), microbial biomass C (MBC), readily oxidizable C (ROC), water-soluble organic N (WSON), and microbial biomass N (MBN) determined. The spectral characteristics of soil organic C were also determined by using nuclear magnetic resonance (NMR) technique. After the conversion from NF to CP, the soil alkalyzable N, available phosphorus, and available potassium contents increased significantly, while the soil WSOC, MBC, ROC, WSON, and MBN were in adverse. The soil organic C in both NF and CP was dominated by alkyl C and O-alkyl C, but the proportions of O-alkyl C and carbonyl C in soil organic C decreased while the proportions of alkyl C and aromatic C as well as the alkyl C/O-alkyl C ratio and the aromaticity of soil organic C all increased significantly after the conversion from NF to CP, indicating that this conversion increased the stability of soil organic C pool significantly. In conclusion, the conversion from NF to CP and the intensive management of CP decreased the contents of soil labile C and soil N but increased the stability of soil C pool significantly.

Based on the 1974-2008 forest fire and meteorological data in Tahe County of Great Xing' an Mountains region, and with the help of Canadian Forest Fire Weather Index System (CFFWIS), this paper qualitatively and quantitatively analyzed the dynamics of forest fire weather indices in the region. In 1974-2008, the mean annual fire occurrence in the region showed an increasing trend, and the increment in 2000-2008 was 72.2%, compared with that in 1974-1999. The fuel moisture codes, fire behavior indices, and fire severity indices in 1974-2008 had an overall increasing trend, which was more evident with time extended. In the future, the probability of forest fire occurrence in the region would be increasing, fuels would be getting drier, fire intensity would be increased, fire weather would be more serious, and fire control would be more difficult. Therefore, more efforts should be made to improve the capability of forest fire control, especially in summer. As a fundamental technique, prescribed burning should be applied to reduce the fuel load. From this doing, the probability of fire occurrence and the fire potential intensity could be reduced.

Aiming at the deficiencies in the researches about the probability distribution model for mixed forests tree measurement factors, a joint maximum entropy probability density function was put forward, based on the maximum entropy principle. This function had the characteristics of 1) each element of the function was linked to the maximum entropy function, and hence, could integrate the information about the probability distribution of measurement factors of main tree species in mixed forests, 2) the function had a probability expression of double-weight, being possible to reflect the characteristics of the complex structure of mixed forests, and accurately and completely reflect the probability distribution of tree measurement factors of mixed forests based on the fully use of the information about the probability distribution of measurement factors of main tree species in mixed forests, and 3) the joint maximum entropy probability density function was succinct in structure and excellent in performance. The model was applied and tested in two sampling plots in Tianmu Mountain Nature Reserve. The fitting precision (R2 = 0.9655) and testing accuracy (R2 = 0.9772) were both high, suggesting that this model could be used as a probability distribution model for mixed forests tree measurement factors, and provided a feasible method to fully understand the comprehensive structure of mixed forests.

Adhesion pili (fimbriae) play a critical role in initiating the events that lead to intestinal colonization and diarrheal disease by enterotoxigenic Escherichia coli (ETEC), an E. coli pathotype that inflicts an enormous global disease burden. We elucidate atomic structures of an ETEC major pilin subunit, CfaB, from colonization factor antigen I (CFA/I) fimbriae. These data are used to construct models for 2 morphological forms of CFA/I fimbriae that are both observed in vivo: the helical filament into which it is typically assembled, and an extended, unwound conformation. Modeling and corroborative mutational data indicate that proline isomerization is involved in the conversion between these helical and extended forms. Our findings affirm the strong structural similarities seen between class 5 fimbriae (from bacteria primarily causing gastrointestinal disease) and class 1 pili (from bacteria that cause urinary, respiratory, and other infections) in the absence of significant primary sequence similarity. They also suggest that morphological and biochemical differences between fimbrial types, regardless of class, provide structural specialization that facilitates survival of each bacterial pathotype in its preferred host microenvironment. Last, we present structural evidence for bacterial use of antigenic variation to evade host immune responses, in that residues occupying the predicted surface-exposed face of CfaB and related class 5 pilins show much higher genetic sequence variability than the remainder of the pilin protein.

With two rice genotypes (cv. Zhongbu 51 and Azucena) differing in their root-secreted acid phosphatase activity (APA) under low P condition as test materials, an agar culture experiment was conducted to investigate their P utilization from myo-inositol hexaphosphate (IHP) under conditions of sterilization and inoculating with soil microorganisms. Under sterilization, the shoot dry mass, P absorption and P concentration of test genotypes supplied with hexaphosphate (IHP) as the P source were significantly higher than those with no P supply, but significantly lower than those supplied with inorganic P, indicating that rice plants could partly acquire P from IHP under sterilized condition. The root-secreted APA of the two genotypes was significantly higher under low P than under sufficient P supply, and the higher APA of Zhongbu 51 under low P could be one of the mechanisms of higher P utilization capability of Zhongbu 51 than Azucena under deficient P supply. When supplied with high level IHP (0.96 mmol P x L(-1)), the shoot dry mass, P absorption and P concentration of both Zhongbu 51 and Azucena were higher than those when supplied with low level IHP (0.16 mmol P x L(-1)), indicating that substrate availability could be one of the limiting factors to the utilization of phytate by rice. Inoculation with soil microorganisms had no significant effects on the shoot dry mass, P absorption and P concentration of rice plants, regardless of IHP level and rice genotypes, implying that the utilization of IHP by rice could not be improved via inoculation with soil microorganisms.

Enterotoxigenic Escherichia coli (ETEC), a major global cause of diarrhea, initiates the pathogenic process via fimbriae-mediated attachment to the small intestinal epithelium. A common prototypic ETEC fimbria, colonization factor antigen I (CFA/I), consists of a tip-localized minor adhesive subunit CfaE and the stalk-forming major subunit CfaB, both of which are necessary for fimbrial assembly. To elucidate the structure of CFA/I at atomic resolution, three recombinant proteins were generated consisting of fusions of the minor and major subunits (CfaEB) and of two (CfaBB) and three (CfaBBB) repeats of the major subunit. Crystals of CfaEB diffracted X-rays to 2.1 A resolution and displayed the symmetry of space group P2(1). CfaBB exhibited a crystal diffraction limit of 2.3 A resolution and had the symmetry of space group P2(1)2(1)2. CfaBBB crystallized in the monoclinic space group C2 and diffracted X-rays to 2.3 A resolution. These structures were determined using the molecular-replacement method.

Safe and efficient in vivo delivery of Morpholino antisense oligos was probably the last and most difficult challenge for the broad application of antisense in animal research and therapeutics. Several arginine-rich peptides effective for in vivo delivery of Morpholino antisense oligos require rather complex and expensive procedures for synthesis and conjugation. This work describes the design and synthesis of a dendritic transporter in a most concise manner where the selection of the core scaffold, functional group multiplication, orthogonal protecting group manipulation, solid phase conjugation, and off-resin perguanidinylation of the transporter structure are all orchestrated for efficient assembly. We utilized triazine as a core to provide a site for on-column conjugation to the Morpholino oligo and to anchor functional side arms which, after extension, multiplication, and deprotection, are subsequently converted from primary amines to the eight guanidinium headgroups that serve for transport across cell membranes. Intravenous administration of the delivery-enabled Morpholino into a splice-reporter strain of transgenic living mice results in de novo expression of splice-corrected green fluorescent protein in a broad range of tissues and organs in those treated mice. This rigorously demonstrates that this new dendritic transporter achieves effective delivery of a Morpholino oligo into the cytosol/nuclear compartment of cells systemically in vivo. The practical conjugation process may overcome any availability limitation for routine use by the scientific community, and the efficient delivery ability of this transporter may advance the application of Morpholino antisense technology in animals.

CfaE is the minor, tip-localized adhesive subunit of colonization factor antigen I fimbriae (CFA/I) of enterotoxigenic Escherichia coli and is thought to be essential for the attachment of enterotoxigenic E. coli to the human small intestine early in diarrhea pathogenesis. The crystal structure of an in cis donor strand complemented CfaE was determined, providing the first atomic view of a fimbrial subunit assembled by the alternate chaperone pathway. The in cis donor strand complemented variant of CfaE structure consists of an N-terminal adhesin domain and a C-terminal pilin domain of similar size, each featuring a variable immunoglobulin-like fold. Extensive interactions exist between the two domains and appear to rigidify the molecule. The upper surface of the adhesin domain distal to the pilin domain reveals a depression consisting of conserved residues including Arg(181), previously shown to be necessary for erythrocyte adhesion. Mutational analysis revealed a cluster of conserved, positively charged residues that are required for CFA/I-mediated hemagglutination, implicating this as the receptor-binding pocket. Mutations in a few subclass-specific residues that surround the cluster displayed differential effects on the two red cell species used in hemagglutination, suggesting that these residues play a role in host or cell specificity. The C-terminal donor strand derived from the major subunit CfaB is folded as a beta-strand and fits into a hydrophobic groove in the pilin domain to complete the immunoglobulin fold. The location of this well ordered donor strand suggests the positioning and orientation of the subjacent major fimbrial subunit CfaB in the native assembly of CFA/I fimbriae.

The ATP-binding cassette protein ABCG2 is a member of a broad family of ABC transporters with potential clinical importance as a mediator of multidrug resistance. We carried out a homology and knowledge-based, and mutationally improved molecular modeling study to establish a much needed structural framework for the protein, which could serve as guidance for further genetic, biochemical, and structural analyses. Based on homology with known structures of both full-length and nucleotide-binding domains (NBD) of ABC transporters and structural knowledge of integral membrane proteins, an initial model of ABCG2 was established. Subsequent refinement to conform to the lipophilic index distributions in the transmembrane domain (TMD) and to the results of site-directed mutagenesis experiments led to an improved model. The complete ABCG2 model consists of two identical subunits facing each other in a closed conformation. The dimeric interface in the nucleotide-binding domain (NBD) involves a characteristic nucleotide sandwich and the interface in the TMD consists of the TM helices 1-3 of one subunit and the helices 5 and 6 of the other. The interface between the NBD and the TMD is bridged by the conserved structural motif between TM2 and TM3, the intracellular domain 1 (ICD1), and the terminal beta-strand (S6) of the central beta-sheet in the NBD. The apparent flexibility of the ICD1 may play a role in transmitting conformational changes from the NBD to the TMD or from the TMD to the NBD.

ATP-NAD kinase phosphorylates NAD to produce NADP by using ATP, whereas ATP-NADH kinase phosphorylates both NAD and NADH. Three NAD kinase homologues, namely, Utr1p, Pos5p and Utr1p, exist in the yeast Saccharomyces cerevisiae, which were all confirmed as ATP-NADH kinases and found to be important to supply NADP(H) for yeast cells. In S. cerevisiae, fatty acid beta-oxidation is restricted to peroxisomes and peroxisomal NADPH is required for beta-oxidation of unsaturated fatty acids with the double bonds at even positions. Single and double gene disruption strains of NAD kinase genes, i.e., utr1, pos5, yef1, utr1yef1, utr1pos5 and yef1pos5 were constructed by PCR-targeting method. The utilization ability of these mutants for unsaturated fatty acids with the double bonds at even or uneven positions was examined, with wild type BY4742 as positive control cell, and fatty-acyl-CoA oxidase gene deletion mutant (fox1) and peroxisomal NADP-dependent isocitrate dehydrogenase isoenzymes gene deletion mutant (idp3) as negative control cells. The results indicated that the NAD kinase homologues, especially Pos5p, were critical for supplying NADP and then NADPH in peroxisomal matrix. NADP, which was supplied mainly by Utr1p, Pos5p and Yef1p, particularly by Pos5p, was proposed to be able to transfer from outside of peroxisome into peroxisomal matrix and then converted to NADPH by Idp3p.

Three NAD kinase homologs, encoded by UTR1, POS5 and YEF1 genes, are found in the yeast Saccharomyces cerevisiae and proven to be important sources of NADPH for the cell. Pos5p, existing in the mitochondrial matrix, is critical for higher temperature endurance and mitochondrial functions, such as glycerol usability and arginine biosynthesis. Through constructing the high-copy expression plasmids of YEF1 and UTR1, which contained the green fluorescent protein reporter tag at their 3' terminus, and introducing them into POS5 gene deletion mutants (i.e. pos5, utr1pos5, yef1pos5 and utr1yef1pos5), the high-copy YEF1 and UTR1 plasmids carrying transformants for pos5 mutants were obtained. Their temperature sensitivity and growth phenotype on media with glycerol as the sole carbon source, or on media without arginine, were checked. Results showed the partial rescue of mitochondrial dysfunctions and temperature sensitivity of pos5 mutants by the high-copy YEF1 gene, and of glycerol growth defect and temperature sensitivity by the high-copy UTR1 gene, which confirmed the potential supplying ability of Yef1p and Utr1p for mitochondrial NADP(H) and implied the weak transport of NADP from cytosol to mitochondria. However, even through the green fluorescent protein reporter label, the subcellular localization of Yef1p and Utr1p in yeast cells could not be observed, which indicated the low expression level of these two NAD kinase homologs.

Enterotoxigenic Escherichia coli (ETEC) represents a formidable food and waterborne diarrheal disease threat of global importance. The first step in ETEC pathogenesis is bacterial attachment to small-intestine epithelial cells via adhesive fimbriae, many of which are genetically related to the prototype colonization factor antigen I (CFA/I). The minor fimbrial subunit CfaE is required for initiation of CFA/I fimbrial assembly and mediates bacterial attachment to host cell-surface receptors. A donor-strand complemented variant of CfaE (dscCfaE) was expressed with a hexahistidine tag, purified to homogeneity and crystallized using the hanging-drop vapor-diffusion method. X-ray diffraction data sets were collected to 2.4 A resolution for both native and derivatized crystals and showed the symmetry of space group P6(2)22, with unit-cell parameters a = b = 142.9, c = 231.9 A. Initial phases were derived from the SIRAS approach and electron density showed two molecules in the crystallographic asymmetric unit. Sequence assignments were aided by anomalous signals from the selenium of an SeMet-derivatized crystal and from S atoms of a native crystal.