73 resultados para Early Stage


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In plants and less-advanced animal species, such as C.elegans, introduction of exogenous double-stranded RNA (dsRNA) into cells would trigger degradation of the mRNA with homologous sequence and interfere with the endogenous gene expression. It might represent an ancient anti-virus response which could prevent the mutation in the genome that was caused by virus infection or mobile DNA elements insertion. This phenomenon was named RNA interference, or RNAi. In this study, RNAi was used to investigate the function of basonuclin gene during oogenesis. Microinjection of dsRNA directed towards basonuclin into mouse germinal-vesicle-intact (GV) oocytes brought down the abundance of the cognate mRNA effectively in a time- and concentration-dependent manner. This reduction effect was sequence-specific and showed no negative effect on other non-homologous gene expression in oocytes, which indicated that dsRNA can recognize and cause the degradation of the transcriptional products of endogenous basonuclin gene in a sequence-specific manner. Immunofluorescence results showed that RNAi could reduce the concentration of basonuclin protein to some extent, but the effect was less efficient than the dsRNA targeting towards tPA and cMos which was also expressed in oocytes. This result might be due to the long half life of basonuclin protein in oocytes and the short reaction time which was posed by the limited life span of GV oocytes cultured in vitro. In summary, dsRNA could inhibit the expression of the cognate gene in oocytes at both mRNA and protein levels. The effect was similar to Knock-out technique which was based on homologous recombination. Furthermore, hairpin-style dsRNA targeting basonuclin gene could be produced by transcription from a recombinant plasmid and worked efficiently to deplete the cognate mRNA in oocytes. This finding offered a new way to study the function of basonuclin in the early stage of oogenesis by infection of primordial oocytes with the plasmid expressing hairpin-style basonuclin dsRNA.

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In the interferon-induced antiviral mechanisms, the Mx pathway is one of the most powerful. Mx proteins have direct antiviral activity and inhibit a wide range of viruses by blocking an early stage of the viral genome replication cycle. However, antiviral activity of piscine Mx remains unclear in vivo. In the present study, an Mx-like gene was cloned, characterized and gene-transferred in rare minnow Gobiocypris rarus, and its antiviral activity was confirmed in vivo. The full length of the rare minnow Mx-like cDNA is 2241 bp in length and encodes a polypeptide of 625 amino acids with an estimated molecular mass of 70.928 kDa and a predicted isoelectric point of 7.33. Analysis of the deduced amino acid sequence indicated that the mature peptide contains an amino-terminal tripartite GTP-binding motif, a dynamin family signature sequence, a GTPase effector domain and two carboxy-terminal leucine zipper motifs, and is the most similar to the crucian carp (Carassius auratus) Mx3 sequence with an identity of 89%. Both P0 and F1 generations of Mx-transgenic rare minnow demonstrated very significantly high survival rate to GCRV infection (P < 0.01). The mRNA expression of Mx gene was consistent with survival rate in F1 generation. The virus yield was also concurrent with survival time using electron microscope technology. Rare minnow has Mx gene(s) of its own but introducing more Mx gene improves their resistance to GCRV. Mx-transgenic rare minnow might contribute to control the GCRV diseases. (C) 2008 Published by Elsevier Ltd.

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Previous study and analysis of cytochrome b suggested that polyploidization event in the genus Tor occurred about 10 Mya ago. In order to understand evolutionary fates of Sox gene in the early stage of genome duplication at the nucleotide level, PCR surveys for Sox genes in three closely related cyprinid fishes T douronensis (2n = 100), T qiaojiensis (2n = ?), T sinensis (2n = 100) and their relative T brevifilis (2n = 50) were performed. Totally, 52 distinct Sox genes were obtained in these four species, representing SoxB, SoxC, and SoxE group. As expected, isoforms of some Sox genes correspond with the ploidy of species, such as two copies of Sox9a exist in tetraploid species. Analysis indicated that duplicated Sox gene pairs caused by polyploidization evolved independently of each other within polyploid species. Results of substitution rate showed nearly equal rate of nonsynonymous substitution of duplicated Sox orthologs among different polyploid species and their diploid relative orthologs, suggesting at the early stage of genome duplicated Sox orthologs are under similar selective constraints in different polyploidy species and their diploid relative at the amino acid level. All PCR fragments of Sox genes obtained in this study are not accompanied by obvious increase in mutations and pseudogene formation which means that they are under strong purifying selection, suggesting that they are functional at the DNA level. Cenealogical analysis revealed that T qiaojiensis was tetraploid, and T douronensis, T qiaojiensis as well as T sinensis had an allotetraploid ancestor. (C) 2009 Elsevier B.V. All rights reserved.

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A novel multi-cell device made of organic glass was designed to study morphological and physiological characteristics of Microcystis population trapped in simulated sediment conditions. Changes of colonial morphology and antioxidant activities of the population were observed and measured over the range of 31-day incubation. During the incubation, the antioxidant enzyme activities fluctuated significantly in sediment environments. The activities of catalase (CAT), glutathione peroxidase (GPx) and malondialdehyde (NIDA) reached the highest on the 11(th) day, 6(th) day and 6(th) day. respectively, and then dropped down remarkably in the following days. The ratios of Fv/Fm and the maximal electron transfer rate (ETRm) declined during the initial days (1 similar to 11(th) day), but rebounded on the 16(th) day, which were consistent with the variations of total protein. In the end of incubation. gas vacuoles were hard]), observed and the gelatinous sheath was partly disappeared in the population of Microcystis. Nevertheless, the remaining populations. upon transferred to culture medium, were able to grow though experiencing a longer lag phase of nine days. The results indicated that the sediment environments were able to cause negative effects on M. aeruginosa cells. The cells, however, responded to against the possible damage afterwards. It is thus proposed the acute responses in the population during the early stage of sedimentation could be of importance in aiding the long-term survivor of Microcystis and recruitment in lake sediments. The present study also demonstrated the utility of the device in simulating the sediment environments for further investigation.

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The genus Sarcocheilichthys is a group of small cyprinid fishes comprising 10 species/sub-species widely distributed in East Asia, which represents a valuable model for understanding the speciation of freshwater fishes in East Asia. In the present study, the molecular phylogenetic relationship of the genus Sarcocheilichthys was investigated using a 1140 bp section of the mitochondrial cytochrome b gene. Two different tree-building methods, maximum parsimony (MP) and Bayesian methods, yielded trees with almost the same topology, yielding high bootstrap values or posterior probabilities. The results showed that the genus Sarcocheilichthys consists of two large clades, clades I and II. Clade I contains Sarcocheilichthys lacustris, Sarcocheilichthys sinensis and Sarcocheilichthys parvus, with S. parvus at a basal position. In clade II, Sarcocheilichthys variegatus microoculus is at a basal position; samples of the widespread species, Sarcocheilichthys nigripinnis, form a large subclade containing another valid species Sarcocheilichthys czerskii. Sarcocheilichthys kiangsiensis is retained at an intermediate position. Since S. czerskii is a valid species in the S. nigripinnis clade, remaining samples of S. nigripinnis form a paraphyly. This speciation process is attributed to geographical isolation and special environmental conditions experienced by S. czerskii and stable environments experienced by the other S. nigripinnis populations. This type of speciation process was suggested to be very common. Samples of Sarcocheilichthys sinensis sinensis and Sarcocheilichthys sinensis fukiensis that did not form their own monophyletic groups suggest an early stage of speciation and support their sub-species status. Molecular clock analysis indicates that the two major lineages of the genus Sarcocheilichthys, clades I and II diverged c. 8.89 million years ago (mya). Sarcocheilichthys v. microoculus from Japan probably diverged 4.78 mya from the Chinese group. The northern-southern clades of S. nigripinnis began to diverge c. 2.12 mya, while one lineage of S. nigripinnis evolved into a new species, S. czerski, c. 0.34 mya. (C) 2008 The Authors Journal compilation (C) 2008 The Fisheries Society of the British Isles.

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The androgen role in the maintenance of prostate epithelium is subject to conflicting opinions. While androgen ablation drives the regression of normal and cancerous prostate, testosterone may cause both proliferation and apoptosis. Several investigators note decreased proliferation and stronger response to chemotherapy of the prostate cancer cells stably expressing androgen receptor (AR), however no mechanistic explanation was offered. In this paper we demonstrate in vivo anti-tumor effect of the AR on prostate cancer growth and identify its molecular mediators. We analyzed the effect of AR on the tumorigenicity of prostate cancer cells. Unexpectedly, the AR-expressing cells formed tumors in male mice at a much lower rate than the AR-negative controls. Moreover, the AR-expressing tumors showed decreased vascularity and massive apoptosis. AR expression lowered the angiogenic potential of cancer cells, by increasing secretion of an anti-angiogenic protein, thrombospondin-1. AR activation caused a decrease in RelA, a subunit of the pro-survival transcription factor NF kappa B, reduced its nuclear localization and transcriptional activity. This, in turn, diminished the expression of its anti-apoptotic targets, Bcl-2 and IL-6. Increased apoptosis within AR-expressing tumors was likely due to the NF kappa B suppression, since it was restricted to the cells lacking nuclear (active) NF kappa B. Thus we for the first time identified combined decrease of NF kappa B and increased TSP1 as molecular events underlying the AR anti-tumor activity in vivo. Our data indicate that intermittent androgen ablation is preferable to continuous withdrawal, a standard treatment for early-stage prostate cancer. (C) 2007 Wiley-Liss, Inc.

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The microbiotic crust study is among new focuses in investigating on the desertification control. Based on determination of algal crusts with different successive ages (4-, 8-, 17-, 34-, 42-year-old) and unconsolidated sand in the desert area, species composition and clustering analyses were carried out in this study. Results on successional orientation revealed that (1) the abundance of Cyanophyta, specially of Scytonema javanicum gradually decreased; (2) the abundance of Chlorophyta, Bacillariophyta and a species of Cyanophyta, Phormidium tenue increased; (3) the biodiversity increased gradually with the community succession; and (4) biomass of microalgae increased at the early stage, but decreased at the later stage due to the abundance of lichens and mosses. But, the speed of natural succession was so slow that the community-building species was still the first dominant species after 42 years, except that its dominant degree decreased just slightly. However, successive speed and trend were affected by water, vegetation coverage, terrain, time and soil physico-chemical properties as well, especially Mn content in the soil appeared to have a threshold effect.

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Nanostructured FeAl intermetallics were prepared directly by mechanical alloying (MA) in a high-energy planetary ball-mill. The phase transformations and structural changes occurring in the studied material during mechanical alloying were investigated by X-ray diffraction (XRD). Transmission electron microscopy (TEM) was employed to examine the morphology of the powders. Thermal behavior of the milled powders was examined by differential scanning calorimetry (DSC). Disordered Fe(Al) solid solution was formed at the early stage. After 30 h of milling, Fe(Al) solid solution transformed into an ordered FeAl phase. The average crystallite size reduction down to about 12 nm was accompanied by the introduction of the average lattice strain up to 1.7%. The TEM picture showed that the size of milled powders was less than 30 nm. (c) 2007 Elsevier B.V. All rights reserved.

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We present a comparative study of InAs quantum dots grown on Si-doped GaAs (10 0) substrates, Si-doped GaAs (10 0) vicinal substrates, and semi-insulating GaAs (10 0) substrates. The density and size distribution of quantum dots varied greatly with the different substrates used. While dots on exact substrates showed only one dominant size, a clear bimodal size distribution of the InAs quantum dots was observed on GaAs vicinal substrates, which is attributed to the reduced surface diffusion due to the presence of multiatomic steps. The emission wavelength is blueshifted during the growth of GaAs cap layer with a significant narrowing of FWHM. We found that the blueshift is smaller for QDs grown on GaAs (10 0) vicinal substrates than that for dots on exact GaAs (100) substrates. This is attributed to the energy barrier formed at the multiatomic step kinks which prohibits the migration of In adatoms during the early stage of cap layer growth. (c) 2005 Elsevier B.V. All rights reserved.

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Novel hydrogen dilution profiling (HDP) technique was developed to improve the uniformity in the growth direction of mu c-Si:H thin films prepared by hot wire chemical vapor deposition (HWCVD). It was found that the high H dilution ratio reduces the incubation layer from 30 nm to less than 10 nm. A proper design of hydrogen dilution profiling improves the uniformity of crystalline content, X-c, in the growth direction and restrains the formation of micro-voids as well. As a result the compactness of mu c-Si:H films with a high crystalline content is enhanced and the stability of mu c-Si:H thin film against the oxygen diffusion is much improved. Meanwhile the HDP mu c-Si:H films exhibit the low defect states. The high nucleation density from high hydrogen dilution at early stage is a critical parameter to improve the quality of mu c-Si:H films. (c) 2006 Published by Elsevier B.V.

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Novel hydrogen dilution profiling (HDP) technique was developed to improve the uniformity in the growth direction of mu c-Si:H thin films prepared by hot wire chemical vapor deposition (HWCVD). It was found that the high H dilution ratio reduces the incubation layer from 30 nm to less than 10 nm. A proper design of hydrogen dilution profiling improves the uniformity of crystalline content, X-c, in the growth direction and restrains the formation of micro-voids as well. As a result the compactness of mu c-Si:H films with a high crystalline content is enhanced and the stability of mu c-Si:H thin film against the oxygen diffusion is much improved. Meanwhile the HDP mu c-Si:H films exhibit the low defect states. The high nucleation density from high hydrogen dilution at early stage is a critical parameter to improve the quality of mu c-Si:H films. (c) 2006 Published by Elsevier B.V.

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An important concept proposed in the early stage of robot path planning field is the shrinking of the robot to a point and meanwhile expanding of the obstacles in the workspace as a set of new obstacles. The resulting grown obstacles are called the Configuration Space (Cspace) obstacles. The find-path problem is then transformed into that of finding a collision free path for a point robot among the Cspace obstacles. However, the research experiences obtained so far have shown that the calculation of the Cspace obstacles is very hard work when the following situations occur: 1. both the robot and obstacles are not polygons and 2. the robot is allowed to rotate. This situation is even worse when the robot and obstacles are three dimensional (3D) objects with various shapes. Obviously a direct path planning approach without the calculation of the Cspace obstacles is strongly needed. This paper presents such a new real-time robot path planning approach which, to the best of our knowledge, is the first one in the robotic community. The fundamental ideas are the utilization of inequality and optimization technique. Simulation results have been presented to show its merits.

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In the present research, two Chinese rhesus monkeys were inoculated intravenously with 5000 TCID50 of SIVmac239. The changes in the numbers of CD4+ T lymphocyte in peripheral blood, plasma viral loads, proviral DNA and humoral antibodies against virus were periodically monitored during 121 days. At the early stage of infection, proviral DNA had been detected in PBMCs, and infectious SIVmac239 virus had been isolated from PBMCs. At the same period, the numbers of CD4+ T lymphocytes were significantly decreased, and maintained at low level during the 121-day period of infection. Plasma viral loads reached the peak at week 2 post-inoculation and kept at a steady state subsequently. Moreover, antibodies against viral proteins were detected from plasma. All the results showed that the two Chinese rhesus monkeys had been infected with SIVmac239 successfully. This animal model can be applied for further AIDS researches.

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A polycrystalline silicon thin film was fabricated on glass substrate by means of aluminum induced crystallization (AIC). Al and alpha-Si layers were deposited by magnetron sputtering respectively and annealed at 480A degrees C for 1 h to realize layer exchange. The polycrystalline silicon thin film was continuous and strongly (111) oriented. By analyzing the structure variation of the oxidation membrane and lattice mismatch between gamma-Al2O3 and Si, it was concluded that aluminum promoted the formation of (111) oriented silicon nucleus by controlling the orientation of gamma-Al2O3, which was formed at the early stage of annealing.

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土壤可溶性有机物质(Dissolved organic matter,DOM)作为土壤有机质的活性组分,在陆地生态系统物质循环中扮演非常重要的角色。土壤DOM的主要成分可溶性有机碳(Dissolved organic carbon,DOC)和氮(Dissolved organic nitrogen,DON)参与C、N循环过程。为深入揭示全球C、N循环过程机制,在未开展DOC和DON的地区进行相关研究是有必要的。森林土壤(包括枯枝落叶层)DOC、DON动态及调控机理的研究是目前国际上森林生态系统C、N循环研究热点之一。本研究立足于暖温带岷江上游茂县地区人工林植被,对土壤DOC和DON的库容量,季节动态及其与其它养分之间的关系进行了系统研究,旨在了解DOC和DON在该区生态系统中的重要作用,并探讨作为DOM主要来源的叶凋落物对DOC和DON的动态影响,研究有助于更加详细地了解该区生态系统C和N循环过程。本论文主要研究结论如下: 1研究了岷江上游地区两大主要土壤类型(棕壤和褐土)不同植物群落下土壤的DOC和DON含量及特征,结果表明:DOC和DON在两种土壤类型中均有库容量存在,DOC在0-10cm和10-20cm土层的含量幅度分别111.96~159.95 mg kg-1和69.02~100.84 mg kg-1。DON在0-10cm和10-20cm土层的含量幅度分别11.88~23.08 mg kg-1和4.70~10.77 mg kg-1。游离氨基酸在0-10cm和10-20cm土层的含量幅度分别0.84~1.66 mg kg-1和0.39~0.73 mg kg-1。DOC、DON与土壤中的一些养分因子表现出了显著的相关关系,共同反映了土壤的状况和质量,在该区开展DOC和DON的系统研究是有必要的。 2 对油松与连香树林地土壤DOC、DON以及其它化学指标的季节动态进行了研究,结果表明:油松与连香树林地土壤DOC和DON的季节动态变化表现了类似的规律,DOC和DON的含量均以秋季最高。DOC和DON的季节动态变化主要受凋落物生物因素的影响,但其微生物活力的生物因素以及降雨、温度等非生物因素也是控制土壤DOC和DON含量的重要因素。土壤DON在土壤中的行为不同于矿质氮,其季节动态不同于NO3--N和NH4+-N的季节动态,在研究N循环过程中,应考虑DON的变化情况。 3 对油松与连香树林地分解层和表层土壤(0-10cm)氨基酸周转动态进行了研究,结果表明:油松林地和连香树林地均以分解层的氨基酸含量高于矿质表层土壤的含量。每个取样时期,油松林地内各层次的氨基酸含量高于连香树林地内相应层次的含量。两林地各层次无机氮含量均超过了氨基酸的含量,并且室内培养30天后无机N的含量仍然高于氨基酸的含量,所以可以认为该区立地条件下无论是在有机分解层还是矿质土层植物吸收利用的氮素仍是以无机N为主。 4 松林下松针凋落物易于累积,这与松针凋落物分解缓慢有关,从而导致松林内养分周转缓慢。通过用不同性质凋落物和灌丛地土壤构建微生态系统,比较油松、辐射松、连香树、灌丛虎榛子凋落物分解对C、N循环过程的影响,结果显示油松和辐射松针叶凋落物比连香树、虎榛子凋落物分解更慢,减缓了养分循环过程。然而将针叶凋落物与阔叶凋落物混合后,油松和辐射松针叶凋落物的分解加快,C、N元素的循环过程也加速。此结果表明在松林内维持具有高质量凋落物的灌丛植被或在松林内栽植一些阔叶树种如连香树对维持和增进松树人工林的土壤肥力有重大的作用。室内培养的结果还显示添加凋落物后土壤DOC和DON的含量显著增加,表明凋落物是土壤DOM的直接来源。然而不同物种凋落物处理下土壤DOC和DON的含量有所不同,并随时间发生改变。混合凋落物处理下土壤DOC和DON的含量均高于松针凋落物单独处理下土壤DOC和DON的含量。DON是一个主要的水溶性N库,随时间的变化趋势与无机N的变化趋势不同,在土壤N循环过程中起到了中间N库的作用。 As a labile fraction of soil organic matter, dissolved organic matter (DOM) plays a very important role in material cycling of terrestrial ecosystem. The turnover of DOM is now being considered as main components in nutrient cycling. DOM mainly includes dissolved organic carbon (DOC), -nitrogen (DON), -phosphorous (DOP) and –sulfur (DOS). Among these constituents, DOC and DON directly participate in C and N cycling. It is essential to study DOC and DON dynamics and their controlling factors in the areas where no related study has ever been carried out. Study about them can provide data supports on understanding the mechanism of the global C and N cycling. DOC and DON dynamics and their controlling factors have been focused on in the research of C and N cycling of forest ecosystems. Based on forest plantations of Maoxian, Minjiang River in warm temperate zone, soil DOC and DON pool size, their seasonal dynamics, and the correlation between DOC, DON and other nutrients were studied in order to understand the importance of DOC and DON in the study area. Soil DOC and DON dynamics induced by leaf litter decomposition were also studied. The study contributed to comprehensively understanding C and N cycling processes and providing baseline data for including DOC and DON into the indices system of evaluating nutrient conditions. The results were as follows: 1 Several different plant communities under brown soil and Cinnamon soil were chosen as sampling plots. The contents and features of soil DOC and DON were evaluated. The results showed that DOC and DON were present under the two soil types. DOC contents in the top soil (0-10 cm) and the subsoil (10-20 cm) respectively varied from 111.96 mg kg-1to 159.95 mg kg-1, and 69.02 mg kg-1 to 100.84 mg kg-1. DON contents in the top soil (0-10 cm) and the subsoil (10-20 cm) respectively varied from 11.88 mg kg-1to 23.08 mg kg-1, and 4.70 mg kg-1 to 10.77 mg kg-1. Free amino acid contents in the top soil (0-10 cm) and the subsoil (10-20 cm) respectively varied from 0.84 mg kg-1to 1.66 mg kg-1, and 0.39 mg kg-1 to 0.73 mg kg-1. Significant correlations were found between DOC, DON and some nutrient indices, which together reflected soil condition and quality. It was hence essential to study DOC and DON in the study area. 2 Seasonal dynamics of DOC, DON, inorganic N, microbial biomass C and N were studied under Pinus tabulaeformis and Cercidiphyllum japonicum plantation. The results indicated that seasonal dynamics of soil DOC and DON under the two plantations performed similar change pattern, with the highest values in autumn. The seasonal dynamics of soil DOC and DON were mainly influenced by the litterfall. However, biotic factors such as soil microbial activities and abiotic factors such as precipitation and temperature also controlled the dynamics of soil DOC and DON. The seasonal dynamic of DON was different from that of NO3--N and NH4+-N, which showed that the behavioral differences between DON and inorganic nitrogen. And hence, it was proposed to include DON into soil N cycling in the study area. 3 Amino acid dynamics in Oa and topsoil (0-10 cm) under P. tabulaeformis and C. japonicum plantation were studied. The results showed that amino acid content in Oa was significantly higher than that in mineral soil. At each sampling time, significantly higher amino acid contents were found in P. tabulaeformis plantation than in C. japonicum plantation. The content of inorganic nitrogen was much higher than the content of amino acid in each sampling layer at each sampling time. After a 30-days laboratory incubation the content of amino acid was still lower than the content of inorganic nitrogen. The results implicated that the form of N absorbed by plants in these study sites were mainly inorganic nitrogen. 4 Usually needle litter is more resistant to decomposition, which leads to needle litter accumulation in pure coniferous stands and slows down the rate of nutrient circulation. By constructing microcosms with local shrubland soil and containing the four single-species (P. tabulaeformis, P. radiata, C. japonicum, Ostryopsis davidiana) litters, the decomposition rates and related C and N dynamics of needle litters and broadleaved litters during the early stage were compared. The results showed that the decomposition rates of pine needles were lower than those of broadleaved litters, which descended C and N cycling processes. However, the presence of C. japonicum or O. davidiana litter into pine needles increased the decomposition rates of pine needles and also dramatically promoted C and N cycling processes. It should be appropriate for plantation managers to consider C. japonicum as an ameliorative species or remain O. davidiana in pine plantations to improve soil conditions and help maintain soil fertility. The laboratory incubation still showed that DOC and DON contents in all litter-amended treatments were significantly higher than no litter-amended treatment, which proved that litter could be a direct source of DOM in soils. Different species litters induced different soil DOC and DON contents, which correspondingly changed over time. DOC and DON contents in mixed litter treatments were higher than those in pine needle litter treatments. As a major soluble N pool, DON developed a different changing pattern over time compared with inorganic N and played a role of interim N pool in soil N cycling.