[Intensive Citrus Cultivation Suppresses Soil Phosphorus Cycling Microbial Activity].

Soil microbes are key drivers in regulating the phosphorus cycle. Elucidating the microbial mineralization process of soil phosphorus-solubilizing bacteria is of great significance for improving nutrient uptake and yield of crops. This study investigated the mechanism by which citrus cultivation affects the soil microbial acquisition strategy for phosphorus by measuring the abundance of the phoD gene, microbial community diversity and structure, and soil phosphorus fractions in the soils of citrus orchards and adjacent natural forests. The results showed that citrus cultivation could lead to a decrease in soil pH and an accumulation of available phosphorus in the soil, with a content as high as 112 mg·kg-1, which was significantly higher than that of natural forests (3.7 mg·kg-1). Citrus cultivation also affected the soil phosphorus fractions, with citrus soil having higher levels of soluble phosphorus (CaCl2-P), citrate-extractable phosphorus (Citrate-P), and mineral-bound phosphorus (HCl-P). The phosphorus fractions of natural forest soils were significantly lower than those of citrus soils, whereas the phoD gene abundance and alkaline phosphatase activity were significantly higher in natural forest soils than in citrus soils. High-throughput sequencing results showed that the Shannon diversity index of phosphate-solubilizing bacteria in citrus soils was 4.61, which was significantly lower than that of natural forests (5.35). The microbial community structure in natural forests was also different from that of citrus soils. In addition, the microbial community composition of phosphate-solubilizing bacteria in citrus soils was also different from that of natural forests, with the relative abundance of Proteobacteria being lower in natural forest soils than in citrus soils. Therefore, citrus cultivation led to a shift of soil microbial acquisition strategy for phosphorus, with external phosphorus addition being the main strategy in citrus soils, whereas microbial mineralization of organic phosphorus was the main strategy in natural forest soils to meet their growth requirements.

Aureobasidium pullulans from the Fire Blight Biocontrol Product, Blossom Protect, Induces Host Resistance in Apple Flowers.

Fire blight, caused by Erwinia amylovora, is a devastating disease of apple. Blossom Protect, a product that contains Aureobasidium pullulans as the active ingredient, is one of the most effective biological controls of fire blight. It has been postulated that the mode of action of A. pullulans is to compete against and antagonize epiphytic growth of E. amylovora on flowers, but recent studies have found that flowers treated with Blossom Protect harbored similar to or only slightly reduced E. amylovora populations compared with nontreated flowers. In this study, we tested the hypothesis that A. pullulans-mediated biocontrol of fire blight is the result of induced host resistance. We found that PR genes in the systemic acquired resistance pathway, but not genes in the induced systemic resistance pathway, were induced in hypanthial tissue of apple flowers after the Blossom Protect treatment. Additionally, the induction of PR gene expression was coupled with an increase of plant-derived salicylic acid in this tissue. After inoculation with E. amylovora, PR gene expression was suppressed in nontreated flowers, but in flowers pretreated with Blossom Protect, the heightened PR expression offset the immune repression caused by E. amylovora, and prevented infection. Temporal and spatial analysis of PR gene induction showed that induction of PR genes occurred 2 days after the Blossom Protect treatment, and required direct flower-yeast contact. Finally, we observed deterioration of the epidermal layer of the hypanthium in some of the Blossom Protect-treated flowers, suggesting that PR gene induction in flowers may be a result of pathogenesis by A. pullulans.

Phosphorus limitation induced by nitrogen addition changed soil microbial community structure in a subtropical Pinus taiwanensis forest.

Soil microorganisms play an important role in the biogeochemical cycles of terrestrial ecosystems. How-ever, it is still unclear how the amount and duration of nitrogen (N) addition affect soil microbial community structure and whether there is a correlation between the changes in microbial community structure and their nutrient limi-tation status. In this study, we conducted an N addition experiment in a subtropical Pinus taiwanensis forest to simulate N deposition with three treatments: control (CK, 0 kg N·hm-2·a-1), low N (LN, 40 kg N·hm-2·a-1), and high N (HN, 80 kg N·hm-2·a-1). Basic soil physicochemical properties, phospholipid fatty acids content, and carbon (C), N and phosphorus (P) acquisition enzyme activities were measured after one and three years of N addition. The relative nutrient limitation status of soil microorganisms was analyzed using ecological enzyme stoichiometry. The results showed that one-year N addition did not affect soil microbial community structure. Three-year LN treatment significantly increased the contents of Gram-positive bacteria (G+), Gram-negative bacteria (G-), actinomycetes (ACT), and total phospholipid fatty acids (TPLFA), whereas three-year HN treatment did not significantly affect soil microbial community, indicating that bacteria and ACT might be more sensitive to N addition. Nitrogen addition exacerbated soil C and P limitation. Phosphorus limitation was the optimal explanatory factor for the changes in soil microbial community structure. It suggested that P limitation induced by N addition might be more beneficial for the growth of certain oligotrophic bacteria (e.g. G+) and the microorganisms participating in the P cycling (e.g. ACT), with consequences on soil microbial community structure of subtropical Pinus taiwanensis forest.

[Responses of soil microbial carbon use efficiency to short-term nitrogen addition in Castanopsis fabri forest]. / ç½—æµ®æ ²æž—åœŸå£¤å¾®ç”Ÿç‰©ç¢³åˆ©ç”¨æ•ˆçŽ‡å¯¹çŸ­æœŸæ°®æ·»åŠ çš„å“åº”.

As an important parameter regulating soil carbon mineralization, microbial carbon use efficiency (CUE) is essential for the understanding of carbon (C) cycle in terrestrial ecosystems. Three nitrogen supplemental levels, including control (0 kg N·hm-2·a-1), low nitrogen (40 kg N·hm-2·a-1), and high nitrogen (80 kg N·hm-2·a-1), were set up in a Castanopsis fabri forest in the Daiyun Mountain. The basic physical and chemical properties, organic carbon fractions, microbial biomass, and enzyme activities of the soil surface layer (0-10 cm) were measured. To examine the effects of increasing N deposition on microbial CUE and its influencing factors, soil microbial CUE was measured by the 18O-labelled-water approach. The results showed that short-term N addition significantly reduced microbial respiration rate and the activities of C and N acquisition enzymes, but significantly increased soil microbial CUE. ß-N-acetyl amino acid glucosidase (NAG)/microbial biomass carbon (MBC), microbial respiration rate, ß-glucosidase (BG)/MBC, cellulose hydrolase (CBH)/MBC, and soil organic carbon content were the main factors affecting CUE. Moreover, CUE significantly and negatively correlated with NAG/MBC, microbial respiration rate, BG/MBC, and CBH/MBC, but significantly and positively correlated with soil organic carbon. In summary, short-term N addition reduced the cost of soil microbial acquisition of C and N and microbial respiration, and thus increased soil microbial CUE, which would increase soil carbon sequestration potential of the C. fabri forest.

[Effects of nitrogen addition on the kinetic parameters of soil acid phosphomonoesterase in a Moso bamboo forest]. / æ°®æ·»åŠ å¯¹æ¯›ç«¹æž—åœŸå£¤é ¸æ€§ç£·é ¸å•é ¯é ¶åŠ¨åŠ›å­¦å‚æ•°çš„å½±å“.

Soil phosphatases are important in the mineralization of organophosphates and in the phosphorus (P) cycle. The kinetic mechanisms of phosphatases in response to nitrogen (N) deposition remain unclear. We carried out a field experiment with four different concentrations of N: 0 g N·hm-2·a-1(control), 20 g N·hm-2·a-1(low N), 40 g N·hm-2·a-1(medium N), and 80 g N·hm-2·a-1(high N) in a subtropical Moso bamboo forest. Soil samples were then collected from 0 to 15 cm depth, after 3, 5 and 7 years of N addition. We analyzed soil chemical properties and microbial biomass. Acid phosphatase (ACP) was investigated on the basis of maximum reaction velocity (Vm), Michaelis constant (Km), and catalytic efficiency (Ka). Results showed that N addition significantly decreased soil dissolved organic carbon (DOC), available phosphorus, and organophosphate content, but significantly increased soil ammonium, nitrate-N content, and Vm. There was a significant relationship between Vm and the concentrations of available phosphorus, organophosphate, and soil DOC. In general, N addition substantially increased Ka, but did not affect Km. The Km value in the high N treatment group was higher than that in the control group after five years of N addition. Km was significantly negatively associated with both available phosphorus and organophosphate. Medium and high N treatments had stronger effects on the kinetic parameters of ACP than low N treatment. Results of variation partition analysis showed that changes in soil chemical properties, rather than microbial biomass, dominated changes in Vm(47%) and Km(33%). In summary, N addition significantly affected substrate availability in Moso bamboo forest soil and modulated soil P cycle by regulating ACP kinetic parameters (especially Vm). The study would improve the understanding of the mechanisms underlying soil microorganisms-regulated soil P cycle under N enrichment. These mechanisms would identify the important parameters for improving soil P cycling models under global change scenarios.

A Functional Screening Identifies a New Organic Selenium Compound Targeting Cancer Stem Cells: Role of c-Myc Transcription Activity Inhibition in Liver Cancer.

Cancer stem cells (CSCs) are reported to play essential roles in chemoresistance and metastasis. Pathways regulating CSC self-renewal and proliferation, such as Hedgehog, Notch, Wnt/ß-catenin, TGF-ß, and Myc, may be potential therapeutic targets. Here, a functional screening from the focused library with 365 compounds is performed by a step-by-step strategy. Among these candidate molecules, phenyl-2-pyrimidinyl ketone 4-allyl-3-amino selenourea (CU27) is chosen for further identification because it proves to be the most effective compound over others on CSC inhibition. Through ingenuity pathway analysis, it is shown CU27 may inhibit CSC through a well-known stemness-related transcription factor c-Myc. Gene set enrichment analysis, dual-luciferase reporter assays, expression levels of typical c-Myc targets, molecular docking, surface plasmon resonance, immunoprecipitation, and chromatin immunoprecipitation are conducted. These results together suggest CU27 binds c-Myc bHLH/LZ domains, inhibits c-Myc-Max complex formation, and prevents its occupancy on target gene promoters. In mouse models, CU27 significantly sensitizes sorafenib-resistant tumor to sorafenib, reduces the primary tumor size, and inhibits CSC generation, showing a dramatic anti-metastasis potential. Taken together, CU27 exerts inhibitory effects on CSC and CSC-associated traits in hepatocellular carcinoma (HCC) via c-Myc transcription activity inhibition. CU27 may be a promising therapeutic to treat sorafenib-resistant HCC.

[Soil enzyme stoichiometry revealed the changes of soil microbial carbon and phosphorus limitation along an elevational gradient in a Pinus taiwanensis forest of Wuyi Mountains, Southeast China]. / åœŸå£¤é ¶è®¡é‡æ­ç¤ºäº†æ­¦å¤·å±±é»„å±±æ¾æž—åœŸå£¤å¾®ç”Ÿç‰©æ²¿æµ·æ‹”æ¢¯åº¦çš„ç¢³ç£·é™åˆ¶å˜åŒ–.

Understanding changes in soil enzyme activities and ecoenzymatic stoichiometry is important for assessing soil nutrient availability and microbial nutrient limitation in mountain ecosystems. However, the variations of soil microbial nutrient limitation across elevational gradients and its driving factors in subtropical mountain forests are still unclear. In this study, we measured soil properties, microbial biomass, and enzyme activities related to carbon (C), nitrogen (N), and phosphorus (P) cycling in Pinus taiwanensis forests at different altitudes of Wuyi Mountains. By analyzing the enzyme stoichiometric ratio, vector length (VL), and vector angle (VA), the relative energy and nutrient limitation of soil microorganisms and its key regulatory factors were explored. The results showed that ß-glucosaminidase (BG) activities increased along the elevational gradient, while the activities of ß-N-acetyl glucosaminidase (NAG), leucine aminopeptidase (LAP), acid phosphatase (AcP) and (NAG+LAP)/microbial biomass carbon (MBC) and AcP/MBC showed the opposite trend. Enzyme C/N, enzyme C/P, enzyme N/P, and VL were enhanced with increasing elevation, while VA decreased, indicating a higher degree of microbial P limitation at low elevation and higher C limitation at high elevation. In addition, our results suggested that dissolved organic carbon and microbial biomass phosphorus are critical factors affecting the relative energy and nutrient limitation of soil microorganisms at different elevations. The results would provide a theoretical basis for the responses of soil carbon, nitrogen, and phosphorus availability as well as the relative limitation of microbial energy and nutrition to elevational gradients, and improve our understanding of soil biogeochemical cycle process in subtropical montane forest ecosystems.

Enzyme stoichiometry evidence revealed that five years nitrogen addition exacerbated the carbon and phosphorus limitation of soil microorganisms in a Phyllostachys pubescens forest.

The activity and stoichiometry of soil extracellular enzyme can provide a good indication for changes in soil nutrient availability and microbial demands for nutrients. However, it remains unclear how would nitrogen (N) deposition affect nutrient limitation of microbes in subtropical forest soils. We conducted a 5 years N addition experiment in a subtropical Phyllostachys pubescens forest. The soil nutrients and enzyme activities associated with carbon (C), N, and phosphorus (P) cycles were measured. We also examined the nutrient distribution of microorganisms using enzyme stoichiometry and vector analysis. The results showed that N addition significantly decreased the contents of soil soluble organic C and available P and increased that of available N. Furthermore, N addition significantly decreased ß-N-acetyl-glucosaminidase (NAG) activity and NAG/ microbial biomass carbon (MBC), and increased acid phosphatase (ACP) and ACP/MBC. The low and moderate N addition levels significantly increased enzyme C/P, vector length, and vector angle, but significantly decreased enzyme N/P. Results of redundancy analysis showed that the change in soil enzyme activity and enzymatic stoichiometry were mainly driven by soil available P content under N addition. In summary, N addition altered the microbial nutrient acquisition strategy, which increased nutrient allocation to P-acquiring enzyme production but reduced that to N-acquiring enzyme production. Moreover, N addition exacerbated the C and P limitation of soil microorganisms. Appropriate amount of P fertilizer could be applied to improve soil fertility of subtropical P. pubescens forest in the future.

[Responses of soil phosphorus fractions and microorganisms to nitrogen application in a subtropical Phyllostachys pubescen forest]. / 亚热带毛竹林土壤磷组分和微生物对施氮的响应.

Phosphorus (P) is an important nutrient for plant and microbial growth. Soil P availabi-lity is poor in subtropical areas. Long-term heavy nitrogen (N) deposition might further reduce P availability. The experiment was performed in a Phyllostachys pubescens forest in Daiyun Mountain. The effects of N application on soil basic physical and chemical properties, soil P fractions, microbial biomass, and acid phosphomonoesterase activity were analyzed after three years of N application. The results showed that N application significantly increased NO3--N content and thus soil N availability, while it significantly reduced the percentage of decomposable organic P to total P, with the ratio of carbon (C) to organic P being over 200. The soil microbial biomass C, microbial biomass P, acid phosphomonoesterase, and the ratio of microbial biomass N to microbial biomass P and microbial biomass C to microbial biomass P were increased as the N application rate increased. There was a significant negative correlation between the percentage of decomposable organic P to total P and microbial biomass P. Consequently, N application enhanced soil P limitation and increased microbial P demand.

Cell-length heterogeneity: a population-level solution to growth/virulence trade-offs in the plant pathogen Dickeya dadantii.

Necrotrophic plant pathogens acquire nutrients from dead plant cells, which requires the disintegration of the plant cell wall and tissue structures by the pathogen. Infected plants lose tissue integrity and functional immunity as a result, exposing the nutrient rich, decayed tissues to the environment. One challenge for the necrotrophs to successfully cause secondary infection (infection spread from an initially infected plant to the nearby uninfected plants) is to effectively utilize nutrients released from hosts towards building up a large population before other saprophytes come. In this study, we observed that the necrotrophic pathogen Dickeya dadantii exhibited heterogeneity in bacterial cell length in an isogenic population during infection of potato tuber. While some cells were regular rod-shape (<10µm), the rest elongated into filamentous cells (>10µm). Short cells tended to occur at the interface of healthy and diseased tissues, during the early stage of infection when active attacking and killing is occurring, while filamentous cells tended to form at a later stage of infection. Short cells expressed all necessary virulence factors and motility, whereas filamentous cells did not engage in virulence, were non-mobile and more sensitive to environmental stress. However, compared to the short cells, the filamentous cells displayed upregulated metabolic genes and increased growth, which may benefit the pathogens to build up a large population necessary for the secondary infection. The segregation of the two subpopulations was dependent on differential production of the alarmone guanosine tetraphosphate (ppGpp). When exposed to fresh tuber tissues or freestanding water, filamentous cells quickly transformed to short virulent cells. The pathogen adaptation of cell length heterogeneity identified in this study presents a model for how some necrotrophs balance virulence and vegetative growth to maximize fitness during infection.

Systems Pharmacology and Microbiome Dissection of Shen Ling Bai Zhu San Reveal Multiscale Treatment Strategy for IBD.

Generally, inflammatory bowel disease (IBD) can be caused by psychology, genes, environment, and gut microbiota. Therefore, IBD therapy should be improved to utilize multiple strategies. Shen Ling Bai Zhu San (SLBZS) adheres to the aim of combating complex diseases from an integrative and holistic perspective, which is effective for IBD therapy. Herein, a systems pharmacology and microbiota approach was developed for these molecular mechanisms exemplified by SLBZS. First, by systematic absorption-distribution-metabolism-excretion (ADME) analysis, potential active compounds and their corresponding direct targets were retrieved. Then, the network relationships among the active compounds, targets, and disease were built to deduce the pharmacological actions of the drug. Finally, an "IBD pathway" consisting of several regulatory modules was proposed to dissect the therapeutic effects of SLBZS. In addition, the effects of SLBZS on gut microbiota were evaluated through analysis of the V3-V4 region and multivariate statistical methods. SLBZS significantly shifted the gut microbiota structure in a rat model. Taken together, we found that SLBZS has multidimensionality in the regulation of IBD-related physiological processes, which provides new sights into herbal medicine for the treatment of IBD.

[Qilin Pills improve sperm quality in patients with oligozoospermia].

OBJECTIVE: To observe the therapeutic effect of Qilin Pills (QLP) on oligoasthenospermia. METHODS: This prospective study included 168 patients diagnosed with oligozoospermia in our hospital between September 2015 and September 2017, and all of them failed to achieve pregnancy. We randomly divided them into a QLP group (n = 82) and a placebo control group (n = 86) to receive oral QLPs or placebo pills, respectively, both at 6 g tid for 6 months. We followed up the patients and recorded their routine semen parameters every month during the medication and the rate of pregnancy in their spouses. RESULTS: After 6 months of treatment, the QLP group, as compared with the placebo controls, showed significantly improved sperm concentration (25.13 ×106/ml vs 11.62 ×106/ml, P < 0.01), grade a+b sperm (33.81% vs 17.32%, P < 0.01), grade a sperm (22.84% vs 13.56%, P < 0.01) and sperm motility (56.33% vs 26.23%, P < 0.01); and 26 pregnancies were achieved in the QLP group (32.91%), remarkably more than 13 in the placebo control group (15.85%) (P < 0.01). CONCLUSIONS: Qilin Pills can effectively improve sperm quality and increase the rate of pregnancy.

[oil microbial biomass and enzyme activities among different artificial forests in Ziwuling, Northwest China.] / å­åˆå²­äººå·¥æž—åœŸå£¤å¾®ç”Ÿç‰©ç”Ÿç‰©é‡åŠé ¶æ´»æ€§.

Decades of ecological restoration on the Loess Plateau has achieved significant on-site benefits to reduce soil erosion and improve soil quality, with remarkable off-site effects of reducing sediment delivery to Yellow River. However, regional forest community succession is still far from being adequately developed. The Ziwuling forest region and its highly developed forest community, as an advanced eco-zone, can lend practical experience to other regions on the Loess Plateau and help to identify the most suitable tree species for a better regional restoration in the future. With the aim to systematically understand the potential effects of typical local tree species to soil properties, three typical and well-established artificial forests in the Ziwuling region, Robinia pseudoacacia, Pinus tabuliformis and Platycladus orientalis were investigated in this study, with the climax community Quercus wutaishanica as a reference. All the four forest type had comparable stand age (25 years). Soil samples from 0-20 cm layers were collected from those four plantations. The soil microbial biomass (carbon and nitrogen), soil enzyme (invertase, urease and alkaline phosphatase) activities and their correlations were measured and analyzed. The results showed that: 1) soil invertase activity ranged from 16.94 to 64.49 mg·g-1·24 h-1, the soil urease activity from 0.15 to 0.26 mg·g-1·24 h-1, and the alkaline phosphatase activity from 0.65 to 1.23 mg·g-1·24 h-1. The activities of those three enzymes were significantly higher in the P. orientalis soil that in the R. pseudoacacia and P. tabuliformis soils. The geometric average values in the P. orientalis soil were even greater than that in the Q. wutaishanica soil. 2) The soil microbial biomass carbon and nitrogen varied from 247.37 to 529.84 mg·kg-1 and 41.48 to 77.91 mg·kg-1, respectively. Both of them were significantly greater in the P. orientalis soil than that in the R. pseudoacacia and P. tabu-liformis soils. Even though the soil microbial biomass carbon in the P. orientalis soil remained lower than that in the Q. wutaishanica soil, its soil microbial biomass nitrogen was greater than in the Q. wutaishanica soil. 3) The dissolved organic carbon and nitrogen in the P. tabuliformis soil were much greater than that in other species, even greater than their own soil microbial biomass carbon and nitrogen. Such a result indicated that dissolved organic matter might play a more important role in providing plant available nutrients than microbial biomass in the P. tabuliformis soil. 4) The microbial biomass carbon and nitrogen were significantly positively correlated with the total organic carbon and the total nitrogen, particularly for the R. pseudoacacia and P. tabuliformis soils. There were significantly positive relationships between the soil invertase activity, urease activity and alkaline phosphatase activity, and their soil organic carbon, total nitrogen and total phosphorus contents. 5) Based on the results of principal component analysis, we concluded that the artificial forests types had obvious effects on soil microbial carbon and nitrogen, soil organic carbon, total nitrogen, the ratio of carbon to phosphorus, the ratio of nitrogen to phosphorus and urease activity. Overall, our findings suggested that P. orientalis is better than R. pseudoacacia and P. tabuliformis in term of improving soil properties in the south forest zone on the Chinese Loess Plateau.

[Ecological stoichiometry of leaf and litter of herbaceous plants in loess hilly and gully regions, China]. / 黄土丘陵区草本植物叶片与枯落物生态化学计量学特征.

This study was conducted in the forest, forest-steppe and steppe vegetation zones along the Yanhe River Basin, where the leaf and litter samples from four dominant herbaceous plants including Lespedeza davurica, Stipa bungeana, Artemisia sacrorum, Artemisia giraldii were taken. By measuring the concentrations of carbon (C), nitrogen (N), phosphorus (P) and potassium (K), we measured the concentrations and their ratios to explore the limit and resorption of nutrient in the herbaceous plants. The results showed that the leaf N/P of four herbaceous plants was all lower than 14, suggesting their growth was mainly limited by N content. Except for L. davurica, the mean nutrient resorption efficiency of K, N and P in the other three plants was 79.9%, 48.7% and 32.5%, respectively. The higher nutrient resorption efficiency and K concentration in the leaf were beneficial for soil water competition of A. sacrorum and A. giraldii. The litter C/N in A. sacrorum was significantly lower than that in S. bungeana and A. giraldii, which was easy to decompose to benefit the nutrient recycling. This resulted in the wide distribution of A. sacrorum in the three vegetation areas.

[Effects of stem and leaf decomposition in typical herbs on soil enzyme activity and microbial diversity in the south Ningxia loess hilly region of Northwest China]. / å®å—å±±åŒºå ¸åž‹è‰æœ¬æ¤ç‰©èŒŽå¶åˆ†è§£å¯¹åœŸå£¤é ¶æ´»æ€§åŠå¾®ç”Ÿç‰©å¤šæ ·æ€§çš„å½±å“.

With the method of litter bags, the characteristics of soil enzyme activities, soil microbial diversity at later stage of decomposition, and the relationships between soil enzyme activity and initial soil property were investigated in the process of stem and leaf decomposition of three typical herbs, i.e., Stipa bungeana, Artemisia sacrorum and Thymus mongolicus in the south Ningxia loess hilly region, Northwest China. The results showed that soil enzyme activity increased under different treatments after 480 d during stem and leaf decomposition. Soil sucrose activity (32.40 mg·g-1·24 h-1) and alkaline phosphatase activity (1.99 mg·g-1·24 h-1) were the highest in S. bungeana treatment. Soil urease activity (2.66 mg·g-1·24 h-1) was the highest in T. mongolicus treatment, and soil cellulase activity (1.42 mg·g-1·72 h-1) was the highest in A. sacrorum treatment. Soil cellulose activity at later stage of decomposition had significant positive correlation with initial microbial biomass carbon of soil. Soil cellulose activity at later stage of decomposition had significant negative correlation with initial nitrate nitrogen content of soil. Ace index, Chao index and Shannon index of soil bacteria and fungi in plant tissue addition treatments were higher than in the control. However, Simpson index was opposed. The stem and leaf decomposition significantly promoted the abundance and diversity of soil bacteria and fungi, accelerated the decomposition rate of stems and leaves, and promoted the cycle and transformation of soil nutrients.

[Ecological stoichiometry characteristics of leaf-litter-soil in different plantations on the Loess Plateau, China]. / 黄土高原不同人工林叶片-凋落叶-土壤生态化学计量特征.

In order to research into the influence of Grain to Green Project in Ziwuling forest region, this paper took three typical plantations which were Robinia pseudoacacia plantation, Pinus tabuliformis plantation, and Platycladus orientalis plantation in the Ziwuling forest region of Shaanxi Province as research objects and analyzed the carbon, nitrogen and phosphorus contents of leaf, litter and soil among the three plantations. The results showed that the contents of C, N and P in the three plantations were in order of leaf > litter > soil, the contents of N and P in leaf of R. pseu-doacacia plantation were significantly higher than that of P. tabuliformis plantation and P. orientalis. Leaf N:P was 12.21, 5.36 and 6.09 in R. pseudoacacia plantation, P. tabuliformis plantation and P. orientalis plantation, respectively. It was indicated that the three species were all subject to N deficiency. C:N and C:P showed the trend of litter > leaf > soil, and N:P demonstrated the trend of leaf > litter > soil. There were highly significant positive relationships in C:N between leaf and litter in P. tabuliformis plantation. N and P in the leaf development process of R. pseu-doacacia plantation were proportionally absorbed, and proportionally remained in the litter after N and P resorption. R. pseudoacacia was the better plantation species than P. tabuliformis and P. orientalis in the south fo-rest zone on the Loess Plateau.

Discovery of plant phenolic compounds that act as type III secretion system inhibitors or inducers of the fire blight pathogen, Erwinia amylovora.

Erwinia amylovora causes a devastating disease called fire blight in rosaceous plants. The type III secretion system (T3SS) is one of the important virulence factors utilized by E. amylovora in order to successfully infect its hosts. By using a green fluorescent protein (GFP) reporter construct combined with a high-throughput flow cytometry assay, a library of phenolic compounds and their derivatives was studied for their ability to alter the expression of the T3SS. Based on the effectiveness of the compounds on the expression of the T3SS pilus, the T3SS inhibitors 4-methoxy-cinnamic acid (TMCA) and benzoic acid (BA) and one T3SS inducer, trans-2-(4-hydroxyphenyl)-ethenylsulfonate (EHPES), were chosen for further study. Both the T3SS inhibitors (TMCA and BA) and the T3SS inducer (EHPES) were found to alter the expression of T3SS through the HrpS-HrpL pathway. Additionally, TMCA altered T3SS expression through the rsmBEa-RsmAEa system. Finally, we found that TMCA and BA weakened the hypersensitive response (HR) in tobacco by suppressing the T3SS of E. amylovora. In our study, we identified phenolic compounds that specifically targeted the T3SS. The T3SS inhibitor may offer an alternative approach to antimicrobial therapy by targeting virulence factors of bacterial pathogens.

Hepatocellular carcinoma-associated mesenchymal stem cells promote hepatocarcinoma progression: role of the S100A4-miR155-SOCS1-MMP9 axis.

UNLABELLED: Cancer-associated mesenchymal stem cells (MSCs) play a pivotal role in modulating tumor progression. However, the interactions between liver cancer-associated MSCs (LC-MSCs) and hepatocellular carcinoma (HCC) remain unreported. Here, we identified the presence of MSCs in HCC tissues. We also showed that LC-MSCs significantly enhanced tumor growth in vivo and promoted tumor sphere formation in vitro. LC-MSCs also promoted HCC metastasis in an orthotopic liver transplantation model. Complementary DNA (cDNA) microarray analysis showed that S100A4 expression was significantly higher in LC-MSCs compared with liver normal MSCs (LN-MSCs) from adjacent cancer-free tissues. Importantly, the inhibition of S100A4 led to a reduction of proliferation and invasion of HCC cells, while exogenous S100A4 expression in HCC cells resulted in heavier tumors and more metastasis sites. Our results indicate that S100A4 secreted from LC-MSCs can promote HCC cell proliferation and invasion. We then found the expression of oncogenic microRNA (miR)-155 in HCC cells was significantly up-regulated by coculture with LC-MSCs and by S100A4 ectopic overexpression. The invasion-promoting effects of S100A4 were significantly attenuated by a miR-155 inhibitor. These results suggest that S100A4 exerts its effects through the regulation of miR-155 expression in HCC cells. We demonstrate that S100A4 secreted from LC-MSCs promotes the expression of miR-155, which mediates the down-regulation of suppressor of cytokine signaling 1, leading to the subsequent activation of STAT3 signaling. This promotes the expression of matrix metalloproteinases 9, which results in increased tumor invasiveness. CONCLUSION: S100A4 secreted from LC-MSCs is involved in the modulation of HCC progression, and may be a potential therapeutic target. (HEPATOLOGY 2013).

[Analysis of acute stroke trials conducted in mainland China in the past 17 years].

OBJECTIVE: To evaluate the current status and trend of acute stroke randomized controlled trials (RCTs) in Mainland China. METHODS: Acute stroke RCTs were retrieved from 8 databases published in Chinese or English language from January 1996 to May 2013. Those with transient ischemic attack (TIA) or traumatic hemorrhage were excluded. Methodological items were referred to the Cochrane Collaboration's tool for assessing risk of bias. The definitions of Wade were used to assess the outcome measures. The data were processed by SPSS 16.0. Data were summarized as frequency (and %) or median and interquartile range. Difference in proportions was assessed with χ(2) test. RESULTS: A total of 13 493 RCTs were identified. The number of acute stroke RCTs increased by years, but only 52 multi-center RCTs were published. There were 13 multi-center placebo controlled drug trials. Among them, only 3 had a sample size of over 500 cases. In multi-center non-drug trials, only 3 used allocation concealment. The studied types of stroke included ischemic stroke (63.9%, 8623/13 493), intracerebral hemorrhage (30.8%, 4157/13 493) and subarachnoid hemorrhage (5.3%, 713/13 495). There were 61.5% (32/52) multi-center drug trials, including 65.6% (21/32) in Western drug treatment and 34.4% (11/32) in traditional Chinese medicine. There were 38.5% (20/52) non-drug trials including 8 physical therapy, 7 surgery and 5 acupuncture treatment. There were 2 multi-center placebo controlled drug trials and 2 multi-center non-drug trials used mortality/disability for assessing outcome measures over a follow-up period of 90 days or more. CONCLUSION: In the past 17 years in mainland China, the number of acute stroke RCTs has increased dramatically, but the high-quality trials are scarce. The future acute stroke trials should pay more attention to true randomization, blinding and better patient outcome measures.

Derivatives of plant phenolic compound affect the type III secretion system of Pseudomonas aeruginosa via a GacS-GacA two-component signal transduction system.

Antibiotic therapy is the most commonly used strategy to control pathogenic infections; however, it has contributed to the generation of antibiotic-resistant bacteria. To circumvent this emerging problem, we are searching for compounds that target bacterial virulence factors rather than their viability. Pseudomonas aeruginosa, an opportunistic human pathogen, possesses a type III secretion system (T3SS) as one of the major virulence factors by which it secretes and translocates T3 effector proteins into human host cells. The fact that this human pathogen also is able to infect several plant species led us to screen a library of phenolic compounds involved in plant defense signaling and their derivatives for novel T3 inhibitors. Promoter activity screening of exoS, which encodes a T3-secreted toxin, identified two T3 inhibitors and two T3 inducers of P. aeruginosa PAO1. These compounds alter exoS transcription by affecting the expression levels of the regulatory small RNAs RsmY and RsmZ. These two small RNAs are known to control the activity of carbon storage regulator RsmA, which is responsible for the regulation of the key T3SS regulator ExsA. As RsmY and RsmZ are the only targets directly regulated by GacA, our results suggest that these phenolic compounds affect the expression of exoS through the GacSA-RsmYZ-RsmA-ExsA regulatory pathway.