PdmIRD: missense variants pathogenicity prediction for inherited retinal diseases in a disease-specific manner.

Accurate discrimination of pathogenic and nonpathogenic variation remains an enormous challenge in clinical genetic testing of inherited retinal diseases (IRDs) patients. Computational methods for predicting variant pathogenicity are the main solutions for this dilemma. The majority of the state-of-the-art variant pathogenicity prediction tools disregard the differences in characteristics among different genes and treat all types of mutations equally. Since missense variants are the most common type of variation in the coding region of the human genome, we developed a novel missense mutation pathogenicity prediction tool, named Prediction of Deleterious Missense Mutation for IRDs (PdmIRD) in this study. PdmIRD was tailored for IRDs-related genes and constructed with the conditional random forest model. Population frequencies and a newly available prediction tool were incorporated into PdmIRD to improve the performance of the model. The evaluation of PdmIRD demonstrated its superior performance over nonspecific tools (areas under the curves, 0.984 and 0.910) and an existing eye abnormalities-specific tool (areas under the curves, 0.975 and 0.891). We also demonstrated the submodel that used a smaller gene panel further slightly improved performance. Our study provides evidence that a disease-specific model can enhance the prediction of missense mutation pathogenicity, especially when new and important features are considered. Additionally, this study provides guidance for exploring the characteristics and functions of the mutated proteins in a greater number of Mendelian disorders.

A critical thermal transition driving spring phenology of Northern Hemisphere conifers.

Despite growing interest in predicting plant phenological shifts, advanced spring phenology by global climate change remains debated. Evidence documenting either small or large advancement of spring phenology to rising temperature over the spatio-temporal scales implies a potential existence of a thermal threshold in the responses of forests to global warming. We collected a unique data set of xylem cell-wall-thickening onset dates in 20 coniferous species covering a broad mean annual temperature (MAT) gradient (-3.05 to 22.9°C) across the Northern Hemisphere (latitudes 23°-66° N). Along the MAT gradient, we identified a threshold temperature (using segmented regression) of 4.9 ± 1.1°C, above which the response of xylem phenology to rising temperatures significantly decline. This threshold separates the Northern Hemisphere conifers into cold and warm thermal niches, with MAT and spring forcing being the primary drivers for the onset dates (estimated by linear and Bayesian mixed-effect models), respectively. The identified thermal threshold should be integrated into the Earth-System-Models for a better understanding of spring phenology in response to global warming and an improved prediction of global climate-carbon feedbacks.

Photoperiod and temperature as dominant environmental drivers triggering secondary growth resumption in Northern Hemisphere conifers.

Wood formation consumes around 15% of the anthropogenic CO2 emissions per year and plays a critical role in long-term sequestration of carbon on Earth. However, the exogenous factors driving wood formation onset and the underlying cellular mechanisms are still poorly understood and quantified, and this hampers an effective assessment of terrestrial forest productivity and carbon budget under global warming. Here, we used an extensive collection of unique datasets of weekly xylem tissue formation (wood formation) from 21 coniferous species across the Northern Hemisphere (latitudes 23 to 67°N) to present a quantitative demonstration that the onset of wood formation in Northern Hemisphere conifers is primarily driven by photoperiod and mean annual temperature (MAT), and only secondarily by spring forcing, winter chilling, and moisture availability. Photoperiod interacts with MAT and plays the dominant role in regulating the onset of secondary meristem growth, contrary to its as-yet-unquantified role in affecting the springtime phenology of primary meristems. The unique relationships between exogenous factors and wood formation could help to predict how forest ecosystems respond and adapt to climate warming and could provide a better understanding of the feedback occurring between vegetation and climate that is mediated by phenology. Our study quantifies the role of major environmental drivers for incorporation into state-of-the-art Earth system models (ESMs), thereby providing an improved assessment of long-term and high-resolution observations of biogeochemical cycles across terrestrial biomes.

Application of Ceriporia lacerata HG2011 as biocontrol agent against multiple phytopathogenic fungi and oomycetes.

Overuse of fungicides to control crop diseases results in ecological damage, environmental pollution, and human health risks. Biocontrol is an increasingly popular alternative in plant disease management due to sustainability and environmental friendliness. Herein, antagonistic tests and greenhouse experiments were conducted to investigate the antagonism of a self-isolated white-rot fungus Ceriporia lacerata HG2011 against phytopathogens in vitro, the underlying mechanism exerted by this fungus, and disease control efficiency in the greenhouse. The results demonstrated that both soluble and volatile substances produced by this fungus suppressed the growth of all test phytopathogen fungi and oomycetes in vitro, with the inhibitory rates of 10.4-60.6% for soluble metabolites and 30.3-52.9% for volatiles. C. lacerata HG2011 could grow in and gradually spread on living phytopathogenic colonies, concurrently deformed and lysed pathogenic hyphae in dual culture, which were associated with the release of hydrolase (cellulose, chitinase, ß-glucanase, and protease) from this biocontrol fungus for the use of the pathogens as nutrient sources. The chitinolytic and cellulolytic production by C. lacerata HG2011 presents the specific response to the cell wall of pathogenic fungi and oomycetes, and ß-glucanase was triggered by carbon competition. Consequently, C. lacerata HG2011 successfully controlled eggplant stem blight and cucumber vine blight (control efficacy 67.9-70.9%) in the greenhouse experiments. C. lacerata HG2011 showed multiple antagonistic mechanisms against the phytopathogenic fungi and oomycetes concurrently. Our results provided information about a new potential use of this fungus as a biocontrol agent to control plant diseases in modern agriculture beyond medical purposes, wastewater treatment, and biofuel production.

Climate warming leads to advanced fruit development period of temperate woody species but divergent changes in its length.

Climate warming has significantly altered the phenology of plants in recent decades. However, in contrast to the widely reported warming-induced extension of vegetative growing season, the response of fruit development period (FDP) from flowering to fruiting remains largely unexplored, particularly for woody plants. Analyzing >560,000 in situ observations of both flowering and fruiting dates for six temperate woody species across 2958 European phenological observations sites during 1980-2013, we found that in all species both flowering and fruiting phenology, that is, the FDP, advanced with climate warming. However, the advancing rates of the two events were not necessarily equal for any given species, resulting in divergent changes in the length of FDP among species with climate warming. During 1980-2013, not only the temperature during FDP but also the forcing requirement for fruit development increased, both affecting the length of FDP. The shortened FDP was mainly due to elevated temperature, thus accelerating the accumulation of forcing, whereas the prolonged FDP was primarily caused by the substantial increase of the forcing requirement of fruiting, which could be fulfilled only in a longer time and thus slowed down the advance of fruiting. This study provides large-scale empirical evidence of warming-induced advances of FDP but divergent changes in its length in temperate woody species. Our findings demonstrate the contrasting reproductive phenological strategies among temperate woody species under the pressure of warming climate, contrary to the lengthening of vegetative growing season, which is by and largely similar with different woody species.

Divergent trends in the risk of spring frost damage to trees in Europe with recent warming.

Frost events during the active growth period of plants can cause extensive frost damage with tremendous economic losses and dramatic ecological consequences. A common assumption is that climate warming may bring along a reduction in the frequency and severity of frost damage to vegetation. On the other hand, it has been argued that rising temperature in late winter and early spring might trigger the so called "false spring", that is, early onset of growth that is followed by cold spells, resulting in increased frost damage. By combining daily gridded climate data and 1,489 k in situ phenological observations of 27 tree species from 5,565 phenological observation sites in Europe, we show here that temporal changes in the risk of spring frost damage with recent warming vary largely depending on the species and geographical locations. Species whose phenology was especially sensitive to climate warming tended to have increased risk of frost damage. Geographically, compared with continental areas, maritime and coastal areas in Europe were more exposed to increasing occurrence of frost and these late spring frosts were getting more severe in the maritime and coastal areas. Our results suggest that even though temperatures will be elevated in the future, some phenologically responsive species and many populations of a given species will paradoxically experience more frost damage in the future warming climate. More attention should be paid to the increased frost damage in responsive species and populations in maritime areas when developing strategies to mitigate the potential negative impacts of climate change on ecosystems in the near future.

Long-term changes in the impacts of global warming on leaf phenology of four temperate tree species.

Contrary to the generally advanced spring leaf unfolding under global warming, the effects of the climate warming on autumn leaf senescence are highly variable with advanced, delayed, and unchanged patterns being all reported. Using one million records of leaf phenology from four dominant temperate species in Europe, we investigated the temperature sensitivities of spring leaf unfolding and autumn leaf senescence (ST , advanced or delayed days per degree Celsius). The ST of spring phenology in all of the four examined species showed an increase and decrease during 1951-1980 and 1981-2013, respectively. The decrease in the ST during 1981-2013 appears to be caused by reduced accumulation of chilling units. As with spring phenology, the ST of leaf senescence of early successional and exotic species started to decline since 1980. In contrast, for late successional species, the ST of autumn senescence showed an increase for the entire study period from 1951 to 2013. Moreover, the impacts of rising temperature associated with global warming on spring leaf unfolding were stronger than those on autumn leaf senescence. The timing of leaf senescence was positively correlated with the timing of leaf unfolding during 1951-1980. However, as climate warming continued, the differences in the responses between spring and autumn phenology gradually increased, so that the correlation was no more significant during 1981-2013. Our results further suggest that since 2000, due to the decreased temperature sensitivity of leaf unfolding the length of the growing season has not increased any more. These finding needs to be addressed in vegetation models used for assessing the effects of climate change.

Chilling and forcing temperatures interact to predict the onset of wood formation in Northern Hemisphere conifers.

The phenology of wood formation is a critical process to consider for predicting how trees from the temperate and boreal zones may react to climate change. Compared to leaf phenology, however, the determinism of wood phenology is still poorly known. Here, we compared for the first time three alternative ecophysiological model classes (threshold models, heat-sum models and chilling-influenced heat-sum models) and an empirical model in their ability to predict the starting date of xylem cell enlargement in spring, for four major Northern Hemisphere conifers (Larix decidua, Pinus sylvestris, Picea abies and Picea mariana). We fitted models with Bayesian inference to wood phenological data collected for 220 site-years over Europe and Canada. The chilling-influenced heat-sum model received most support for all the four studied species, predicting validation data with a 7.7-day error, which is within one day of the observed data resolution. We conclude that both chilling and forcing temperatures determine the onset of wood formation in Northern Hemisphere conifers. Importantly, the chilling-influenced heat-sum model showed virtually no spatial bias whichever the species, despite the large environmental gradients considered. This suggests that the spring onset of wood formation is far less affected by local adaptation than by environmentally driven plasticity. In a context of climate change, we therefore expect rising winter-spring temperature to exert ambivalent effects on the spring onset of wood formation, tending to hasten it through the accumulation of forcing temperature, but imposing a higher forcing temperature requirement through the lower accumulation of chilling.

Responses of bud-break phenology to daily-asymmetric warming: daytime warming intensifies the advancement of bud break.

There is evidence that the ongoing climate change is happening through nighttime rather than daytime warming. How such a daily-asymmetric warming modifies plant phenology is still unclear. We investigated the effects of asymmetric warming on bud break by daily monitoring seedlings belonging to 26 black spruce [Picea mariana (Mill.) BSP.] and 15 balsam fir [Abies balsamea (L.) Mill.] provenances from the native range in Canada. Seedlings were subjected to either daytime or nighttime warming in three growth chambers at temperatures ranging between 10 and 24 °C. On average, a warming of 4 °C advanced the timings of bud break in both species by 2.4 days, with the later phases being more sensitive to the treatment. Bud break of both species responded more strongly to daytime warming, with the bud break occurred 1.2 and 3.2 days earlier under daytime than nighttime warming in black spruce and balsam fir, respectively. A marked ecotypic differentiation was only observed in black spruce that originated from provenances distributed broadly across Canada, with seedlings from the warmest provenance completing bud break 8.3 days later than those from the coldest one. However, no significant effect of provenance was observed for balsam fir, the narrowly distributed species. Overall, the above results suggest that a higher temporal resolution such as temperatures during daytime and nighttime, and higher spatial resolution should be taken into account to improve the accuracy of phenological model predictions under global change scenarios. Phenological models based on daily average temperature should take into account the diverging impacts of asymmetric warming on plant phenology. Our findings may indicate that the influence of warming on plant phenology may be less dramatic than expected.

[Effect of different fertilization treatments on growth,secondary metabolites,and seed yield and quality of Perilla frutescens].

Perilla frutescens,an annual plant in Labiatae family,is grown throughout China and can be used for medicine purposes and as food additives. The present field experiment was carried out to study the effects of different fertilizer treatments on the concentrations and accumulations of antioxidant components,including flavonoids and polyphenols,growth,seed yields and qualities of this plant.The main aim of this study is to provide farmers some advice for improving the yields and qualities of P. frutescens in theory and practice.Five treatments were set up,including a no fertilizer control(CK),chemical fertilizers(CF),organic fertilizers(M),organic fertilizers plus chemical fertilizers at the rates of 1 ∶1 and 1 ∶3 in terms of nitrogen(50 M,25 M). Plant growth parameters were recorded and total flavonoids and polyphenols were determined in three key growth stages of P. frutescens. At the fast growth period,samples of roots,leaves,and stems were collected for determining a total of flavonoids and polyphenols as well as DPPH removal rate of ethanol extracts. Seed yields and qualities were also recorded at harvest. The results showed fertilization enhanced growth and seed yields although no significant difference was observed in growth and seed yields in inorganic-organic fertilizer treatments. The total flavonoids,polyphenols,and DPPH removal rate of ethanol extracts followed the sequence leaves>stems>roots,indicating synthesis of these metabolites in the leaves. DPPH removal rate showed a positive linear correlation with total flavonoid and polyphenol concentrations. In addition,organic-inorganic fertilization significantly increased the numbers of both effective panicles and paniclegrains. Fertilizer treatments had no effect on seed qualities of P. frutescens,while 50 M achieved the highest yield,which increased by 14. 73% compared to CF alone. In general,50 M increased antioxidant components,biomass,and seed yield of P. frutescens,meriting advocate in cultivation.

Spring phenology at different altitudes is becoming more uniform under global warming in Europe.

Under current global warming, high-elevation regions are expected to experience faster warming than low-elevation regions. However, due to the lack of studies based on long-term large-scale data, the relationship between tree spring phenology and the elevation-dependent warming is unclear. Using 652k records of leaf unfolding of five temperate tree species monitored during 1951-2013 in situ in Europe, we discovered a nonlinear trend in the altitudinal sensitivity (SA , shifted days per 100 m in altitude) in spring phenology. A delayed leaf unfolding (2.7 ± 0.6 days per decade) was observed at high elevations possibly due to decreased spring forcing between 1951 and 1980. The delayed leaf unfolding at high-elevation regions was companied by a simultaneous advancing of leaf unfolding at low elevations. These divergent trends contributed to a significant increase in the SA (0.36 ± 0.07 days 100/m per decade) during 1951-1980. Since 1980, the SA started to decline with a rate of -0.32 ± 0.07 days 100/m per decade, possibly due to reduced chilling at low elevations and improved efficiency of spring forcing in advancing the leaf unfolding at high elevations, the latter being caused by increased chilling. Our results suggest that due to both different temperature changes at the different altitudes, and the different tree responses to these changes, the tree phenology has shifted at different rates leading to a more uniform phenology at different altitudes during recent decades.

Contributions of insects and droughts to growth decline of trembling aspen mixed boreal forest of western Canada.

Insects, diseases, fire and drought and other disturbances associated with global climate change contribute to forest decline and mortality in many parts of the world. Forest decline and mortality related to drought or insect outbreaks have been observed in North American aspen forests. However, little research has been done to partition and estimate their relative contributions to growth declines. In this study, we combined tree-ring width and basal area increment series from 40 trembling aspen (Populus tremuloides Michx.) sites along a latitudinal gradient (from 52° to 58°N) in western Canada and attempted to investigate the effect of drought and insect outbreaks on growth decline, and simultaneously partition and quantify their relative contributions. Results indicated that the influence of drought on forest decline was stronger than insect outbreaks, although both had significant effects. Furthermore, the influence of drought and insect outbreaks showed spatiotemporal variability. In addition, our data suggest that insect outbreaks could be triggered by warmer early spring temperature instead of drought, implicating that potentially increased insect outbreaks are expected with continued warming springs, which may further exacerbate growth decline and death in North America aspen mixed forests.

Linc-RoR promotes MAPK/ERK signaling and confers estrogen-independent growth of breast cancer.

BACKGROUND: The conversion from estrogen-dependent to estrogen-independent state of ER+ breast cancer cells is the key step to promote resistance to endocrine therapies. Although the crucial role of MAPK/ERK signaling pathway in estrogen-independent breast cancer cell growth is well established, the underlying mechanism is not fully understood. METHODS: In this study, we profiled lncRNA expression against a focused group of lncRNAs selected from lncRNA database. CRISPR/Cas9 was employed to knockout (KO) linc-RoR in MCF-7 cells, while rescue experiments were carried out to re-express linc-RoR in KO cells. Colony formation and MTT assays were used to examine the role of linc-RoR in estrogen-independent growth and tamoxifen resistance. Western blot and qRT-PCR were used to determine the change of protein and lncRNA levels, respectively. The expression of DUSP7 in clinical specimens was downloaded from Oncomine ( www.oncomine.org ) and the dataset from Kaplan-Meier Plotter ( http://kmplot.com ) was used to analyze the clinical outcomes in relation to DUSP7. RESULTS: We identified that linc-RoR functions as an onco-lncRNA to promote estrogen-independent growth of ER+ breast cancer. Under estrogen deprivation, linc-RoR causes the upregulation of phosphorylated MAPK/ERK pathway which in turn activates ER signaling. Knockout of linc-RoR abrogates estrogen deprivation-induced ERK activation as well as ER phosphorylation, whereas re-expression of linc-RoR restores all above phenotypes. Moreover, we show that the ERK-specific phosphatase Dual Specificity Phosphatase 7 (DUSP7), also known as MKP-X, is involved in linc-RoR KO-induced repression of MAPK/ERK signaling. Interestingly, linc-RoR KO increases the protein stability of DUSP7, resulting in repression of ERK phosphorylation. Clinical data analysis reveal that DUSP7 expression is lower in ER+ breast cancer samples than that in ER- breast cancer. Moreover, downregulation of DUSP7 expression is associated with poor patient survival. CONCLUSION: Taken together, these results suggest that linc-RoR promotes estrogen-independent growth and activation of MAPK/ERK pathway of breast cancer cells by regulating the ERK-specific phosphatase DUSP7. Thus, this study might help not only in establishing a role for linc-RoR in estrogen-independent and tamoxifen resistance of ER+ breast cancer, but also suggesting a link between linc-RoR and MAPK/ERK pathway.

The Contribution of Carbon and Water in Modulating Wood Formation in Black Spruce Saplings.

Nonstructural carbohydrates (NSCs) play a crucial role in xylem formation and represent, with water, the main constraint to plant growth. We assessed the relationships between xylogenesis and NSCs in order to (1) verify the variance explained by NSCs and (2) determine the influence of intrinsic (tissue supplying carbon) and extrinsic (water availability and temperature) factors. During 2 years, wood formation was monitored in saplings of black spruce (Picea mariana) subjected to a dry period of about 1 month in June and exposed to different temperature treatments in a greenhouse. In parallel, NSC concentrations were determined by extracting the sugar compounds from two tissues (cambium and inner xylem), both potentially supplying carbon for wood formation. A mixed-effect model was used to assess and quantify the potential relationships. Total xylem cells, illustrating meristematic activity, were modeled as a function of water, sucrose, and d-pinitol (conditional r(2) of 0.79). Water availability was ranked as the most important factor explaining total xylem cell production, while the contribution of carbon was lower. Cambium stopped dividing under water deficit, probably to limit the number of cells remaining in differentiation without an adequate amount of water. By contrast, carbon factors were ranked as most important in explaining the variation in living cells (conditional r(2) of 0.49), highlighting the functional needs during xylem development, followed by the tissue supplying the NSCs (cambium) and water availability. This study precisely demonstrates the role of carbon and water in structural growth expressed as meristematic activity and tissue formation.

Drought causes reduced growth of trembling aspen in western Canada.

Adequate and advance knowledge of the response of forest ecosystems to temperature-induced drought is critical for a comprehensive understanding of the impacts of global climate change on forest ecosystem structure and function. Recent massive decline in aspen-dominated forests and an increased aspen mortality in boreal forests have been associated with global warming, but it is still uncertain whether the decline and mortality are driven by drought. We used a series of ring-width chronologies from 40 trembling aspen (Populus tremuloides Michx.) sites along a latitudinal gradient (from 52° to 58°N) in western Canada, in an attempt to clarify the impacts of drought on aspen growth by using Standardized Precipitation Index (SPI) and Standardized Precipitation Evapotranspiration Index (SPEI). Results indicated that prolonged and large-scale droughts had a strong negative impact on trembling aspen growth. Furthermore, the spatiotemporal variability of drought indices is useful for explaining the spatial heterogeneity in the radial growth of trembling aspen. Due to ongoing global warming and rising temperatures, it is likely that severer droughts with a higher frequency will occur in western Canada. As trembling aspen is sensitive to drought, we suggest that drought indices could be applied to monitor the potential effects of increased drought stress on aspen trees growth, achieve classification of eco-regions and develop effective mitigation strategies to maintain western Canadian boreal forests.

Pattern of xylem phenology in conifers of cold ecosystems at the Northern Hemisphere.

The interaction between xylem phenology and climate assesses forest growth and productivity and carbon storage across biomes under changing environmental conditions. We tested the hypothesis that patterns of wood formation are maintained unaltered despite the temperature changes across cold ecosystems. Wood microcores were collected weekly or biweekly throughout the growing season for periods varying between 1 and 13 years during 1998-2014 and cut in transverse sections for assessing the onset and ending of the phases of xylem differentiation. The data set represented 1321 trees belonging to 10 conifer species from 39 sites in the Northern Hemisphere and covering an interval of mean annual temperature exceeding 14 K. The phenological events and mean annual temperature of the sites were related linearly, with spring and autumnal events being separated by constant intervals across the range of temperature analysed. At increasing temperature, first enlarging, wall-thickening and mature tracheids appeared earlier, and last enlarging and wall-thickening tracheids occurred later. Overall, the period of wood formation lengthened linearly with the mean annual temperature, from 83.7 days at -2 °C to 178.1 days at 12 °C, at a rate of 6.5 days °C-1 . April-May temperatures produced the best models predicting the dates of wood formation. Our findings demonstrated the uniformity of the process of wood formation and the importance of the environmental conditions occurring at the time of growth resumption. Under warming scenarios, the period of wood formation might lengthen synchronously in the cold biomes of the Northern Hemisphere.

Basigin-2 upregulated by receptor activator of NF-κB ligand enhances lung cancer-induced osteolytic lesions.

BACKGROUND: Lung cancer bone metastasis causes poor prognosis. Basigin-2, a novel cancer-associated biomarker, is upregulated in lung cancer and has been linked with tumor progression. But little is known about the role of basigin-2 in lung cancer bone metastasis and osteolytic lesion. METHODS: Basigin-2 expression was evaluated in biopsy tissue specimens of 20 lung cancer patients with bone metastases via immunohistochemistry. Invasion assay and MTT proliferation assay were performed to test the invasion and proliferation of lung cancer cell after modulated basigin-2 expression. The osteoclastic activity of basigin-2 was detected in tibia cancer model by injected of lung cancer cells. The regulation role of receptor activator of NF-κB ligand (RANKL) on basigin-2 and its downstream molecules were measured by real-time quantitative RT-PCR, gelatin zymography and western blot analysis. RESULTS: We found that basigin-2 was highly expressed in lung cancer bone metastases. Then, we demonstrated that basigin-2 could promote lung cancer cells invasion, metastasis and proliferation through upregulating metalloproteinases-2 (MMP-2), MMP-9 and vascular endothelial growth factor (VEGF) expression. The lung cancer cells overexpressing basigin-2 strongly induced the osteolytic lesions in immunodeficient mice, which were reduced by treatment with basigin-2 blocking antibody. Furthermore, we explored the enhanced basigin-2 molecular mechanism in lung cancer bone metastasis. Our results indicated the RANKL, pivotal for the control of bone resorption, could increase basigin-2 and its downstream molecules MMP-2, MMP-9 and VEGF expression in vitro. CONCLUSIONS: Basigin-2 upregulated by RANKL induces MMPs and VEGF, which may increase lung cancer cell metastasis ability and support osteoclastic activity. Thus, our data suggest important roles for basigin-2 in lung cancer-induced osteolytic lesion and implicate this protein potential application as a target for lung cancer bone metastasis therapy.

[Influence of continuous cropping on growth of Artemisia annua and bacterial communities in soil].

In this study, several types of Artemisia annua in soil, including the soil which had not been planted, or planted for one year, or continuously planted for three or five years were collected, in order to study the influences of continuous cropping on the growth of A. annua, content of artemisinin, available nutrient of soil, and bacterial community structure through adopting routine analysis and Illumina MiSeq high-throughput sequencing. The results showed that continuous cropping inhibited significantly the growth of A. annua and reduced leaf biomass, content and yield of artemisinin, with the maximum decreasing amplitude of 30.20%, 7.70% and 35.58% respectively. The content of soil organic matter, available nitrogen, available phosphorus and 16S rRNA sequence number were increased to different extents after continuous cropping of A. annua. According to the results of high-throughput sequencing, 634-812 types of common bacteria belonged to 21 categories were planted in different soil of A. annua with different planting years, which represented that the distribution distance of the point of bacterial community with different years among coordinate system of principal component was relative distant, and community structure had significant changes (P<0.05). As the planting years increased, the abundance of Actinobacteria, Chloroflexi, Gemmatimonadetes decreased in contrast to Proteobacteria, Acidobacteria and Verrucomicrobia. In the top 20 types of predominant bacteria,Nitrospira japonica and Nitrospira disappeared, among which, only Gemmatimonadaceae, Micromonosporaceae, Nitrosomonadaceae, Xanthobacteraceae, and unculture bacterium JG30-KF-AS9 were similar, indicating that the planting and continuous cropping of A. annua selectively inhibited the growth and reproduction of soil bacteria, and influenced the supply and transform of soil nutrient, leading to a poor growth and resulting in reduction of artemisinin content and yield. Therefore, it is necessary to advocate crop rotation in the process of planting A. annua.

miR-10b is overexpressed in hepatocellular carcinoma and promotes cell proliferation, migration and invasion through RhoC, uPAR and MMPs.

BACKGROUND: Recently, miR-10b is identified as a miRNA highly expressed in many human cancers, promoting cell migration and invasion. However, the specific function of miR-10b in hepatocellular carcinoma (HCC) is unclear at this point. METHODS: The miR-10b expression levels in 60 paired different TNM Stage HCC tumor tissues compared with adjacent non-tumor (ANT) tissues, normal tissue control (8 benign tumor and 7 normal liver tissues), 3 normal liver and 7 HCC cell lines were measured by real-time quantitative RT-PCR and to evaluate their association with HCC clinicopathologic features. Invasion assay, MTT proliferation assay and wound-healing assay were performed to test the invasion and proliferation of HCC cell after transfection. The effect of miR-10b on HCC in vivo was validated by murine xenograft model. RESULTS: We found that miR-10b expression was increased in human HCC tissues and cell lines compared with normal control, respectively. The expression of miR-10b was correlated with HCC metastatic ability. Overexpression of miR-10b in MHCC-97L cells increased cell motility and invasiveness, whereas inhibition of miR-10b in MHCC-97H cells reduced cell motility and invasiveness in vitro and in vivo. We also showed that HOXD10 was negatively regulated by miR-10b at the posttranscriptional level, via a specific target site within the 3'UTR by luciferase reporter assay. Furthermore, we found that miR-10b induced HCC cell invasion and migration by modulating the HOXD10 target gene RhoC, uPAR, MMP-2 and MMP-9 expression. CONCLUSIONS: Our results suggested that miR-10b was overexpressed in HCC and promoted HCC cell migration and invasion through the HOXD10/ RhoC/ uPAR/ MMPs pathway which may provide a novel bio-target for HCC therapy.