Analysis of microbial community composition and diversity in the rhizosphere of Salvia miltiorrhiza at different growth stages.

Salvia miltiorrhiza is a kind of medicinal plant with various pharmacological activities. Few studies on the composition and diversity of rhizosphere microbial communities at different growth stages have been conducted on Salvia miltiorrhiz; in particular, salviorrhiza grows in soil that has been continuously planted for 3 years. The purpose of this study was to understand the changes of soil physicochemical properties of Salvia miltiorrhiza at different growth stages, and to study the composition and diversity of rhizosphere microbial community at different growth stages. Illumina NovaSeq sequencing technology was used to analyze the bacterial 16S rRNA gene and the fungal ITS region in the rhizosphere soil of Salvia miltiorrhiza at different growth stages. The results showed that the dominant bacterial phyla in the Salvia miltiorrhiza rhizosphere were Proteobacteria, Bacteroidetes, Acidobacteria, Firmicutes, Actinobacteria, and Chloroflexi. The dominant fungal phyla were Ascomycota, Mortierellomycota, Basidiomycota, and Rozellomycota. During the growth of Salvia miltiorrhiza, the physical and chemical properties of soil changed. As the Salvia miltiorrhiza grew, the content of available phosphorus, available potassium, pH, nitrate nitrogen, and ammonium nitrogen significantly decreased. Ammonium nitrogen and nitrate nitrogen had a greater impact on the bacterial community structure in the rhizosphere than on the fungal community structure. The work was to reveal differences in the rhizosphere bacterial and fungal community structure during different growth stages of Salvia miltiorrhiza, further understand the changes of rhizosphere microbial ecological characteristics and soil physicochemical properties during the cultivation of Salvia miltiorrhiza.

The mechanical properties measurement could be affected by forceps: a technical note on sampling human annulus fibrosus.

PURPOSE: To compare the mechanical properties of human annulus fibrosus obtained by forceps versus bistoury and observe whether the measurement could be affected by forceps sampling method. METHODS: In this study, the mechanical properties of the the extracellular matrix (ECM) of human annulus fibrosus, including elastic modulus and stiffness, were investigated using atomic force microscope (AFM). Tissue was obtained from patients during operation using a bistoury or nucleus pulposus forceps. Tissues obtained with the nucleus pulposus forceps were considered as the forceps group and those obtained with a bistoury were considered as the bistoury group. RESULTS: There was no significant difference observed between the forceps and bistoury group according to histological staining. The elastic modulus of the forceps group was 0.41 ± 0.08 MPa, and that of bistoury group was 0.53 ± 0.13 MPa, and the difference between the two groups was statistically significant (p < 0.05). The stiffness of the forceps group was 0.024 ± 0.003 N/m, and that of the bistoury group was 0.037 ± 0.003 N/m, and the difference between the two groups was statistically significant (p < 0.05). CONCLUSION: The results indicate that the forceps sampling method has a substantial negative effect on the micromechanical properties of the ECM of the annulus fibrosus. Bistoury sampling method is recommended as the experimental subject for exploring the micromechanics mechanisms of cervical degenerative disease.

The effect of suboccipital muscle dysfunction on the biomechanics of the upper cervical spine: a study based on finite element analysis.

OBJECTIVE: Muscle dysfunction caused by repetitive work or strain in the neck region can interfere muscle responses. Muscle dysfunction can be an important factor in causing cervical spondylosis. However, there has been no research on how the biomechanical properties of the upper cervical spine change when the suboccipital muscle group experiences dysfunction. The objective of this study was to investigate the biomechanical evidence for cervical spondylosis by utilizing the finite element (FE) approach, thus and to provide guidance for clinicians performing acupoint therapy. METHODS: By varying the elastic modulus of the suboccipital muscle, the four FE models of C0-C3 motion segments were reconstructed under the conditions of normal muscle function and muscle dysfunction. For the two normal condition FE models, the elastic modulus for suboccipital muscles on both sides of the C0-C3 motion segments was equal and within the normal range In one muscle dysfunction FE model, the elastic modulus on both sides was equal and greater than 37 kPa, which represented muscle hypertonia; in the other, the elastic modulus of the left and right suboccipital muscles was different, indicating muscle imbalance. The biomechanical behavior of the lateral atlantoaxial joint (LAAJ), atlanto-odontoid joint (ADJ), and intervertebral disc (IVD) was analyzed by simulations, which were carried out under the six loadings of flexion, extension, left and right lateral bending, left and right axial rotation. RESULTS: Under flexion, the maximum stress in LAAJ with muscle imbalance was higher than that with normal muscle and hypertonia, while the maximum stress in IVD in the hypertonic model was higher than that in the normal and imbalance models. The maximum stress in ADJ was the largest under extension among all loadings for all models. Muscle imbalance and hypertonia did not cause overstress and stress distribution abnormalities in ADJ. CONCLUSION: Muscle dysfunction increases the stress in LAAJ and in IVD, but it does not affect ADJ.

Soil prokaryotic community shows no response to 2 years of simulated nitrogen deposition in an arid ecosystem in northwestern China.

An arid ecosystem might be sensitive to nitrogen (N) deposition, but the associated ecosystem-specific response of soil microbes is not well studied. To assess the N enrichment effects on plant and prokaryotic community diversity, we performed a two-year NH4 NO3 treatment in a desert steppe in northwestern China. Results showed that N addition increased plant aboveground biomass and decreased plant Shannon diversity. A C4 herb (Salsola collina) became dominant, and loss of legume species was observed. The concentrations of soil NH4 + -N, NO3 - -N, microbial biomass N, and the plant aboveground biomass N pool increased in contrast to total N, suggesting that the N input into the arid ecosystem might mainly be assimilated by plants and exit the ecosystem. Remarkably, the α-diversity and structure of the soil prokaryotic community did not vary even at the highest N addition rate. Structural equation modelling further found that the plant aboveground N pool counteracted the acidification effect of N deposition and maintained soil pH thus partially stabilizing the composition of prokaryotic communities in a desert steppe. These findings suggested that the plants and N loss might contribute to the lack of responsiveness of soil prokaryotic community to N deposition in a desert steppe.

An improved level set method for vertebra CT image segmentation.

BACKGROUND: Clinical diagnosis and therapy for the lumbar disc herniation requires accurate vertebra segmentation. The complex anatomical structure and the degenerative deformations of the vertebrae makes its segmentation challenging. METHODS: An improved level set method, namely edge- and region-based level set method (ERBLS), is proposed for vertebra CT images segmentation. By considering the gradient information and local region characteristics of images, the proposed model can efficiently segment images with intensity inhomogeneity and blurry or discontinuous boundaries. To reduce the dependency on manual initialization in many active contour models and for an automatic segmentation, a simple initialization method for the level set function is built, which utilizes the Otsu threshold. In addition, the need of the costly re-initialization procedure is completely eliminated. RESULTS: Experimental results on both synthetic and real images demonstrated that the proposed ERBLS model is very robust and efficient. Compared with the well-known local binary fitting (LBF) model, our method is much more computationally efficient and much less sensitive to the initial contour. The proposed method has also applied to 56 patient data sets and produced very promising results. CONCLUSIONS: An improved level set method suitable for vertebra CT images segmentation is proposed. It has the flexibility of segmenting the vertebra CT images with blurry or discontinuous edges, internal inhomogeneity and no need of re-initialization.

Long-term phosphorus addition alters plant community composition but not ecosystem stability of a nitrogen-enriched desert steppe.

Under ongoing global change, whether grassland ecosystems can maintain their functions and services depends largely on their stability. However, how ecosystem stability responds to increasing phosphorus (P) inputs under nitrogen (N) loading remains unclear. We conducted a 7-year field experiment to examine the influence of elevated P inputs (ranging from 0 to 16 g P m-2 yr-1) on the temporal stability of aboveground net primary productivity (ANPP) under N addition of 5 g N·m-2·yr-1 in a desert steppe. We found that under N loading, P addition altered plant community composition but did not significantly affect ecosystem stability. Specifically, with the increase in the P addition rate, declines in the relative ANPP of legume could be compensated for by an increase in the relative ANPP of grass and forb species, yet community ANPP and diversity remained unchanged. Notably, the stability and asynchrony of dominant species tended to decrease with increasing P addition, and a significant decrease in legume stability was observed at high P rates (>8 g P m-2 yr-1). Moreover, P addition indirectly affected ecosystem stability by multiple pathways (e.g., species diversity, species asynchrony, dominant species asynchrony, and dominant species stability), as revealed by structural equation modeling results. Our results suggest that multiple mechanisms work concurrently in stabilizing the ecosystem stability of desert steppes and that increasing P inputs may not alter desert steppe ecosystem stability under future N-enriched scenarios. Our results will help improve the accuracy of vegetation dynamics assessments in arid ecosystems under future global change.

Factors driving the assembly of prokaryotic communities in bulk soil and rhizosphere of Torreya grandis along a 900-year age gradient.

Excessive nutrient inputs imperil the stability of forest ecosystems via modifying the interactions among soil properties, microbes, and plants, particularly in forests composed of cash crops that are under intensive disturbances of agricultural activities, such as Torreya grandis. Understanding the potential drivers of soil microbial community helps scientists develop effective strategies for balancing the protection and productivity of the ancient Torreya forest. Here, we assayed the link between plant and soil parameters and prokaryote communities in bulk soil and T. grandis rhizosphere in 900-year-old stands by detecting plant and soil properties in two independent sites in southeastern China. Our results showed no apparent influence of stand age on the compositions of prokaryote communities in bulk soil and T. grandis rhizosphere. In contrast, soil abiotic factors (i.e., soil pH) overwhelm plant characteristics (i.e., height, plant tissue carbon, nitrogen, and phosphorus content) and contribute most to the shift in prokaryote communities in bulk soil and T. grandis rhizosphere. Soil pH leads to an increase in microbiota alpha diversity in both compartments. With the help of a random forest, we found a critical transition point of pH (pH = 4.9) for the dominance of acidic and near-neutral bacterial groups. Co-occurrence network analysis further revealed a substantially simplified network in plots with a pH of <4.9 versus samples with a pH of ≥4.9, indicating that soil acidification induces biodiversity loss and disrupts potential interactions among soil microbes. Our findings provide empirical evidence that soil abiotic properties nearly completely offset the roles of host plants in the assembly and potential interactions of rhizosphere microorganisms. Hence, reduction in inorganic fertilization and proper liming protocols should be seriously considered by local farmers to protect ancient Torreya forests.

A Novel Construct Incorporating C2 Unilateral Pedicle and Contralateral Translaminar Screws for Occipitocervical Internal Fixation: An In Vitro Biomechanical Study.

BACKGROUND: Occipitocervical fixation using bilateral C2 pedicle screws (C0-C2BiPS) and occipitocervical fixation using bilateral C2 translaminar screws (C0-C2BiLS) provide satisfactory stability. Bilateral fixation is not feasible for cases of C2 unilateral pedicle morphology abnormality and ipsilateral laminectomy. This study proposed and evaluated novel occipitocervical fixation using C2 unilateral pedicle screw and contralateral translaminar screws (C0-C2PSLS). METHODS: In 6 human cadaveric specimens, an in vitro experiment was performed with 2.0-Nm moment control in flexion-extension, lateral bending, and axial rotation to investigate biomechanical stability. Neutral zone and range of motion (ROM) between the occiput (C0) and C2 were measured in the intact state, after destabilization, and after sequential stabilization using C0-C2BiPS, C0-C2BiLS, and C0-C2PSLS constructs. RESULTS: Flexion-extension ROM of the intact specimens at C0-C2 was 27.4° ± 2.4°. Instrumentation with C0-C2PSLS, C0-C2BiPS, and C0-C2BiLS reduced flexion-extension ROM to 3.7° ± 1.3°, 4.7° ± 1.4°, and 4.5° ± 1.4°, respectively. In lateral bending, ROM values were 7.0° ± 0.6°, 4.5° ± 1.4°, 4.2° ± 1.4°, 2.7° ± 1.0°, respectively. In axial rotation, ROM values were 65.3° ± 5.7°, 2.5° ± 0.5°, 1.4° ± 0.5°, and 0.9° ± 0.6°, respectively. Comparing destabilized and intact specimens, all 3 constructs significantly reduced ROM and neutral zone values in flexion-extension, lateral bending, and axial rotation (P < 0.05). Direct comparisons between the 3 constructs revealed no significant difference (P > 0.05). CONCLUSIONS: Novel C0-C2PSLS provides similar stabilization effect as C0-C2BiPS and C0-C2BiLS constructs and has potential for clinical use, especially for cases of C2 unilateral pedicle morphology abnormality and ipsilateral laminectomy.

Water supply changes N and P conservation in a perennial grass Leymus chinensis.

Changes in precipitation can influence soil water and nutrient availability, and thus affect plant nutrient conservation strategies. Better understanding of how nutrient conservation changes with variations in water availability is crucial for predicting the potential influence of global climate change on plant nutrient-use strategy. Here, green-leaf nitrogen (N) and phosphorus (P) concentrations, N- and P-resorption proficiency (the terminal N and P concentration in senescent leaves, NRP and PRP, respectively), and N- and P-resorption efficiency (the proportional N and P withdrawn from senescent leaves prior to abscission, NRE and PRE, respectively) of Leymus chinensis (Trin.) Tzvel., a typical perennial grass species in northern China, were examined along a water supply gradient to explore how plant nutrient conservation responds to water change. Increasing water supply at low levels (< 9000 mL/year) increased NRP, PRP and PRE, but decreased green-leaf N concentration. It did not significantly affect green-leaf P concentration or NRE. By contrast, all N and P conservation indicators were not significantly influenced at high water supply levels (> 9000 mL/year). These results indicated that changes in water availability at low levels could affect leaf-level nutrient characteristics, especially for the species in semiarid ecosystems. Therefore, global changes in precipitation may pose effects on plant nutrient economy, and thus on nutrient cycling in the plant-soil systems.

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Soil samples from four vegetation mini-patches (Artemisia scoparia, Glycyrrhiza uralensis, Sophora alopecuroides, Astragalus melilotoides) in a desert steppe in central Ningxia were collected. Soil physico-chemical properties including soil particle-size distribution, organic matter, pH, EC, total N, total K, total P of three depths were measured. The fractal dimension of particle size distribution characteristics of soils derived from four different vegetation mini-patches and their correlations with soil physico-chemical properties were examined. The results showed that patch vege-tation distribution affected the distribution of soil particle size, with the A. melilotoides mini-patch being the highest (D=2.51) and G. uralensis mini-patch being the lowest (D=2.46). There were significant positive correlation between fractal dimensions and the contents of clay and silt, and nega-tive correlation between fractal dimensions and sand content. Fractal dimensions were positively correlated with pH value and EC, negatively correlated with the contents of soil organic matter and total N, and had no correlation with the contents of soil total K and total P. The patchy vegetation distribution had potential trends of salinization and degradation.

Effects of arbuscular mycorrhizal fungi on growth and nitrogen uptake of Chrysanthemum morifolium under salt stress.

Soil salinity is a common and serious environmental problem worldwide. Arbuscular mycorrhizal fungi (AMF) are considered as bio-ameliorators of soil salinity tolerance in plants. However, few studies have addressed the possible benefits of AMF inoculation for medicinal plants under saline conditions. In this study, we examined the effects of colonization with two AMF, Funneliformis mosseae and Diversispora versiformis, alone and in combination, on the growth and nutrient uptake of the medicinal plant Chrysanthemum morifolium (Hangbaiju) in a greenhouse salt stress experiment. After 6 weeks of a non-saline pretreatment, Hangbaiju plants with and without AMF were grown for five months under salinity levels that were achieved using 0, 50 and 200 mM NaCl. Root length, shoot and root dry weight, total dry weight, and root N concentration were higher in the mycorrhizal plants than in the non-mycorrhizal plants under conditions of moderate salinity, especially with D. versiformis colonization. As salinity increased, mycorrhizal colonization and mycorrhizal dependence decreased. The enhancement of root N uptake is probably the main mechanism underlying salt tolerance in mycorrhizal plants. These results suggest that the symbiotic associations between the fungus D. versiformis and C. morifolium plants may be useful in biotechnological practice.

Phosphorus addition changes belowground biomass and C:N:P stoichiometry of two desert steppe plants under simulated N deposition.

Many studies have reported that increasing atmospheric nitrogen (N) deposition broadens N:phosphorus (P) in both soils and plant leaves and potentially intensifies P limitation for plants. However, few studies have tested whether P addition alleviates N-induced P limitation for plant belowground growth. It is also less known how changed N:P in soils and leaves affect plant belowground stoichiometry, which is significant for maintaining key belowground ecological processes. We conducted a multi-level N:P supply experiment (varied P levels combined with constant N amount) for Glycyrrhiza uralensis (a N fixing species) and Pennisetum centrasiaticum (a grass) from a desert steppe in Northwest China during 2011-2013. Results showed that increasing P addition increased the belowground biomass and P concentrations of both species, resulting in the decreases in belowground carbon (C):P and N:P. These results indicate that P inputs alleviated N-induced P limitation and hence stimulated belowground growth. Belowground C:N:P stoichiometry of both species, especially P. centrasiaticum, tightly linked to soil and green leaf C:N:P stoichiometry. Thus, the decoupling of C:N:P ratios in both soils and leaves under a changing climate could directly alter plant belowground stoichiometry, which will in turn have important feedbacks to primary productivity and C sequestration.

Changes in C:N:P stoichiometry modify N and P conservation strategies of a desert steppe species Glycyrrhiza uralensis.

Numerous studies have concluded that carbon (C):nitrogen (N):phosphorus (P) stoichiometry in both soils and plants tends to be decoupled under global change. We consequently hypothesized that plants will adjust nutrient conservation strategies to balance the altered elemental stoichiometry accordingly. To test our hypothesis, we conducted two pot-cultured experiments (with 8-level water and 6-level N addition treatments) using N-fixing species Glycyrrhiza uralensis Fisch from a desert steppe in northwestern China. We observed that high water availability lowered total N content and the N:P ratio in soils, further promoting both N and P resorption from senescing leaves of G. uralensis. High N addition enhanced soil N availability and the N:P ratio, thereby reducing N resorption, but increasing P resorption of G. uralensis. Comparatively, there were also great changes in senescing leaf C:N:P stoichiometry while no clear changes were observed in either green leaf or root C:N:P stoichiometry of G. uralensis. As expected, the altered C:N:P stoichiometry may, in turn, modify N and P conservation strategies through their close linkages with N and P uptake in green leaves of G. uralensis. This modification may also further exert effects on N and P cycling of the desert steppe.

Numerical analysis of the influence of nucleus pulposus removal on the biomechanical behavior of a lumbar motion segment.

Nucleus replacement was deemed to have therapeutic potential for patients with intervertebral disc herniation. However, whether a patient would benefit from nucleus replacement is technically unclear. This study aimed to investigate the influence of nucleus pulposus (NP) removal on the biomechanical behavior of a lumbar motion segment and to further explore a computational method of biomechanical characteristics of NP removal, which can evaluate the mechanical stability of pulposus replacement. We, respectively, reconstructed three types of models for a mildly herniated disc and three types of models for a severely herniated disc based on a L4-L5 segment finite element model with computed tomography image data from a healthy adult. First, the NP was removed from the herniated disc models, and the biomechanical behavior of NP removal was simulated. Second, the NP cavities were filled with an experimental material (Poisson's ratio = 0.3; elastic modulus = 3 MPa), and the biomechanical behavior of pulposus replacement was simulated. The simulations were carried out under the five loadings of axial compression, flexion, lateral bending, extension, and axial rotation. The changes of the four biomechanical characteristics, i.e. the rotation degree, the maximum stress in the annulus fibrosus (AF), joint facet contact forces, and the maximum disc deformation, were computed for all models. Experimental results showed that the rotation range, the maximum AF stress, and joint facet contact forces increased, and the maximum disc deformation decreased after NP removal, while they changed in the opposite way after the nucleus cavities were filled with the experimental material.