Non-flooding conditions caused by water table drawdown alter microbial network complexity and decrease multifunctionality in alpine wetland soils.

Several soil functions of alpine wetland depend on microbial communities, including carbon storage and nutrient cycling, and soil microbes are highly sensitive to hydrological conditions. Wetland degradation is often accompanied by a decline in water table. With the water table drawdown, the effects of microbial network complexity on various soil functions remain insufficiently understood. In this research, we quantified soil multifunctionality of flooded and non-flooded sites in the Lalu Wetland on the Tibetan Plateau. We employed high-throughput sequencing to investigate the microbial community responses to water table depth changes, as well as the relationships between microbial network properties and soil multifunctionality. Our findings revealed a substantial reduction in soil multifunctionality at both surface and subsurface soil layers (0-20 cm and 20-40 cm) in non-flooded sites compared to flooded sites. The α-diversity of bacteria in the surface soil of non-flooded sites was significantly lower than that in flooded sites. Microbial network properties (including the number of nodes, number of edges, average degree, density, and modularity of co-occurrence networks) exhibited significant correlations with soil multifunctionality. This study underscores the adverse impact of non-flooded conditions resulting from water table drawdown on soil multifunctionality in alpine wetland soils, driven by alterations in microbial community structure. Additionally, we identified soil pH and moisture content as pivotal abiotic factors influencing soil multifunctionality, with microbial network complexity emerging as a valuable predictor of multifunctionality.

Soil temperature and fungal diversity jointly modulate soil heterotrophic respiration under short-term warming in the Zoige alpine peatland.

Global warming has changed carbon cycling in terrestrial ecosystems, but it remains unclear how climate warming affects soil heterotrophic respiration (Rh). We conducted a field experiment in the Zoige alpine peatland to investigate the mechanism of how short-term warming affects Rh by examining the relationships between plant biomass, soil properties, soil microbial diversity, and functional groups and Rh. Our results showed that warming increased Rh after one growing season of warming. However, warming barely changed the bacterial functional groups involved in the carbon cycle predicted by the functional annotation analysis. According to the Mantel test, NO3- was found to be the primary determinant for bacterial and fungal communities. The results of the Structural Equation Model (SEM) indicate that soil temperature and fungal diversity jointly modulate Rh, suggesting that short-term warming may not affect Rh by altering the structural and functional composition of microorganisms, which provides new insight into the mechanisms of the effects of warming on Rh in terrestrial ecosystems.

Effect of 9-year water and nitrogen additions on microbial necromass carbon content at different soil depths and its main influencing factors.

Microbial necromass carbon (MNC) is a major source of soil organic carbon (SOC) pool, significantly influencing soil carbon sequestration. The effects of long-term water and nitrogen addition on MNC in soils at different depths, as well as their interactions, remain poorly understood. In this study, we examined the influence of water addition (+51.67 mm), nitrogen addition (25, 50, 100 kg N ha-1 yr-1), and their interactions on MNC accumulation at different soil depths in temperate grasslands. The addition of water and nitrogen over nine years has been observed to exhibit a decreasing trend in the MNC at different soil depths. Notably, MNC in the topsoil layer (0-10 cm) decreased significantly by 18.56 % under low nitrogen addition treatment, while MNC in the subsoil layer (40-60 cm) declined significantly by 27.19 % under high nitrogen addition treatment. Fungal microbial necromass carbon (FNC) contributed 3.25 times more to SOC than bacterial microbial necromass carbon (BNC). In the 0-10 cm soil layer, MNC is primarily governed by both microbial attributes and the physicochemical properties of the soil, in the 20-40 cm soil layer, the physicochemical properties of the soil predominantly control MNC, in the 40-60 cm layer, microbial characteristics exert a more significant influence on MNC. Collectively, our observations suggest that soil MNC decreased with the addition of water and nitrogen in the 0-60 cm soil slope, which could enable the accurate prediction of global change impacts on the accumulation of soil carbon, thus facilitating the implementation of strategies to augment soil carbon sequestration.

Changes in soil oxidase activity induced by microbial life history strategies mediate the soil heterotrophic respiration response to drought and nitrogen enrichment.

Drought and nitrogen deposition are two major climate challenges, which can change the soil microbial community composition and ecological strategy and affect soil heterotrophic respiration (Rh). However, the combined effects of microbial community composition, microbial life strategies, and extracellular enzymes on the dynamics of Rh under drought and nitrogen deposition conditions remain unclear. Here, we experimented with an alpine swamp meadow to simulate drought (50% reduction in precipitation) and multilevel addition of nitrogen to determine the interactive effects of microbial community composition, microbial life strategy, and extracellular enzymes on Rh. The results showed that drought significantly reduced the seasonal mean Rh by 40.07%, and increased the Rh to soil respiration ratio by 22.04%. Drought significantly altered microbial community composition. The ratio of K- to r-selected bacteria (BK:r) and fungi (FK:r) increased by 20 and 91.43%, respectively. Drought increased hydrolase activities but decreased oxidase activities. However, adding N had no significant effect on microbial community composition, BK:r, FK:r, extracellular enzymes, or Rh. A structural equation model showed that the effects of drought and adding nitrogen via microbial community composition, microbial life strategy, and extracellular enzymes explained 84% of the variation in Rh. Oxidase activities decreased with BK:r, but increased with FK:r. Our findings show that drought decreased Rh primarily by inhibiting oxidase activities, which is induced by bacterial shifts from the r-strategy to the K-strategy. Our results highlight that the indirect regulation of drought on the carbon cycle through the dynamic of bacterial and fungal life history strategy should be considered for a better understanding of how terrestrial ecosystems respond to future climate change.

The driving effects of nitrogen deposition on nitrous oxide and associated gene abundances at two water table levels in an alpine peatland.

Alpine peatlands are recognized as a weak or negligible source of nitrous oxide (N2O). Anthropogenic activities and climate change resulted in the altered water table (WT) levels and increased nitrogen (N) deposition, which could potentially transition this habitat into a N2O emission hotspot. However, the underlying mechanism related with the effects is still uncertain. Hence, we conducted a mesocosm experiment to address the response of growing-season N2O emissions and the gene abundances of nitrification (bacterial amoA) and denitrification (narG, nirS, norB and nosZ) to the increased N deposition (20 kg N ha-1 yr-1) at two WT levels (WT-30, 30 cm below soil surface; WT10, 10 cm above soil surface) in the Zoige alpine peatland, Qinghai-Tibetan Plateau. The results showed that the WT did not affect N2O emissions, and this was attributed with the limitation of soil NO3-. The higher WT level increased denitrification (narG and nirS gene abundance) resulting in the depletion of soil NO3-, but the consequent NO3- deficiency further limited denitrification, while the WT did not affect nitrification (bacterial amoA gene abundance). Meanwhile, the N deposition increased N2O emissions, regardless of WT levels. This was associated with the N-deposition induced increase in denitrification-related gene abundances of narG, nirS, norB and nosZ at WT-30 and narG at WT10. Additionally, the N2O emission factor assigned to N deposition was 1.3 % at WT-30 and 0.9 % at WT10, respectively. Our study provided comprehensive understanding of the mechanisms referring N2O emissions in response to the interactions between climate change and human disturbance from this high-altitude peatland.

Soil Water Content Shapes Microbial Community Along Gradients of Wetland Degradation on the Tibetan Plateau.

Soil microbes are important components in element cycling and nutrient supply for the development of alpine ecosystems. However, the development of microbial community compositions and networks in the context of alpine wetland degradation is unclear. We applied high-throughput 16S rRNA gene amplicon sequencing to track changes in microbial communities along degradation gradients from typical alpine wetland (W), to wet meadow (WM), to typical meadow (M), to grassland (G), and to desert (D) in the Zoige alpine wetland region on the Tibetan Plateau. Soil water content (SWC) decreased as wetland degradation progressed (79.4 and 9.3% in W and D soils, respectively). Total organic carbon (TOC), total nitrogen (TN), and total phosphorus (TP) increased in the soils of WM, and then decreased with alpine wetlands degradation from WM to the soils of M, G, and D, respectively. Wetland degradation did not affect microbial community richness and diversity from W soils to WM, M, and G soils, but did affect richness and diversity in D soils. Microbial community structure was strongly affected by wetland degradation, mainly due to changes in SWC, TOC, TN, and TP. SWC was the primary soil physicochemical property influencing microbial community compositions and networks. In wetland degradation areas, Actinobacteriota, Acidobacteriota, Cholorflexi, and Proteovacteria closely interacted in the microbial network. Compared to soils of W, WM, and M, Actinobacteriota played an important role in the microbial co-occurrence network of the G and D soils. This research contributes to our understanding of how microbial community composition and networks change with varied soil properties during degradation of different alpine wetlands.

Changes in precipitation regime lead to acceleration of the N cycle and dramatic N2O emission.

Alpine meadows on the Qinghai-Tibetan Plateau are sensitive to climate change. The precipitation regime in this region has undergone major changes, "repackaging" precipitation from more frequent, smaller events to less frequent, larger events. Nitrous oxide (N2O) is an important indicator of responses to global change in alpine meadow ecosystems. However, little information is available describing the mechanisms driving the response of N2O emissions to changes in the precipitation regime. In this study, a manipulative field experiment was conducted to investigate N2O flux, soil properties, enzyme activity, and gene abundance in response to severe and moderate changes in precipitation regime over two years. Severe changes in precipitation regime led to a 12.6-fold increase in N2O fluxes (0.0068 ± 0.0018 mg m-2 h-1) from Zoige alpine meadows relative to natural conditions (0.0005 ± 0.0029 mg m-2 h-1). In addition, severe changes in precipitation regime significantly suppressed the activities of leucine amino peptidase (LAP) and peroxidase (PEO), affected ecoenzymatic stoichiometry, and increased the abundances of gdhA, narI and nirK genes, which significantly promoted organic nitrogen (N) decomposition, denitrification, and anammox processes. The increase in abundance of these genes could be ascribed to changes in the abundance of several dominant bacterial taxa (i.e., Actinobacteria and Proteobacteria) as a result of the altered precipitation regime. Decreases in nitrate and soil moisture caused by severe changes in precipitation may exacerbate N limitation and water deficit, lead to a suppression of soil enzyme activity, and change the structure of microorganism community. The N cycle of the alpine meadow ecosystem may accelerate by increasing the abundance of key N functional genes. This would, in turn, lead to increased N2O emission. This study provided insights into how precipitation regimes changes affect N cycling, and may also improve prediction of N2O fluxes in response to changes in precipitation regime.

The divergent vertical pattern and assembly of soil bacterial and fungal communities in response to short-term warming in an alpine peatland.

Soil microbial communities are crucial in ecosystem-level decomposition and nutrient cycling processes and are sensitive to climate change in peatlands. However, the response of the vertical distribution of microbial communities to warming remains unclear in the alpine peatland. In this study, we examined the effects of warming on the vertical pattern and assembly of soil bacterial and fungal communities across three soil layers (0-10, 10-20, and 20-30 cm) in the Zoige alpine peatland under a warming treatment. Our results showed that short-term warming had no significant effects on the alpha diversity of either the bacterial or the fungal community. Although the bacterial community in the lower layers became more similar as soil temperature increased, the difference in the vertical structure of the bacterial community among different treatments was not significant. In contrast, the vertical structure of the fungal community was significantly affected by warming. The main ecological process driving the vertical assembly of the bacterial community was the niche-based process in all treatments, while soil carbon and nutrients were the main driving factors. The vertical structure of the fungal community was driven by a dispersal-based process in control plots, while the niche and dispersal processes jointly regulated the fungal communities in the warming plots. Plant biomass was significantly related to the vertical structure of the fungal community under the warming treatments. The variation in pH was significantly correlated with the assembly of the bacterial community, while soil water content, microbial biomass carbon/microbial biomass phosphorous (MBC/MBP), and microbial biomass nitrogen/ microbial biomass phosphorous (MBN/MBP) were significantly correlated with the assembly of the fungal community. These results indicate that the vertical structure and assembly of the soil bacterial and fungal communities responded differently to warming and could provide a potential mechanism of microbial community assembly in the alpine peatland in response to warming.

Asynchronous responses of microbial CAZymes genes and the net CO2 exchange in alpine peatland following 5 years of continuous extreme drought events.

Peatlands act as an important sink of carbon dioxide (CO2). Yet, they are highly sensitive to climate change, especially to extreme drought. The changes in the net ecosystem CO2 exchange (NEE) under extreme drought events, and the driving function of microbial enzymatic genes involved in soil organic matter (SOM) decomposition, are still unclear. Herein we investigated the effects of extreme drought events in different periods of plant growth season at Zoige peatland on NEE and microbial enzymatic genes of SOM decomposition after 5 years. The results showed that the NEE of peatland decreased significantly by 48% and 26% on average (n = 12, P < 0.05) under the early and midterm extreme drought, respectively. The microbial enzymatic genes abundance of SOM decomposition showed the same decreasing trend under early and midterm extreme drought, but an increasing trend under late extreme drought. The microbial community that contributes to these degradation genes mainly derives from Proteobacteria and Actinobacteria. NEE was mainly affected by soil hydrothermal factors and gross primary productivity but weakly correlated with SOM enzymatic decomposition genes. Soil microbial respiration showed a positive correlation with microbial enzymatic genes involved in the decomposition of labile carbon (n = 18, P < 0.05). This study provided new insights into the responses of the microbial decomposition potential of SOM and ecosystem CO2 sink function to extreme drought events in the alpine peatland.

Different grassland managements significantly change carbon fluxes in an alpine meadow.

Alpine meadow plays vital roles in regional animal husbandry and the ecological environment. However, different grassland managements affect the structure and function of the alpine meadow. In this study, we selected three typical grassland managements including free grazing, enclosure, and artificial grass planting and conducted a field survey to study the effects of grassland managements on carbon fluxes in an alpine meadow. The carbon fluxes were observed by static chamber and environmental factors including vegetation and soil characteristics were measured simultaneously. Our results show that the alpine meadow was a CO2 and CH4 sink, and grassland managements had a significant effect on all CO2 fluxes, including gross ecosystem production (GEP, P< 0.001), net ecosystem production (NEP, P< 0.001) and ecosystem respiration (ER, P< 0.001) but had no significant effect on CH4 fluxes (P > 0.05). The ranking of GEP under the different grassland managements was enclosure > free grazing > artificial grass planting. Furthermore, NEP and ER at enclosure plots were significantly higher than those of the free grazing and artificial grass planting plots. In addition, different grassland managements also affected the vegetation and soil characteristics of the alpine meadow. The aboveground biomass of artificial grass planting was significantly higher than that of the free grazing and enclosure plots. The vegetation coverage under three different grassland managements was ranked in the order of enclosure > artificial grass planting > free grazing and significant differences were observed among them. Moreover, significant differences in the number of species (P< 0.01) and the Margalef richness index (P< 0.05) were detected under three different grassland managements. Further analysis of the relationship between environmental factors and carbon fluxes revealed that GEP and NEP of the alpine meadow were positively correlated with vegetation coverage, the number of species, and the Margalef richness index. Therefore, grassland restoration should be configured with multiple species, which could improve carbon sink capacity while considering the functions of grassland restoration and production.

Plant and Soil Enzyme Activities Regulate CO2 Efflux in Alpine Peatlands After 5 Years of Simulated Extreme Drought.

Increasing attention has been given to the impact of extreme drought stress on ecosystem ecological processes. Ecosystem respiration (Re) and soil respiration (Rs) play a significant role in the regulation of the carbon (C) balance because they are two of the largest terrestrial C fluxes in the atmosphere. However, the responses of Re and Rs to extreme drought in alpine regions are still unclear, particularly with respect to the driver mechanism in plant and soil extracellular enzyme activities. In this study, we imposed three periods of extreme drought events based on field experiments on an alpine peatland: (1) early drought, in which the early stage of plant growth occurred from June 18 to July 20; (2) midterm drought, in which the peak growth period occurred from July 20 to August 23; and (3) late drought, in which the wilting period of plants occurred from August 23 to September 25. After 5 years of continuous extreme drought events, Re exhibited a consistent decreasing trend under the three periods of extreme drought, while Rs exhibited a non-significant decreasing trend in the early and midterm drought but increased significantly by 58.48% (p < 0.05) during the late drought compared with the ambient control. Plant coverage significantly increased by 79.3% (p < 0.05) in the early drought, and standing biomass significantly decreased by 18.33% (p < 0.05) in the midterm drought. Alkaline phosphatase, polyphenol oxidase, and peroxidase increased significantly by 76.46, 77.66, and 109.60% (p < 0.05), respectively, under late drought. Structural equation models demonstrated that soil water content (SWC), pH, plant coverage, plant standing biomass, soil ß-D-cellobiosidase, and ß-1,4-N-acetyl-glucosaminidase were crucial impact factors that eventually led to a decreasing trend in Re, and SWC, pH, ß-1,4-glucosidase (BG), ß-1,4-xylosidase (BX), polyphenol oxidase, soil organic carbon, microbial biomass carbon, and dissolved organic carbon were crucial impact factors that resulted in changes in Rs. Our results emphasize the key roles of plant and soil extracellular enzyme activities in regulating the different responses of Re and Rs under extreme drought events occurring at different plant growth stages.

Divergent responses of CO2 and CH4 fluxes to changes in the precipitation regime on the Tibetan Plateau: Evidence from soil enzyme activities and microbial communities.

Carbon fluxes (CO2 and CH4) are important indicators of the response of alpine meadow ecosystems to global climate change. Alpine meadows on the Qinghai-Tibet Plateau are sensitive to climate change. Although the temporal allocation of precipitation can vary, its intensity is expected to increase, and its frequency is expected to decrease in the future. In this study, a manipulative field experiment was conducted to investigate how carbon fluxes are altered in response to moderate and severe changes in the precipitation regime. Fluctuations in CH4 flux were large under a severely altered precipitation regime (range of -0.048-0.038 mg m-2 h-1). Severe changes in the precipitation regime significantly reduced soil CH4 uptake by approximately 54.3%. This was probably affected by the decrease in the dissolved organic carbon concentration and changes in the microbial community (mainly Gammaproteobacteria), which were induced by variation in soil water conditions under various precipitation regimes. Under moderate changes in the precipitation regime, the average value of CO2 fluxes (ecosystem respiration) was 698.21 ± 35.19 mg m-2 h-1, which was significantly decreased by 20.7% compared with the control. This likely stems from the suppression of enzyme activity (particularly α-1,4-glucosidase and ß-1,4-glucosidase) and the alteration of microbial community structure in this treatment, which led to a decrease in organic matter breakdown and a reduction in the release of CO2 to the atmosphere. However, CO2 fluxes were slightly (i.e., not significantly) decreased under the severely altered precipitation regime. Such different responses of CO2 flux are probably driven by differences in microbial strategies. This study not only increases our understanding of the mechanisms underlying the adaptation of alpine meadow ecosystems to global climate change but also provides new insight into the carbon source/sink functions of alpine meadows.

Drought-induced reduction in methane fluxes and its hydrothermal sensitivity in alpine peatland.

Accurate estimation of CH4 fluxes in alpine peatland of the Qinghai-Tibetan Plateau under extreme drought is vital for understanding the global carbon cycle and predicting future climate change. However, studies on the impacts of extreme drought on peatland CH4 fluxes are limited. To study the effects of extreme drought on CH4 fluxes of the Zoige alpine peatland ecosystem, the CH4 fluxes during both extreme drought treatment (D) and control treatment (CK) were monitored using a static enclosed chamber in a control platform of extreme drought. The results showed that extreme drought significantly decreased CH4 fluxes in the Zoige alpine peatland by 31.54% (P < 0.05). Extreme drought significantly reduced the soil water content (SWC) (P < 0.05), but had no significant effect on soil temperature (Ts). Under extreme drought and control treatments, there was a significant negative correlation between CH4 fluxes and environmental factors (Ts and SWC), except Ts, at a depth of 5cm (P < 0.05). Extreme drought reduced the correlation between CH4 fluxes and environmental factors and significantly weakened the sensitivity of CH4 fluxes to SWC (P < 0.01). Moreover, it was found that the correlation between subsoil (20 cm) environmental factors and CH4 fluxes was higher than with the topsoil (5, 10 cm) environmental factors under the control and extreme drought treatments. These results provide a better understanding of the extreme drought effects on CH4 fluxes of alpine peatland, and their hydrothermal impact factors, which provides a reliable reference for peatland protection and management.

The Association Between Phosphorylated Neurofilament Heavy Chain (pNF-H) and Small Fiber Neuropathy (SFN) in Patients with Impaired Glucose Tolerance.

INTRODUCTION: Small fiber neuropathy (SFN)-the early stage of diabetic peripheral neuropathy (DPN)-progresses gradually and is difficult to diagnose using neurophysiological tests. To facilitate the early diagnosis of SFN, biomarkers for SFN must be identified. The purpose of this study was to investigate the characteristics of SFN in prediabetic patients and the relationship between pNF-H and SFN. METHODS: 44 IGT patients (inpatients and outpatients) were selected at random. 33 healthy subjects served as controls. Data on clinical characteristics and laboratory parameters were collected. Quantitative sensory testing (QST), electromyography (EMG), and Sudoscan were performed, and pNF-H was measured by ELISA. RESULTS: 24 of the 44 patients with impaired glucose tolerance (IGT) were diagnosed with SFN according to the modified Toronto Diabetic Neuropathy Expert Group consensus criteria. The thermal sensory thresholds of the IGT-SFN group were significantly different from those of the CTRL group (p < 0.05), except for the heat pain threshold. The sensory nerve action potential (SNAP) of the sural nerve was 12.39 in the IGT-SFN group, which was significantly lower than those in the other groups. No significant difference in nerve conduction velocity (NCV) was observed among the three groups. The electrochemical skin conductance (ESC) in the IGT-SFN group was 69.78 ± 14.03uS, which was significantly lower than that in the CTRL group. The pNF-H in the IGT-SFN group was 170.6 (140.0, 223.6) pg/ml, which was significantly higher than those in the CTRL and IGT-non-SFN groups (76.55 and 64.7 pg/ml, respectively). Multivariate regression analysis demonstrated that pNF-H and 2h plasma glucose were independently correlated with SFN; the ORs (95% CI) were 1.429 (1.315, 1.924) and 2.375 (1.157, 4.837), respectively. CONCLUSIONS: Serum pNF-H may be associated with SFN in IGT patients, and serum pNF-H could therefore serve as a sensitive biomarker for the detection of SFN.

Correction to: The Association Between Phosphorylated Neurofilament Heavy Chain (pNF-H) and Small Fiber Neuropathy (SFN) in Patients with Impaired Glucose Tolerance.

In the original article, there was some error in Table 2. The correct table is given below.

Plant biomass and soil organic carbon are main factors influencing dry-season ecosystem carbon rates in the coastal zone of the Yellow River Delta.

Coastal wetlands are considered as a significant sink of global carbon due to their tremendous organic carbon storage. Coastal CO2 and CH4 flux rates play an important role in regulating atmospheric CO2 and CH4 concentrations. However, the relative contributions of vegetation, soil properties, and spatial structure on dry-season ecosystem carbon (C) rates (net ecosystem CO2 exchange, NEE; ecosystem respiration, ER; gross ecosystem productivity, GEP; and CH4) remain unclear at a regional scale. Here, we compared dry-season ecosystem C rates, plant, and soil properties across three vegetation types from 13 locations at a regional scale in the Yellow River Delta (YRD). The results showed that the Phragmites australis stand had the greatest NEE (-1365.4 µmol m-2 s-1), ER (660.2 µmol m-2 s-1), GEP (-2025.5 µmol m-2 s-1) and acted as a CH4 source (0.27 µmol m-2 s-1), whereas the Suaeda heteroptera and Tamarix chinensis stands uptook CH4 (-0.02 to -0.12 µmol m-2 s-1). Stepwise multiple regression analysis demonstrated that plant biomass was the main factor explaining all of the investigated carbon rates (GEP, ER, NEE, and CH4); while soil organic carbon was shown to be the most important for explaining the variability in the processes of carbon release to the atmosphere, i.e., ER and CH4. Variation partitioning results showed that vegetation and soil properties played equally important roles in shaping the pattern of C rates in the YRD. These results provide a better understanding of the link between ecosystem C rates and environmental drivers, and provide a framework to predict regional-scale ecosystem C fluxes under future climate change.

Facilitating effects of berberine on rat pancreatic islets through modulating hepatic nuclear factor 4 alpha expression and glucokinase activity.

AIM: To observe the effect of berberine on insulin secretion in rat pancreatic islets and to explore its possible molecular mechanism. METHODS: Primary rat islets were isolated from male Sprague-Dawley rats by collagenase digestion and treated with different concentrations (1, 3, 10 and 30 micromol/L) of berberine or 1 micromol/L Glibenclamide (GB) for 24 h. Glucose-stimulated insulin secretion (GSIS) assay was conducted and insulin was determined by radioimmunoassay. 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay was performed to evaluate cytotoxicity. The mRNA level of hepatic nuclear factor 4 alpha (HNF4alpha) was determined by reverse transcription polymerase chain reaction (RT-PCR). Indirect immunofluorescence staining and Western blot analysis were employed to detect protein expression of HNF4alpha in the islets. Glucokinase (GK) activity was measured by spectrophotometric method. RESULTS: Berberine enhanced GSIS rather than basal insulin secretion dose-dependently in rat islets and showed no significant cytotoxicity on islet cells at the concentration of 10 mumol/L. Both mRNA and protein expressions of HNF4alpha were up-regulated by berberine in a dose-dependent manner, and GK activity was also increased accordingly. However, GB demonstrated no regulatory effects on HNF4alpha expression or GK activity. CONCLUSION: Berberine can enhance GSIS in rat islets, and probably exerts the insulinotropic effect via a pathway involving HNF4alpha and GK, which is distinct from sulphonylureas (SUs).

[Effects of berberine on expression of hepatocyte nuclear factor 4alpha and glucokinase activity in mouse primary hepatocytes].

OBJECTIVE: To observe the expression of hepatocyte nuclear factor 4alpha (HNF4alpha) and the activity of key enzyme glucokinase (GK) in glucose metabolism, and further to investigate the possible mechanism of berberine in treating type 2 diabetes. METHOD: Mouse primary hepatocytes were isolated by an improved single two-step perfusion method. The murine hepatocytes were cultured and incubated with berberine (0, 1, 3, 10, 30, 100 micromol x L(-1)) and 1 mmol x L(-1) metformin for 24 h respectively. The mRNA expression of HNF4alpha were quantified by RT-PCR and the protein expression of HNF4alpha were quantified by Western-blot. And the activity of GK were detected with enzyme kinetics method. RESULT: As compared with the negative control group, at a certain concentration range, the expression of HNF4alpha mRNA and protein and the activity of GK were promoted by berberine. Both of them reached the top at the concentration of 30 micromol x L(-1) (P<0.01). But the metformin made no difference with the negative control group on the expression of HNF4alpha and the activity of GK. CONCLUSION: It is suggested that the effects of berberine on improving glucose metabolism can be mechanically associated with its up-regulating the HNF4a expression and inducing the activity of hepatic glucokinase.

[Effects of Short-term Exogenous Nitrogen and Carbon Input on Soil Respiration Under Changing Precipitation Pattern].

The responses of soil respiration to exogenous carbon (C) and nitrogen (N) inputs under changing precipitation patterns were explored via in-situ field experiments. In 2014, a typical temperate grassland on the Xilin River of Inner Mongolia was taken as the research site, and soil respiration was measured in the following treatments:addition of water alone (CK), addition of water + N fertilizer[CN, 2.5 g·(m2·a)-1], addition of water + labile C[CG, 24 g·(m2·a)-1], and addition of water + N fertilizer+ labile C[CNG, 2.5 g·(m2·a)-1+24 g·(m2·a) -1], and the correlations of soil respiration with soil temperature, soil moisture, soil dissolved organic C (DOC), and soil microbial biomass C (MBC) were analyzed. During the first water application event (FWE) with the frequency of natural precipitation, cumulative CO2 efflux over 168 hours significantly increased in the CG and CNG treatments, whereas there was no such change in the CN treatment. In addition, soil MBC contents in the CG and CNG treatments were significantly higher than that in the CK and CN treatments, and the correlation of average soil respiration rate with soil MBC content among these treatments was positively significant (P<0.05). In contrast with during the FWE, cumulative CO2 efflux over 168 hours and soil MBC content significantly decreased during the second water application event (SWE) with no natural precipitation (P<0.05), whereas soil DOC content significantly increased (P<0.05). The cumulative CO2 efflux over 168 hours significantly decreases in the CN and CG treatments (P<0.05).During both the water application events, soil respiration rate had a positive relationship with soil temperature and soil volume water content (P<0.05). Therefore, it is proposed that the distribution of natural precipitation influences soil water content, which controls the effects of exogenous C and N on soil respiration in semiarid grassland ecosystems.

[Effect of Irrigation Patterns on Soil CO2 and N2O Emissions from Winter Wheat Field in North China Plain].

The water-saving irrigation is the trend of modernized agriculture. This paper aimed to study the effect of water-saving irrigation on soil CO2 and N2O emissions. The field experiments were conducted under micro sprinkler irrigation of integrated water and fertilizer (MSI) and conventional flooding irrigation (FI) in winter wheat growth season in the west of North China Plain during 2013- 2014 using the static chamber method. This paper analyzed the seasonal variation of soil CO2and N2O emissions under MSI and FI, and then compared the soil CO2 and N2O emissions from treatments located in different vertical distance away from micro sprinkler pipe. Root exclusion was used to estimate the components of soil respiration and agricultural carbon sequestration intensity under MSI and FI in winter wheat field. The results indicated that: (1) The average soil CO2 emissions under MSI and FI were 418.19 mg (m² · h)⁻¹ and 372.14 mg · (m² · h)⁻¹ respectively with no significant difference, and cumulative CO2 emissions under MSI and FI were 2 150.6 g · m⁻² and 1 904.6 g · m⁻², respectively. (2) During returning green stage to harvest stage of winter wheat, the highest soil CO2 cumulative emissions were found at the closest site to the micro sprinkler irrigated pipes under MSI. However, there were no significant differences among spatial treatments. (3) Under MSI and FI, soil heterotrophic respiration (C) was 468.49 g · m⁻² and 427.31 g · m⁻², and the net primary productivity (3) was 1988.21 g · m⁻² and 1770.54 g · m⁻²; the carbon sink (C) during winter wheat growing season was 1 519.72 g · m⁻² and 1 343.24 g · m⁻², respectively. (4) The average N2O emissions under MSI and FI were 50.77 µg · (m² · h)⁻¹ and 28.81 µg · (m² · h)⁻¹ respectively with no significant difference. Cumulative N2O emission under MSI and FI was 272.67 mg · m⁻² and 154.08 mg · m⁻², respectively. (5) During returning green stage to harvest stage of winter wheat, the farther the distance away from the micro sprinkler irrigated pipes, the smaller the soil N2O emissions. Moreover, there were no significant differences among sptial treatment under MSI. Therefore, despite of the increase of soil CO2and N2O emissions, the intensity of carbon sink increased during the transformation from traditional flood irrigation to micro sprinkler irrigation in winter wheat fields.