Clinical and molecular characterization of 10 Chinese children with congenital adrenal hyperplasia due to 11beta-hydroxylase deficiency.

BACKGROUND: The clinical manifestations of nonclassical 11beta-hydroxylase deficiency are very similar to those of non-classical 21-hydroxylase deficiency. For this study, we investigated the relationship between the clinical and molecular features of congenital adrenal hyperplasia caused by 11beta-hydroxylase deficiency and reviewed the related literature, which are expected to provide assistance for the clinical diagnosis and analysis of congenital adrenal hyperplasia. METHODS: Clinical data for 10 Chinese patients diagnosed with congenital adrenal hyperplasia in our hospital from 2018 to 2022 were retrospectively analyzed. We examined the effects of gene mutations on protease activity and constructed three-dimensional structure prediction models of proteins. RESULTS: We describe 10 patients with 11beta-hydroxylase gene mutations (n = 5, 46,XY; n = 5, 46,XX), with 10 novel mutations were reported. Female patients received treatment at an early stage, with an average age of 2.08 ± 1.66 years, whereas male patients received treatment significantly later, at an average age of 9.77 ± 3.62 years. The most common CYP11B1 pathogenic variant in the Chinese population was found to be c.1360C > T. All mutations lead to spatial conformational changes that affect protein stability. CONCLUSIONS: Our study found that there was no significant correlation between each specific mutation and the severity of clinical manifestations. Different patients with the same gene pathogenic variant may have mild or severe clinical manifestations. The correlation between genotype and phenotype needs further study. Three-dimensional protein simulations may provide additional support for the physiopathological mechanism of genetic mutations.

Impact of bentonite on greenhouse gas emissions during pig manure composting and its subsequent application.

Additives were widely investigated to retain the nutrients and mitigate the greenhouse gas emissions (GHGs) during manure composting. However, the sustained effects of additives on the GHGs emissions following incorporation of composts to soil were scarcely explored. This study evaluated the effects of bentonite added at the beginning of pig manure composting on the GHGs emissions during two successive processes, i.e., composting and soil incubation amended with composting products. Addition of bentonite did not hinder the composting process and alter the total CO2 emission. On the other hand, reduction by about 17% and 29% for CH4 and N2O emission, respectively, was achieved in the presence of bentonite during composting. Incorporation of the final composting products to soil enhanced significantly the soil C and N of various forms, and gas emissions of CO2 and N2O. However, no significant differences were observed between bentonite-manure co-compost and manure-only compost application except for the N2O emission. Compared to the manure-only compost, compost amended with bentonite reduced N2O loss by around 6.8%, but not statistically significant. This study confirmed that addition of bentonite at the composting stage can mitigate the GHGs emission considering both composting and compost application stages, with all reductions occurring at the composting stage.

[Effects of Microplastics Addition on Soil Organic Carbon Mineralization in Citrus Orchard].

To investigate the effects of microplastics on soil organic carbon mineralization and the changes in soil enzyme activities, an incubation experiment was conducted whereby single applications of either microplastics or straw, and combined application of both, were added to Dangyang citrus orchard soil. The results showed that the combined application of straw and microplastics significantly affected organic carbon mineralization in the soil, but the single addition of microplastics had no significant effect. Compared with straw alone, the application of a small combined amount of microplastics and straw significantly increased soil organic carbon mineralization by 8.20%, while medium and high amounts of the combined application significantly inhibited soil organic carbon mineralization. The lowest amount of organic carbon mineralization occurred with the highest amount of combined microplastics and straw, 10.13% lower than with straw alone. The addition of microplastics significantly reduced the activity of ß-glucosidase. In particular, a high amount of microplastics significantly decreased the activity of ß-glucosidase, compared with the control, by 20.52%, 43.93%, and 17.79% on the day 1, 6, and 35, respectively. However, straw application alleviated the inhibition effect of microplastic application on soil ß-glucosidase activity. The soil organic carbon mineralization rate was significantly positively correlated with DOC, MBC and ß-glucosidase activity.

Autophagy receptor OPTN (optineurin) regulates mesenchymal stem cell fate and bone-fat balance during aging by clearing FABP3.

Senile osteoporosis (OP) is often concomitant with decreased autophagic activity. OPTN (optineurin), a macroautophagy/autophagy (hereinafter referred to as autophagy) receptor, is found to play a pivotal role in selective autophagy, coupling autophagy with bone metabolism. However, its role in osteogenesis is still mysterious. Herein, we identified Optn as a critical molecule of cell fate decision for bone marrow mesenchymal stem cells (MSCs), whose expression decreased in aged mice. Aged mice revealed osteoporotic bone loss, elevated senescence of MSCs, decreased osteogenesis, and enhanced adipogenesis, as well as optn-/ - mice. Importantly, restoring Optn by transplanting wild-type MSCs to optn-/ - mice or infecting optn-/ - mice with Optn-containing lentivirus rescued bone loss. The introduction of a loss-of-function mutant of OptnK193R failed to reestablish a bone-fat balance. We further identified FABP3 (fatty acid binding protein 3, muscle and heart) as a novel selective autophagy substrate of OPTN. FABP3 promoted adipogenesis and inhibited osteogenesis of MSCs. Knockdown of FABP3 alleviated bone loss in optn-/ - mice and aged mice. Our study revealed that reduced OPTN expression during aging might lead to OP due to a lack of FABP3 degradation via selective autophagy. FABP3 accumulation impaired osteogenesis of MSCs, leading to the occurrence of OP. Thus, reactivating OPTN or inhibiting FABP3 would open a new avenue to treat senile OP.Abbreviations: ADIPOQ: adiponectin, C1Q and collagen domain containing; ALPL: alkaline phosphatase, liver/bone/kidney; BGLAP/OC/osteocalcin: bone gamma carboxyglutamate protein; BFR/BS: bone formation rate/bone surface; CALCOCO2/NDP52: calcium binding and coiled-coil domain 2; CDKN1A/p21: cyclin-dependent kinase inhibitor 1A; CDKN2A/p16: cyclin dependent kinase inhibitor 2A; CDKN2B/p15: cyclin dependent kinase inhibitor 2B; CEBPA: CCAAT/enhancer binding protein (C/EBP), alpha; COL1A1: collagen, type I, alpha 1; Ct. BV/TV: cortical bone volume fraction; Ct. Th: cortical thickness; Es. Pm: endocortical perimeter; FABP4/Ap2: fatty acid binding protein 4, adipocyte; H2AX: H2A.X variant histone; HE: hematoxylin and eosin; MAP1LC3B: microtubule-associated protein 1 light chain 3 beta; MAR: mineral apposition rate; MSCs: bone marrow mesenchymal stem cells; NBR1: NBR1, autophagy cargo receptor; OP: osteoporosis; OPTN: optineurin; PDB: Paget disease of bone; PPARG: peroxisome proliferator activated receptor gamma; Ps. Pm: periosteal perimeter; qRT-PCR: quantitative real-time PCR; γH2AX: Phosphorylation of the Serine residue of H2AX; ROS: reactive oxygen species; RUNX2: runt related transcription factor 2; SA-GLB1: senescence-associated (SA)-GLB1 (galactosidase, beta 1); SP7/Osx/Osterix: Sp7 transcription factor 7; SQSTM1/p62: sequestosome 1; TAX1BP1: Tax1 (human T cell leukemia virus type I) binding protein 1; Tb. BV/TV: trabecular bone volume fraction; Tb. N: trabecular number; Tb. Sp: trabecular separation; Tb. Th: trabecular thickness; µCT: micro computed tomography.

[Effect of Phosphorus Addition on N2O Emissions from Rice-Rapeseed Rotation Soils].

The red soils in southern China are generally classified as phosphorus-deficient, and therefore planting crops in these regions usually requires high applications of phosphate fertilizer. However, the effect of phosphorus addition on N2O emissions in rice-rapeseed rotation soils is not clear. We carried out an incubation experiment with the rice-rapeseed rotation soil from Qianjiang and Xianning to explore the effect of different concentrations of phosphorus (0, 15, and 30 mg·kg-1) and different concentrations of nitrogen (0 and 100 mg·kg-1) on N2O emission. Studies have shown that the addition of phosphorus has a significant effect on soil N2O emissions, but the pathways of impact are varied:in the case of low nitrogen soil, the addition of phosphorus promotes the fixation of nitrogen in the soil by microorganisms and thus reduces N2O emissions; in case of sufficient nitrogen content in soil, adding less phosphorus promotes the activity of nitrifies and thereby promotes the emission of N2O, while adding more phosphorus also promotes fixation by microorganisms in the soil; when there is a high content of available phosphorus in the soil, whether the nitrogen is sufficient or not, the addition of phosphorus will inhibit the emission of N2O.

[Effects of UV-B Radiation on Soil Carbon and Nitrogen Transformation under Different Soil Moisture Contents from Two Paddy Fields].

Carbon and nitrogen in soils play an important role in the global carbon and nitrogen cycle. The enhancement of ultraviolet radiation (predominantly UV-B) resulting from the depletion of stratospheric ozone has raised significant concern. The effects of UV-B radiation on soil carbon and nitrogen transformation is connected directly to the physical and chemical properties of the soil. In order to clearly understand the effects of soil moisture on UV-B radiation, this study collected soil samples from two paddy fields with different levels of organic matter in a subtropical region of China. The response of the total organic carbon (TOC), dissolved organic carbon (DOC), ammonia nitrogen (NH4+ -N), nitrate nitrogen (NO3- -N) and cumulative net nitrogen mineralization to UV-B radiation under three different moisture gradients (W1=25%, W2=50%, and W3=100%) were monitored in laboratory for 120 h. After this period, the results were compared with a control treatment (CK) and it was found that:the TOC content had significantly decreased under UV-B radiation (p<0.05). From low to high moisture content (W1, W2 and W3), the TOC decreased by 9.9%, 4.5% and 6.3%, respectively for soil with low organic matter (L), and by 10.9%, 5.6% and 6.3%, respectively for soil with high organic matter (H), under UV-B radiation. However, UV-B radiation was found to enhance the DOC content in the soil compared with the CK. Furthermore, the DOC for soil moisture contents under 100% (W3) was higher than for other moisture contents (W1, W2). The measured DOC increased by 21.5% (W1), 9.4% (W2), and 26.3% (W3) for soil with L. In addition, the measured DOC increased by 26.7% (W1), 14.2% (W2) and 33.8% (W3) for soil with H under UV-B radiation after 120 h. Compared with control treatment (CK), UV-B radiation decreased the NH4+ -N content significantly, but there was an increased NO3- -N content. The decrease of the NH4+ -N content was largest for W3 and smallest for W1. The increase in NO3- -N content was largest for W2 and smallest for W1 for the two soil samples under UV-B radiation. UV-B radiation demonstrated an obvious effect on the cumulative net nitrogen mineralization (p<0.05) after 24 h compared with the CK and the effect of different soil moisture treatment was also significant (p<0.05). Overall, light degradation played a major role in the stabilization of soil organic matter, soil moisture, and UV-B radiation could accelerate the loss of soil organic carbon and has a major impact on the transformation of mineral nitrogen in the soil. Therefore, in agricultural production systems, completely bare surfaces should be avoided. For example, paddy rice-upland crop rotation systems could reduce the use of fallow periods.

Degradation of the Separase-cleaved Rec8, a Meiotic Cohesin Subunit, by the N-end Rule Pathway.

The Ate1 arginyltransferase (R-transferase) is a component of the N-end rule pathway, which recognizes proteins containing N-terminal degradation signals called N-degrons, polyubiquitylates these proteins, and thereby causes their degradation by the proteasome. Ate1 arginylates N-terminal Asp, Glu, or (oxidized) Cys. The resulting N-terminal Arg is recognized by ubiquitin ligases of the N-end rule pathway. In the yeastSaccharomyces cerevisiae, the separase-mediated cleavage of the Scc1/Rad21/Mcd1 cohesin subunit generates a C-terminal fragment that bears N-terminal Arg and is destroyed by the N-end rule pathway without a requirement for arginylation. In contrast, the separase-mediated cleavage of Rec8, the mammalian meiotic cohesin subunit, yields a fragment bearing N-terminal Glu, a substrate of the Ate1 R-transferase. Here we constructed and used a germ cell-confinedAte1(-/-)mouse strain to analyze the separase-generated C-terminal fragment of Rec8. We show that this fragment is a short-lived N-end rule substrate, that its degradation requires N-terminal arginylation, and that maleAte1(-/-)mice are nearly infertile, due to massive apoptotic death ofAte1(-/-)spermatocytes during the metaphase of meiosis I. These effects ofAte1ablation are inferred to be caused, at least in part, by the failure to destroy the C-terminal fragment of Rec8 in the absence of N-terminal arginylation.

Recurrent gain-of-function USP8 mutations in Cushing's disease.

Cushing's disease, also known as adrenocorticotropic hormone (ACTH)-secreting pituitary adenomas (PAs) that cause excess cortisol production, accounts for up to 85% of corticotrophin-dependent Cushing's syndrome cases. However, the genetic alterations in this disease are unclear. Here, we performed whole-exome sequencing of DNA derived from 12 ACTH-secreting PAs and matched blood samples, which revealed three types of somatic mutations in a candidate gene, USP8 (encoding ubiquitin-specific protease 8), exclusively in exon 14 in 8 of 12 ACTH-secreting PAs. We further evaluated somatic USP8 mutations in additional 258 PAs by Sanger sequencing. Targeted sequencing further identified a total of 17 types of USP8 variants in 67 of 108 ACTH-secreting PAs (62.04%). However, none of these mutations was detected in other types of PAs (n = 150). These mutations aggregate within the 14-3-3 binding motif of USP8 and disrupt the interaction between USP8 and 14-3-3 protein, resulting in an elevated capacity to protect EGFR from lysosomal degradation. Accordingly, PAs with mutated USP8 display a higher incidence of EGFR expression, elevated EGFR protein abundance and mRNA expression levels of POMC, which encodes the precursor of ACTH. PAs with mutated USP8 are significantly smaller in size and have higher ACTH production than wild-type PAs. In surgically resected primary USP8-mutated tumor cells, USP8 knockdown or blocking EGFR effectively attenuates ACTH secretion. Taken together, somatic gain-of-function USP8 mutations are common and contribute to ACTH overproduction in Cushing's disease. Inhibition of USP8 or EGFR is promising for treating USP8-mutated corticotrophin adenoma. Our study highlights the potentially functional mutated gene in Cushing's disease and provides insights into the therapeutics of this disease.

[Effect of carbon substrate concentration on N2, N2O, NO, CO2, and CH4 emissions from a paddy soil in anaerobic condition].

Understanding the effects of carbon and nitrogen substrates concentrations on the emissions of denitrification gases including nitrogen (N2) , nitrous oxide (N2O) and nitric oxide (NO), carbon dioxide (CO2) and methane (CH4) from anaerobic paddy soils is believed to be helpful for development of greenhouse gas mitigation strategies. Moreover, understanding the quantitative dependence of denitrification products compositions on carbon substrate concentration could provide some key parameters or parameterization scheme for developing process-oriented model(s) of nitrogen transformation. Using a silt loam soil collected from a paddy field, we investigated the influence of carbon substrate concentration on the emissions of the denitrification gases, CO2 and CH4 from anaerobically incubated soils by setting two treatments: control (CK) with initial soil nitrate and dissolved organic carbon (DOC) concentrations of ~ 50 mg.kg-1 and -28 mg kg-1 , respectively; and DOC added (C + ) with initial soil nitrate and DOC concentrations of ~50 mg.kg-1 and ~300 mg.kg-1 , respectively. The emissions of denitrification gases, CO2 and CH4, as well as concentrations of carbon and nitrogen substrates for each treatment were dynamically measured, using the gas-flow-soil-core technique and a paralleling substrate monitoring system. The results showed that CH4 emission was not observed in CK treatment while observed in C treatment. Aggregate emission of greenhouse gases for C + treatment was significantly higher comparing with the CK treatment (P <0. 01). The mass fractions of NO, N20 and N2 emissions in total nitrogen gases emissions were approximately 9% , 35% and 56% for CK treatment, respectively; and approximately 31% , 50% and 19% for C+ treatment, respectively, with significant differences between these two treatments (P < 0.01). The results indicated that carbon substrate concentrations can significantly change the composition of nitrogen gas emissions. The results also implicated that organic fertilizer should not be applied to nitrate-rich paddy soils prior to or during flooding so as to mitigate greenhouse gases emissions.

[Characteristics of soil organic carbon mineralization at different temperatures in paddy soils under long-term fertilization].

Dynamics of soil organic carbon mineralization affected by long-term fertilizations and temperature in relation to different soil carbon fractions were investigated in paddy soils. Soil samples were collected from the plough layer of 3 long-term national experimental sites in Xinhua, Ningxiang and Taojiang counties of Hunan Province. Mineralization of soil organic C was estimated by 33-day aerobic incubation at different temperatures of 10, 20 and 30 degrees C. The results showed that the rates of CO2 production were higher during the earlier phase (0-13 d) in all treatments, and then decreased according to a logarithm function. Higher incubation temperature strengthened C mineralization in the different treatments. The quantities of cumulative CO2 production in NPK with manure or straw treatments were greater than in inorganic fertilizers treatments. The Q10 values in the different soil treatments ranged from 1.01-1.53. There were significantly positive correlations between the Q10 values and soil total organic carbon (TOC), easy oxidation organic carbon (EOOC), humic acid carbon (C(HA)), fulvic acid carbon (CFA). The cumulative amount of mineralized C was significantly positively correlated with microbial biomass carbon (MBC) at 10 and 20 degrees C, but not significantly at 30 degrees C. Significant correlations were found between the cumulative amount of mineralized C and different soil carbon fractions and C(HA)/C(FA). The correlations of differ- ent soil carbon fractions with the ratio of cumulative mineralized C to TOC were negatively correlated at 10 degrees C, but not significantly at 20 and 30 degrees C. These results suggested that the application of NPK with manure or straw would be helpful to increase the sequestration of C in paddy soils and reduce its contribution of CO2 release in the atmosphere.

[Influence of preoperative chemoradiotherapy on pulmonary function and postoperative pulmonary complication in esophageal cancer patients].

OBJECTIVE: To study the influence of preoperative chemoradiotherapy (CRT) on pulmonary function and postoperative pulmonary complications in esophageal cancer patients. METHODS: Pulmonary function and postoperative pulmonary complications of 63 esophageal cancer patients undergoing preoperative CRT and operation in Cancer Center of Sun Yat-sen University between 2002 and 2013 were collected retrospectively. The influence of preoperative CRT on pulmonary functional indexes and postoperative pulmonary complications were analyzed. RESULTS: After preoperative CRT, DLco% decreased significantly (83.7±17.7 vs. 96.4±17.8, P<0.01), while no obvious changes in other indexes were found. Postoperative pulmonary complication rate was 34.9% (22/63), including 19 cases of pneumonia and 3 cases of acute pulmonary injury/acute respiratory distress syndrome. Differences in postoperative pulmonary complication rates were not statistically significant between patients with DLco% <80 and those with DLco% ≥80 patients (29.7% vs. 41.7%, P>0.05), and between patients with DLco% decline ≥15% and those with DLco% decline <15% patients (31.6% vs. 37.8%, P>0.05). CONCLUSION: Preoperative CRT can damage the diffusion function but not ventilation function of esophageal cancer patients, and does not increase the postoperative pulmonary complication rate.

High expression of transient potential receptor C6 correlated with poor prognosis in patients with esophageal squamous cell carcinoma.

TRPC6 plays a crucial role in the tumor progression of various cancers. The relation between the expression of TRPC6 and clinical prognosis has not been studied yet. Our study was to elucidate the role of TRPC6 in predicting outcomes of patients with esophageal squamous cell carcinoma (ESCC). Fresh frozen samples were collected immediately from 172 patients with ESCC after surgical resection from 2003 to 2008 at Sun Yat-sen University Cancer Center, including 45 pairs of tumor tissues and nontumor tissues. TRPC6 expression was measured by quantitative real-time PCR and Western blotting analyses. TRPC6 mRNA and protein were up-regulated in ESCC tissues when compared with the paired nontumor tissues. High expression of TRPC6 mRNA was associated with the higher pT status (P = 0.016) and pathological staging (P = 0.040). The 5-year disease-specific survival in the high expression of TRPC6 mRNA group (>188.98, n = 81) is poorer than that in low-level expression group (≤188.98, n = 91) (42.1 vs. 62.7 %, P = 0.004). Stratified analysis according to the pathological stage revealed its discernibility on DSS was only pronounced in patients with pStage III (P = 0.015). Cox multivariate analysis revealed that pN category (P < 0.001; Relative risk, 2.897, 95 % CI 1.830-4.585) and the expression of TRPC6 mRNA (P = 0.006; Relative risk, 1.863, 95 % CI 1.196-2.902) were independent prognostic factors. TRPC6 mRNA overexpression correlated with poor prognosis in patients with ESCC and might serve as a novel prognostic biomarker for resected ESCC patients in advanced stage.

[Response of black soil organic carbon, nitrogen and its availability to longterm fertilization].

Based on the long-term fertilization experiments, effects of various fertilization practices on the soil organic carbon (SOC) and total nitrogen (TN) in the surface (0-20 cm) and subsurface (20-40 cm) black soil in northeast China were studied. Results showed that, compared with the CK, long-term application of organic manure, especially the combination of mineral fertilizers and organic manure significantly increased the organic SOC and TN in the surface soil. Application of mineral fertilizers plus organic manure with conventional (NPM) and high application (N2P2M2) rate increased SOC significantly by 24. 6% and 25.1% , and TN by 29.5% and 32.8%, respectively. However, there was no significant difference among the treatments for SOC and TN at the subsurface. Compared with the CK (CKh), mineral fertilizer plus organic manure (NPM and N2P2M2) did not only increase the soil microbial biomass carbon (SMBC) and nitrogen (SMBN) , dissolved organic carbon (DOC) and nitrogen (DN), but also significantly increased the ratio of SMBC and DOC to SOC, SMBN and TN to TN. Application of the NPM and N2P2M2 increased the value of SMBC/SOC by 0.36 to 0.59 and SMBN/TN by 1.21 to 1.95 percentage points, respectively. The value of DOC/SOC and DN/TN ranged from 0.53% to 0.72% and 1.41% to 1.78%, respectively. This result indicated that SMBC, SMBN, DOC, DN and SMBC/ SOC, SMBN/TN, DOC/SOC, DN/TN were more sensitive than SOC and TN to long-term fertilization in the soil profile, and were better indicators for the impact of long-term fertilization soil fertility. The concluded that the application of manure especially manure plus mineral fertilizers can increase soil nutrients activity in the surface and subsurface black soil, acting as a helpful practice to improve soil fertility and the ability of nutrient supply, while it may cause potential environment pollution on carbon and nitrogen loss in the agroecosystem.

[Influence of land use structure on nitrogen output in the watershed of suburban agriculture regions].

This study was conducted in Jintuo watershed of Changsha City, Hunan Province, in the suburban agriculture regions, by selecting 8 sub-basins to examine the effect of land use on watershed runoff nitrogen output. The land use map of watershed was interpreted from Spot-5 image of 2009, and by using hydraulic analysis function and spatial analysis extensions of ArcGIS 9.3, the catchment areas were delineated from DEM. Water sampling was carried out from Dec. 2009 to Nov. 2010, and the relationships between different types of nitrogen export and land use were analyzed. The results showed that nitrogen pollution in the watershed was extremely serious. There was a distinct seasonal variation in the following order: WI > SP > SU approximately = FA in the output concentration of total nitrogen and ammonium nitrogen which was not observed in the nitrate output. Moreover, land use was a dominant factor that determined the export of nitrogen, and especially a significant correlation was figured out between the nitrate output concentration and the land use structure. Forest and water body had a negative impact on nitrate output concentration while dry land, paddy field and habitation road had a positive effect. However, the effects varied with time. Dry land had the most significant important effect on nitrate output concentration in winter and fall, but in spring and summer was the forest land. The correlation between land use structure and the output concentration of total nitrogen and ammonium nitrogen was not found. The resident number, pig number and fertilization also had a major impact on nitrogen output quantity.

[Effects of enhanced UV-B radiation on terrestrial ecosystem carbon cycle: a review].

As one of the most important phenomena of global climate change, the enhancement of ultraviolet-B radiation (UV-B, 280-320 nm) could have critical impact on the carbon cycle in terrestrial ecosystem. Through the impacts on plant photosynthesis, litter decomposition, and soil respiration, the enhanced UV-B radiation can affect the carbon input, turnover, and output of terrestrial ecosystem. Other climatic factors (ambient CO2 concentration, air temperature, and precipitation) may promote or mitigate the impact of enhanced UV-B radiation on terrestrial ecosystem carbon cycle. This paper introduced the background of UV-B radiation enhancement, reviewed the impacts of enhanced UV-B radiation and its interactions with other climatic factors on terrestrial ecosystem carbon cycle, summarized the existing problems in related researches, and discussed the priorities and directions of future researches.

pVHL mediates K63-linked ubiquitination of nCLU.

pVHL, product of von Hippel-Lindau (VHL) tumor suppressor gene, functions as the substrate recognition component of an E3-ubiquitin ligase that targets proteins for ubiquitination and proteasomal degradation. Hypoxia-inducible factor α (HIFα) is the well-known substrate of pVHL. Besides HIFα, pVHL also binds to many other proteins and has multiple functions. In this manuscript, we report that the nuclear clusterin (nCLU) is a target of pVHL. We found that pVHL had a direct interaction with nCLU. nCLU bound to pVHL at pVHL's ß domain, the site for recognition of substrate, indicating that nCLU might be a substrate of pVHL. Interestingly, pVHL bound to nCLU but did not lead to nCLU destruction. Further studies indicated that pVHL mediated K63-linked ubiquitination of nCLU and promoted nCLU nuclear translocation. In summary, our results disclose a novel function of pVHL that mediates K63-linked ubiquitination and identify nCLU as a new target of pVHL.

[Effects of long-term fertilization on paddy soils organic nitrogen, microbial biomass, and microbial functional diversity].

Soil samples were collected from the plow layers at two long-term experiment sites in Xinhua and Ningxiang counties of Hunan Province, China to study the effects of long-term fertilization on organic nitrogen, microbial biomass, and microbial functional diversity of paddy soils. Long-term fertilization showed great effects on the soil N content. Compared with CK, treatments NPK plus manure or straw increased the contents of soil total acid-hydrolysable N and its fractions amino sugar N, amino acid N, and ammonium N. Treatment NPK had no significant effects on soil microbial biomass C and N, but treatments NPK plus manure increased the contents of soil microbial biomass C and N significantly. BIOLOG test showed that treatments NPK plus manure enhanced the carbon utilization efficiency of soil microbes, and improved the functional diversity of soil microbial communities, compared with CK. Long-term different fertilizer treatments resulted in the differences of carbon substrate utilization patterns of soil microbial communities.

[Effect of long-term fertilization on the diversity of nitrite reductase genes (nirK and nirS) in paddy soil].

In order to investigate the effects of long-term application of nitrogen fertilizer on soil denitrifying communities, the diversities of nir genes (nirK and nirS) were studied using molecular approaches in the long-term paddy field experiment (started in 1990) located in Taoyuan. Analysis of clone sequences indicated that the nirK fragments from paddy soil showed close similarity (90.7%) to the nirK sequences registered in GenBank database, but were not related to any known strain. Whereas, most of the airS clones showed low similarity (74.7%) to the nirS gene fragments registered in GenBank. The Chao1 estimates showed that the diversity of nirK gene 13) OTUs] than in N treatment [(49 +/- 9) OTUs], but the difference was not significant. However, application of nitrogen fertilizer resulted in significant difference of nirS-community compared to CK. Nitrogen fertilizer had obvious effect on tbe community structure of nirK-denitrifiers (p < 0.022), but the nirS-containing community was not affected. Based on phylogenetic analysis, nirK clones grouped into three clusters with aggregations of some OTUs cloned from N treatment. Although nirS clones grouped into four clusters, the majority of the clones were attributed in one cluster. The results suggested that application of nitrogen fertilizer had a greater influence on the diversity of nirS-containing bacterial community than that of the nirK. However, the community structure of nirK-containing denitrifiers was more sensitive to nitrogen fertilization than that of the mrS.