[Application of Modified DTT Methods to Reduce the Interference of Daratumumab on Serological Detection].

OBJECTIVE: To modified the classic dithiothreitol (DTT) method for treating red blood cells (RBCs) in Technical Manual of American Association of Blood Banks（AABB） and evaluate its application value in pre-transfusion examination of patients treated with daratumumab. METHODS: The classic 0.2 mol/L DTT method was improved in terms of PBS, DTT concentration, donor RBCs concentration (suspended/packed) and sample processing time. The modified DTT methods and AABB classic DTT method were applied to the blood matching tests of 12 multiple myeloma patients treated with daratumumab. The effect of treating panel RBCs with modified DTT methods on the detection of other irregular antibodies was evaluated by using antiserum and antibody reagents with known antibody properties. RESULTS: Two modified DTT methods were established (method 1: changed the concentration of DTT to 0.01 mol/L; method 2: changed the concentration of DTT to 0.02 mol/L and replaced the packed RBCs with 3% RBCs suspension). The optimal treatment time was 35 min for the modified DTT methods. At this time, the pan-agglutination caused by daratumumab was eliminated, but the detection of antibodies such as anti-E, anti-JKa, anti-M were not affected, and the titer of anti-K antibodies was only slightly decreased. CONCLUSION: The modified DTT methods were effective, which can eliminate the interference of daratumumab while retaining the activity of the Kell blood group system, and can replace the current classic DTT method in AABB Technical Manual.

[Annual Nitrogen Removal Efficiency and Change in Abundance of Nitrogen Cycling Microorganisms in Swine Wastewater Treated by Crop Straw Materials].

Based on previous research, using straw material to treat swine wastewater can effectively reduce the concentration of nitrogen (N); however, the annual N-removal efficiency and change in the abundance of N-cycling functional genes remain unclear. In this study, four treatments (wheat straw, rice straw, corn stalk, and CK) were set up, with the aim of studying the annual N-removal efficiency and change in the abundance of functional genes. Our results showed that:① the total nitrogen (TN) removal and NH4+-N removal efficiency were the best in the first six months and were significantly reduced in the following six months. In addition, the TN removal and NH4+-N efficiency in straw and wheat straw were better than those in corn straw. The TN-removal efficiency in straw and wheat straw were 32.81%±11.34% and 32.99%±9.60%, respectively. The NH4+-N removal efficiency in straw and wheat straw were 35.3%±13.23% and 34.97%±12.00%, respectively. ② The abundance of N-cycling functional genes significantly increased by the addition of straw materials, compared with that of the CK (P<0.05). The average abundances of nirK, nirS, and hzsB were 6.45×109 copies·L-1, 6.18×109 copies·L-1, and 2.31×109 copies·L-1, respectively. The average abundances of ammonia-oxidizing archaea (AOA) and ammonia-oxidizing bacteria (AOB) were 6.12×1010 copies·L-1 and 4.93×109 copies·L-1, respectively. The average hzsB gene abundance was 2.31×109 copies·L-1. The average abundance of 16S rRNA in the treatment was 8.90×1010 copies·L-1. The abundances of hzsB and nirS genes in the straw and wheat straw were higher than those in the other treatment, indicating that the activities of anaerobic ammonia oxidation and denitrifying microorganisms were significantly increased by the addition of straw and wheat straw (P<0.05). In addition, the abundance of AOA and AOB genes were increased in wheat straw, suggesting that wheat straw could promote nitrification. The results provided data supporting the molecular mechanism of nitrogen removal in swine wastewater treatment with straw materials.

Farnesoid X receptor (FXR) inhibits coagulation process via inducing hepatic antithrombin III expression in mice.

Farnesoid X receptor (FXR) has been identified as an inhibitor of platelet function and an inducer of fibrinogen protein complex. However, the regulatory mechanism of FXR in hemostatic system remains incompletely understood. In this study, we aimed to investigate the functions of FXR in regulating antithrombin III (AT III). C57BL/6 mice and FXR knockout (FXR KO) mice were treated with or without GW4064 (30 mg/kg per day). FXR activation significantly prolonged prothrombin time (PT) and activated partial thromboplastin time (APTT), lowered activity of activated factor X (FXa) and concentrations of thrombin-antithrombin complex (TAT) and activated factor II (FIIa), and increased level of AT III, whereas all of these effects were markedly reversed in FXR KO mice. In vivo, hepatic AT III mRNA and protein expression levels were up-regulated in wild-type mice after FXR activation, but down-regulated in FXR KO mice. In vitro study showed that FXR activation induced, while FXR knockdown inhibited, AT III expression in mouse primary hepatocytes. The luciferase assay and ChIP assay revealed that FXR can bind to the promoter region of AT III gene where FXR activation increased AT III transcription. These results suggest FXR activation inhibits coagulation process via inducing hepatic AT III expression in mice. The present study reveals a new role of FXR in hemostatic homeostasis and indicates that FXR might act as a potential therapeutic target for diseases related to hypercoagulation.

[Role of prostaglandin E2 in the modulation of renal water transport].

Prostaglandin E2 (PGE2), a bioactive lipid mediator, is one of the most important locally acting factors involved in a variety of physiological and pathophysiological processes. PGE2 binds with four EP receptors (EP1-4) to activate G protein-coupled receptor signaling responses. Recent functional and molecular studies have revealed that PGE2 plays an essential role in regulation of renal fluid transport via a variety of mechanisms. The water balance mainly depends on the regulation of aquaporin-2 (AQP2) by arginine vasopressin (AVP) in renal collecting duct principal cells. In recent years, increasing evidence suggests that PGE2 plays an important role in renal water reabsorption in the collecting ducts. In this paper, we reviewed the role of PGE2 and its receptors in the regulation of water reabsorption in the kidney, which may provide a new therapeutic strategy for many diseases especially nephrogenic diabetes insipidus.

[Nitrogen and Phosphorus Removal in Surface Flow Constructed Wetland Planted with Myriophyllum elatinoides Treating Swine Wastewater in Subtropical Central China].

The loss of nitrogen (N) and phosphorus (P) from aquaculture has caused eutrophication of freshwater systems. Here, surface flow constructed wetland (SFCW) planted with Myriophyllum elatinoides were used to treat swine wastewater from a medium-sized hoggery in subtropical Central China. Inflow concentrations of NH4+-N, TN, TP, and COD ranged from 535.4 to 591.09, 682.09 to 766.96, 57.73 to 82.29, and 918.4 to 1940.43 mg·L-1, respectively. The mean removal efficiencies of NH4+-N, TN, TP, and COD were 97.4%, 97.1%, 91.0%, and 90.2%, respectively, and CW1 had the largest contributions of 37.3%, 38.4%, 43.3%, and 27.4%, respectively. Plant N and P uptake ranged 23.87-79.96 g·m-2 and 5.34-18.98 g·m-2, accounting for 19.1% and 20.2% of removal, respectively. Sediment N and P accumulation ranged 19.17-56.62 g·m-2 and 10.59-26.62 g·m-2, accounting for 19.8% and 61.7% of removal, respectively. Multiple linear regression showed that environmental factors explained 79.9% of the N removal and 70.1% of the P removal; DO was the main factor affecting N removal, and sediment adsorption was the key process in P removal. These results show that M. elatinoides constructed wetland can efficiently treat swine wastewater, thereby reduce the discharge of pollutants downstream.

The roles and interaction of FXR and PPARs in the pathogenesis of nonalcoholic fatty liver disease.

BACKGROUND AND STUDY AIMS: To identify the roles and interaction of farnesoid X receptor (FXR) and peroxisome proliferator activated receptors (PPARs) in Non-alcoholic fatty liver disease (NAFLD) pathogenesis. MATERIAL AND METHODS: 16 C57/BL male FXR knockout (KO) mice and sex- and age-matched C57/BL wild type mice were received either standard rodent chow or high-fat and sucrose diet (Blank control, NAFLD, FXR KO and FXR KO NAFLD) for 8 weeks. After that, all mice were sacrificed. Liver tissues and blood samples were collected for laboratory and RT-PCR examination. RESULTS: NAFLD, FXR KO and FXR KO NAFLD mouse models were successful established. Compared with blank control, FXR and PPAR-α mRNA expression decreased significantly (P < 0.05), PPAR-ß expression increased slightly (P > 0.05), PPAR-γ expression increased significantly in NAFLD (P < 0.05). Slight increased PPAR-α mRNA expression (P > 0.05) and markedly decreased PPAR-ß and PPAR-γ expression (P < 0.05) were found in FXR KO. Compared with FXR KO group, there was a slight increase in PPAR-αand PPAR-ßmRNA expression (P > 0.05) and significant increase in PPAR-γ expression (P < 0.05) in FXR KO NAFLD group. Comparison with NAFLD, PPAR-α mRNA expression increased slightly (P > 0.05), PPAR-ß and PPAR-γ expression decreased significantly (P < 0.05) in FXR KO NAFLD. CONCLUSION: FXR and PPARs interaction may play important roles in NAFLD pathogenesis.

[Treatment of Highly Concentrated Swine Wastewater and Its Degradation Processes Using Three Matrix Materials].

The direct discharge of wastewater from pig farms can restrict wetland plant growth meaning that constructed wetlands (CWs) have generally low treatment efficiency. The treatment of farming wastewater using pre-ecological treatment technologies can be used to ensure that effluent concentrations reach the tolerable limits of wetland plants. This study focused on the efficient use of crop straw for reducing the pollution load of swine wastewater. Using field-scale pilot tests, wheat straw, straw, and corn stalk were used as test biological matrix pool fillers to treat the farming wastewater. Continuous intake of wastewater and a hydraulic retention time of 7 days was adopted. When the average concentrations of COD, TN, NH4+-N, NO3--N, and TP in the influent were 1652.83 mg·L-1, 371.31 mg·L-1, 303.51 mg·L-1, 0.67 mg·L-1, and 65.22 mg·L-1, respectively, wheat straw had the greatest removal effect on COD, TN, and TP, achieving a removal rate of 32.1%, 40.9%, and 33.3%, respectively. The removal efficiency of straw on NH4+-N was highest, reaching 43.4%. After 180 days, the lignin, cellulose, and hemicellulose of the three matrix materials were not completely decomposed. The degradation rate of lignin was lower than for cellulose and hemicellulose; the degradation of lignin and cellulose in the straw was fastest; and the degradation hemicellulose in wheat straw was fastest. The results show that wheat straw and straw offer a higher efficiency treatment for swine wastewater than corn stalk, and the suggested replacement cycle period is five months. These results provide initial data to support the application of biological matrix materials in the treatment of swine wastewater.

Diagnostic Performance of Intestinal Fusobacterium nucleatum in Colorectal Cancer: A Meta-Analysis.

BACKGROUND: Increasing evidence has supported the link of intestinal Fusobacterium nucleatum infection to colorectal cancer (CRC). However, the value of F. nucleatum as a biomarker in CRC detection has not been fully defined. In order to reduce the random error and bias of individual research, this meta-analysis aimed to evaluate the diagnostic performance of intestinal F. nucleatum in CRC patients and provide evidence-based data to clinical practice. METHODS: An article search was performed from PubMed, Embase, Cochrane Library, and Web of Science databases up to December 2017, using the following key words: "Fusobacterium nucleatum", "Fusobacterium spp.", "Fn", "colorectal cancer(s)", "colorectal carcinoma(s)", "colorectal neoplasm(s)", and "colorectal tumor(s)". Articles on relationships between F. nucleatum and CRC were selected according to the preestablished inclusion and exclusion criteria. This meta-analysis was performed using STATA 12.0 software, which included mapping of forest plots, heterogeneity tests, meta-regression, subgroup analysis, sensitivity analysis, and publication bias. The sensitivity, specificity, positive likelihood ratio (LR), negative LR, diagnostic odds ratio (DOR), and their corresponding 95% confidence interval (CI) of each eligible study were summarized. RESULTS: Finally, data for 1198 participants (629 CRC and 569 healthy controls) in 10 controlled studies from seven articles were included. The summary receiver operator characteristic curve was mapped. The diagnostic performance of intestinal F. nucleatum infection on CRC was as follows: the area under the curve: 0.86 (95% CI: 0.83-0.89), the pooled sensitivity: 0.81 (95% CI: 0.64-0.91), specificity: 0.77 (95% CI: 0.59-0.89), and DOR: 14.00 (95% CI: 9.00-22.00). CONCLUSION: Intestinal F. nucleatum is a valuable marker for CRC diagnosis.

Fatty liver mediated by peroxisome proliferator-activated receptor-α DNA methylation can be reversed by a methylation inhibitor and curcumin.

OBJECTIVE: Our studies in vitro and in vivo aimed to investigate the influence of DNA methylation of peroxisome proliferator activated receptor-α (PPAR-α) gene in non-alcoholic fatty liver disease (NAFLD) pathogenesis and to observe whether the DNA methylation inhibitor 5-Aza-2'-deoxycytidine (5-Aza-CdR) and the herbal medicine curcumin might reverse the effect both in vivo and in vitro. METHODS: Steatotic hepatocyte model of cell lines and NAFLD rat models were established following protocols documented in previous studies. Subsequently, the models received 5-Aza-CdR and curcumin treatment. Morphological, histological and laboratory variables in each group were determined by routine methods, including PPAR-α mRNA expression by polymerase chain reaction (PCR), PPAR-α protein expression by Western blot and DNA methylation by pyrosequencing. RESULTS: The steatotic hepatocyte model and NAFLD rat model were completely established. The expressions of PPAR-α mRNA and protein were significantly lower in the steatotic hepatocyte and NAFLD rat model groups than in the controls (P < 0.05). The mean DNA methylation levels of the PPAR-α gene were significantly higher in the two steatotic model groups than in the controls, especially at several CpG sites (P < 0.05). 5-Aza-CdR and curcumin treatment significantly reversed the DNA methylation levels, increased PPAR-α mRNA and protein expression, and improved lipid accumulation in the two steatotic models (P < 0.05). CONCLUSIONS: DNA methylation at the PPAR-α gene is involved in the pathogenesis of NAFLD and is possibly reversible by 5-Aza-CdR and curcumin. Curcumin may be a promising candidate for NAFLD therapy.

Evolution of correlation between Helicobacter pylori infection and autoimmune liver disease.

The present study was planned to evaluate correlation between Helicobacter pylori (HP) infection and autoimmune liver disease (AILD). A total of 60 patients diagnosed with AILD in Affiliated Hospital of Binzhou Medical College were continuously enrolled in the present study. HP infection was detected by 13C-urea breath test. The levels of anti-myeloperoxidase were tested by ELISA. The positive rate of anti-nuclear antibody (ANA), anti-mitochondrial antibody (AMA), anti-smooth muscle antibody (SMA) and anti-neutrophil cytoplasm antibody (ANCA) were tested by indirect immunofluorescence. The positive rates of anti-mitochondrial antibody (AMA-M2), anti-liver-kidney microsomal antibody (LKM-1), anti-liver cytoplasm antibody I (LC-1) and anti-soluble liver antigen/liver-pancreas antigen (SLA/LP) were tested by immunoblotting. Liver function indexes including alanine transaminase, aspartate transaminase, alkaline phosphatase and glutamyltransferase, were analyzed with a fully automatic biochemical analyzer. The levels of serum cytokine IFN-γ, interleukin-6 (IL-6), IL-10 and tumor necrosis factor-α (TNF-α) were tested by ELISA. A total of 37 patients (61.67%) were observed to be HP-positive. MPO-positive rate, positive rate of ANA, AMA, SMA and ANCA and positive rate of AMA-M2, LKM-1, LC-1 and SLA/LP in patients with positive HP infection were significantly higher than those of patients with negative HP infection. On the other hand, the levels of liver function indices did not showed any significant differences among HP-positive cases or HP-negative cases. However, the levels of IFN-γ, IL-6, IL-10 and TNF-α in patients with positive HP infection were significantly higher than those of patients with negative HP infection. In conclusion, the positive infection rate of HP infection in patients with AILD is high and is closely associated with various positive immune antibodies as well as cytokine levels.

[Characteristic of Abundances and Diversity of Carbon Dioxide Fixation Microbes in Paddy Soils].

To get a better understanding of the microbial autotrophic carbon sequestration potential of paddy fields and its mechanisms, soil incubation experiment was conducted for four representative paddy soils. The molecular biological methods[quantitative PCR (qPCR), clone library and terminal-restriction fragment length polymorphism (T-RFLP) technique] based on cbbL and cbbM genes encoding the key enzymes[ribulose-1,5-bisphosphate carboxylase/oxygenase (RubisCO)] of Calvin cycle were used to determine the abundance and diversity of autotrophic microbes. The results showed that, after 45 days of incubation, carbon dioxide fixation autotrophic microbial abundances were generally increased compared with those before incubation, and cbbL gene abundances were approximately three magnitudes higher than those of cbbM. Dominant microbial populations varied among the four paddy soils, and most of these OTUs were distantly related to known sequences, only part of them could be grouped into Proteobacteria and Actinobacteria. RDA analysis results showed that soil organic carbon (SOC), cation exchange capacity (CEC), pH, clay, silk and sand content had significant effects on the CO2 fixation microbial community. Consequently, the results of this study provide significant reference to understand the role of microorganisms in carbon cycle process. The results are helpful for providing a scientific basis for scientific management of paddy soil fertility and low carbon agriculture construction.

The role of ADIPOQ methylation in curcumin-administrated experimental nonalcoholic fatty liver disease.

OBJECTIVE: To investigate the role of adiponectin precursor (ADIPOQ) DNA methylation in the pathogenesis of nonalcoholic fatty liver disease (NAFLD) and the effect of curcumin on the development of NAFLD using rat models. METHODS: Male Sprague-Dawley rats were divided into the control, NAFLD and curcumin-treated groups. The genetic and epigenetic features of each rat were measured and recorded. Real-time polymerase chain reaction (PCR), Western blot and bisulfite sequencing PCR (BSP) were used to quantify the ADIPOQ mRNA and protein expressions, and DNA methylation status, respectively. RESULTS: Serum levels of alanine aminotransferase (ALT), triglyceride (TG), total cholesterol (TC) and fasting blood glucose in the NAFLD group were significantly increased compared with the control group. The genetic and epigenetic features were reversed after curcumin treatment. The ADIPOQ mRNA and protein expressions in the livers of the NAFLD rats was lower compared with the control and the curcumin-treated groups. ADIPOQ methylation rate in the NAFLD group was significantly higher than in the control group, which was declined slightly following curcumin treatment. A negative correlation was found between the degrees of DNA methylation and ADIPOQ mRNA expression. ALT, TC, TG and homeostatic model assessment insulin resistance index had a positive correlation with ADIPOQ DNA methylation, showing that curcumin might affect the gene expression involved in lipid and glucose metabolism by influencing ADIPOQ DNA methylation modifications, which contributed to alleviation of NAFLD. CONCLUSION: Altering the DNA methylation of ADIPOQ is one of the mechanisms by which curcumin executes its hepatoprotective function in NAFLD.

Expression and inhibitory role of TIMP-3 in hepatocellular carcinoma.

As a member of the tissue inhibitor of metallo-proteinases (TIMP) family, it has been reported that TIMP-3 is involved in human cancer development. However, the function of TIMP-3 in hepatocellular carcinoma (HCC) development is unclear. We aimed to determine the biological role of TIMP-3 in HCC by evaluating the effects of its methylation status and expression on HCC cell function. TIMP-3 expression in HCC tissues was visibly analyzed by immunohistochemistry. Methylation of the TIMP-3 promoter was evaluated by methylation-specific PCR. Effects of TIMP-3 on HCC cell growth, apoptosis, migration, and invasion were examined by transfecting the TIMP-3-expressing plasmid, pCMV6. TIMP-3 was expressed in non-tumorous live tissue, but silenced or downregulated in 60% of HCC cases (P<0.05). Reduced protein expression of TIMP-3 was associated with reduced tumor differentiation (P=0.003) and increased metastatic activity (P=0.005) in HCC cell lines. Promoter methylation contributed to the TIMP-3 inactivation. Overexpression of TIMP-3 in HCC cell lines suppressed cell proliferation, induced apoptosis, and inhibited migration and invasion in vitro. TIMP-3 expression is suppressed by promoter methylation in HCC. This inhibitory protein acts as a functional tumor suppressor by inhibiting HCC cell proliferation, invasion, and migration and by inducing apoptosis and cell cycle arrest at the G2/M phase.

Association of Fusobacterium nucleatum infection with colorectal cancer in Chinese patients.

AIM: To investigate Fusobacterium nucleatum (F. nucleatum) abundance in colorectal cancer (CRC) tissues and its association with CRC invasiveness in Chinese patients. METHODS: The resected cancer and adjacent normal tissues (10 cm beyond cancer margins) from 101 consecutive patients with CRC were collected. Fluorescent quantitative polymerase chain reaction (FQ-PCR) was applied to detect F. nucleatum in CRC and normal tissues. The difference of F. nucleatum abundance between cancer and normal tissues and the relationship of F. nucleatum abundance with clinical variables were evaluated. Fluorescence in situ hybridization (FISH) analysis was performed on 22 CRC tissues with the highest F. nucleatum abundance by FQ-PCR testing to confirm FQ-PCR results. RESULTS: The median abundance of F. nucleatum in CRC tissues [0.242 (0.178-0.276)] was significantly higher than that in normal controls [0.050 (0.023-0.067)] (P < 0.001). F. nucleatum was over-represented in 88/101 (87.1%) CRC samples. The abundance of F. nucleatum determined by 2(-ΔCT) was significantly greater in tumor samples [0.242 (0.178, 0.276)] than in normal controls [0.050 (0.023, 0.067)] (P < 0.001). The frequency of patients with lymph node metastases was higher in the over-abundance group [52/88 (59.1%)] than in the under-abundance group [0/13 (0%)] (P < 0.005). No significant association of F. nucleatum with other clinico-pathological variables was observed (P > 0.05). FISH analysis also found more F. nucleatum in CRC than in normal tissues (median number 6, 25(th) 3, 75(th) 10 vs 2, 25(th) 1, 75(th) 5) (P < 0.01). CONCLUSION: F. nucleatum was enriched in CRC tissues and associated with CRC development and metastasis.

Association of adiponectin gene variation with progression of nonalcoholic fatty liver disease: A 4-year follow-up survey.

OBJECTIVE: To explore the role of tagging single nucleotide polymorphisms (tagSNPs) in the adiponectin gene in the natural course of nonalcoholic fatty liver disease (NAFLD). METHODS: The participants were chosen from our previous survey containing 3543 individuals. Finally, a total of 696 participants who had been followed up for a median of 4 years were included. Each participant was administered with an interview, physical examination, blood tests and ultrasonic examination at both baseline and end-point. Polymerase chain reaction-restriction fragment length polymorphism was applied to determine seven tagSNPs in the adiponectin gene, namely, rs182052, rs16861205, rs822396, rs7627128, rs1501299, rs2241767 and rs3774261. Ordinal logistic regression was used to screen risk factors of NAFLD progression as well as the susceptibility to the disease. Haplotypes analyses were performed to confirm the results. RESULTS: After adjusting for age and gender, rs1501299 (G276T), rs2241767 (A45G) and rs3774261 (A712G) were found to be risk factors of both susceptibility (OR 5.040, 7.471 and 3.546, respectively) and progression (OR 3.83, 3.51 and 3.30, respectively) to NAFLD. Nevertheless, rs182052, rs16861205, rs822396 and rs7627128 had no impact on them. These findings were confirmed by haplotype analysis. CONCLUSION: The tagSNPs rs2241767, rs1501299 and rs3774261 in the adiponectin gene are risk factors for the individuals' susceptibility to and progression of NAFLD.

[Impact of rice agriculture on nitrogen and phosphorus exports in streams in hilly red soil earth region of central subtropics].

The research selected the Tuojia catchment and Jianshan catchment in Changsha County, Hunan Province, to comparatively study the effects of rice agriculture on the nitrogen and phosphorus concentrations and exports in streams in the typical agricultural catchments of the hilly red soil earth region. The monitoring of 16 months suggested that, there was a moderate stream nutrient pollution in both Tuojia and Jianshan catchments, especially for nitrogen pollution. Comparing the two catchments, the nitrogen and phosphorus concentrations were higher and the water quality was worse in the Tuojia catchment than that in the Jianshan catchment. From the nutrient composition of view, ammonia nitrogen was the main species of total nitrogen in the Tuojia catchment (accounting for 58.5% of total nitrogen), while it was nitrate nitrogen in the Jianshan catchment (accounting for 76. 1% of total nitrogen). The proportion of dissolved phosphorus in total phosphorus was 47. 1% in the Tuojia catchment, higher than the proportion of 37.5% in the Jianshan catchment. From temporal variations of nutrient dynamics of view, concentrations of all forms of nitrogen were higher during January to February and in July, respectively, and total phosphorus and dissolved phosphorus were higher during May to June and during October to December. Since the stream discharge in the catchments concentrated during the rice growing period from April to October, the higher nutrient concentrations during the period suggested potential risks of nitrogen and phosphorus losses. The total nitrogen mass flux was 1.67 kg x (hm2 x month)(-1) and TP was 0.06 kg x (hm2 x month)(-1) in the Tuojia catchment, which were greater than the 0.44 kg x (hm2 x month)(-1) and 0.02 kg x (hm2 x month)(-1) in the Jianshan catchment. Given the similar climate, geomorphology, soil type and cultivation patterns but the different area proportion of rice agriculture between two catchments, results suggested that, under the traditional crop management in hilly red soil earth region of central subtropics, the higher area proportion of rice agriculture has the potential to degrade stream aquatic environment.

The effect of miRNA-122 in regulating fat deposition in a cell line model.

Accumulating evidence supports the role of miR-122 in fatty liver disease. We investigated miR-122 expression in a steatotic hepatocyte model, the effect of miR-122 over-expression and inhibition in the pathogenesis. Human hepatic cell line L02 was induced with oleic acid to establish the steatotic hepatocyte model. Intracellular lipid content was observed with laser scanning confocal microscope (LSCM), and triglyceride content was determined with kits. Total RNA was extracted and reversely transcribed into cDNA. miR-122 expression was measured using qRT-PCR. Subsequently, miR-122 mimic and miR-122 inhibitor were transfected into steatotic hepatocytes to observe their effect on intracellular lipid content. The lipid fluorescence intensity and triglyceride content within the steatotic hepatocytes were significantly higher than those in normal control (860.01 ± 26.52 vs. 257.77 ± 29.69 and 3.47 ± 0.12 vs. 1.85 ± 0.02 at 24 h) (P < 0.01). miR-122 expression in steatotic hepatocytes was down-regulated compared with that in control (2-ΔCt value: 0.0286 ± 0.0078 vs. 0.0075 ± 0.0012) (P ≪ 0.01). After transfection, miR-122 expression (2-ΔCt value) in the miR-122 mimic group increased 2.96-fold compared with that in control, and its lipid fluorescence intensity was significantly lower than that in control (790.92 ± 46.72 vs. 1,022.16 ± 49.66) (P < 0.01). Nevertheless, miR-122 expression decreased 3.45-fold in the miR-122 inhibitor group compared with that in control, and its fluorescence intensity was significantly higher than that in control (1,386.49 ± 40.34 vs 1,022.16 ± 49.66)(P ≪ 0.01). We concluded that miR-122 was down-regulated in steatotic hepatocytes model. The pathogenesis of hepatocyte steatosis was enhanced by miR-122 mimic and reduced with miR-122 inhibitor.

Association between metabolic syndrome and the development of non-alcoholic fatty liver disease.

The aim of this study was to examine the effects of metabolic syndrome (MS) and the number of MS components on the development of non-alcoholic fatty liver disease (NAFLD). A total of 1,343 males and 574 females aged ≥50 years without NAFLD at baseline were included. Information on lifestyle, including alcohol use and personal history, was collected by face-to-face interviews. Biochemical parameters were assayed using fasting blood samples. NAFLD was diagnosed by abdominal ultrasonography. During follow-up at an average of 4.8 years, 223 patients developed NAFLD. Following adjustment for multiple covariates, age was an independent protective predictor [hazard ratio (HR), 0.96; 95% confidence interval (CI), 0.95-0.98], while the independent risk predictors were obesity (HR, 2.81; 95% CI, 2.14-3.69), higher triglycerides (HR, 2.56; 95% CI, 1.95-3.32) and alanine aminotransferase (HR, 1.004; 95% CI, 1.000-1.008). Participants with a diagnosis of MS had a significantly increased risk of developing NAFLD (HR, 3.17; 95% CI, 2.42-4.14). A greater number of MS components was significantly associated with a higher risk of NAFLD (all adjusted P for trend <0.001). Compared with those without any components of MS, participants with only one component of MS had a 3.6-fold higher risk of developing NAFLD (adjusted HR, 3.64; 95% CI, 1.50-8.88). The diagnosis and the number of components of MS were prospectively associated with the risk of developing NAFLD. Even in those with only one component of MS, the risk increased by 2.6-fold compared with that for the individuals without any components, suggesting a beneficial effect of intervention at the very early stage of MS on the prevention of NAFLD.

Expression of survivin and its four splice variants in colorectal cancer and its clinical significances.

The aim of this study was to investigate the clinical significances of the mRNA expression of survivin gene and its four splice variants in the pathogenesis of colorectal cancer (CRC). CRC samples, matched adjacent tissues, and normal tissues were collected from surgical resections of 39 patients with histologically confirmed diagnosis. The mRNA expression of survivin and its four splice variants, that is, survivin-△Ex3, survivin-2B, survivin-3B, and survivin-2α, was detected using semiquantitative PCR and RT-PCR. Carcinoembryonic antigen (CEA) CAM5 was determined as control. The mRNA expression rates of survivin, survivin-△Ex3, survivin-2B, survivin-3B, surviving-2α, and CEA CAM5 in CRC samples were significantly higher than those in adjacent tissues (P < 0.01) and those in normal tissues (P < 0.01). The mRNA levels of the above variants in CRC samples were also significantly higher than those in adjacent tissues (P < 0.01) and those in normal tissues (P < 0.01). The mRNA levels of survivin, survivin-2B, and survivin-2α were not associated with any clinical variable of patients, while the levels of survivin-△Ex3 and survivin-3B were associated with lymphoid metastasis and Dukes grade (P < 0.05), and survivin-△Ex3 was associated with invasiveness. We concluded that mRNA expression rates and levels of survivin and its four splice variants elevated in CRC tissues, and expression levels of survivin-△Ex3 and survivin-3B were positively associated with tumor aggression.