The effect of different combinations of antibiotic cocktails on mice and selection of animal models for further microbiota research.

The gut microbiota is closely related to host health and disease. However, there are no suitable animal models available at present for exploring its functions. We analyzed the effect of 3 different antibiotic cocktails (ABx) via two administration routes on the composition of murine gut microbiota, as well as on the general physiological and metabolic indices. High-throughput 16S rRNA sequencing showed that ABx treatment altered the gut microbiota community structure, and also caused low-degree inflammation in the colon. In addition, ad libitum administration of antibiotics depleted the gut microbiota more effectively compared to direct oral gavage, especially with 3ABx. The ABx treatment also had a significant impact on renal and liver functions, as indicated by the altered serum levels of creatinine, urea, total triglycerides, and total cholesterol. Finally, Spearman's correlation analysis showed that the predominant bacterial genera resulting from ABx intervention, including Lactobacillus, Roseburia, and Candidatus-Saccharimonas, were negatively correlated with renal function indices. Taken together, different antibiotic combinations and interventions deplete the gut microbiota and induce physiological changes in the host. Our findings provide the basis for developing an adaptive animal model for studying gut microbiota. KEY POINTS: • Ad libitum administration of 3ABx can effectively deplete intestinal microbiota. • ABx treatment may have slight effect on renal and liver function. • The levels of urea and creatinine correlated with the growth of Roseburia.

Evolutionary agroecology: individual fitness and population yield in wheat (Triticum aestivum).

Although the importance of group selection in nature is highly controversial, several researchers have argued that plant breeding for agriculture should be based on group selection, because the goal in agriculture is to optimize population production, not individual fitness. A core hypothesis behind this claim is that crop genotypes with the highest individual fitness in a mixture of genotypes will not produce the highest population yield, because fitness is often increased by "selfish" behaviors, which reduce population performance. We tested this hypothesis by growing 35 cultivars of spring wheat (Triticum aestivum L.) in mixtures and monocultures, and analyzing the relationship between population yield in monoculture and individual yield in mixture. The relationship was unimodal, as predicted. The highest-yielding populations were from cultivars that had intermediate fitness, and these produced, on average, 35% higher yields than cultivars with the highest fitness. It is unlikely that plant breeding or genetic engineering can improve traits that natural selection has been optimizing for millions of years, but there is unutilized potential in traits that increase crop yield by decreasing individual fitness.

Response of chickpea (Cicer arietinum L.) to terminal drought: leaf stomatal conductance, pod abscisic acid concentration, and seed set.

Flower and pod production and seed set of chickpea (Cicer arietinum L.) are sensitive to drought stress. A 2-fold range in seed yield was found among a large number of chickpea genotypes grown at three dryland field sites in south-western Australia. Leaf water potential, photosynthetic characteristics, and reproductive development of two chickpea genotypes with contrasting yields in the field were compared when subjected to terminal drought in 106kg containers of soil in a glasshouse. The terminal drought imposed from early podding reduced biomass, reproductive growth, harvest index, and seed yield of both genotypes. Terminal drought at least doubled the percentage of flower abortion, pod abscission, and number of empty pods. Pollen viability and germination decreased when the fraction of transpirable soil water (FTSW) decreased below 0.18 (82% of the plant-available soil water had been transpired); however, at least one pollen tube in each flower reached the ovary. The young pods which developed from flowers produced when the FTSW was 0.50 had viable embryos, but contained higher abscisic acid (ABA) concentrations than those of the well-watered plants; all pods ultimately aborted in the drought treatment. Cessation of seed set at the same soil water content at which stomata began to close and ABA increased strongly suggested a role for ABA signalling in the failure to set seed either directly through abscission of developing pods or seeds or indirectly through the reduction of photosynthesis and assimilate supply to the seeds.

Effect of CYP2C19 Gene Polymorphisms on Proton Pump Inhibitor, Amoxicillin, and Levofloxacin Triple Therapy for Eradication of Helicobacter Pylori.

BACKGROUND The effects of PPI are variable owing to the CYP2C19 polymorphisms. However, whether the polymorphisms could affect the Hp eradication efficacy of triple therapy is still not clear. The present study aimed to assess the effects of CYP2C19 gene polymorphisms on proton pump inhibitor (PPI), amoxicillin, and levofloxacin triple therapy for Helicobacter pylori (Hp) eradication. MATERIAL AND METHODS We randomly assigned 160 Hp-positive patients with chronic gastritis to 2 groups to receive either 20 mg bid omeprazole (OAL group, n=80) or 10 mg bid rabeprazole (RAL group, n=80), combined with 1000 mg bid amoxicillin and 500 mg qd levofloxacin. The 2 groups were treated for 10 days. The CYP2C19 genotypes included wild-type, M1 mutant gene (*2, the mutation of exon 5), and M2 mutant gene (*3, the mutation of exon 4) identified by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFIP). According to CYP2C19 genotype combinations, the patients were divided into extensive metabolizer (EM), intermediate metabolizer (IM), and poor metabolizer (PM) subgroups. The eradication efficacy of Hp was evaluated by 14C-UBT at 28 days after treatment. RESULTS The trial was completed by 155 patients. Hp eradication rates in OAL and RAL groups were 78.2% and 88.3%, respectively, on per-protocol (PP) analysis, indicating no significant difference (P>0.05). Regarding CYP2C19 genotypes, eradication rates of 60.7%, 84.2%, and 100% were obtained for EM, IM, and PM subgroups, respectively, of the OAL group. EM group eradication rates were significantly lower than IM and PM group values (P<0.05). In the RAL group, no such difference was observed (P>0.05). Hp eradication rates were significantly lower in the EM subgroup of the OAL group compared with that of the RAL group. CONCLUSIONS Hp eradication rates were higher in the RAL group than in OAL-treated patients. Interestingly, omeprazole-based therapy was significantly affected by the CYP2C19 genotype, unlike the rabeprazole-based therapy.

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.

ß-Aminobutyric acid increases abscisic acid accumulation and desiccation tolerance and decreases water use but fails to improve grain yield in two spring wheat cultivars under soil drying.

A pot experiment was conducted to investigate the effect of the non-protein amino acid, ß-aminobutyric acid (BABA), on the homeostasis between reactive oxygen species (ROS) and antioxidant defence during progressive soil drying, and its relationship with the accumulation of abscisic acid (ABA), water use, grain yield, and desiccation tolerance in two spring wheat (Triticum aestivum L.) cultivars released in different decades and with different yields under drought. Drenching the soil with 100 µM BABA increased drought-induced ABA production, leading to a decrease in the lethal leaf water potential (Ψ) used to measure desiccation tolerance, decreased water use, and increased water use efficiency for grain (WUEG) under moderate water stress. In addition, at severe water stress levels, drenching the soil with BABA reduced ROS production, increased antioxidant enzyme activity, and reduced the oxidative damage to lipid membranes. The data suggest that the addition of BABA triggers ABA accumulation that acts as a non-hydraulic root signal, thereby closing stomata, and reducing water use at moderate stress levels, and also reduces the production of ROS and increases the antioxidant defence enzymes at severe stress levels, thus increasing the desiccation tolerance. However, BABA treatment had no effect on grain yield of wheat when water availability was limited. The results suggest that there are ways of effectively priming the pre-existing defence pathways, in addition to genetic means, to improve the desiccation tolerance and WUEG of wheat.

Allometry and Yield Stability of Cereals.

Crop plants grow, and then, they allocate resources to different structures, including seeds and fruits, which represent yield in most crops. We define the yield stability of a genotype as its ability to reduce the effects of temporal variation in resources and conditions on yield production, and we argue that yield stability can be understood in terms of two processes: (1) crop survival and growth (biomass production): the ability of the crop plants to survive and produce biomass under the range of conditions to which it is exposed and (2) the pattern of allocation of this biomass to yield across this range of conditions. Plant breeders and crop physiologists have focused on (1), but much less attention has been paid to (2). We hypothesize that (2) is primarily the result of reproductive allometry: the quantitative relationship between vegetative and reproductive biomass. Ecological theory and the allometric models we present predict a tradeoff between (a) the ability of a genotype to produce yield over a wide variety of conditions and (b) its ability to produce very high yields under optimal or near-optimal conditions. We reanalyze the data from two recent studies, and the results are consistent with this hypothesis. Yield stability in crops corresponds to bet-hedging in evolutionary ecological theory. It is the most appropriate strategy for smallholder farmers in developing countries, a group that comprises most of the farmers in the world. Researchers and crop breeders need to rethink their objectives if they want to develop optimal varieties for these farmers.

Inhibition of PD-1 Protects against TNBS-Induced Colitis via Alteration of Enteric Microbiota.

METHODS: Colitis was induced in mice using 2,4,6-trinitrobenzene-sulfonic acid (TNBS), and mice were subsequently treated with either a PD-1 inhibitor or 5-amino-salicylic acid (ASA) as a positive control. Body weight, disease activity index (DAI), colon length, and tissue damage were evaluated, and the enteric microbiota was profiled using high-throughput 16S rRNA sequencing of fecal samples from the experimental mice. RESULTS: TNBS caused mice to experience IBD-like symptoms, which were attenuated by the PD-1 inhibitor, as indicated by a decrease in DAI scores (p = 0.0002). Furthermore, in this mouse model of IBD, PD-1 inhibition improved the alpha diversity as well as restored the beta diversity of the enteric microbiome. It also significantly enriched the abundance of short-chain fatty acid- (SCFA-) producing bacteria of the Firmicutes (p < 0.05) and Bacteroidetes (p < 0.05) phyla but depopulated Proteobacteria (p < 0.05). CONCLUSION: PD-1 inhibition can partly mitigate TNBS-induced colitis and restore the enteric microbiota by enriching the abundance of SCFA-producing bacteria.

[Association of hepatic lipase gene promoter polymorphism -514C/T with nonalcoholic fatty liver disease].

OBJECTIVE: To study the prevalence of the hepatic lipase gene (LIPC) promoter polymorphism (at position -514) in patients with nonalcoholic fatty liver disease (NAFLD), and its relationship with the susceptibility to NAFLD. METHODS: Genotype of LIPC promoter was detected with PCR-RFLP in 106 patients with NAFLD. Body mass index, waist-to-hip ratio (WHR), blood pressure, CHOL, HDL, LDL, TG, FPG and FINS of the patients were measured. Index of insulin resistance was determined using the homeostasis model assessment (HOMA) method. One hundred six healthy subjects matched for age and sex served as controls. RESULTS: The frequency of CC genotype and C allele in the NAFLD group were significantly higher than those in the control group (31.1% vs 26.4%, 62.7% vs 54.2%, P<0.05). Compared with TT genotype, both CC genotype and CT genotypes had higher relative risk of NAFLD (OR: 3.73, 95% CI: 1.31, 10.63; OR: 3.60, 95% CI: 1.35, 9.60). At the same time, the non-carriers of T allele in -514 had higher WHR than the T carriers (0.877+/-0.06 vs 0.848+/-0.06, t=2.072, P<0.05)). Logistic regression analysis showed that T substitution in LIPC-514 position (OR: 1.28, 95% CI 0.10-0.74) had a lower susceptibility to NAFLD. CONCLUSION: The LIPC-514C/T polymorphism is associated with WHR, and the T substitution of LIPC-514 may lower the susceptibility to NAFLD.

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.

[Effects of RNA interference on expression of tumor necrosis factor-alpha in lipopolysaccharide-activated mouse macrophages].

OBJECTIVE: To investigate the effects of siRNAs targeting mouse tumor necrosis factor (TNF)-alpha gene on the expression of TNF-alpha mRNA and protein in the LPS activated macrophages. METHODS: Mouse macrophages of the line RAW264.7 were cultured. Three siRNA sequences targeting different sites of mouse TNF-alpha gene (siRNA 1 - 3) and a fluorescein-labeled double-stranded RNA (dsRNA) oligomer (siRNA 4), as negative control, were designed and chemically synthesized. All siRNAs were transfected into the mouse macrophages. 0, 3, 6, 9, 12, 15, and 18 hours after the transfection samples of supernatant were collected. Inverse microscopy was used to observe the efficacy of transfection. Then the macrophages were co-incubated with LPS for 9 hours. Macrophages stimulated by LPS only and macrophages without transfection and LPS stimulation were used as controls. Then samples of supernatant were collected. TNF-alpha protein expression was detected by ELISA. Real time PCR was used to detect the TNF-alpha mRNA expression. RESULTS: The transfection rate was 72% - 80%. The TNF-alpha mRNA expression levels of the LPS + siRNA2 and LPS + siRNA3 groups were 0.158 +/- 0.031 and 0.114 +/- 0.028 respectively, both significantly lower than that of the LPS control group (0.294 +/- 0.147, P < 0.05 and P < 0.01 respectively), however, the TNF-alpha mRNA expression level of the LPS + siRNA1 and LPS + siRNA4 groups were not significantly different from that of the LPS control group. The TNF-alpha mRNA expression inhibition rates of the LPS + siRNA and LPS + siRNA3 groups were 46.0% and 61.2% respectively. The TNF-alpha protein expression levels of the LPS + siRNA2 and LPS + siRNA3 group were (1358 +/- 348) pg/ml and (817 +/- 138) pg/ml, both significantly lower than that of the LPS control group [(2104 +/- 32) pg/ml, P < 0.05 and P < 0.01], and the TNF-alpha mRNA expression levels of the LPS + siRNA1 and LPS + siRNA4 groups were not significantly different from that of the LPS control group. CONCLUSION: LPS time-dependently increases the expression of TNF-alpha in macrophages. SiRNAs targeting TNF-alpha inhibit the expression levels of TNF-alpha mRNA and protein in macrophages treated with LPS.

[The relationship between insulin resistance and adiponectin gene expression in nonalcoholic fatty liver disease].

OBJECTIVE: To investigate the relationship between insulin resistance (IR) and adiponectin gene expression in patients with nonalcoholic fatty liver disease (NAFLD). METHODS: Subcutaneous (SC) and omental (OM) adipose tissues were obtained from 21 NAFLD patients with obesity (n=10) and nonobesity (n=11) and also from 24 subjects (without NAFLD) with obesity (n=11) and nonobesity (n=13) who served as controls. Adiponectin mRNA expression levels in subcutaneous and omental adipose tissues were measured using SYBR Green I quantitative real-time PCR. The levels of plasma adiponectin and insulin were measured with ELISA. IR was estimated using the homeostasis assessment (HOMA). RESULTS: The scores of HOMA-IR levels of the NAFLD patients and the controls with obesity and nonobesity were: 3.0+/-0.8, 2.8+/-0.9, 2.0+/-0.6, 1.2+/-0.5 respectively. The relative mRNA expression of adiponectin and blood adiponectin levels in NAFLD patients differed significantly from those of the controls. The HOMA-IR negatively correlated with the adiponectin mRNA expression levels of adipose tissues (r = -0.5) and blood adiponectin; it positively correlated with body mass index (r = 0.4), waist-hip-ratio (r = 0.4) and serum triglyceride (r = 0.3), but did not correlate with serum total cholesterol (r = 0.2). CONCLUSION: IR of NAFLD patients was linked to low adiponectin gene expression in their adipose tissues. This finding suggests that low adiponectin gene expression may play a role in the pathogenesis of insulin resistance and NAFLD.

Pattern of Water Use and Seed Yield under Terminal Drought in Chickpea Genotypes.

Drought, particularly terminal drought, reduces the yield of chickpea (Cicer arietinum L.). Terminal drought tolerance and water use patterns were evaluated under controlled conditions in 10 genotypes of desi chickpea. Withholding water from early podding reduced vegetative growth, reproductive growth, seed yield, and water use efficiency for seed yield in all genotypes. The genotype Neelam, which produced the highest seed yield when water was withheld, used the least water when well-watered; however, its aboveground biomass at maturity did not differ significantly from six of the nine other genotypes. Indeed, the water-stressed Neelam had the lowest daily transpiration rate during the early stages of water stress and the highest during the later stages, thereby maintaining the highest soil water content in the first 16 days after water was withheld, which enabled higher pod production, lower pod abortion, and better seed filling. Genotypes differed in the threshold value of the fraction of transpirable soil water when flowering and seed set ceased in the water-stress treatment. We conclude that a conservative water use strategy benefits seed yield of chickpea exposed to water shortage during early podding.

Genotypic Variation in Yield, Yield Components, Root Morphology and Architecture, in Soybean in Relation to Water and Phosphorus Supply.

Water shortage and low phosphorus (P) availability limit yields in soybean. Roots play important roles in water-limited and P-deficient environment, but the underlying mechanisms are largely unknown. In this study we determined the responses of four soybean [Glycine max (L.) Merr.] genotypes [Huandsedadou (HD), Bailudou (BLD), Jindou 21 (J21), and Zhonghuang 30 (ZH)] to three P levels [applied 0 (P0), 60 (P60), and 120 (P120) mg P kg-1 dry soil to the upper 0.4 m of the soil profile] and two water treatment [well-watered (WW) and water-stressed (WS)] with special reference to root morphology and architecture, we compared yield and its components, root morphology and root architecture to find out which variety and/or what kind of root architecture had high grain yield under P and drought stress. The results showed that water stress and low P, respectively, significantly reduced grain yield by 60 and 40%, daily water use by 66 and 31%, P accumulation by 40 and 80%, and N accumulation by 39 and 65%. The cultivar ZH with the lowest daily water use had the highest grain yield at P60 and P120 under drought. Increased root length was positively associated with N and P accumulation in both the WW and WS treatments, but not with grain yield under water and P deficits. However, in the WS treatment, high adventitious and lateral root densities were associated with high N and P uptake per unit root length which in turn was significantly and positively associated with grain yield. Our results suggest that (1) genetic variation of grain yield, daily water use, P and N accumulation, and root morphology and architecture were observed among the soybean cultivars and ZH had the best yield performance under P and water limited conditions; (2) water has a major influence on nutrient uptake and grain yield, while additional P supply can modestly increase yields under drought in some soybean genotypes; (3) while conserved water use plays an important role in grain yield under drought, root traits also contribute to high nutrient uptake efficiency and benefit yield under drought.

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.

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.

Overexpression of activated leukocute cell adhesion molecule in gastric cancer is associated with advanced stages and poor prognosis and miR-9 deregulation.

Activated leukocyte cell adhesion molecule (ALCAM) has been identified as a novel potential molecular marker of human tumors. The present study aimed to assess ALCAM as a prognostic marker for gastric cancer (GC), and to explore the mRNA deregulation underlying the abnormal expression of ALCAM. The mRNA and protein expression of ALCAM in GC and adjacent non­tumor tissues from 66 patients with GC were analyzed. The association between miR­9 and ALCAM mRNA expression was determined by quantitative polymerase chain reaction. Serum soluble ALCAM (sALCAM) was analyzed by ELISA in 72 patients with GC, 82 patients with gastric precancerous lesions and 73 controls. ALCAM and sALCAM levels were associated with certain clinicopathological variables, including overall survival. Compared with the non­tumor tissues, the expression of ALCAM mRNA in the GC tissues was significantly upregulated (P=0.013). The expression of miR­9 was reduced and inversely correlated with ALCAM mRNA levels in GC tissues and cell lines. The ALCAM mRNA level was reduced following ectopic overexpression of miR­9 in SGC­7901 human gastric cancer cells. The rates of membranous and cytoplasmic expression of ALCAM in GC tissues were 59.1 and 48.48%, respectively, and the serum sALCAM levels were significantly elevated in patients with GC. Elevated ALCAM mRNA, membranous ALCAM expression in GC tissues and high sALCAM levels are associated with advanced tumor stage, lymphatic invasion and shorter overall survival duration. The results of the current study indicated that membranous ALCAM expression and high serum sALCAM levels are independent prognostic markers of poor survival for patients with GC, and that the overexpression of ALCAM may be due to the downregulation of miR­9.

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.

Exogenous abscisic acid reduces water loss and improves antioxidant defence, desiccation tolerance and transpiration efficiency in two spring wheat cultivars subjected to a soil water deficit.

The effect of soil drenching with 10µM abscisic acid (ABA) on the physiological responses of two spring wheat (Triticum aestivum L.) cultivars released in different decades was evaluated when subjected to a water deficit at jointing or at booting. Exogenous ABA application increased the ABA concentration in the leaves, reduced the stomatal conductance (gs), slowed the rate of water use, decreased the lethal leaf water potential (ψ) used to measure desiccation tolerance and lowered the soil water content (SWC) at which leaf relative water content (RWC) began to decrease and wilting was observed. Exogenous ABA application also reduced reactive oxygen species (ROS) formation and increased antioxidant enzyme activity, leading to a reduction in the oxidative damage to lipid membranes in both cultivars exposed to water stress at jointing and booting. The decrease in leaf RWC and wilting occurred at lower values of SWC in the recently-released cultivar than in the earlier-released cultivar. The recently-released cultivar also had higher grain yield than the earlier-released cultivar at moderate water stress, but the grain yield in both cultivars was reduced by water stress and by the exogenous ABA treatment. However, exogenous ABA treatment increased transpiration efficiency for grain (TEG) of both cultivars under moderate water stress. These results indicate that ABA played an important role in slowing water use and enhancing the antioxidant defence during soil drying, but this did not result in increased yields under drought stress.

Nur-related receptor 1 gene polymorphisms and alcohol dependence in Mexican Americans.

AIM: To investigate the association of polymorphisms of nur-related receptor 1 (Nurr1) and development of alcohol dependence in Mexican Americans. METHODS: Peripheral blood samples were collected from 374 alcoholic and 346 nonalcoholic Mexican Americans; these two groups were sex- and age-matched. Sample DNA was extracted and genomic DNA was amplified by polymerase chain reaction. The -2922(C) 2-3 polymerase chain reaction products were digested with Sau96I, alleles of 1345(G/C), and -1198(C/G) in the regulatory region as well as Ex+132 (G/T/A/C) and Ex+715(T/-) in exon 3 were studied by sequencing. RESULTS: The C2/C2, C2/C3, C3/C3 genotype distribution of -2922(C) 2-3 was 34.4%, 38.2% and 27.5% in the nonalcoholic group compared to 23.3%, 51.2% and 25.4% in the alcoholic group (P = 0.001). The C/C, C/G, G/G genotype distribution of -1198(C/G) was 23.5%, 46.1% and 30.3% in the nonalcoholic group compared to 13.9%, 50.9% and 35.3% in the alcoholic group (P = 0.007). However, the -1345 (G/C), Ex3+132(G/T/A/C) and Ex3+715(T/-) alleles were not polymorphic in Mexican Americans, and all those studied had G/G, G/G and T/T genotype for these three alleles, respectively. The -2922(C) 2-3 did not show allele level difference between alcoholic and nonalcoholic individuals, but -1198 (C/G) showed a significant allele frequency difference between alcoholic (39.3%) and nonalcoholic (46.6%) populations (P = 0.005). Excluding obese individuals, significant differences were found at both genotypic and allelic levels for the -2922(C) 2-3 polymorphism (P = 0.000 and P = 0.049) and the -1198 (C/G) polymorphism (P = 0.008 and P = 0.032) between nonobese alcoholics and nonobese controls. Excluding smokers, a significant difference was found only at the genotypic level for the -2922(C) 2-3 polymorphism (P = 0.037) between nonsmoking alcoholics and nonsmoking controls, but only at the allelic level for the -1198(C/G) polymorphism (P = 0.034). CONCLUSION: Polymorphisms in the regulatory region of Nurr1 are implicated in pathogenesis of alcohol dependence and the Nurr1/dopamine signaling pathway might be important for this dependence development in Mexican Americans.