Baicalein, an enteric microbial metabolite, suppresses gut inflammation and cancer progression in ApcMin/+ mice.

OBJECTIVE: Chronic inflammation is recognized as a risk factor for colorectal cancer (CRC) development. Baicalin (BI), a major constituent in an anti-inflammatory herb Scutellaria baicalensis, can be biotransformed into baicalein (BE) by the intestinal microbiota. We evaluated the anti-inflammation and anti-CRC effects of the metabolite BE. METHODS: The in vitro biotransformation by human intestinal microbiota from BI into BE has been determined with HPLC. Using a gut-specific ApcMin/+ mouse model, the effects of oral BE on the life span, organ index, and tumor multiplicity were evaluated. The expressions of inflammatory cytokines were determined using ELISA. To verify the in vivo data, the anti-inflammatory and antiproliferative effects of BE were determined with an in vitro cell model. RESULTS: HPLC analysis showed that BI was quickly transformed into BE by the intestinal microbiota. Oral BE (30 mg/kg/day) significantly increased the life span, from 125.2 to 218.4 days (P < 0.01%). BE treatment also decreased intestine index and increased spleen index. Compared with the model group, following BE treatment, tumor numbers were significantly reduced in the small intestine and colon (P < 0.01, P < 0.05, respectively). In the gut tissues, BE treatment significantly reduced inflammatory cytokine levels such as IL-1ß, IL-2, IL-6, IL-10, G-CSF, and GM-CSF. In vitro data supported our in vivo results that the anti-CRC effects of BE were via the inhibition of gut inflammation and induction of cancer cell death. CONCLUSION: Our results suggest that the parent compound BI can be quickly converted into its microbial metabolite BE, which has stronger bioactive effects than BI. Baicalein is an active chemopreventive metabolite for inflammatory associated CRC.

Association between XRCC1 polymorphisms and laryngeal cancer susceptibility in a Chinese sample population.

Laryngeal cancer is the major malignant tumor affecting the upper respiratory tract. Previous studies have reported on the association between XRCC1 genetic polymorphisms and risk of laryngeal cancer, but with conflicting results. In this study, we attempted to assess the association between XRCC1 Arg194Trp, Arg280His and Arg399Gln polymorphisms and risk of laryngeal cancer in a Chinese population. A total of 126 laryngeal cancer patients and 254 control subjects were recruited to this study from the Second Medical College of Jinan University between December 2013 and May 2015. The XRCC1 Arg194Trp, Arg280His, and Arg399Gln polymorphic sites were genotyped by polymerase chain reaction-restriction fragment length polymorphism. Our results revealed a significant association between the AA genotype of XRCC1 Arg280His [odds ratio (OR) = 2.51, 95% confidence interval (CI) = 1.29-4.87, P = 0.01] and an increased risk of laryngeal cancer susceptibility compared to the GG genotype. Moreover, the A allele showed a higher risk of laryngeal cancer susceptibility compared to the G allele (OR = 1.63, 95%CI = 1.19-2.50, P = 0.002). In conclusion, the results of our study suggest that the AA genotype and A allele of the XRCC1 Arg280His polymorphism are associated with an increased laryngeal cancer risk in a Chinese population.

Relationship between serum GAD-Ab and the genetic polymorphisms of GAD2 and type 2 diabetes mellitus.

In this study, we investigated the relationship between serum glutamic acid decarboxylase (GAD) autoantibody (Ab) levels and single nucleotide polymorphisms (SNPs) of the glutamic acid de-carboxylase 2 (GAD2) 5'-untranslated region and the susceptibility to type 2 diabetes in the Han population. The distributions of patients with SNPs in the GAD2 5'-untranslated region (rs2236418, rs185649317, and rs8190590) and type 2 diabetes and that of the healthy group were genotyped and analyzed using Sequenom MassArray SNP genotyp-ing. GAD-Ab levels were also detected. The frequency distributions of the AA, AG, and GG genotypes in the polymorphic site rs2236418 in the diabetes GAD-Ab-positive group were 45.9, 42.8, and 11.4%, respectively, whereas those in the control group were 36.6, 43.7, and 19.8%, respectively. The difference between the 2 groups was statis-tically significant (P < 0.05). Unlike the GG genotype, the AA and AA + AG genotypes increased the risk of GAD-Ab (odds ratios (95% confidence intervals) = 2.623 (1.351-4.937) and 2.152 (1.375-4.202), respectively). The associations of the 3 SNPs of the GAD2 gene 5'-un-translated region polymorphisms with susceptibility to type 2 diabe-tes in the Chongqing Han population were significant. The SNP of rs2236418 in the Chongqing Han population of diabetic patients with serum GAD-Ab levels was significantly correlated with the SNPs rs185649317 and rs8190590.

Effect of Ginkgo biloba extract on matrix metalloproteinase-3 expression in a rat model of chondrocyte injury.

A rat model with cartilage chondrocyte injury was established using interleukin-1ß (IL-1ß) to investigate the effect of Ginkgo biloba extract (EGb) on matrix metalloproteinase-3 (MMP-3) expression. Rat chondrocytes were extracted and randomly divided into six groups: control group, IL-1ß (model) group, IL-1ß + dexamethasone group, and IL-1ß + EGb group (both high and low dose groups). Reverse transcriptase polymerase chain reaction and enzyme-linked immunosorbent assay were used to detect MMP-3 expression. Compared to the MMP-3 mRNA level in the control group, MMP-3 mRNA level significantly increased in the model group (P < 0.05). The application of dexamethasone or EGb significantly decreased the MMP-3 mRNA level (P < 0.05). MMP-3 mRNA and protein levels decreased in the EGb-treated group, especially in the high-dose group, compared to those in the dexamethasone group (P < 0.05). EGb may reduce MMP-3 production during IL-1ß-induced chondrocyte damage and protect chondrocytes to some extent, with better efficacy at high doses.

Relationship between the angle of vertebral screws and spinal lateral angulation after fixation of thoracolumbar fractures via an anterior approach.

This study investigated possible contributors to lateral spinal angulation after surgical fixation of thoracolumbar fractures via an anterior approach. We retrospectively examined lateral angulation in 172 cases of thoracolumbar fractures treated in this manner. The coronal Cobb angle and angles of the screws relative to the endplates were determined from radiographs. The patients completed the Short Form 36, Oswestry Disability Index, Japanese Orthopaedic Association Back Pain Evaluation Questionnaire, and Visual Analogue Scale at the final follow-up visit. The mean coronal Cobb angle was 0.75° ± 3.91° (-14.25° to 14.55°) preoperatively, 3.17° ± 4.07° (-8.18° to 14.01°) immediately postoperatively, and 3.46° ± 4.21° (-1.05° to 17.27°) at the final follow-up visit. The superior posterior and inferior anterior screws were more parallel to their respective endplates when the approach was made ≥2 vs ≤1 vertebral levels above the fracture (P < 0.001). Lateral angulation was more likely when the approach was made ≤1 vs ≥2 levels above the fracture (P < 0.001). The coronal Cobb angle differed significantly (P < 0.01) between patients with lumbar and thoracic fractures. The immediate postoperative coronal Cobb angle correlated tightly with the sum of the screw angles (superior plus inferior posterior and/or inferior plus superior anterior). Lateral angulation may occur after surgical fixation of thoracic and lumbar fractures via an anterior approach. Non-parallelism between the vertebral screws and their corresponding endplates may predict postoperative lateral spinal angulation. Postoperative lateral angulation does not correlate with low back pain, quality of life, or preoperative lateral angulation.

Prevalence and risk factors of overweight and obesity among individuals over 40 years old in Luzhou city.

In this study, a survey was conducted through questionnaire distribution and physical examinations were performed in 10,150 residents that were over 40 years old in Luzhou city. Respondents were selected by the multi-stage sampling method. The mean body mass index (BMI) of the sample population was 23.9 ± 3.3 kg/m(2). Among men, BMI showed a negative relationship with increasing age (P < 0.05), whereas among women, it showed a positive relationship (P < 0.001). The rates of overweight and obesity increased with age and reached a peak between 60 to 70 years of age (P < 0.001). The rates of overweight and obesity varied with different working conditions, training situations, educational levels, marital status, and other factors (P < 0.05). Age, educational level, daily sitting time, and family history of diabetes were factors that influenced the prevalence of overweight and obesity through multivariate logistic regression analysis (P < 0.05). The incidences of overweight and obesity among the middle-aged population were found to be significantly high. Therefore, prevention and control measures should be adopted as soon as possible.

Association between developmental defects of enamel and dental caries in schoolchildren.

Despite improvement, dental caries is still the main public oral health problem worldwide and the major cause of pain, tooth loss and chewing difficulties in children and adolescents; and it impacts negatively on oral health-related quality of life. A cross-sectional study of a multistage representative sample of 8-12-year-old Brazilian school children was carried out in order to investigate the association between enamel defects and dental caries. Children's mothers completed a questionnaire about socio-demographic and behavioural characteristics at home. Firth's bias reduced logistic regression models were undertaken to assess the association between the main exposure (enamel defects) and caries experience. The prevalence of any enamel defect was 64.0%; the prevalence of diffuse opacities, demarcated opacities and enamel hypoplasia was 35.0%, 29.5% and 3.7%, respectively. The prevalence of dental caries was 32.4%, with mean DMFT of 0.6 (SD, 1.2). Dental caries experience was more common among children who had enamel hypoplasia in their posterior teeth (OR=2.79; 95% CI: 1.05, 6.51) than among those with none. In anterior teeth, there was no association. Enamel hypoplasia appears to be an important risk factor for dental caries.

Bright solitons from the nonpolynomial Schrödinger equation with inhomogeneous defocusing nonlinearities.

Extending the recent work on models with spatially nonuniform nonlinearities, we study bright solitons generated by the nonpolynomial self-defocusing (SDF) nonlinearity in the framework of the one-dimensional (1D) Muñoz-Mateo-Delgado (MM-D) equation (the 1D reduction of the Gross-Pitaevskii equation with the SDF nonlinearity), with the local strength of the nonlinearity growing at |x|→∞ faster than |x|. We produce numerical solutions and analytical ones, obtained by means of the Thomas-Fermi approximation, for nodeless ground states and for excited modes with one, two, three and four nodes, in two versions of the model, with steep (exponential) and mild (algebraic) nonlinear-modulation profiles. In both cases, the ground states and the single-node ones are completely stable, while the stability of the higher-order modes depends on their norm (in the case of the algebraic modulation, they are fully unstable). Unstable states spontaneously evolve into their stable lower-order counterparts.

Histone demethylase retinoblastoma binding protein 2 regulates the expression of α-smooth muscle actin and vimentin in cirrhotic livers.

Liver cirrhosis is one of the most common diseases of Chinese patients. Herein, we report the high expression of a newly identified histone 3 lysine 4 demethylase, retinoblastoma binding protein 2 (RBP2), and its role in liver cirrhosis in humans. The siRNA knockdown of RBP2 expression in hepatic stellate cells (HSCs) reduced levels of α-smooth muscle actin (α-SMA) and vimentin and decreased the proliferation of HSCs; and overexpression of RBP2 increased α-SMA and vimentin levels. Treatment with transforming growth factor ß (TGF-ß) upregulated the expression of RBP2, α-SMA, and vimentin, and the siRNA knockdown of RBP2 expression attenuated TGF-ß-mediated upregulation of α-SMA and vimentin expression and HSC proliferation. Furthermore, RBP2 was highly expressed in cirrhotic rat livers. Therefore, RBP2 may participate in the pathogenesis of liver cirrhosis by regulating the expression of α-SMA and vimentin. RBP2 may be a useful marker for the diagnosis and treatment of liver cirrhosis.

Rituximab regulates the expression of the Raf kinase inhibitor protein via NF-κB in renal tissue of rats with diabetic nephropathy.

This study aimed to investigate the expression levels of the Raf kinase inhibitor protein (RKIP) and NF-κB in renal tissues of diabetic nephropathy (DN) rats, and to determine the underlying molecular targets of rituximab (RTX), with the goal of developing new clinical treatment selection for DN. Sprague-Dawley rats were randomly divided into a normal group (N), a DN group (M), and an RTX treatment group (D). Blood glucose and 24-h urine protein levels of rats were determined. The expression levels of RKIP and NF-κB in glomerular tissues were determined by immunohistochemistry staining and Western blotting. Comparisons between the M and N groups revealed that the concentrations of blood glucose and 24-h urine protein were significantly increased by DN (P < 0.01), and the expression levels of RKIP and NF-κB were significantly decreased and increased (P < 0.05), respectively. In the D group, the expression levels of RKIP and NF-κB were, respectively, upregulated and downregulated by RTX, and the concentrations of 24-h urine protein were also decreased by RTX. These results suggest that expression levels of RKIP might be regulated by RTX via NF-κB. This pathway could play an important role in the development and pathogenesis of DN. Therefore, RTX could be selected for clinical treatment of DN.

Histone demethylase retinoblastoma binding protein 2 regulates the expression of -smooth muscle actin and vimentin in cirrhotic livers

Liver cirrhosis is one of the most common diseases of Chinese patients. Herein, we report the high expression of a newly identified histone 3 lysine 4 demethylase, retinoblastoma binding protein 2 (RBP2), and its role in liver cirrhosis in humans. The siRNA knockdown of RBP2 expression in hepatic stellate cells (HSCs) reduced levels of α-smooth muscle actin (α-SMA) and vimentin and decreased the proliferation of HSCs; and overexpression of RBP2 increased α-SMA and vimentin levels. Treatment with transforming growth factor β (TGF-β) upregulated the expression of RBP2, α-SMA, and vimentin, and the siRNA knockdown of RBP2 expression attenuated TGF-β-mediated upregulation of α-SMA and vimentin expression and HSC proliferation. Furthermore, RBP2 was highly expressed in cirrhotic rat livers. Therefore, RBP2 may participate in the pathogenesis of liver cirrhosis by regulating the expression of α-SMA and vimentin. RBP2 may be a useful marker for the diagnosis and treatment of liver cirrhosis.

Target replacement strategy for selection of DNA aptamers against the Fc region of mouse IgG.

Aptamers that recognize the IgG Fc region are of great interest because of their wide application as an immunology probing tool, for diagnostics, and as affinity agents for antibody purification. We developed a target replacement strategy as a modification of conventional Systematic Evolution of Ligands by EXponential enrichment (SELEX) in order to efficiently select and identify novel DNA aptamers against the Fc region of mouse IgG. In this new approach, multiple IgG subclasses (IgG1, IgG2a, mouse IgG Fc, and anti-HBs IgG) were sequentially used to select aptamers in one continuous SELEX. After 8 rounds of selection, the aptamers were analyzed using dot blot and an electrophoretic mobility shift assay, which showed universal binding capability to different IgG subclasses. Secondary structure analysis of the aptamers indicated that the stem-loop structure of the aptamers play an important role in binding to the common site in different mouse IgG subclasses. This demonstrated the feasibility of using multiple target replacement SELEX for the selection of aptamers. This target replacement strategy is also expected to be useful for selecting aptamers that bind common regions of molecules other than antibodies.

Genetic background of Citrus nobilis Lour. 'Gonggan' based on the chloroplast trnL gene.

The genetic backgrounds of many Citrus varieties are quite complex. Classifications and phylogenetic relationships of Citrus species have become the focus of researchs. Some conserved genes of chloroplast genome's research have been proven effective in determining the biosources of hybrids and phylogenetic analysis. Thus, we studied variations among the chloroplast trnL gene sequences of 10 Citrus species, including C. nobilis Lour. 'Gonggan'. The amplification results of different trnL target genes and identification of the double-enzyme cut after cloning show that lengths of all trnL sequences were within 895 to 935 bp and a total of 24 variation sites were detected among the 10 material samples. Clustering analysis revealed differences in trnL genes caused by systematic evolution and allowed the determination of variations among Citrus varieties. Variation sites of trnL sequences can be used in the phylogenetic classification and species identification of Citrus, and the results agreed with random amplified polymorphic DNA marker results. C. nobilis Lour. 'Gonggan' is closely associated with the other two varieties in Zhaoqing area, and C. nobilis Lour. 'Gonggan' and C. haniana Hort. ex Tseng 'Sihuihanggan' can be classified into the same category. C. nobilis Lour. 'Gonggan' as a natural hybrid is probably a hybrid with C. haniana Hort. ex Tseng 'Sihuihanggan' as its female parent.

Mitotic and meiotic behavior of rye chromosomes in wheat - Psathyrostachys huashanica amphiploid x triticale progeny.

The dynamics of rye chromosomes during mitosis and meiosis was analyzed in a subset comprising 33 F3 lines from the cross of wheat, Psathyrostachys huashanica amphiploid (AABBDDNsNs) and hexaploid triticale (AABBRR), as visualized by genomic in situ hybridization. The results indicated that 31 of the total lines contained 4-14 rye chromosomes. Twenty-eight combinations had more rye chromosomes than the F1 hybrids, suggesting the occurrence of spontaneous quantitative increment. No P. huashanica chromosomes were detected in all of the combinations tested. Mitotic analysis showed that rye chromosomes progressed normally with the wheat counterparts without loss. However, abnormal meiosis was found in almost all lines. Similar progression between wheat and rye genomes appeared from interphase to metaphase I. It was at anaphase I that many rye univalents lagged behind those of wheat, followed by equational division. This resulted in the formation of chromosomal segments and micronuclei at telophase I or II. Micronuclei could also be generated from the immobilized univalents in the periphery of cells. Synapsis and translocations between wheat and rye genomes, chromosome bridges, and unreduced gametes were detected. Therefore, it is proposed that rye chromosome elimination may involve chromatid lagging, fragmentation and micronucleation, or the immobilization of certain univalents during meiosis instead of mitosis in the relatively advanced generations. This mechanism, together with spontaneous incremental increase of rye chromosome number, permitted the generation of various germplasms for wheat improvement.

RXR agonists inhibit high glucose-induced upregulation of inflammation by suppressing activation of the NADPH oxidase-nuclear factor-κB pathway in human endothelial cells.

An inflammatory response induced by high glucose is a cause of endothelial dysfunction in diabetes and is an important contributing link to atherosclerosis. Diabetes is an independent risk factor of atherosclerosis and activation of retinoid X receptor (RXR) has been shown to exert anti-atherogenic effects. In the present study, we examined the effects of the RXR ligands 9-cis-retinoic acid (9-cis-RA) and SR11237 on high glucose-induced inflammation in human umbilical endothelial vein endothelial cells (HUVECs) and explored the potential mechanism. Our results showed that the inflammation induced by high-glucose in HUVECs was mainly mediated by the activation of nuclear factor-B (NF- κB). High glucose-induced expression of intercellular adhesion molecule-1 (ICAM-1) and vascular cell adhesion molecule-1 (VCAM-1) were in comparison, significantly decreased by treatment with RXR. The effect of RXR agonists was mainly due to the inhibition of NF-κB activation. Using pharmacological inhibitors and siRNA, we confirmed that nicotinamide adenine dinucleotide phosphate (NADPH) oxidase was an upstream activator of NF-κB. Furthermore, RXR agonists significantly inhibited high glucose-induced activation of NADPH oxidase and significantly decreased the production of reactive oxygen species (ROS). To explore whether the rapid inhibitory effects of RXR agonists were in fact mediated by RXR, we examined the effect of RXR downregulation by RXR siRNA. Our results showed that RXR siRNA largely abrogated the effects of RXR agonists, suggesting the requirement of RXR expression. Therefore, we have shown that RXR is involved in the regulation of NADPH oxidase- NF-κB signal pathway, as the RXR ligands antagonized the inflammatory response in HUVECs induced by high glucose.