Dual antioxidant activity and the related mechanisms of a novel pentapeptide GLP4 from the fermented mycelia of Ganoderma lingzhi.

Oxidative stress causes chronic inflammation, and mediates various diseases. The discovery of antioxidants from natural sources is important to research. Here we identified a novel antioxidant peptide (GLP4) from Ganoderma lingzhi mycelium and investigated its antioxidant type and potential protective mechanisms. Through free radical scavenging assay, active site shielding validation, superoxide dismutase (SOD) activity assay, and lipid peroxidation assay, we demonstrated that GLP4 was a novel protective agent with both direct and indirect antioxidant activities. GLP4 could directly enter human umbilical vein endothelial cells (HUVECs) as an exogenous substance. Meanwhile, GLP4 promoted the nuclear translocation of nuclear factor erythroid-2-related factor 2 (Nrf2) and activated the Nrf2/antioxidant response element (ARE) signaling pathway, exhibiting antioxidant and anti-apoptotic cytoprotective effects on hydrogen peroxide (H2O2)-induced HUVECs. Pull-down experiments of GLP4 target proteins, bioinformatics analysis and molecular docking further revealed that GLP4 mediated Nrf2 activation through binding to phosphoglycerate mutase 5 (PGAM5). The results suggested that GLP4 is a novel peptide with dual antioxidant activity and has promising potential as a protective agent in preventing oxidative stress-related diseases.

NR4A1 counteracts JNK activation incurred by ER stress or ROS in pancreatic ß-cells for protection.

Sustained hyperglycaemia and hyperlipidaemia incur endoplasmic reticulum stress (ER stress) and reactive oxygen species (ROS) overproduction in pancreatic ß-cells. ER stress or ROS causes c-Jun N-terminal kinase (JNK) activation, and the activated JNK triggers apoptosis in different cells. Nuclear receptor subfamily 4 group A member 1 (NR4A1) is an inducible multi-stress response factor. The aim of this study was to explore the role of NR4A1 in counteracting JNK activation induced by ER stress or ROS and the related mechanism. qPCR, Western blotting, dual-luciferase reporter and ChIP assays were applied to detect gene expression or regulation by NR4A1. Immunofluorescence was used to detect a specific protein expression in ß-cells. Our data showed that NR4A1 reduced the phosphorylated JNK (p-JNK) in MIN6 cells encountering ER stress or ROS and reduced MKK4 protein in a proteasome-dependent manner. We found that NR4A1 increased the expression of cbl-b (an E3 ligase); knocking down cbl-b expression increased MKK4 and p-JNK levels under ER stress or ROS conditions. We elucidated that NR4A1 enhanced the transactivation of cbl-b promoter by physical association. We further confirmed that cbl-b expression in ß-cells was reduced in NR4A1-knockout mice compared with WT mice. NR4A1 down-regulates JNK activation by ER stress or ROS in ß-cells via enhancing cbl-b expression.

NR4A1 retards adipocyte differentiation or maturation via enhancing GATA2 and p53 expression.

Nuclear receptor subfamily 4 group A member 1 (NR4A1) is an orphan nuclear receptor with diverse functions. It has been reported that NR4A1, as a transcriptional activator, is implicated in glucose and lipid metabolism. The aim of this study was to investigate the regulatory role of NR4A1 in adipogenesis and explore the underlying mechanisms. Quantitative real-time PCR and Western blotting were used to analyse the expression of genes involved in synthesis and mobilization of fats in vivo and in vitro. Dual-luciferase reporter assay was conducted to study the regulatory mechanisms of NR4A1. Our data from in vivo study confirmed that NR4A1 knockout (KO) mice fed with high-fat diet were more prone to obesity, and gene expression levels of PPARγ and FAS were increased in KO mice compared to controls; our data from in vitro study showed that NR4A1 overexpression in 3T3-L1 pre-adipocytes inhibited adipogenesis. Moreover, NR4A1 enhanced GATA binding protein 2 (GATA2) expression, which in turn inhibited peroxisome proliferator-activated receptor γ (PPARγ); NR4A1 inhibited sterol regulatory element binding transcription factor 1 (SREBP1) and its downstream gene fatty acid synthase (FAS) by up-regulating p53. NR4A1 inhibits the differentiation and lipid accumulation of adipocytes by enhancing the expression of GATA2 and p53.

Renal Interstitial Arteriosclerotic Lesions in Lupus Nephritis Patients: A Cohort Study from China.

OBJECTIVE: The aim of this study was to evaluate renal arteriosclerotic lesions in patients with lupus nephritis and investigate their associations with clinical and pathological characteristics, especially cardio-vascular features. DESIGN: A retrospective cohort study. PARTICIPANTS: Seventy-nine patients with renal biopsy-proven lupus nephritis, diagnosed between January 2000 and June 2008 from Peking University First Hospital. RESULTS: In clinico-pathological data, patients with arteriosclerosis had higher ratio of hypertension and more severe renal injury indices compared with patients with no renal vascular lesions. More importantly, patients with renal arteriosclerosis had worse cardiac structure and function under transthoracic echocardiographic examination. Patients with renal arteriosclerosis tend to have higher ratios of combined endpoints compared with those of no renal vascular lesions, although the difference didn't reach statistical meanings (P = 0.104). CONCLUSION: Renal arteriosclerotic lesion was common and associated with vascular immune complex deposits in lupus nephritis. It might have a certain degree of association with poor outcomes and cardiovascular events, which needs further explorations.

[Effects of batroxobin and electric cauterization on vascular remodeling of rabbit with a removal of carotid arterial adventitia].

OBJECTIVE: To explore the topical hemostatic effects of batroxobin (BX) and electric cauterization (EC) on capillary hemorrhage of rabbit with a removal of carotid arterial adventitia. METHODS: A total of 27 New Zealand rabbits were randomly divided into control, BX and EC groups. Each group received BX (2 kU/L), EC (power = 40 W) and saline for topical hemostasis after a removal of carotid arterial adventitia and blunt dissection. The animals were euthanized by 0, 14 and 28 d post-operation. The specimens of adventitia removal section were divided into three parts for histology (hematoxylin and eosin, MASSON & transmission electron microscope), immunohistochemistry (IHC) [monocyte chemoattractant protein-1 (MCP-1), transforming growth factor-ß1 (TGF-ß1) and vascular endothelial growth factor (VEGF)] and reverse transcription-polymerase chain reaction (RT-PCR). RESULTS: At Day 28 post-operation, the remodeling index, stenotic rate and collagen fiber density of BX and EC groups were (0.753 ± 0.0739) and (0.618 ± 0.0989), (0.298 ± 0.030)% and (0.363 ± 0.039)%, (15.4 ± 3.5)% and (23.4 ± 5.1)% respectively. There was statistically significant difference between two groups (P < 0.05) while no difference existed between hemocoagulase and control groups. The results of electron microscopy showed that the atrial fibroblasts of EC group increased markedly versus BX group. As demonstrated by RT-PCR and IHC, the expressions of MCP-1, TGF-ß1 and VEGF in BX group were lower than those in EC group (P < 0.05) while no difference versus the control group. CONCLUSION: As a safe and effective topical hemostatic method, BX can effectively decrease inflammation response and reduce vascular remodeling and narrowing in rabbits with a removal of carotid arterial adventitia. And its effect mimic closely natural conditions.

Hemocoagulase atrox reduces vascular modeling in rabbit carotid artery adventitia.

OBJECTIVE: This study aimed to compare the effects of hemocoagulase atrox and cauterization hemostasis on intimal hyperplasia and explore the effect of hemocoagulase atrox on vascular modeling in rabbit carotid artery adventitia. METHODS: A total of 27 rabbits were randomly divided into 3 groups (0d, 14d, 28d). They were anaesthetized using an intramuscular injection of phenobarbital sodium (1 ml/kg). The left and right common carotid arteries were exposed and capillary hemorrhaged after blunt dissection of the adventitia layers of common carotid arteries. Nine rabbits in each group were again randomly divided into 3 groups, in which animals were respectively treated with hemocoagulase (2 U/ml), cauterization (power = 40 w) and saline (as control). Groups of animals were euthanized at 0, 14 and 28 days after surgery. The samples were equally divided in the middle of the adventitia removal section to obtain equal parts for histologic, immunohistochemical and molecular biologic analysis. The vascular repair after adventitial stripping was observed by HE staining, Masson staining and transmission electron microscopy. The expression of carotid MCP-1, PCNA, TGF-ß1, α-SMA and VEGF were measured at different time points by RT-PCR and immunohistochemical staining. RESULTS: HE staining and Masson staining showed that hemocoagulase atrox had a significantly stronger effect on reducing intimal hyperplasia than the cauterization after 14 and 28 days. The results of RT-PCR showed that the expression of MCP-1, TGF-ß1, α-SMA and VEGF in hemocoagulase atrox-treated animals were lower than that of cauterization-treated animals. CONCLUSION: Our results suggested that hemocoagulase atrox as a topical hemostatic is safety and efficiently and it can accelerate adventitia restoration and decrease intimal proliferation.

[The topically hemostatic effects of batroxobin on the carotid arteries adventitia removal rabbit].

OBJECTIVE: To observe the topically hemostatic effects of batroxobin (BX) in different concentrations on the carotid arteries adventitia removal rabbit. METHODS: 18 rabbits were removed vascular adventitia by collagenase digestion and mechanical dissection, causing capillary hemorrhage. Then all of them were randomly divided into 6 groups: blank control, negative control group, 0.5, 1, 2 and 4 kU/L (U/ml) BX group. The hemostatic time and bleeding volume were observed to compare the hemostatic effect of each group. Haematoxylin-eosin, Masson staining and immunohistochemistry were performed to assure adventitia removed. RESULTS: It was feasible to remove vascular adventitia with collagenase digestion and mechanical dissection. The hemostatic time and bleeding volume were significantly different (P < 0.05) from 0.5 U/ml BX group [(97 ± 20)s,(0.102 ± 0.013)g/cm(2)] of the negative control group[(143 ± 33)s,(0.130 ± 0.023) g/cm(2)]. With the increase of BX concentration, there was a significant difference (P < 0.05) between 2 U/ml BX group (32 ± 13,0.056 ± 0.015) and 1 U/ml BX group (32 ± 13,0.056 ± 0.015), but there was no statistical significance (P > 0.05) between 2 U/ml BX group and 4 U/ml BX group (28 ± 14,0.053 ± 0.012). Thus, the best topical hemostatic concentration of BX was 2 U/ml. CONCLUSION: The topical hemostatic effect of batroxobin is reliable in small area of blood oozing.

Highly enantioselective Henry reactions of aromatic aldehydes catalyzed by an amino alcohol-copper(II) complex.

Amino alcohol-Cu(II) catalyst: Highly enantioselective Henry reactions between aromatic aldehydes and nitromethane have been developed. The reactions were catalyzed by an easily available and operationally simple amino alcohol-copper(II) catalyst. In total, 38 substrates were tested and the R-configured products were obtained in good yields with excellent enantioselectivities.

[Integrating genetic and gene expression data: methods and applications of eQTL mapping].

The availability of high-throughput genotyping technologies and microarray assays has allowed researchers to investigate genetic variations that influence levels of gene expression. Expression Quantitative Trait Locus (eQTL) mapping methods have been used to identify the genetic basis of gene expression. Similar to traditional QTL studies, the main goal of eQTL is to identify the genomic locations to which the expression traits are linked. Although microarrays provide the expression data of thousands of transcripts, standard QTL mapping methods, which are able to handle at most tens of traits, cannot be applied directly. As a result, it is necessary to consider the statistical principles involved in the design and analysis of these experiments. In this paper, we reviewed individual selection, experimental design of microarray, normalization of gene expression data, mapping methods, and explaining of results and proposed potential methodological problems for such analyses. Finally, we discussed the applications of this integrative genomic approach to estimate heritability of transcripts, identify candidate genes, construct gene networks, and understand interactions between genes, genes and environments.

Do reproductive hormones explain the association between body mass index and semen quality?

AIM: To examine whether reproductive hormones play a role in the association between body mass index (BMI) and semen quality. METHODS: Semen quality and testosterone (T), luteinizing hormone (LH), follicle-stimulating hormone (FSH) and estradiol (E(2)) were evaluated in 990 fertile males with age 38.9 +/- 9.7 (mean +/- SD) years recruited from the Chinese general population in 2001 and 2002. RESULTS: Semen quality was reduced among underweight (BMI < 18.5) compared with normal (BMI 18.5-24.9) and overweight (BMI 25.0-29.9), but the associations were independent of reproductive hormones. After adjustment for the potential confounders, underweight men had reductions in sperm concentration (22.4 X 10(6)/mL), total sperm count (52.9 X 10(6)) and percentage of normal sperm forms (6.9%) compared with men with normal BMI. Being underweight may be a risk factor for low sperm concentration (OR: 4.68, 95% confidence intervals [CI]: 2.01-10.91). Otherwise, being overweight may be a protected factor for low sperm concentration (OR: 0.25; 95% CI: 0.08-0.83) and low total sperm count (OR: 0.37, 95% CI: 0.15-0.87). CONCLUSION: Low BMI was associated with reduced semen quality. The associations between BMI and semen quality were found statistically significant even after adjustment for reproductive hormones. Reproductive hormones cannot explain the association between BMI and semen quality.

[A case-control study on the risk factor of perinatals' congenital malformations in seven cities of Guangxi].

OBJECTIVE: To investigate the possible risk factors of congenital malformations in cities of Guangxi. METHODS: A case-control study was carried out on 281 cases of congenital malformations and 730 controls. Analysis of simple factor and multiple factors unconditional logistic regression were done. RESULTS: The analysis of simple factor and multiple factors showed that main risk factors of congenital malformations as multiple pregnancies (OR = 2.6), pregnancy complications (OR = 3.2), exposure to chemical substances before or during pregnancy (OR = 3.0), taking sedatives (OR = 10.2), hormone drug (OR = 9.4) or Chinese herbal medicines (OR = 2.5) during the early stage of pregnancy, mothers' blood type as AB (OR = 3.5) or A (OR = 2.2), mothers' emotion being nervous and melancholy (OR = 2.6), mothers' occupation being workers (OR = 3.8) or peasants (OR = 3.0), fathers' exposure to noise (OR = 5.7) or suffering from chronic diseases (OR = 2.8). CONCLUSIONS: Some risk factors were identified as having important effect on perinatal congenital malformations, including taking sedatives, hormone drug or Chinese herbal medicines during the early stage of pregnancy, mothers' emotion being nervous and melancholy, multiple pregnancies, pregnancy complications, exposure to chemical substances before or during pregnancies, mothers' blood type as AB or A, mothers' occupation being workers or peasants, fathers' exposure to noise or suffering from chronic diseases.