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ObjectiveTo explore the effect of okra extract on gestational diabetes mellitus (GDM) rats and its probable molecular mechanism.MethodsA total of 30 female SD rats were caged with male rats for pregnancy, 27 pregnant rats were obtained and weighed. The pregnant rats were equally randomized into the control group, GDM group and intervention group. Once the pregnancy was verified, GDM group and intervention group were given 45 mg/kg streptozotocin by peritoneal injection for inducing GDM, control group was given equal volume of citrate buffer. Once the model was established successfully, intervention group was administered orally the solution containing 200 mg/kg/d okra extract, the other groups were given the diet and water only. On the 19th day of pregnancy, the blood samples and fetal rats of all groups were collected, fetal rats weight and placental weight was recorded and the serum glucose, lipids, serum insulin and C-peptide of pregnant rats before the delivery were determined.ResultsThe pregnant rats weight before the delivery, fetal rats weight and placental weight of GDM group were lower than control group and intervention group (P 0.05). Antioxidant enzymes levels of GDM group in liver and pancreas tissues were lower than the other groups, and after treatment of okra extract, antioxidant enzymes levels in liver and pancreas tissues were equivalent to control group (P > 0.05).ConclusionsOkra extract, rich in antioxidant substances, could avoid the excessive consuming of antioxidant enzymes, then, suppresses the oxidative stress and insulin resistance, thereby improving blood glucose level of GDM rats.

Human papillomavirus (HPV) is the most common viral infection of the reproductive tract, which can increase the risk of cervical cancer. Development of HPV-specific vaccines that can be recognized and presented by human leukocyte antigen (HLA) system is considered as an important approach to prevention and treatment of cervical cancer. In this study, a combination of bioinformatics analysis and peptide binding assay was described to identify potential cytotoxic T lymphocyte epitopes from the HPV genome. In the procedure, a structure-based quantitative structure–activity relationship (SB-QSAR) was used to analyze the intermolecular interactions between HLA proteins and peptide epitopes, and the obtained nonbonded descriptors were then correlated linearly and nonlinearly with HLA–peptide affinities by using partial least squares regression and support vector machine, respectively. Several SB-QSAR models were established and evaluated rigorously, from which the best predictor was applied to virtual screening against thousands of nonapeptide segments generated from the HPV-16 genome. With this protocol a number of peptide ligands were inferred to have high affinity for a variety of frequently observed HLA-A alleles, including 0101, 0201, 0202, 0203, 0206 and 0301, from which several promising candidates were selected and tested in vitro to determine their binding strength toward HLA proteins. Consequently, several peptides were identified as potent HLA binders with half-maximal binding BL50 at micromolar level. The molecular mechanism underlying the high-affinity binding of identified peptide ligands to HLA proteins was also investigated in detail.

Selenium is an essential element with antioxidant roles in immune regulation, but there is little understanding of how Se acts in apoptosis in the immune organs of birds. The aim of study was to evaluate the influence of Se deficiency on oxygen free radicals, NO and apoptosis in immune organ of chickens. 160 1-day-old chickens were randomly assigned to two groups of 80 each and were fed on a low-Se diet (0.032 mg/kg Se) or a control diet (0.282 mg/kg Se), respectively. OFR production in blood was determined on days 30, 45, 60 and 75, respectively. The iNOS-NO system activity in immune organ (thymus, spleen, bursa of fabricius) was identified by NO content and NOS activity assay on days 30, 45, 60 and 75, respectively. Apoptosis was measured by DNA ladder analysis, ultrastructural observations, TdT-mediated dUTP nick end labeling TUNEL assay and flow cytometric analysis of apoptotic DNA. The transcription of factor-associated suicide, caspase-3 mRNA was tested by fluorescence quantitative PCR. The results showed that OFR production, NO and inducible NO synthases (iNOS) activity in the low-Se group were significantly increased (p < 0.05) than in the control group. In addition, apoptosis was observed in chicken immune organ in the low-Se group. The degree and the number of apoptotic cells rose in a time-dependent manner. The expression of Fas and caspase-3 mRNA increased (p < 0.05) than in the control group. It indicated that the oxidative stress and NO played a causative role in the apoptosis of immune tissues induced by selenium deficiency.

Selenium (Se) is an important nutritional trace element possessing immune-stimulatory properties. The aim of this 75-day study was to investigate effect of oxidative stress on immunosuppression induced by selenium deficiency by determining antioxidative function, morphological changes, DNA damage, and immune function in immune organ of chickens. One hundred sixty 1-day-old chickens (egg-type birds) were randomly assigned to two groups of 80 each and were fed on a low-Se diet (0.032 mg/kg Se) or a control diet (0.282 mg/kg Se, sodium selenite), respectively. Se contents in blood and immune organ (thymus, spleen, bursa of Fabricius) were determined on days 30, 45, 60, and 75, respectively. Antioxidative function was examined by total antioxidant capacity (T-AOC), superoxide dismutase (SOD), glutathione peroxidase (GSH-Px), and xanthine oxidase (XOD), and oxidative damage was examined by malondialdehyde (MDA) detection. DNA damage was measured by comet assay, and immune function was examined by determining serum interleukin-1β (IL-1β), interleukin-2 (IL-2), and tumor necrosis factor (TNF) contents. The results showed that Se concentrations in the low-Se group were significantly lower (P

To investigate the value of metalloproteinase-3 (MMP-3) levels in assessing efficacy of etanercept treatment in patients with ankylosing spondylitis (AS).The serum and synovial fluid levels of MMP-3 were measured by enzyme linked immunosorbent assay (ELISA) in 48 patients with AS in week 0, 6 and 12; and also measured in 30 serum samples and 10 synovial fluid samples from healthy controls.The serum levels of MMP-3 in AS patients were significantly higher than those in controls. In AS patients, the MMP-3 levels in synovial fluid were significantly higher than those in serum levels. The serum MMP-3 levels in AS patients with peripheral arthritis were higher than those with exclusively axial involvement; while C-reactive protein (CRP) levels and erythrocyte sedimentation rate (ESR) did not differ between these 2 groups of AS. At week 6 and week 12 of etanercept treatment, the serum MMP-3 levels were significantly decreased (p0.01) with the declining trend of ESR, CRP, Bath Ankylosing Spondylitis Disease Activity Index (BASDAI) and Bath Ankylosing Spondylitis Functional Index (BASFI) (all p0.01). Before the etanercept treatment (week 0), serum levels of MMP-3 were correlated with ESR, CRP, BASDAI and BASFI (p0.05). ESR was also correlated with CRP and BASFI, but not with BASDAI (r=0.361, P=0.071). At weeks 12, serum MMP-3 levels were still correlated with ESR, CRP and BASDAI (P0.05), but not with BASFI (P=0.339); ESR was correlated with CRP, but not with BASDAI and BASFI. There was a significant correlation between BASDAI and BASFI (r=0.818,P=0.001).Serum MMP-3 levels are closely related to disease activity and may serve as an useful indicator for efficacy of etanercept treatment in AS patients.

To investigate the effects of alendronate (Alen) on the prevention of systemic glucocorticoid-induced osteoporosis in patients with rheumatic diseases.140 patients suffering from rheumatic diseases, including systemic lupus erythematosus, polymyositis, dermatomyositis, and Sjögren's syndrome, with normal bone mineral density (BMD) and treated with oral glucocorticoids were randomly divided into 2 groups: Alen + calcium group (n = 74) receiving Alen 10 mg once a day and castrate D 600 0.6 g once a day for 24 weeks and control group (n = 66) receiving castrate D 600 0.6 g once a day for 24 weeks. The BMD and biomarkers of bone turnover were measured at baseline and 24 weeks after initiating glucocorticoid therapy.After 24 weeks, the BMD values at lumbar spine, femoral neck, major trochanter, and Ward' s triangle increased by 6.1%, 6.3%, 3.3%, and 2.2% respectively compared with those at baseline (all P0.05), however, those of the control group decreased by 8.7%, 9.1%, 7.7%, and 6.4% respectively (P0.01, P0.05), and the BMD levels at lumbar spine and femoral neck 24 weeks later of the Alen + calcium group were both higher than those of the control group (P0.01, P0.05). 24 weeks later the level of urine cross linked N-telopeptides of type I collagen (NTX) of the Alen + calcium group decreased (P0.05), and the blood osteocalcin (BGP) of the Alen + calcium group increased, however, not significantly (P0.05). There were no significant differences in serum AKP and BGP and urine NTX 24 weeks later between these 2 groups.Improving BMD, alendronate plays an important role in the prevention of glucocorticoid-induced osteoporosis. However, calcium treatment alone fails to prevent the loss of bone.

To investigate the expressions of C(4)D(+)CD(25)(+) T cells in the peripheral blood of patients with systemic lupus erythematosus (SLE) and to identify the relationship between the levels of CD(4)(+)CD(25)(+) T cells and the disease activity and progression of SLE.Fifty-three SLE patients were enrolled in the study. Flow-cytometric assay was employed for detection of CD(4)(+)CD(25)(+) T cells and CD(4)(+)CD(25)(bright) T cells were defined according to fluorescence intensity of CD(25) expression exceeding 100. Meanwhile, correlation analysis was performed between their expression and the scores of SLE disease active index (SLEDAI), C(3), dsDNA and antinuclear antibody titles.The levels of peripheral blood CD(4)(+)CD(25)(+) T cells in SLE were (7.84 +/- 1.85)%, which were significantly lower than those in a group of healthy control [(9.18 +/- 2.01)%, P0.05]. The levels of CD(4)(+)CD(25)(+) T cells in a group of active SLE [(6.72 +/- 1.16)%] were higher than those in a group of stable SLE [(8.57 +/- 1.91)%, P0.01]. There was no difference between the active and stable groups of SLE in peripheral blood CD(4)(+)CD(25)(bright) T cells [(0.85 +/- 0.24)% vs (0.91 +/- 0.25)%, P = 0.686], but they were significantly lower than those in the group of healthy controls [(1.43 +/- 1.08)%, P0.01]. With the reduction of the SLEDAI scores in SLE patients after relevant treatment, the levels of peripheral blood CD(4)(+)CD(25)(bright) T cells did not change. No correlation was found between the levels of CD(4)(+)CD(25)(bright) T cells and SLEDAI scores, antinuclear antibody titles, dsDNA and C(3), respectively (rho = -0.188, P = 0.178; rho = -0.216, P = 0.121; rho = 0.082, P = 0.560; rho = 0.010, P = 0.944).The reduction of CD(4)(+)CD(25)(+) T cells in peripheral blood may participate in the pathogenesis of SLE.

Cell-surface glycoproteins are regarded as candidates for involvement in the spread of tumor cells. N-linked beta1-6 branched oligosaccharides may contribute directly to the malignant or metastatic phenotypes of tumor cells. Increased beta1-6 branching has been associated with an increased level of N-acetylglucosaminyltransferase V (GlcNAc transferase V), the glycosyltransferase that initiates the beta1-6 branching. In this report, 33 pathologically verified hepatocellular carcinoma (HCC) specimens, six non-cancerous tissues surrounding HCC and five normal liver specimens have been studied. We have quantified N-linked beta1-6 branched oligosaccharides indirectly by measuring GlcNac transferase V activity. The average GlcNac transferase V activities in hepatocellular carcinoma (HCC), noncancerous tissues surrounding HCC and normal liver tissues were 324.2 +/- 269.8, 84.8 +/- 20.7 and 7.0 +/- 6.2 pmol product h(-1) mg protein(-1) (P < 0.05) respectively. In addition, the activity was correlated with the TNM classification of HCC. The average activities of GlcNAc transferase V in stages T1, T2-3 and T4 were 77.6 +/- 57.8, 369.0 +/- 294.7 and 329.9 +/- 205.9 pmol product h(-1) mg protein h(-1) respectively (P < 0.05), showing that the activity of the enzyme in advanced HCC was higher than that in early HCC. Our preliminary results indicated that GlcNAc transferase V activity increased in human HCC and was correlated with its progression.

A fluorescence assay method for beta1-4galactosyltransferase (beta1-4GT) has been developed involving a pyridylaminated sugar as an acceptor substrate, a fluorescent sugar chain, with the reducing end of the Gnbeta1 - 2Malpha1 - 6(Gnbeta1-2Malpha1-3)Mbeta1 - 4Gnbeta1 - 4Gn - PA aminated with 2-aminopyridine. Microsome was prepared from the liver of normal male rats as an enzyme sample. Then the fluorescent reaction product was separated by reverse-phase HPLC. The kinetic experiments were carried out using crude enzyme extracts of the Golgi complex from the rat liver. The enzyme has a pH optimum of 6.5,and optimal concentration of Triton X-100 of 0.5%. The K(m) and V(max) values for the sugar acceptor substrates were found to be 2.31x10(-3)M(-1) and 5.75x10(-2)mgr;mol/(min.mg) respectively. Furthermore, our research revealed that beta1-4GT had branch specificity. The Gn of alpha1-3 mannose branch of the acceptor substrate was more susceptible to galactosylation than that of the alpha1-6 branch by 2.10 times.