早发性卵巢功能不全患者血清25-OH-VD、DBP、SIRT1与性激素和氧化应激的关系研究

摘要 目的：探讨早发性卵巢功能不全（POI）患者血清25-羟基维生素D（25-OH-VD）、邻苯二甲酸二丁酯（DBP）、沉默信息调节因子2相关酶1（SIRT1）与性激素和氧化应激的关系。方法：选取2019年1月～2020年12月攀枝花学院附属医院妇产科收治的97例POI患者（POI组）,另选取同期54名体检健康女性（对照组）。检测两组血清性激素[卵泡刺激素（FSH）、黄体生成素（LH）、雌二醇（E2）、抗缪勒管激素（AMH）]、氧化应激[超氧化物岐化酶（SOD）、谷胱甘肽过氧化物酶（GSH-Px）、丙二醛（MDA）]、25-OH-VD、DBP、SIRT1水平。采用Pearson/Spearman相关系数分析POI患者血清25-OH-VD、DBP、SIRT1与性激素、氧化应激指标的相关性,Logistic回归分析血清25-OH-VD、DBP、SIRT1与POI的关系。结果：POI组FSH、LH、MDA、DBP水平高于对照组,E2、AMH、SOD、GSH-Px、25-OH-VD、SIRT1水平低于对照组（P＜0.05）。Pearson/Spearman相关系数显示,POI患者血清25-OH-VD、SIRT1与FSH、LH、MDA呈负相关,与E2、AMH、SOD、GSH-Px呈正相关（P＜0.05）;血清DBP与FSH、LH、MDA呈正相关,与E2、AMH、SOD、GSH-Px呈负相关（P＜0.05）。Logistic回归分析显示,调整混杂因素后,25-OH-VD（OR＝0.825,95％CI：0.741～0.919）、SIRT1（OR＝0.872,95％CI：0.810～0.938）是POI发生的保护因素,DBP（OR＝1.173,95％CI：1.074～1.282）是危险因素（P＜0.05）。结论：POI患者血清25-OH-VD、SIRT1水平下调,DBP水平上调,与性激素和氧化应激相关,可能成为POI的辅助预测因子。     

Identification of a novel duplication CFTRdup2 and functional impact of large rearrangements identified in the CFTR gene

The aim of this study was to investigate associations of two candidate gene SNPs of the endocannabinoid receptor type 1 gene (CNR1) with overweight, obesity and obesity-related traits in Chinese retired women. The study subjects were a subsample of the Taizhou Retiree Women Cohort, consisting of 2812 retired women aged 50-64 years recruited from Taizhou, Jiangsu, China. Neither rs2023239 nor rs806381 polymorphism was significantly associated with body mass index-defined overweight and obesity or waist-to-hip-ratio-defined obesity. For obesity-related traits, rs2023239 was significantly associated with glutamate pyruvate transaminase (GPT) (median, 18.00 vs 17.00 for TT and TC genotypes, respectively, P=0.043). The rs806381 also showed significant association with triglyceride (TG) (mean±SD, 1.46±0.20 vs 1.53±0.20 for GA and GG+AA genotypes, respectively, P=0.013) under the dominant genetic model. In conclusion, the rs2023239 and rs806381 polymorphisms of CNR1 were not associated with increased overweight and obesity risk. But the rs2023239 polymorphism was significantly associated with GPT, and the rs806381 polymorphism was significantly associated with TG.     

Functional analysis of an auxin-inducible DNA-binding protein gene

Over the past decades, several studies indicate a correlation between the phytohormone auxin and cell division. The molecular players of this signaling pathway are now being uncovered. DNA Binding Protein1 from Arabidopsis (AtDBP1) is an auxin-inducible gene able to bind DNA non-specifically. In this work the tissue-expression pattern of this gene was investigated. Promoter-GUS analysis demonstrated that the AtDBP1 promoter is active in regions exhibiting intense cell division such as meristems and nematode feeding sites. Also, the promoter expression was modulated upon incubation with cell cycle blockers, indicating a potential role in cell division for this gene. Lastly, AtDBP1 antisense plants presented a higher insensitivity to auxin, and interfered negatively with auxin–induced callus formation and reduced apical dominance.     

Plasticizer DBP Activates NLRP3 Inflammasome through the P2X7 Receptor in HepG2 and L02 Cells

Ditubyl phthalate (DBP), one of the most widely used plasticizers, can migrate out to contaminate our bodies and environment. A number of studies have showed that DBP is closely related to liver pathological changes and diseases. Inflammasomes are multiprotein complexes composed of procaspase and pattern recognition receptors such as Nucleotide oligomerization domain (NOD) like receptor family, pyrin domain containing 3 (NLRP3). Activation of NLRP3 inflammasome is implicated in the pathogeneses of liver damage. The aim of this study was to determine the effects of DBP on NLRP3 inflammasome. We found that DBP triggered the activation of NLRP3 inflammasome in hepatocyte cell lines. By using Ca‐074‐Me, N‐acetylcysteine and KN‐62, we observed that the P2X₇ receptor participated in the DBP‐induced activation of NLRP3 inflammasome. DBP could also trigger the ATP release. In conclusion, we demonstrated that DBP is one of the activator of NLRP3 inflammasome and may play an important role in liver damage.     

环境激素DBP对拟南芥试管形态发生的影响

报道了酞酸酯类化合物DBP对拟南芥试管形态发生的影响。利用不同浓度的DBF处理拟南芥茎段外植体,切片观察拟南芥茎段试管形态发生过程中脱分化、分化及再分化的变化情况。结果发现：拟南芥茎段外植体在对照组培养基上生长良好,首先是愈伤组织产生,而后愈伤组织块增大,颜色呈淡绿色,生长旺盛,并有分化出芽现象;添加0．01mg／L、0．1mg／L和1．0mg／LDBP,茎段愈伤组织诱导率低,且高浓度时愈伤组织出现褐化、活力差的现象;培养12h、24h、2d、3d、7d、14d、21d和28d茎段显微切片结果显示,DBP具有抑制拟南芥试管形态发生的作用,随着DBP浓度的增加呈现剂量抑制效应。     

邻苯二甲酸二丁酯对多刺裸腹溞生长繁殖的影响

在实验室条件下研究了邻苯二甲酸二丁酯对多刺裸腹溞的急性毒性和连续作用四个世代的慢性毒性。研究结果表明:邻苯二甲酸二丁酯对多刺裸腹溞的生长和繁殖具有显著的毒性效应;邻苯二甲酸二丁酯对多刺裸腹溞的24 h半致死浓度为8.44mg·L^-1;持续暴露在邻苯二甲酸二丁酯下的多刺裸腹溞子一代和子二代受到的伤害最为显著;不同世代多刺裸腹溞对邻苯二甲酸二丁酯具有一定的耐受性,子一代和子二代耐受性下降,敏感性增强,子三代敏感性下降。研究结果有助于了解环境激素类物质对枝角类动物生长和繁殖的长期毒性效应及其规律。     

Serum paraoxonase-1 (PON1) activities (PONase/AREase) and polymorphisms in patients with type 2 diabetes mellitus in a North-West Indian population

Objective

Paraoxonase-1 (PON1), an HDL-C associated enzyme, protects lipoproteins from oxidation. There is evidence that PON1 enzyme activity is reduced in the patients with type 2 diabetes mellitus (T2DM). North-West Indian Punjabis, a distinct ethnic group has high incidence of T2DM. However till date there is no information regarding PON1 enzyme activities and PON1 polymorphisms in T2DM patients of this ethnic group.

Methods

We identified polymorphisms in the coding Q192R, L55M and promoter − 909G/C, − 162A/G, − 108C/T of the PON1 gene by using PCR-RFLP, multiplex PCR and allele specific oligonucleotide PCR assays in 250 T2DM patients and 300 healthy controls. We also assessed paraoxonase (PONase) and arylesterase (AREase) activities of PON1 enzyme.

Results

The serum PONase (114.2 vs. 178.0 nmol/min/ml) and AREase (62.7 vs. 82.5 μmol/min/ml) activities were significantly lower (p < 0.0001) in patients as compared to controls. PONase activity was affected by all the studied PON1 polymorphisms. However, AREase activity was not affected by any of these polymorphisms. Coding Q192R and promoter − 909G/C polymorphisms showed significant differences in genotypic distribution. QR, RR (Q192R) and GC, CC (− 909G/C) genotypes and L-C-A-R-G, L-T-A-R-G, L-T-G-Q-C haplotypes showed significant association with type 2 diabetes. No significant linkage disequilibrium was observed among the five polymorphisms.

Conclusion

Both PONase and AREase activities are lower in patients and this could lead to increased lipid peroxidation and accelerated atherosclerosis in them. PONase activity, but not AREase activity is influenced by PON1 polymorphisms. QR, RR, GC, CC genotypes and L-C-A-R-G, L-T-A-R-G, L-T-G-Q-C haplotypes are commoner in diabetics as compared to controls and may be related to genetic susceptibility to type 2 diabetes.     

Kinetics and pathway of biodegradation of dibutyl phthalate by Pleurotus ostreatus

Dibutyl phthalate (DBP) is a plasticizer, whose presence in the environment as a pollutant has attained a great deal of attention due to its reported association with endocrine system disturbances on animals. Growth parameters, glucose uptake, percentage of removal efficiency (%E) of DBP, biodegradation constant of DBP (k) and half-life of DBP biodegradation (t_1/2) were evaluated for Pleurotus ostreatus grown on media containing glucose and different concentrations of DBP (0, 500 and 1000 mg l^?1). P. ostreatus degraded 99.6 % and 94 % of 500 and 1000 mg of DBP l^?1 after 312 h and 504 h, respectively. The k was 0.0155 h^?1 and 0.0043 h^?1 for 500 and 1000 mg of DBP l^?1, respectively. t_1/2 was 44.7 h and 161 h for 500 and 1000 mg of DBP l^?1, respectively. Intermediate compounds of biodegraded DBP were identified by GC-MS and a DBP biodegradation pathway was proposed using quantum chemical calculation. DBP might be metabolized to benzene and acetyl acetate, the first would be oxidated to muconic acid and the latter would enter into the Krebs cycle. P. ostreatus has the ability to degrade DBP and utilizes it as source of carbon and energy.     

Nup159 Weakens Gle1 Binding to Dbp5 But Does Not Accelerate ADP Release

Dbp5, DDX19 in humans, is an essential DEAD-box protein involved in mRNA export, which has also been linked to other cellular processes, including rRNA export and translation. Dbp5 ATPase activity is regulated by several factors, including RNA, the nucleoporin proteins Nup159 and Gle1, and the endogenous small-molecule inositol hexakisphosphate (InsP₆). To better understand how these factors modulate Dbp5 activity and how this modulation relates to in vivo RNA metabolism, a detailed characterization of the Dbp5 mechanochemical cycle in the presence of those regulators individually or together is necessary. In this study, we test the hypothesis that Nup159 controls the ADP-bound state of Dbp5. In addition, the contributions of Mg²⁺ to the kinetics and thermodynamics of ADP binding to Dbp5 were assessed. Using a solution based in vitro approach, Mg²⁺ was found to slow ADP and ATP release from Dbp5 and increased the overall ADP and ATP affinities, as observed with other NTPases. Furthermore, Nup159 did not accelerate ADP release, while Gle1 actually slowed ADP release independent of Mg²⁺. These findings are not consistent with Nup159 acting as a nucleotide exchange factor to promote ADP release and Dbp5 ATPase cycling. Instead, in the presence of Nup159, the interaction between Gle1 and ADP-bound Dbp5 was found to be reduced by ~ 18-fold, suggesting that Nup159 alters the Dbp5–Gle1 interaction to aid Gle1 release from Dbp5.     

Effect of eldecalcitol on articular cartilage through the regulation of transcription factor Erg in a murine model of knee osteoarthritis

Clinical studies have reported an association between low blood levels of 25-hydroxyvitamin D and the progression of osteoarthritis (OA), but the mechanism and effects of vitamin D signaling on articular chondrocytes and cartilage remains unclear. The purpose of this study was to investigate the effects of vitamin D on articular cartilage degeneration using eldecalcitol (ED-71), which is an active vitamin D₃ analog. Eight-week old male C57BL/6NCrSlc mice were subjected to experimental surgery to induce OA and local treatments with 10 μL ED-71 (0.5 μg/mL) were administered weekly. Four and 12 weeks after surgery, joints were evaluated using histological scoring systems. In addition, gene expression was analyzed in chondrocytes that were isolated from wildtype neonatal mice, cultured, and treated with ED-71 (10^?8 M). Joints treated with ED-71 demonstrated slowed progression of OA at 4 weeks after surgery, but few effects were observed at 12 weeks after surgery. Ets-related gene (Erg) expression was upregulated in OA articular cartilage, and further increased by ED-71 treatment. In primary chondrocytes cultured with ED-71, the gene expression of Erg and lubricin/proteoglycan 4 significantly increased, as compared to that of cells cultured without ED-71. Local treatment with ED-71 reduced degenerative changes to the articular cartilage during the early phase of experimental OA. Regulation of Erg by ED-71 in articular cartilage could confer resistance to early osteoarthritic changes.