脱氧雪腐镰刀菌烯醇毒理学研究进展

脱氧雪腐镰刀菌烯醇是一种单端孢霉烯族毒素，主要是由某些镰刀菌产生。它对人类和不同种属动物具有广泛的毒性效应。本文综述了脱氧雪腐镰刀菌烯醇近年的研究进展，就其生态学、污染状况、毒理学及其机制等作一简要介绍。     

谷物中脱氧雪腐镰刀菌烯醇和雪腐镰刀菌烯醇的气相色谱测定

用乙腈—水（３＋１）提取毒素，经硅镁型吸附剂柱层析和活性炭柱净化后，用具电子捕获检测器的气相色谱仪测定。当小麦和玉米中添加毒素的量为０．１～０．５ｍｇ／ｋｇ时，小麦中脱氧雪腐镰刀菌烯醇的回收率为８７％～９６％，雪腐镰刀菌烯醇为７７％～８４％，玉米中脱氧雪腐镰刀菌烯醇的回收率为９１％～１０２％，雪腐镰刀菌烯醇为９３％，精密度小于８％，方法的检测限为０．０１０ｍｇ／ｋｇ。     

河北省玉米、面粉中脱氧雪腐镰刀菌烯醇污染调查

用薄层层析法测定了105份玉米和50份小麦面粉中脱氧雪腐镰刀菌烯醇(DON)含量,结果玉米中DON检出率为61.9%,平均含量为208.8μg/kg;小麦面粉中DON捡出率为54%,平均含量为75.2μg/kg。     

脱氧雪腐镰刀菌烯醇对幼鼠关节软骨代谢影响

目的 观察脱氧雪腐镰刀菌烯醇(DON)对Wistar幼鼠关节软骨胶原合成和分解代谢的影响.方法 24只健康Wistar幼鼠,随机分为对照组、DON低剂量和高剂量组,低剂量和高剂量组隔日灌胃染毒,处理80 d后,免疫组织化学法检测软骨内Ⅱ型胶原、诱导型一氧化氮合酶(iNOS)、基质金属蛋白酶13(MMP-13)表达;ELISA法检测鼠尿液中脱氧吡啶啉(DPD)含量;电镜观察胶原纤维变化情况.结果 对照组、低、高剂量DON组Ⅱ型胶原表达量分别为(7.42±1.12)%、(4.79±0.96)%、(2.82±0.68)%,组间差异有统计学意义(P<0.05);对照组、低、高剂量DON组iNOS与MMP-13表达量分别为(6.90±1.61)%、(8.52±1.90)%、(11.78±2.51)%和(3.06±1.13)%、(6.45±1.56)%、(12.47±2.45)%,随DON浓度增高而增加(P<0.05);ELISA结果显示,DPD表达量随DON浓度增高呈上升趋势;电镜显示,胶原纤维空隙增大,稀疏,呈老化现象,网络结构破坏较明显.结论 DON可增加软骨细胞iNOS表达,提高NO合成,促进MMP-13合成并增加Ⅱ型胶原降解,进而导致软骨损伤.     

面粉加工工艺对小麦脱氧雪腐镰刀菌烯醇的影响

目的　了解加工工艺对小麦中脱氧雪腐镰刀菌烯醇 (DON)含量影响的规律 ,探讨降低面粉中DON含量的有效方法。方法　用薄层层析法[1 ] 检测小麦在加工过程不同阶段DON含量并作出分析。结果　原麦中DON含量为 36 2 0 ppb ,经净化仍保持在 344 0 ppb。制粉过程各组分DON含量在 2 90 0～ 1 86 70ppb之间。霉变小麦DON含量高达 1 4 2～ 1 4 4 .2ppm ,外观正常小麦DON含量仅为 0 .8ppm。 结论　小麦在入磨前 ,利用风力、筛理等方法将干瘪、比重轻、易碎的赤霉病麦粒去掉 ,可使面粉中DON含量得到有效控制     

脱氧雪腐镰刀菌烯醇分析方法的现状

旨在综述脱氧雪腐镰刀菌烯醇(DON)的分析方法,比较各自的优、缺点。DON的分析涉及从基质中提取、净化、分离和最终的检测等环节,分析效果包括回收率、最低检出限、时间、费用、对设备的要求和方法学上的简易程度。     

2013年中国小麦粉中脱氧雪腐镰刀菌烯醇污染调查

目的了解2013年中国小麦粉中脱氧雪腐镰刀菌烯醇(DON)的污染状况。方法从中国28个省(自治区、直辖市)的流通环节,以随机抽样的方式采集定型包装或散装小麦粉样品,用高效液相色谱法测定其DON含量,并对结果进行分析、评估。结果共检测5678份小麦粉,DON检出率为58.74%,超标率为4.60%,平均含量为317μg/kg,含量范围0~56.1 mg/kg。结论 2013年中国小麦粉中DON污染比较普遍,但超标率并不高;各地区污染程度不同,总体上华东、西南和西北地区超标率相对较高,与当地温湿度条件相关;2013年污染水平与2010年和2011年基本一致,但低于2012年监测结果。     

毛细气相色谱方法检测谷物及豆类中脱氧雪腐镰刀菌烯醇

目的：建立检测谷物及豆类中脱氧雪腐镰刀菌烯醇的毛细气相色谱方法。并且通过与酶联免疫方法的比较来探讨此方法的优势与缺点。方法：用甲醇-水（3：1）提取毒素,经过硅镁型吸附剂净化,采用DB-5MS毛细柱以及配备微池电子捕获检测器（μECD）的毛细气相色谱仪测定。结果：方法的检测限为1.78μg/kg。在阴性样品中分别添加标准品浓度为0.09、0.15、0.3μg/ml的回收率为86.84%-101.85%,RSD〈10%。结论：此方法有很好的灵敏度、精确度和准确性。并且,通过与酶联免疫方法的比较,显示出了毛细气相色谱方法的显著优势。     

安徽省谷物及制品中脱氧雪腐镰刀菌烯醇的污染调查

用薄层层析法调查厂安徽省１９８３～１９９１年小麦、玉米、大麦及面粉（标准粉）中脱氧雪腐镰刀菌烯醇（ＤＯＮ）的污染水平及其影响因隶。其分析样品６７２件，ＤＯＮ阳性率为６３．２％，平均含量为９０８．７μｇ／ｋｇ，范围在０～１３３３０μｇ／ｋｇ。其中以小麦和面粉污染比较严重，明显高于玉米中ＤＯＮ的水平（Ｐ＜０．０１）。小麦中ＤＯＮ的污染水个个同年度和地区间差异有非常显著性（Ｐ＜０．０１），经方差分析，提示与该年度和地区小麦赤霉病的流行程度有关，但玉米中ＤＯＮ污染水平不同年度和地区间差异均无显著性（Ｐ＞０．１０）。     

脱氧雪腐镰刀菌烯醇检测方法建立

脱氧雪腐镰刀菌烯醇(deoxynivalenol,DON)污染广泛存在于全球,主要污染小麦、大麦、燕麦、玉米等谷类作物,也污染粮食制品,如面包、饼干、麦制点心等。     

脱氧雪腐镰刀菌烯醇对人胚胎膝关节软骨细胞的毒性及作用机制

目的 探讨脱氧雪腐镰刀菌烯醇(DON)对人胚胎膝关节软骨细胞的毒性及作用机制.方法 取原代培养人胚胎膝关节软骨,用DMEM/F12培养基进行培养和传代,在培养基中加入DON,建立以下DON染毒浓度:0.1、0.2、0.4、1.0 μg/ml组和空白对照组.在处理72 h后进行相关研究指标的检测:分别用ELISA法检测培养上清基质金属蛋白酶13(MMP-13)、前列腺素E2(PGE2)含量,硝酸还原酶法检测上清液的一氧化氮(NO)含量,流式细胞术法检测软骨细胞凋亡情况、软骨细胞内诱导型一氧化氮合酶(iNOS)和Ⅱ型胶原表达的情况,RT-PCR法分析iNOS mRNA和Ⅱ型胶原mRNA的表达.结果 DON组凋亡率为6.78%～19.05%,高于对照组的1.20%,凋亡率随DON浓度增高而增加(F=174.761,P<0.05).DON组上清液NO含量为20.8～40.7 μmol/L,高于对照组的10.2 μmol/L(F=91.966,P<0.05);MMP-13含量为0.25～0.56 μmol/L,高于对照组的0 μmol/L(F=78.420,P<0.05);PGE2含量为3.2～20.6 μmol/L,高于对照组的1.6 μmol/L(F=276.453,P<0.05).DON组软骨细胞iNOS表达强度为14.8%～56.8%,高于对照组7.1%(F=214.614,P<0.05).高剂量DON组(0.4 μg/ml和1.0 μg/ml)Ⅱ型胶原表达强度分别为56.7%和52.7%,低于对照组的62.2%(F=5.134,P<0.05).DON组软骨细胞内iNOS mRNA表达的吸光度值(A值)为1.07～1.33,高于对照组的0.62(F=8.358,P<0.05).高剂量DON组(0.4、1.0 μg/ml)软骨细胞内Ⅱ型胶原mRNA表达量分别为0.83和0.82,低于对照组的1.14(F=7.887,P<0.05).结论 DON导致人软骨细胞合成NO增加,并通过NO调节MMP-13和PGE2等促进Ⅱ型胶原等软骨基质的降解和软骨毒性损伤;DON可促进软骨细胞的凋亡.

Abstract：

Objective This study was to explore the cytotoxic effect and the related injury mechanism of deoxynivalenol(DON)on articular chondrocytes in human embryo.Methods Articular cartilage cells were isolated from knees of human embryo and cultured in DMEM/F12 medium.The cells of the 4th generation were divided into five groups and incubated with varying concentrations of DON as the followings: control group and group with DON of 0.1,0.2,0.4,1.0 μg/ml.The effects of DON were observed 72 hours after incubation.Cell apoptosis was assayed by flow cytometry(FCM) with Annexin V-FITC/PI staining; MMP-13 and PGE2 were detected by ELISA kits;NO was measured by Griess assay with spectrophotometer.Inducible nitric oxide synthase(iNOS) and collagen Ⅱ in cells were detected by FCM.The expression levels of iNOS,mRNA and collagen Ⅱ mRNA were measured with RT-PCR.Results The rates of cell apoptosis in DON groups were 6.78%-19.05%,which were significantly higher than that in control(1.20%,F=174.761,P<0.05).The levels of NO in DON groups were 20.8-40.7 μmol/L,which were significantly higher than that in control(10.2 μmol/L,F=91.966,P<0.05).The levels of MMP-13 in DON groups were 0.25-0.56 μmol/L,which were significantly higher than that in control(0 μmol/L,F=78.420,P<0.05).The levels of PGE2 in DON groups were 3.2-20.6 μmol/L,which were significantly higher than that in control(11.6 μmol/L,F=276.453,P<0.05).The proportions of cells with positive iNOS in DON groups were 14.8%-56.8% which were significantly higher than that in controls (7.1%,F=214.614,P<0.05).The proportions of cells with positive collagen Ⅱ in groups with DON of 0.4 μg/ml and 1.0 μg/ml were 56.7% and 52.7%,which were significantly lower than that in control(62.2%,F=5.134,P<0.05).The relative absorbance values of iNOS mRNA in DON groups were 1.07-1.33,which were significantly higher than that in control(0.62,F=8.358,P<0.05).The levels of collagen Ⅱ mRNA in groups with DON of 0.4 μg/ml and 1.0 μg/ml were 0.83 and 0.82,which were significantly lower than that in control (1.14,F=7.887,P<0.05).Conclusion DON could promote anabolism of NO in articular cartilage cells by which up-regulated the expression of PGE2 and MMP-13,which both promoted resolution of articular cartilage matrix such as collagen Ⅱ.DON induced apoptosis in articular cartilage cells.     

硼对过量氟引致骨与软骨损害的治疗作用

目的　探讨硼对过量氟所致实验大鼠骨与软骨损害的治疗作用。方法　 46只 SD大鼠 ,4～ 5周龄 ,体重 5 0～ 90 g,随机分为 3组 :对照组 (饮蒸馏水 )、低过剂量氟组 (饮用含氟化钠 1 1 0 .5mg/L蒸馏水 ,F-5 0 mg/L ) ,高过剂量氟组 (饮用含氟化钠 2 2 1 mg/L蒸馏水 ,F-1 0 0 mg/L)。实验后 3个月 ,过量氟大鼠模型成功。造模后用硼治疗 ,各过剂量氟组大鼠又分为过剂量氟组与过剂量氟加硼治疗组 ,各治疗组大鼠饮水含氟量与摄氟组相同 ,同时摄取补硼饮食。治疗两个月后观察实验大鼠血清游离氟和硼含量、四肢骨氟含量、血清 NO含量 ,股骨生物力学、骨密度及大鼠肋骨、肋软骨交界处的病理形态学变化 ,肋软骨 型胶原表型表达和蛋白多糖变化。结果　 1 )硼治疗后大鼠血清和四肢骨中游离氟及血清 NO含量减少 ;2 )光镜下肋骨生长板的损伤显著恢复 ;3)和对照组及摄氟组相比 ,治疗组大鼠蛋白多糖和 型胶原表达恢复。结论　硼对氟中毒导致自由基代谢异常及软骨基质中蛋白多糖和 型胶原的异常表达有改善作用。     

氟对骨形态发生蛋白诱导软骨和骨形成的影响

为了研究氟对软骨和骨形成的影响，利用骨形态发生蛋白（ＢＭＰ）建立诱导性异位成骨模型，采用３５Ｓ、４０Ｃａ掺入率及碱性磷酸酶（ＡＬＰ）和酸性磷酸酶（ＡＣＰ）活力４项指标，反映软骨和骨的形成情况。结果显示：饮水中２００ｍｇ／Ｌ氟对软骨形成过程中的３５Ｓ掺入率无明显影响，但能使骨形成过程中的４０Ｃａ掺入率升高，氟对ＡＬＰ及ＡＣＰ活力均有刺激作用。提示氟对软骨的形成无明显影响，但对软骨内成骨和钙盐的沉积均有促进作用。     

橄榄油对大鼠去势后骨代谢指标及骨密度的影响

目的:观察橄榄油对去势后大鼠骨代谢和骨密度的影响,探讨并分析橄榄油防治绝经后骨质疏松症(PMOP)的有效性及机理。方法:将30只5～6个月龄清洁型SD雌性大鼠进行随机分成四组:①假手术组(Sham组)、②去卵巢组(OVX组)、③去卵巢+橄榄油组(OVX+Olive)、④去卵巢+雌激素组(OVX+E)。治疗组用药:(OVX+Olive)组及(OVX+E)组均采用经口灌胃方式进行用药,按1 ml/100 g体重,1次/天灌胃,连续12周。12周后分别左心室取血,检测血中雌二醇(E2)、血钙、血碱性磷酸酶(ALP)、白介素-6(IL-6)水平,放血处死后取出腰椎及左侧股骨行双能X线骨密度(DEXA)测定。结果:OVX组中血E2明显低于Sham组和治疗组,差异有统计学意义(P<0.01、P<0.05),ALP、IL-6值显著高于其余三组,差异有统计学意义(P<0.05);治疗组中E2、ALP、IL-6与Sham组差异无统计学意义(P>0.05);四组中血钙值差异均无统计学意义(P>0.05)。去势橄榄油组与雌激素组骨密度平均值均较OVX组高,差异有统计学意义(P<0.05),但该两组间差异无统计学意义(P>0.05)。结论:橄榄油能有效地减轻大鼠卵巢切除术引起的骨质丢失,可能通过补充植物雌激素发挥拟雌激素样作用。     

Oral contraceptive use and bone density in adolescent and young adult women

Background

Most of the millions of oral contraceptive (OC) users are under 30 years of age and in the critical period for bone mass accrual.

Study Design

This cross-sectional study of 606 women aged 14-30 years examined both OC duration and estrogen dose and their association with bone mineral density (BMD) at the hip, spine, and whole body (dual-energy X-ray absorptiometry).

Results

Of 389 OC users and 217 nonusers enrolled, 50% were adolescents (14-18 years). Of OC users, 38% used “low-dose” OCs [<30 mcg ethinyl estradiol (EE)]. In adolescents, mean BMD differed by neither OC duration nor EE dose. However, 19- to 30-year-old women's mean BMD was lower with longer OC use for spine and whole body (p=.004 and p=.02, respectively) and lowest for >12 months of low-dose OCs for the hip, spine and whole body (p=.02, .003 and .002, respectively).

Conclusions

Prolonged use of today's OCs, particularly <30 mcg EE, may adversely impact young adult women's bone density while using these agents.     

Correlates of use of long-acting reversible methods of contraception among adolescent and young adult women

Background

Most pregnancies among adolescent and young adult women are unintended, and adolescent birth rates have risen. Use of long-acting reversible contraception may be an effective strategy to reduce the rate of unintended pregnancy.

Study Design

We conducted a secondary data analysis of nationally representative, cross-sectional data from the 2002 National Survey of Family Growth. Our sample included 1722 sexually active women aged 15-24 years. We used multivariable logistic regression to identify correlates of ever-use of depot-medroxyprogesterone acetate (DMPA) or the intrauterine device (IUD).

Results

One-quarter of our sample had ever used DMPA, and less than 2% had ever used the IUD. In multivariable analysis, increasing parity was associated with ever-use of DMPA (OR 2.07, 95% CI 1.55-2.77) and ever-use of the IUD (OR 4.57, 95% CI 1.60-13.03), but age and measures of socioeconomic status were not. Having ever been married (OR 5.54, 95% CI 1.23-24.82) and current cohabitation (OR 4.89, 95% CI 1.10-21.71) were associated with ever-use of the IUD. A history of an adolescent pregnancy was associated with ever-use of DMPA (OR 1.79, 95% CI 1.19-2.70) but not of the IUD.

Conclusions

While similarities exist between the correlates of use of DMPA and the IUD, we discovered important differences, some of which may reflect provider biases regarding IUD provision.     

甲醛致大鼠离体骨髓细胞DNA-蛋白质交联的研究

目的检测经不同浓度甲醛染毒后所致大鼠骨髓细胞的DNA-蛋白质交联程度。方法采用KCl-SDS沉淀法检测液态甲醛所致DNA-蛋白质交联（DPC）效应。结果低浓度的甲醛（10μmol／L和50μmol／L）能引起DNA-蛋白质的交联;较高浓度的甲醛（250μmol／L和1250μmol／L）可以产生明显的DNA-蛋白质交联作用（P〈0．01）。结论甲醛可造成骨髓细胞DNA-蛋白质交联,而DPC可以对细胞产生严重的遗传毒性,这可能是甲醛导致白血病的分子机制之一。     

燃煤污染型氟中毒人群尿氟与骨代谢损伤指标相关关系及其基准剂量分析

目的将基准剂量法应用于燃煤氟暴露人群,探讨燃煤氟污染致暴露人群骨代谢损伤的生物暴露限值,为燃煤氟致骨损伤的防治提供参考依据。方法以贵州省燃煤污染型氟中毒病区295例氟暴露者为氟暴露组,非氟污染区85例居民为对照组。在知情同意原则下,采集调查对象尿液和血液样本,采用氟离子电极法测定尿氟含量,以酶联免疫吸附试验测定血清骨钙素含量,以微量酶标法检测碱性磷酸酶活力。分析尿氟与骨代谢损伤指标之间关系,并应用BMDS 2.6.0.1软件计算基准剂量(BMD)、95%的可信限下限值(BMDL)。结果按尿氟分组后,随着尿氟浓度的升高,血清骨钙素和碱性磷酸酶水平逐渐升高,存在剂量-效应关系,差异有统计学意义(P0.05)。骨钙素与碱性磷酸酶的尿氟BMD与BMDL分别为0.581、0.412 mg/g Cr和2.374、1.362 mg/g Cr。结论建议燃煤氟暴露引起人群骨代谢损伤的尿氟生物暴露限值为0.412 mg/g Cr;骨钙素可作为氟暴露早期效应指标和氟接触高危人群的敏感筛选指标。