米非司酮对早孕小鼠促性腺激素细胞的形态学影响

研究米非司酮终止早孕时小鼠垂体促性腺激素细胞——FSH细胞、LH细胞的形态学变化。方法:应用免疫组织化学PAP方法,对FSH细胞与LH细胞的阳性细胞密度,细胞质内激素颗粒进行观察。结果:实验组与对照组比较,FSH细胞的阳性细胞密度,细胞质内FSH颗粒无显著性差异;LH细胞的阳性细胞密度,细胞质内LH颗粒均明显减少,有显著性差异(P<0.05)。结论:米非司酮对早孕小鼠的FSH细胞的功能无明显影响;对LH细胞的功能有显著的抑制作用。     

DahlS高血压大鼠卵巢的细胞凋亡研究

目的　通过对DahlS高血压大鼠卵巢颗粒细胞及黄体细胞凋亡的初步研究 ,以期阐明高血压时卵巢的细胞凋亡特征。方法　以日本引进DahlS高血压大鼠为研究对象 ,选用生后 5周的DahlS雌性大鼠随机分为实验组、对照组 ,分别投予 8%食盐饲料、正常饲料 ,于 11、13、15周龄取材。应用光镜 ,电镜 ,原位末端标记试剂盒(TUNELBox) ,流式细胞等技术 ,对凋亡的颗粒细胞及黄体细胞进行定性、定位及定量。结果　随高血压病程延长、病情加重 :⑴光镜下观察实验组生长卵泡中颗粒细胞排列不整齐 ,松散 ,闭锁卵泡数目增多 ,黄体数目减少。⑵TUNEL染色可见凋亡在生长卵泡的颗粒细胞中均有发生 ,闭锁卵泡颗粒细胞凋亡最显著 ,黄体细胞未见TUNEL阳性细胞。⑶电镜下观察颗粒细胞及黄体细胞凋亡的特征为核周间隙增宽 ,核固缩 ,染色质边集 ,凝聚 ,线粒体嵴模糊紊乱 ,腔内出现高电子密度异常颗粒 ,滑面内质网减少 ,类脂呈板层状改变。⑷流式细胞检测显示卵巢细胞凋亡率逐渐上升 ,13、15周龄实验组凋亡率分别为 (6 4 2± 1 0 6 ) %、(6 6 7± 2 11) %均明显高于对照组(t =3 0 2 7、3 0 85 ,P <0 0 1)。⑸卵巢细胞凋亡率与血压升高呈正相关 (γ =0 76 8,P <0 0 1)。结论　高血压能加重卵巢的细胞凋亡并有可能导致生殖功能衰退     

甲基对硫磷对小鼠黄体细胞的细胞间隙连接通讯和孕酮分泌的影响

目的 探索甲基对硫磷(methyl parathion,MP)对小鼠黄体细胞的细胞间隙连接通讯(gapjunctional intercellularcommunication,GJIC)和孕酮分泌的影响.方法 体外培养5周龄SPF级BALB/c小鼠黄体细胞,分别设MP暴露组(0、0.05、0.1、0.2、0.4、0.8、1.6 mmol/L的MP处理3h)和GJIC促进剂——1 μmol/L异丙肾上腺素预处理组及蛋白激酶A(PKA)专一抑制剂——10 μmol/L H89预处理组与蛋白激酶C(PKC)专一抑制剂——10 μmol/L恩扎妥林预处理组(预处理5 min,0.8 mmol/L MP处理3h).采用ELISA法检测培养液中孕酮含量,划痕标记染料转移法测定贴壁细胞的GJIC.结果 高浓度(0.2～1.6 mmol/L)MP可明显抑制小鼠黄体细胞的GJIC和孕酮分泌(P＜0.05);异丙肾上腺素、H89和恩扎妥林均能缓解MP对体外小鼠黄体细胞的GJIC和孕酮分泌的抑制作用(P＜0.05).结论 MP可通过抑制小鼠黄体细胞的GJIC导致孕酮分泌受抑制,而PKA与PKC参与该过程.     

全肠外营养相关肝损害小鼠模型的建立

目的:为进一步研究肠外营养(PN)相关肝损害(PNALD)的患病机制及干预措施,建立有效的小鼠模型。方法:将20只小鼠随机分为对照组和模型组,对照组小鼠正常饮食、颈静脉置管后微量泵持续输注等渗盐水;模型组小鼠禁食后颈静脉置管,用微量泵持续输注PN液。1周后比较两组小鼠体重变化,血清生化指标以及肝组织学改变。结果:模型组小鼠体重明显低于对照组(P<0.05),直接胆红素、总胆红素和胆固醇均显著高于对照组(P<0.05),且肝组织在光镜下可见广泛性肝细胞脂肪变性,细胞质内出现大小不一的空泡,主要集中于中央静脉周围。脂肪变性评分为(3.1±0.5)分,显著高于对照组(1.0±0.0)分。结论:模型组小鼠与成人PNALD病人初期的临床和病理改变相似,可用于该病的患病机制,药物治疗疗效以及具体机制的观察和研究。     

茵陈蒿油对小鼠急性酒精性肝损伤的保护作用

目的:研究茵陈蒿油对小鼠急性酒精肝损伤的影响.方法:小鼠一次性经口给予50%乙醇,建立急性酒精性肝损伤模型,测定丙二醛(MDA)、还原性谷胱甘肽(GSH)和甘油三酯(TG)在肝匀浆中的含量,计算肝脏指数,光镜下观察肝组织病理学变化.结果:茵陈蒿油使酒精肝损伤小鼠的MDA含量明显下降,GSH含量明显升高,TG含量下降,明显改善肝组织损伤的程度,与模型组比较差异有统计学意义(P＜0.01).结论:茵陈蒿油对酒精肝损伤具有一定的保护作用.     

无水化处理在病理组织HE染色中的应用价值

目的探讨无水化处理在病理组织HE染色中的应用价值。方法以结肠组织标本为材料,制作切片。对照组给予常规染色处理操作,观察组在染色前对切片进行无水化处理,比较两组切片的质量。结果观察组切片染色效果较好,镜下可见细胞核为深蓝色着色,且着色细腻;可明显分辨核仁与核膜;细胞质与细胞核着色对比清晰,细胞质为鲜红色,细胞核为深蓝色;未见苏木精沉渣。对照组切片染色效果较差,细胞核与细胞质染色区分不清晰,细胞核呈淡蓝色,细胞质为淡红色;镜下可见较多苏木精沉渣。结论经无水化处理后,组织切片HE染色的质量明显提高,具有重要的应用价值。     

米非司酮治疗更年期功能性子宫出血疗效观察

目的:观察米非司酮治疗更年期功能性子宫出血的临床疗效.方法:60例更年期功能性子宫出血患者口服米非司酮25mg,持续3个月,观察其临床疗效和血清雌二醇(E2)及孕酮(P)水平变化、子宫内膜厚度变化变化情况.结果:治疗后其有效率85%:治疗后E2、P与治疗前相比,有统计学差异(P＜0.05);子宫内膜厚度治疗前后比较有显著性差异(P＜0.05).结论:米非司酮治疗更年期功能性子宫出血效果满意,值得推广应用.     

微管相关蛋白tau在小鼠卵巢颗粒细胞中的定位及功能

目的:探究小鼠卵巢颗粒细胞中微管相关蛋白的表达及功能定位。方法:选取雌性小鼠发生6周成年(成年组,6只)与未成年(未成年组,6只),培养卵巢颗粒细胞,将加入促卵泡刺激素(FSH)培养的颗粒细胞作为FSH组,不加FSH培养的颗粒细胞作为非FSH组。免疫组化染色法观察颗粒细胞中微管相关蛋白tau表达,将tau融合蛋白转染到颗粒细胞上,荧光显微镜下观察tau蛋白在颗粒细胞中的定位。结果:小鼠卵巢颗粒细胞tau蛋白表达阳性数成年组高于未成年组,FSH组高于非FSH组(P0.05);未成年小鼠中FSH组阳性数高于非FSH组(P0.05)。成年组小鼠卵巢颗粒细胞中tau蛋白聚集于细胞核,呈放射状。结论:微管相关蛋白tau在成年小鼠卵巢颗粒细胞中高表达,其定位主要集中于细胞核,影响颗粒细胞增殖分化,通过观察tau蛋白表达及定位,可反映微管形成及形态变化。     

滤泡型甲状腺乳头状癌的针吸细胞学诊断探讨

目的 探讨滤泡型甲状腺乳头状癌(FVPTC)的针吸细胞学(FNAC)特点及其鉴别诊断.方法 对12例FVPTC患者行FNAC检查,HE染色,光镜观察其滤泡细胞数量、细胞排列形态、细胞核的形态、细胞质的改变以及涂片背景特点.结果 本组12例中FNAC诊断为甲状腺乳头状癌7例,其中考虑为FVPTC亚型3例,误诊为滤泡性肿瘤5例.诊断为甲状腺乳头状癌的敏感度为58.3%(7/12),诊断为FVPTC的敏感度为25.0%(3/12).复习后FNAC诊断为FVPTC 11例,敏感度为91.7%(11/12).FVPTC的FNAC特点为以微滤泡团和单层合体团为主,单个散在细胞较少,裸核细胞极少.细胞核普遍较大,核不规则和核沟均易见,包涵体多数为偶见,核膜较厚.细胞质宽呈淡嗜酸性,背景容易见到浓缩的胶质球和单核巨噬细胞.结论 FVPTC具有经典型甲状腺乳头状癌的细胞核特点,但通常不够典型,容易误诊为滤泡性肿瘤.综合分析细胞排列、细胞核形态、细胞质的改变和涂片背景的变化,有助于FVPTC的FNAC诊断和鉴别诊断.     

野生牛肝菌保健功能研究

目的观察野生牛肝菌对昆明种小鼠免疫功能的影响和对SD大鼠的降血脂作用。方法 (1)免疫功能实验:实验设野生牛肝菌组9.0 g/(kg bw)+环磷酰胺(CP)、CP组、正常对照组。灌胃小鼠牛肝菌提取液35 d,CP腹腔注射造成小鼠免疫损伤。观察小鼠胸腺脏器系数、血清溶血素、小鼠足趾DTH、碳粒廓清指数α等指标。(2)降血脂实验:设3个野生牛肝菌剂量组4.0、8.0、16.0 g/(kg bw)、阴性对照组和高脂模型对照组。各剂量组大鼠灌胃牛肝菌提取液30 d,对照组灌胃等量蒸馏水,在试验结束时取大鼠血清测定TG、CHO、LDH 3个指标。结果 (1)免疫功能试验中免疫损伤的小鼠血清溶血素(HC50)、小鼠足趾DTH、碳粒廓清指数α明显高于CP组,差异有统计学意义(P<0.01),其中小鼠足趾DTH恢复至正常对照组水平。(2)各剂量组的血脂指标与高脂对照组差异无统计学意义(P>0.01)。结论在本实验条件下,野生牛肝菌对CP所致的免疫损伤小鼠细胞免疫、体液免疫、非特异性免疫功能有明显改善作用,但不能预防大鼠的高脂血症。     

米非司酮用于排卵期房事后避孕的子宫内膜形态学观察

为了解米非司酮用于排卵期房事后避孕引起的子宫内膜形态学改变,选择21名健康育龄妇女做为受试者,于排卵后72小时内口服300mg米非司酮。排卵后7～8天取内膜做常规光镜下观察。结果发现所有内膜改变都与原周期时相不同步。说明排卵后服用米非司酮使内膜所发生的形态学改变足以防止胚胎种植,故可作为一月一次排卵期房事后有效的避孕药物。     

EURRECA: EURopean micronutrient RECommendations Aligned. Preparing the way

Background There is increasing evidence that higher intakes of carotenoids could protect against oxidative and light damage in premature infants and may promote other health benefits in both mothers during pregnancy and lactation and in newborn infants. Aim of the study To develop and validate a brief quantitative food frequency questionnaire (FFQ) aimed at assessing lutein and zeaxanthin intake in women. Methods In this cross-sectional study, estimates of lutein and zeaxanthin intake from the FFQ were compared with a 7-day dietary record and with plasma concentrations of these carotenoids. This primary care study was conducted in Pavia, Italy. Subjects were all female volunteers, aged 20–25 years (mean age 22.7 ± 2.1 years), university students. Of the 110 women initially recruited, 87 completed diet questionnaires and donated a blood sample. Dietary intake was assessed by the FFQ by interview and 7-day dietary records chosen as a reference standard, using photographic estimations of portion sizes. Plasma concentrations of lutein and zeaxanthin were measured by HPLC. Pearson’s correlation coefficient and Bland Altman Regression analysis were used. Results Mean dietary lutein and zeaxanthin intakes were 1,107 ± 113 μg/day from the FFQ questionnaire and 1,083 ± 116 μg/day from the 7 day dietary records. The mean difference in intake assessed by the two methods (−24.5 ± 38.3 μg/day) did not differ significantly from zero. Dietary intake of lutein and zeaxanthin measured with the FFQ and plasma nutrient concentration among this sample were significantly correlated (r = 0.76, P < 0.0001). Mean plasma lutein and zeaxanthin concentrations were 0.33 ± 0.09 μmol/l. Conclusions This FFQ could be used to assess lutein and zeaxanthin intake in adult women.     

叶黄素对D-半乳糖致小鼠氧化损伤的保护作用

目的研究不同剂量叶黄素对D-半乳糖(D-gal)小鼠氧化应激损伤的影响。方法44只昆明种小鼠随机分为D-gal模型对照组、叶黄素低剂量组、叶黄素高剂量组和正常对照组。D-gal120 mg/(kg.d)腹腔注射制作小鼠氧化应激模型,叶黄素低、高剂量组分别灌胃给予叶黄素10 mg/(kg.d)、40 mg/(kg.d),持续6周。检测小鼠肝组织活性氧(ROS)、丙二醛(MDA)、一氧化氮(NO)含量,超氧化物歧化酶(SOD)、总一氧化氮合酶(TNOS)、诱导型一氧化氮合酶(iNOS)、线粒体ATP酶(ATPase)活性和血清ALT、AST水平。结果叶黄素各剂量组小鼠肝组织MDA、ROS含量低于模型对照组;高剂量组SOD活性高于模型对照组,TNOS、iNOS活性低于模型组,肝线粒体Na+-K+-ATP酶活性高于模型组,差异均有统计学意义(P0.05)。结论叶黄素可降低D-gal致氧化应激小鼠体内ROS和NOS活性,提高SOD和线粒体Na+-K+-ATP酶活性,从而缓解氧化应激损伤。     

Eicosapentaenoic acid inhibits the growth of liver preneoplastic lesions and alters membrane phospholipid composition and peroxisomal beta-oxidation

This study investigates the effect of dietary carotenoids on pim-1 gene expression in mouse splenocytes. Female BALB/c mice were fed 0%, 0.02%, or 0.4% astaxanthin, beta-carotene, and lutein for two weeks. Plasma and liver were obtained for the analysis of carotenoids. Splenocytes were isolated and cultured in the presence of concanavalin A, and the level of pim-1 mRNA was determined by Northern blot analysis. None of the carotenoids were detectable in the plasma and liver of unsupplemented mice. In plasma the concentration of astaxanthin (4.9-54.7 mumol/l) was dramatically higher than that of lutein (1.4-2.0 mumol/l) and beta-carotene (0.1-0.7 mumol/l). Carotenoid uptake by the spleen but not the liver reflected that observed in plasma. In mice fed 0.4% of each carotenoid, the absolute concentration of the carotenoid in the liver was highest for astaxanthin (24 nmol/g) followed by beta-carotene (7.5 nmol/g) and lutein (1.58 nmol/g). Mice fed lutein showed a dose-related increase in pim-1 mRNA expression. The steady-state level of pim-1 mRNA in mice fed 0.4% lutein was sixfold higher than in mice fed 0.02% lutein. In contrast, dietary astaxanthin and beta-carotene did not affect pim-1 expression. Therefore, an increase in pim-1 mRNA was observed in splenocytes stimulated with concanavalin A in lutein-fed mice. This appears to be a unique effect of lutein and may be associated with its antitumor activity observed in vivo.     

超量米非司酮对大鼠卵巢细胞微细结构的作用

目的:光、电镜观察超量米非司酮对大鼠卵巢中卵泡、间质腺等细胞微细结构的改变。方法:给实验大鼠米非司酮10mg/只/d,分别用药7、14、28d,取材,制备标本,光、电镜观察。结果:多数卵泡在生长卵泡期间闭锁、部分闭锁卵泡形成间质腺,腺细胞内含大量脂滴、空泡,散在的溶酶体,线粒体包统脂滴,有板层体出现,巨噬细胞内出现巨大包含体,超微病理改变随用药时间延长有加重趋势。结论:超量米非司酮对卵泡发育有抑制作用,对卵泡的细胞、间质腺细胞有一定的损害。     

Changes in vaginal morphology, steroid receptor and natural antimicrobial content following treatment with low-dose mifepristone

BACKGROUND: We have previously shown that the antigestagen mifepristone is contraceptive when given in a daily dose of 5 mg, po. Epidemiological studies suggest that gestagen-only contraceptives may increase the risk of transmission of human immunodeficiency virus (HIV) due to effects on the vaginal defenses to infection. We investigate the effects of mifepristone on vaginal thickness, steroid receptor and natural antimicrobial content and pharmacokinetics of mifepristone. METHODS: In a pilot study, eight women were given mifepristone 5 mg/day for an average of 33 days. Ovarian function was assessed by measurement of estradiol and progesterone in blood and their metabolites in urine and by serial ultrasound of their ovaries. Vaginal biopsies were collected before (late proliferative) and after taking mifepristone. RESULTS: All subjects showed a similar pattern of descending serum concentrations of mifepristone. The elimination phase half-life was 18+/-5.1 h (mean+/-SD). Mean Cmax measured at 1 h was 641.7 nmol/L (range, 502-740 nmol/L). All eight women reported amenorrhea for the duration of treatment and seven of eight women showed biochemical and ultrasound evidence of anovulation. There was no significant change in vaginal thickness following treatment [342+/-40 microm pretreatment, 303+/-69 microm posttreatment (mean+/-SEM); p>.05]. Estrogen (ERalpha, ERbeta) and androgen receptor were expressed in both vaginal epithelium and subepithelial stroma, whereas progesterone receptor was expressed predominantly in the subepithelial stroma. There was no change in receptor content and distribution following mifepristone treatment. Natural antimicrobial mRNA [secretory leukocyte protease inhibitor, human beta defensins mRNA (HBD1, HBD2, HBD3, HBD5), granulysin and elafin] was extracted from the vaginal tissues, and the content was unaffected by mifepristone treatment. CONCLUSION: The absence of changes in vaginal thickness, steroid receptor and natural antimicrobial content and its distribution in this preliminary study suggests that in contrast to other estrogen-free contraceptives, mifepristone is unlikely to be associated with the increased risk of transmission of HIV and other sexually transmitted infections.     

不同浓度米非司酮对体外培养子宫肌瘤细胞周期的影响及其诱导的细胞凋亡

目的:研究不同浓度米非司酮对体外培养子宫肌瘤细胞凋亡的影响。方法:采用体外培养的子宫肌瘤细胞,用不同浓度米非司酮处理后培养48h,用流式细胞仪分析子宫肌瘤细胞周期和凋亡率的改变。结果:不同浓度的米非司酮均可诱导子宫肌瘤细胞的凋亡,随着米非司酮浓度的增加,子宫肌瘤细胞的凋亡率有增高的趋势(P<0·05),低浓度米非司酮(10-8~10-7mol/L)可使G2/M期子宫肌瘤细胞比例减少(P<0·05),高浓度米非司酮(10-6~10-4mol/L)可使S期细胞比例减少(P<0·05)和G2/M期细胞比例减少(P<0·01)。结论:米非司酮可引起体外培养子宫肌瘤细胞的凋亡,呈现浓度-效应关系,低浓度米非司酮和高浓度米非司酮可能引起不同时期子宫肌瘤细胞的凋亡。     

Carotenoid composition of human milk during the first month postpartum and the response to beta-carotene supplementation

BACKGROUND: Information is lacking regarding normal changes in milk carotenoid concentrations in healthy, well-nourished women during the first month of lactation. OBJECTIVES: This study investigated milk carotenoid concentrations during days 4-32 postpartum and assessed the effects of maternal beta-carotene supplementation. DESIGN: Subjects (n = 21; aged 19-39 y) were randomly assigned to receive beta-carotene (30 mg/d) or placebo from days 4 to 32 postpartum. Each subject provided 8 diet records and 8 milk samples during the study. Diet records were analyzed for energy, macronutrients, vitamins A and E, and carotenoids. Milk samples were analyzed with HPLC for concentrations of carotenoids, retinol, and alpha-tocopherol. Data were analyzed by using repeated-measures analysis and orthogonal contrasts. RESULTS: No significant differences in average dietary intakes, body mass index, age, or parity were found between groups at baseline or after supplementation. Milk carotenoid concentrations decreased over time (P < 0.01), as did retinol and alpha-tocopherol concentrations (P < 0.003). Concentrations of most carotenoids decreased to those reported for mature milk by day 32 postpartum. Milk lutein concentrations remained elevated throughout the study compared with values reported for mature milk, whereas plasma lutein concentrations decreased significantly over time. beta-carotene supplementation did not significantly change the milk concentrations of beta-carotene, the other carotenoids, retinol, or alpha-tocopherol. CONCLUSIONS: The lack of increase in milk beta-carotene despite supplementation suggests that transitional milk may be already nearly saturated with beta-carotene. The elevated milk lutein concentration and simultaneous decrease in plasma lutein suggest that lutein metabolism may be altered during early lactation.     

Lutein and beta-carotene from lutein-containing yellow carrots are bioavailable in humans

BACKGROUND: Lutein is a hydroxy-carotenoid constituting the macular pigment of the human retina. Increasing lutein intake from foods could increase the density of this pigment and decrease the risk of developing macular degeneration. Yellow carrots are a novel food source that could increase lutein consumption. OBJECTIVE: We evaluated and compared lutein uptake and clearance in humans from genetically selected lutein-containing yellow carrots fed chronically and from a lutein supplement. DESIGN: Four women and 5 men aged 23-28 y participated in this randomized, blinded, 3 x 3 crossover intervention. Treatments consisted of yellow carrots (YC treatment, 1.7 mg lutein/d), white carrots as a negative control (WC treatment, 0 mg lutein/d), and a lutein supplement in oil as a positive control (LS treatment, 1.7 mg lutein/d). Each treatment lasted 7 d and was followed by a 7-d washout period. RESULTS: Mean (+/- SD) peak changes in serum lutein concentration from baseline were 0.31 +/- 0.08, 0.19 +/- 0.08, and -0.04 +/- 0.04 micromol/L for the LS, YC, and WC treatments, respectively. The areas under the curve for 0-14 d (AUC(0-14d)) differed significantly (P < 0.0001) between treatments. Lutein from the YC treatment was 65% as bioavailable as that from the LS treatment. The AUC(0-14d) for beta-carotene (-0.01 +/- 0.28 micromol.d/L) also showed that the YC treatment maintained peak serum beta-carotene concentrations at 0.35 +/- 0.30 micromol/L, whereas the LS treatment did not (AUC(0-14d) = -0.71 +/- 0.59 micromol.d/L). CONCLUSION: Lutein from this novel food source significantly increases serum lutein concentrations and does not result in the decrease in beta-carotene concentrations that accompanies administration of lutein supplements.