桑色素对小鼠T淋巴细胞体外活化、增殖和细胞周期的影响

目的:研究桑色素(morin)对小鼠T淋巴细胞活化、增殖和细胞周期的影响。方法:以刀豆蛋白A(ConA)刺激培养的淋巴结来源的小鼠淋巴细胞,再以不同终浓度的morin与T细胞共培养,利用流式细胞术(FCM),检测早期T细胞活化的标志CD69分子的表达,以羧基荧光素双醋酸盐琥珀酰脂(CFDA-SE)染色检测T细胞的增殖;以碘化丙锭(PI)染色分析T细胞的细胞周期。结果:小鼠T细胞培养6 h后,未经ConA刺激的对照组中CD69 T的细胞比率为(2.97±0.12)%,经ConA刺激的CD69 T细胞的比率明显增高,达到(72.52±0.66)%,与对照组相比差别明显(P<0.01)。终浓度为25、50、100μmol/L的morin均下调CD69 T细胞的比率,其中,100μmol/L的morin抑制作用最强,为(48.95±0.81)%,与对照组比较具有统计学意义(P<0.01)。CFDA-SE染色分析显示,ConA组培养48 h和72 h的T细胞的增殖指数(PI)分别为(1.58±0.04)和(1.95±0.02),各浓度的morin对ConA刺激的T细胞增殖,具有明显地抑制作用,以100μmol/L的morin抑制作用最明显。培养48 h的ConA组T细胞的PI为(1.02±0.02)、培养72 h的ConA组T细胞的PI为(1.03±0.01),与相应时间的对照组比较,均有统计学意义(P<0.01)。PI染色后流式细胞术分析的结果表明,ConA组处于S期的T细胞的比率为(27.05±0.39)%,显著高于对照组的比率(5.10±0.07)%。morin组中S期的细胞比率较高。结论:Morin可显著抑制ConA刺激的T细胞活化及增殖;其对增殖的抑制作用主要表现为S期的细胞的阻滞。     

PRL-2基因转染对肝细胞增殖及细胞周期的影响

目的：研究PRL-2基因对肝细胞增殖和细胞周期的影响。方法:采用脂质体转染的方法将重组质粒稳定转染至正常永生化肝细胞系CL1中，G418筛选阳性克隆。应用实时荧光定量聚合酶链反应、Western印迹和免疫组化分析PRL-2在阳性细胞的表达及蛋白定位，MTT法检测细胞的群体倍增时间，流式细胞仪检测细胞周期变化，Western印迹分析细胞周期素A、D1、E以及周期蛋白依赖激酶抑制因子p16、p21WAF1及p27Kip1的变化,实时荧光定量聚合酶链反应检测p21WAF1mRNA的变化。结果: 成功构建PRL-2真核表达载体pcDNA3-PRL-2。脂质体转染细胞经过G418筛选后,获得稳定表达PRL-2的细胞亚系PRL-2-CL1。经实时荧光定量PCR、Western印迹和免疫组化证实,PRL-2-CL1细胞系PRL-2基因及蛋白表达水平高于对照组，流式细胞仪检测细胞周期S期细胞比例明显增高，MTT法检测细胞群体倍增时间缩短。Western印迹及实时荧光定量聚合酶链反应显示p21WAF1在转染后较对照组明显降低，细胞周期素A、D1、E及p16、p27Kip1无明显变化。结论: 重组PRL-2真核表达载体构建正确,并在永生化肝细胞中获得稳定、高效表达。PRL-2基因具有促进细胞增殖的作用，这种作用与其降低p21WAF1蛋白含量相关。     

Superior cycle control with a contraceptive vaginal ring compared with an oral contraceptive containing 30 microg ethinylestradiol and 150 microg levonorgestrel: a randomized trial

BACKGROUND: This trial was conducted to compare cycle control with vaginal ring a combined contraceptive vaginal ring, and a combined oral contraceptive (COC) delivering 30 mug ethinylestradiol (EE) and 150 mug levonorgestrel. METHODS: This open-label, randomized, multi-centre, Phase III study involved adult women from 11 countries. Subjects were treated with either vaginal ring or a COC for 13 cycles (12 months). RESULTS: A total of 1030 subjects (vaginal ring, n=512; COC, n=518) comprised the intention-to-treat (ITT) population. The percentage of women in the ITT population who completed the trial was 70.9% for vaginal ring and 71.2% for the COC group. The incidence of breakthrough bleeding and spotting over cycles 2-13, the primary efficacy parameter, was lower with vaginal ring (range 2.0-6.4%) than the COC (range 3.5-12.6%), and for cycles 2 and 9 the lower incidence with vaginal ring was confirmed as statistically significant (P=0.003 and P=0.002 respectively). The incidence of intended bleeding was significantly higher over all cycles with vaginal ring (58.8-72.8%) than with the COC (43.4-57.9%). CONCLUSIONS: Cycle control with vaginal ring was excellent and superior to that of a COC containing 30 mug EE.     

Auditory brainstem response in premenopausal women taking oral contraceptives

BACKGROUND: The aim of this prospective study was to evaluate the effects of the new monophasic oral contraceptives on the audiological system in premenopausal women. METHODS: The auditory brainstem response (ABR) was measured in 94 women during the follicular, periovular and luteal phases of one menstrual cycle in which ovulation was confirmed using sonography and serum progesterone concentration. The latencies for waves I, III and V were determined, and the inter-peak intervals were calculated for waves I-III, I-V and III-V. All 94 women began taking oral contraceptives: 23 women used 20 microg ethinyl estradiol (EE) plus 150 microg desogestrel, 24 women used 30 microg EE plus 75 microg gestodene, and 47 women used 15 microg EE plus 60 microg gestodene. During the third month of contraceptive intake, each subject was again tested for ABR, as above. RESULTS: The wave latencies and inter-peak intervals showed shorter values during the periovular phase with respect to the luteal phase (P < 0.05), the follicular phase for wave I and for inter-peak interval I-V (P < 0.05) of the menstrual cycle. All of the ABR results in pill users were statistically different from those of the periovular phase (P < 0.05), though similar to those of both the luteal and follicular phases (P = NS). CONCLUSIONS: ABR seems to depend on the variations of ovarian steroids during the menstrual cycle and during oral contraceptive intake.     

互隔交链孢霉提取物对淋巴细胞核仁形成区及细胞增殖周期的影响

本实验应用人类核仁形成区(NORs)选择性银染方法和细胞光度术,观察了互隔交链孢霉两菌株提取物261-B_2-3和C_(12)b_3-2对人淋巴细胞NORs活性的影响以及后者对细胞增殖周期移行的影响。结果显示,应用不同浓度的261-B_2-3和C_(12)b_3-2提取物处理细胞后,细胞携带Ag-NORs的染色体数明显减少,与对照组比较差异极显著(P<0.01)。表明两株提取物能明显抑制淋巴细胞NORs的活性。细胞暴露于不同浓度的C_(12)b_3-2提取物后,G_1%明显增多,S%和G_2+M%显著减少,与对照相比差异极显著(P<0.01)。说明C_(12)b_3-2提取物可能作用于G_1后期或S早期,延缓和阻滞了细胞周期的移行。     

<Emphasis Type="Italic">Saccharomyces cerevisiae</Emphasis> Ats1p interacts with Nap1p,a cytoplasmic protein that controls bud morphogenesis

Saccharomyces cerevisiae ATS1 (-tubulin suppressor 1) was originally identified as a high-copy suppressor of class two -tubulin mutations and was proposed to have a regulatory role in coordinating the microtubule state with the cell cycle. Here, we show that Ats1p interacts with Nap1p, a cytoplasmic protein that regulates the activity of the Cdc28p/Clb2p complex. Loss of Nap1p results in a delayed switch from polar to isotropic bud growth. The delayed switch results in elongated buds. Nap1p and Ats1p interact in two-hybrid and co-immunoprecipitation assays. Both nap1 and ats1 cells have a Clb2p-dependent elongated bud morphology. Deletion of ATS1 partially suppresses the elongated bud morphology and benomyl resistance of nap1 mutants. Our results suggest Ats1p might regulate coordination of the microtubule state with the cell cycle through an interaction with Nap1p.Communicated by S. Hohmann     

Hepatic gluconeogenesis and Krebs cycle fluxes in a CCl4 model of acute liver failure

Acute liver failure was induced in rats by CCl4 administration and its effects on the hepatic Krebs cycle and gluconeogenic fluxes were evaluated in situ by 13C NMR isotopomer analysis of hepatic glucose following infusion of [U-13C]propionate. In fed animals, CCl4 injury caused a significant increase in relative gluconeogenic flux from 0.80+/-0.10 to 1.34 +/-0.24 times the flux through citrate synthase (p<0.01). In 24-h fasted animals, CCl4-injury also significantly increased relative gluconeogenic flux from 1.36+/-0.16 to 1.80+/-0.22 times the flux through citrate synthase (p<0.01). Recycling of PEP via pyruvate and oxaloacetate was extensive under all conditions and was not significantly altered by CCl4 injury. CCl4 injury significantly reduced hepatic glucose output by 26% (42.8+/-7.3 vs 58.1+/-2.4 micromol/kg/min, p=0.005), which was attributed to a 26% decrease in absolute gluconeogenic flux from PEP (85.6+/-14.6 vs 116+/-4.8 micromol/kg/min, p<0.01). These changes were accompanied by a 47% reduction in absolute citrate synthase flux (90.6+/-8.0 to 47.6+/-8.0 micromol/kg/min, p<0.005), indicating that oxidative Krebs cycle flux was more susceptible to CCl4 injury. The reduction in absolute fluxes indicate a significant loss of hepatic metabolic capacity, while the significant increases in relative gluconeogenic fluxes suggest a reorganization of metabolic activity towards preserving hepatic glucose output.     

Single-equation models for the tear film in a blink cycle: realistic lid motion.

We consider model problems for the tear film over multiple blink cycles that utilize a single equation for the tear film; the single non-linear partial differential equation that governs the film thickness arises from lubrication theory. The two models that we consider arise from considering the absence of naturally occurring surfactant and the case when the surfactant is strongly affecting the surface tension. The film is considered on a time-varying domain length with specified film thickness and volume flux at each end; only one end of the domain is moving, which is analogous to the upper eyelid moving with each blink. Realistic lid motion from observed blinks is included in the model with end fluxes specified to more closely match the blink cycle than those previously reported. Numerical computations show quantitative agreement with in vivo tear film thickness measurements under partial blink conditions. A transition between periodic and non-periodic solutions has been estimated as a function of closure fraction and this may be a criterion for what is effectively a full blink according to fluid dynamics.