In-vitro maturation of human germinal vesicle stage oocytes: role of cumulus cells and epidermal growth factor in the culture medium

In-vitro maturation (IVM) of oocytes is a promising technique to reduce the costs and avert the side-effects of gonadotrophin stimulation for in-vitro fertilization (IVF). The pregnancy rates from oocytes matured in vitro are much lower than those of in-vivo stimulation cycles indicating that optimization of IVM remains a challenge. Therefore, we investigated the effect of supplementation of the medium with gonadotrophins, oestradiol and epidermal growth factor (EGF) and the effect of retaining or removing the cumulus cells on nuclear and cytoplasmic maturation of immature oocytes. Human germinal vesicle (GV) oocytes obtained after gonadotrophin stimulation for intracytoplasmic sperm injection (ICSI) were cultured in a complex defined medium either supplemented with gonadotrophins, oestradiol and physiological concentrations of EGF (2 ng/ml) or gonadotrophins and oestradiol alone. The cumulus cells were either removed or kept intact. In GV stage oocytes cultured without cumulus (group I) significantly more oocytes reached the metaphase II (MII) stage at 30 h in media supplemented with EGF (64.3 versus 33.9%, P < 0.003). For oocytes cultured with intact cumulus (group II), more oocytes reached MII at 30 h than in group I, but there was no difference in medium with or without EGF supplementation (81.8 and 79.8% respectively). Cytoplasmic maturation of MII oocytes was judged from their capability to activate and fertilize after ICSI. In group I, the rates of activation and normal fertilization were similar. However, in group II, significantly more oocytes underwent normal fertilization in the EGF-supplemented than the unsupplemented group (71.7 versus 45.6%, P < 0.05). The cleavage rates of the fertilized oocytes were similar in the sibling oocyte subgroups cultured with or without EGF supplementation, but the overall cleavage rates were higher in cumulus-intact compared to cumulus-denuded oocytes (88.9 versus 47.8%, P < 0.001). Thus, supplementation of the maturation medium with EGF and maintenance of the cumulus during culture improve the nuclear and cytoplasmic maturation of human oocytes in vitro.      

荧光定量PCR快速诊断21三体综合征

目的　建立荧光定量PCR技术检测 2 1三体综合征。方法　采用PCR方法同时扩增位于 2 1号染色体上的人肝型磷酸果糖激酶基因 (humanliver typephosphofructokinasegene ,PFKL CH 2 1)和位于 1号染色体上的人肌型磷酸果糖激酶基因 (humanmuscle typephosphofructokinasegene ,PFKM CH1) ,使用SYBRGreenⅠ荧光染料处理产物、琼脂糖电泳后在凝胶成像系统进行分析 ,得出扩增产物的荧光强度对比值。用此方法检测 2 6例 2 1三体综合征患儿及 2 0名正常人。结果　 2 6例 2 1三体综合征患儿PFKL CH2 1/PFKM CH 1扩增产物的荧光强度对比值为 1.5 8± 0 .17,而正常人为 1 0 0± 0 .0 5 ,两者差异有显著性。结论　SYBRGreenⅠ荧光定量PCR技术检测 2 1三体综合征具有准确、快速、安全、实用等特点 ,有较高的临床使用价值。     

以造影增强磁共振血管成像术诊断先天性主动脉弓畸形

目的评价造影增强磁共振血管成像术（CE-MRA）对先天性主动脉弓畸形的诊断准确性并与超声心动图等比较。方法413例先天性主动脉弓畸形病例在1999年4月到2004年12月期间做了CE-MRA检查。年龄5d～11岁，平均年龄2.4岁。检查使用1.5T磁共振扫描仪。对比剂gadolinium-DTPA，剂量0.2mmol／kg体质量，手动静脉注射。磁共振检查前均已做心脏超声检查。结果在413例中，动脉导管未闭（PDA）166例，主动脉缩窄（COA）168例，主动脉弓中断（IAA）31例，双主动脉弓等其他先天性主动脉弓畸形48例。全部413例均经手术和／或X线心血管造影证实。CE-MRA的诊断准确率为97.3％（402／413），心脏超声检查的诊断准确率为87.2％（360／413）。结论对于先天性主动脉弓畸形，CE-MRA是可靠的影像学检查方法，不仅优于心脏超声检查，有时CE-MRA甚至能比传统的X线心血管造影术提供更多的诊断信息。     

圆孔外面的观察和测量及其面积的回归方程

60具成人颅骨颅底外面的圆孔观察和测量结果表明:圆孔多呈圆形,其次是卵圆形,其它形较少见.据圆孔与翼突外侧板根部延长线的位置关系,分圆孔位置为三型.圆孔外面观的各项测值左右间均无统计学差异,但其中管性圆孔管长度个体差异较大.圆孔的面积左右比较对称者占3.33±2.34%,左>右者占65.51±6.30%,右>左者占34.48±6.30%.圆孔面积左右对称性比较,对临床诊断疾病有参考意义.其面积的回归方程可由其长、宽径乘积推算 .     

Fourier transform infra-red spectroscopy of atactic polystyrene in the glass transition region

Fourier transform infra-red (IR) studies on samples of atactic polystyrene with different thermal history, i.e. quenched from the melt (QM), quenched from the rubbery state (QR), slowly cooled (SC), and sub-glass-transition (sub-T_g) annealed (AN), were carried out in the absorption region from 500 to 600 cm^?1 with gradually increasing temperatures through the glass transition region at 100°C. The heights of the positive peak (532 cm^?1) and the negative peak (566 cm^?1) appearing in the temperature difference spectra, T-60°C, are considered to be a measure of the amount of frequency shifts for the 540 cm^?1 and 557 cm^?1 bands in the conformationally sensitive spectral range of atactic polystyrene. These bands show a continuously increasing low-frequency shift with increasing temperature, except for sample QM, the 557 cm^?1 band of which shows an inappreciable frequency shift in the range 60 < T/°C < 100. The different temperature behaviour of these shifts for samples QM, QR, SC and AN is due to changes in their intermolecular interactions. Sub-T_g annealing of the sample QM is shown to shift the frozen high-temperature state of the quenched sample to the temperature-defined equilibrium. For all samples of different thermal history the difference in their IR spectra vanish at temperatures above T_g.     

中药灵芝对健康人甲襞微循环的作用

系统观察５７例健康人口服灵芝（灵芝多糖体５５ｍｇ／片）前后甲襞微循环的变化。结果显示：服药前后大、小动脉血压无显著变化；口服灵芝３天后输入枝口径及输出枝口径均有明显扩张；管袢密度、红细胞流速增加不显著。口服灵芝７天的动态变化显示，服药８ｈ后输入枝、输出枝口径即有显著扩张，第７天管袢密度开始明显增加。服用不同剂量灵芝（２片、４片、８片，３次／日）３ｈ后，管袢密度、输入枝、输出枝口径、流速均有增加趋向，但Ｐ＞０．０５，其中８片组管袢口径、管袢密度有显著增加（Ｐ＜０．０５），但红细胞流速却有显著下降（Ｐ＜０．０５）。结果提示：连续、适量（２片，３次～４次／日）服用灵芝，可显著改善微循环灌流量，从而改善和保护组织及脏器功能。     

Requirements for the development of IL-4-producing T cells during intestinal nematode infections: what it takes to make a Th2 cell in vivo

Summary: Components of the type 2 immune response may mediate host protection against both helminthic parasites and harmful allergic responses. A central player in this response is the T‐helper 2 (Th2) effector cell, which produces interleukin (IL)‐4, IL‐5, IL‐13, and other Th2 cytokines during the primary and memory response. Specific aspects of the parasite that trigger Th2‐cell differentiation are not yet defined. Furthermore, the cell types and cell surface and secreted molecules that provide the immune milieu required for the development of Th2 effector cells and also Th2 memory cells are not well understood. They will probably vary with the particular helminth or other antigen inducing the Th2 response. We have used third stage larvae of intestinal nematode parasites as adjuvants to promote naïve nonparasite antigen‐specific T cells to differentiate into Th2 cells. This model system avoids possible parasite antigen‐specific T‐cell clones or cross‐reactive memory T cells that may preferentially differentiate into Th2 effector cells during the course of infection and confound the stereotypical components of parasite‐induced Th2 cell differentiation. We have found that these parasites have a potent adjuvant effect and have used our model system to begin to investigate the events that lead to the development of polarized Th2 cells in vivo.