Expression of Ceramide Kinase （Cerk） and Related Regulation in Early Pregnant Uterus of Mice

Objective To investigate the expression of ceramide kinase (Cerk) and related regulation in mouse uterus during early pregnancy. Methods Several mouse models, including early pregnancy, pseudopregnancy, delayed or activated implantation, artificial decidualization, and steroid hormonal treatment were performed (n=10). Immunohistochemistry and in situ hybridization were used to detect the expression of Cerk protein and mRNA in uterus. Results Expression of Cerk mRNA and protein was strongly detected in the luminal and glandular epithelium on day 1 of pregnancy. However, Cerk mRNA and protein signals were strongly detected in the subluminal stroma surrounding the implanting blastocyst on day 5 and decidua from day 6 to day 8 whereas not in the luminal epithelium. The expression of Cerk in luminal and glandular epithelium of pseudopregnancy was similar to that of early pregnancy from day 1 to day 4 whereas on day 5 of pseudoprgenancy still with remarkable signals in the luminal epithelium. Under delayed implantation, no obvious Cerk expression was detected in the uterus. After delayed implantation was terminated by estrogen treatment and embryo implantation was initiated, both Cerk mRNA and protein were detected in the subluminal stroma surrounding the implanted blastocyst. A strong Cerk signal was detected in decidualized cells under artificial decidualization, whereas only a basal level of Cerk signal was observed in the control uterus which did not inject sesame oil. Progesterone induced a slight expression of Cerk in the luminal and glandular epithelium. Both estradiol and a combination of progesterone with estradiol strongly increased the level of Cerk signal in the luminal and glandular epithelium. Conclusion Cerk expression is under the regulation of progesterone and estrogen. The strong expression of Cerk in implantation site and decidua suggests that Cerk might play an important role during implantation and decidualization.     

Experiment study on the effect of Sheng-Mai injection on the contractivity of diaphragm and its mechanism.

In this study the effect of Sheng-ai Injection i.e. Red Ginseng-Ophiopogon Root Injection (one kind of traditional Chinese medicines) on the contractivity of diaphragm was observed. The results confirmed that Sheng-ai Injection increased Pdi of the fatigued diaphragm in rabbits and reduced the time needed for the recovery of Pdi of fatigued diaphragm to the normal value. These results suggest that Sheng-Mai Injection can increase the contractive force and promote the recovery of the fatigued diaphragm. The effect of Sheng-ai Injection on the contractivity of the isolated diaphragmatic bundle of rats was also observed and the results confirmed that Sheng-ai Injection increased the diaphragmatic contractive force directly. This effect of increasing the contractive force of diaphragm was attenuated by adding calcium channel blocker isoptin and disappeared when there was no calcium in the extracellular fluid. It is deduced, therefore, that the mechanism of the effect of Sheng-mai Injection is related to the     

Postcoital administration of asoprisnil inhibited embryo implantation and disturbed ultrastructure of endometrium in implantation window in mice

Asoprisnil, a member of the selective progesterone receptor modulators, exerts high progesterone receptor selectivity, endometrial targeted advantages and significant anti-implantation effect in rats. The purpose of this study was to confirm the anti-implantation effect of asoprisil, investigate the ultrastructural changes of the peri-implantation endometrium in mice and explore the effect of asoprisnil on endometrial receptivity and its targeted contraceptive proficiency. Post-coitus mice were administered with different dosages (0.2, 0.1, 0.05 mg·g -1 ·day -1 ) of asoprisnil from day 1 of pregnancy to day 3. Then 3 animals in each group were killed on day 5 of pregnancy, and uteri were collected to examine the ultrastructural changes of endometria under a transmission electron microscope (TEM). A total of 80 animals were sacrificed on day 8 of pregnancy, and the uterine horns were examined for the presence or absence of nidation sites and the number of implantation embryos. The results showed that the implantation rate and the average number of implantation embryos in asoprisnil groups were statistically significantly decreased as compared with the vehicle control group (P<0.05). The TEM results revealed that, in vehicle control group, the tight junction between the luminal epithelia cells was short and straight, the gap was wide; the luminal epithelia cells were covered with plenty of short, clavate and neatly arranged microvilli; the endometril stromal cells were large with plenty of cytoplasm, and showed significant decidual change; there was more than one nucleus in stromal cells, and the karyotheca was integrity. In low dosage and high dosage asoprisnil groups, the tight junction was longer and more curve than in the vehicle control group; microvilli were uneven and asymmetrically distributed in luminal epithelia; the stromal cells were small and the decidual change was not significant; there were karyopyknosis and karyolysis in stromal cells; there were abnormal thick-wall vessels in the endometrium. It was suggested that asoprisnil changed the ultrastructure of the endometrium in implantation window, disturbed the endometrial receptivity and finally resulted in embryo implantation failure.     

Expression of Angiopoietin-1/-2 in the Process of Mouse Embryo Implantation

This study examined the expression and distribution of angiopoietin-1/-2 (Ang-1/-2) in the endometrium of early pregnant mice. The expression of Ang-1/-2 was detected by immunohisto- chemical staining and in situ hybridization respectively. Computerized image analysis system was used to measure the average optical intensity of Ang-1/-2 in endometria at different time points after gestation. Mice were randomly divided into 5 groups: control group, D2 group (2 days after preg- nancy), D4 group (4 days after pregnancy), D6 group (6 days after pregnancy) and D8 group (8 days after pregnancy), each containing 15 mice. The results showed that the expression of Ang-1 and Ang-2 was very different among 4 groups (P<0.01). Immunohistochemical staining revealed that Ang-1 was localized in the cytoplasma of stromal cells 2 days after pregnancy (day 2), and in luminal epithelial cells on day 4. The protein of Ang-2 was mainly expressed in the cytoplasma of glandular epithelia and stromal cells. With gestation time, the positive reactions of Ang-1/-2 were stronger in the endometria of the pregnant mice (P<0.01). In situ hybridization showed Ang-1 mRNA in stromal cells on day 2. Hybridization signal was localized in both stromal cells and vessel epithelial cells on day 4; Ang-2 mRNA was expressed in stromal cells and glandular epithelia on day 2; high mRNA levels appeared in stromal cells, glandular epithelia and vascular endothelia on day 4; an increasing in mRNA expression of Ang-1/-2 was observed on day 6 and day 8 (P<0.01). It is suggested that Ang-1/-2 may play an important role in the cross-talk between blastocyst and maternal endometrium during the process of embryo implantation.     

Expression of Nuclear Factor-κB in Mouse Uterus during Peri-implantation

To investigate the expression of the subunit p65 of NF-κB and inhibitor kappa B alpha (IκBα) in mouse uterus during peri-implantation, thereby investigating whether transient activation of nuclear factor-κB (NF-κB) takes place during embryo implantation in mice. Immunohistochemical technique was used to examine the expression and localization of p65 in endometrium or deciduas, and Western blot analysis was employed to detect the levels of IκBα protein in mouse endometrium or deciduas. P65 protein was detected in stromal cells, epithelial cells of endometrium as well as in myometrium. Staining was predominately seen in the cytoplasm of the cells. Staining intensity for p65 was stronger in the epithelial compartment than the stromal compartment and myometrium. Staining intensity increased slightly during pregnancy, and it reached a high level on pregnancy day 5 and day 8. In contrast to p65, the level of IκBα protein was lowest on pregnancy day 5 in all groups. Our results suggested that NF-κB may regulate embryo implantation by its transient activation in mice.     

Effect of Bushenantai Recipe on the Expression of Endometrial LIF in Mice with Embryonic Implantation Dysfunction

In order to observe the effect of Bushenantai recipe on the expression of endometrial leukemia-inhibitory factor （LIF） in mice with embryonic implantation dysfunction （EID）, 120 Kunming mice post coition were randomized into three groups： normal control group, model group and traditional Chinese medicine group （TCM group） （n=40 in each group）. Uterus was collected on the pregnancy day （Pd） 4, 5, 6 after an intravenous injection of Evan＇s blue. The endometrium was dyed by Evan＇s blue and the mean points of response were observed on Pd 5. The expression of LIF mRNA and protein was detected by RT-PCR and immunohistochemistry respectively and analyzed statistically by image system. The results showed that the number of implantation sites in model group was remarkably less than in normal control group and TCM group. There was no significant difference between normal control group and TCM group. The expression of LIF mRNA and protein in model group was delayed. Bushenantai recipe could increase the expression of LIF mRNA and protein in endometria of mice with EID. It was suggested that Bushenantai recipe could improve embryo implantation of mice with EID by promoting the endometrial LIF expression and endometrial decidualization.     

Antifertility Effect of Sperm Coating Protein in Rat Seminal Vesicle by Isoimmunization

This paper reports that seminal vesicle protein Ⅱ (SVP Ⅱ) secreted by rat was used as an antigen in the isoimmunization of rat for the observation of its antifertitity effect. The pregnancy rate was decreased to 7.7% in rats immunized passively with rabbit anti-SVP Ⅱ serum, as compared to 76.9% in rats injected with normal rabbit serum. The antifertility effect of active immunization was found to be related to the titer number of anti-SVP Ⅱ serum. The high anti-SVP Ⅱ liter and its significant antifertility effect demonstrate that the sperm coating protein can be used as an vaccine for fertility inhibition.     

The effect of spleen on the change of Treg cells during mouse pregnancy

Objective： To analyze the effect of spleen on the Treg cells during pregnancy. Methods： The mononuclear cells were separated from the peripheral, spleen and uterus or placenta blood. Flow cytometry was employed to analyze the percent of Treg cells in total T cells in different stages of pregnancy. Immunohistochemical staining was used to make sure the distribution of Treg in spleen in different stages of pregnancy. Results： The results of immunohistochemical staining showed that compared with spleen Treg cells in normal unpregnant mice, spleen Treg cells on day 7 and 14 of pregnancy significantly increased. After splenectomy, peripheral blood and placenta Treg cells on day 7 of pregnancy markedly decreased as compared with the normal pregnancy （P〈0.01）. And the cells on day 14 of pregnancy were markedly recovered as compared with the normal pregnancy. Conclusion： Our study indicated that the spleen and its Treg cells might play important roles in transient tolerance during pregnancy.     

Distribution of materno-fetus materno-suckling transfer of tritium after single injection of tritiated water in mice.

The metabolism and transfer of tritium from pregnant mice to fetus after injection of tritiated water intraperitoneally was investigated in this paper. The study was composed of three experiments. 1. Pregnant mice were injected with tritium water on the first day of pregnancy, in order to obtain a transfer coefficient of tritium from pregnant mice to fetus through placenta. 2. Pregnant mice were injected with tritiated water on the first day of parturition to study the transfer of tritium via mother milk from nursing mice to the baby-animals. 3. Pregnant mice were injected with tritiated water in different periods of gestation. The results showed that tritiated water was uniformly distributed in all of the tissues measured, including placenta, fetal membrane and amniotic fluid in experiment 1. No effect of placentas on tritium transfer from pregnant mice to fetus was found. Concentration of tritium in the baby's tissues was evidently higher than that in the pregnant mice in experiments 2 and 3, and the transfer coefficient in experiments 2 and 3 was generally higher than that in experiment 1.

     

宫内用醋酸棉酚对雌性大鼠抗着床作用的机理分析

在大鼠妊娠的第4天,宫内一次性注入醋酸棉酚0.5mg(0.1ml),4～11天时实验组每天注射孕酮5mg,对照组注射等量生理盐水,第12天处死动物剖检。结果,孕酮能有效地对抗醋酸棉酚的抗着床作用。在切除卵巢延缓着床模型的大鼠,相应实验组宫内一次注入抗生育剂量棉酚,对照组注入等量溶剂,结果表明外源性雌激素不能逆转宫内用醋酸棉酚的抗着床作用。实验组子宫液用SDS-聚丙烯酰胺凝胶电泳分析发现宫内注入醋酸棉酚可使孕酮诱导的分子量7万以下的蛋白带减少,但对雌二醇诱导的分子量大于9万的蛋白带无作用,这可能是其抗着床作用的机理之一。     

钙调素阻滞剂三氟拉嗪对原代肺癌细胞作用的研究

应用细胞培养技术对２８例手术切除标本行原代肺癌细胞培养，并测定了钙调素（ＣａＭ）阻滞剂三氟拉嗪对原代肺癌细胞的作用，结果显示，三氟拉嗪能抑制肺癌细胞进入增殖周期，使细胞阻滞于Ｇ０／Ｇ１期。在长春新碱（ＶＣＲ），阿霉素（ＡＤＲ）和足叶乙甙（ＶＰ１６）各药物组加入三氟拉嗪后能使癌细胞活性（Ａ值）明显降低（Ｐ〈０．０１或Ｐ〈０．０５），单用三氟拉嗪组亦能使癌细胞活性降低，但无统计学意义（Ｐ〉０．０５）。     

酪氨酸及酪氨酰肼对小鼠的抗生育作用

酪氨酸(0.1mg或1mg/只)能使小鼠的妊娠率及胚胎数明显降低,但不能完全阻断妊娠;酪氨酰肼的抗生育作用比酪氨酸强,一定剂量可完全阻断妊娠。酪氨酸与酪氨酰肼对妊娠小鼠血中孕酮含量无明显影响。酪氯酰肼对~3H-Leu掺入小鼠妊娠子宫组织有明显抑制作用。     

CaM抑制剂对LPS/IFN-γ诱导的RAW264.7细胞NO生成及iNOS表达的影响

目的观察钙调蛋白(Calmodulin,CaM)抑制剂对脂多糖/干扰素γ(LPS/IFN-γ)诱导的RAW264.7细胞一氧化氮(NO)的生成及诱生型一氧化氮合酶(iNOS)表达的作用及其可能的作用机制。方法采用LPS/IFN-γ诱导RAW264.7巨噬细胞建立细胞炎症反应模型,钙调蛋白抑制剂(W-7,TFP)预处理细胞后,采用Griess试剂法测定NO释放量;采用Western blot法测定目的蛋白的表达;采用反转录聚合酶链反应法(RT-PCR)分析iNOS mRNA表达的变化。结果 CaM抑制剂可抑制LPS/IFN-γ诱导的RAW264.7细胞NO的释放(P<0.01);可抑制iNOS蛋白和mRNA的表达,早期可抑制IκBα的降解、磷酸化IKK(PIKK)和STAT1的蛋白(PSTAT1)表达。结论 CaM抑制剂可明显降低LPS诱导的RAW264.7细胞NO、iNOS蛋白和mRNA表达;可以通过抑制IκBα降解和磷酸化IKK蛋白表达而发挥抗炎作用。     

三种氨基酸对兔血浆皮质酮浓度的影响

侧脑室注射甘氨酸使健康家兔和烧伤家兔血浆皮质酮浓度降低;注射L—谷氨酸和L—天冬氨酸使健康家兔血浆皮质酮浓度升高,但不能使烧伤后升高的血浆皮质酮浓度发生改变。提示这三种氨基酸作为神经递质参与下丘脑—垂体—肾上腺皮质轴的调控。烧伤刺激通过下丘脑引起内分泌紊乱,这种紊乱与L—谷氨酸和L—天冬氨酸失控有关。     

参附注射液治疗慢性心力衰竭临床疗效分析

目的对参附注射液治疗慢性心力衰竭(CHF)的临床疗效进行分析。方法将72例心力衰竭患者随机分为治疗组和对照组,对照组给予常规西医治疗,使用血管紧张素转换酶抑制剂、β-受体阻滞剂、洋地黄制剂、利尿剂、硝酸酯制剂等,治疗组在对照组治疗的基础上,加用参附注射液50 mL+5%葡萄糖注射液100 mL中,静滴,1次/d,14 d为1个疗程。结果治疗组慢性心力衰竭患者应用参附注射液后,有效率为94.44%,对照组有效率为83.33%,治疗组总有效率明显优于对照组(P<0 05);而治疗组的心功能改善也明显优于对照组(P<0 05)。结论慢性心力衰竭在常规治疗基础上,加用参附注射液的治疗具有更满意的临床疗效,未发现不良反应,值得临床推广。     

薄荷油对小白鼠终止妊娠作用的初步观察

本文报道两种不同给药途径的薄荷油对小白鼠终止妊娠作用的初步观察结果:于孕d_6,将4μl薄荷油或橄榄油分别注入右侧宫角,左侧宫角不给药,于孕d_(11)剖检。给薄荷油侧与橄榄油侧宫角妊娠终止率分别为100%与41. 67％,差别显著。于孕d_4～10,各组分别肌注薄荷油一次,于孕d_(11)剖检。薄荷油不同剂量皆有一定抗着床与抗早孕作用,作用强度随剂量增加,0. 035ml/只,抗着床率达100％。终止妊娠的原因可能是子宫收缩加强,也不能排除蜕膜组织等的直接损伤。     

侧脑室微量注射apelin对大鼠痛阈的影响

观察侧脑室微量注射apelin对大鼠痛阈的影响。方法将30只成年雄性SD大鼠随机分为NS对照组、吗啡（Morphine）组和apelin组。采用辐射热刺激甩尾测痛法测量痛阈,以大鼠甩尾反应的潜伏期（tail-flick la-tency,TFL）作为痛阈指标。侧脑室微量注射药物后分别10,20,30,40,50,60（min）时测TFL。结果侧脑室微量注射apelin后10~30 min大鼠痛阈与NS对照组相比显著降低,差异有统计学意义（P〈0.01）,30 min后大鼠痛阈逐渐回升,至60 min时大鼠痛阈仍低正常水平。结论在中枢神经系统内,apelin参与痛与痛觉调制过程,脑内apelin含量增加,有明显的痛敏作用。     

经输精管向附睾内注射甲酸棉酚的抗生育研究

对72只Wistar大鼠经输精管向附睾内注射甲酸棉酚,观察附睾中精子数量、形态、移动距离和附睾、睾丸的组织学改变。结果:注射12h后精子移动距离缩短,24h后精子计数下降,组织学观察见附睾管呈无精状态,睾丸曲细精管各级生精细胞出现变性等损伤性改变,精母细胞脱落入曲细精管管腔,注射甲酸棉酚后2～40d内交配的雌鼠全部不孕。     

黄芪和灯盏花素注射液治疗充血性心力衰竭临床观察

目的 :观察黄芪和灯盏花素注射液治疗充血性心力衰竭的疗效。方法 :将 80例病例随机分成两组。对照组 4 0例 ,给予常规治疗 ,治疗组 4 0例 ,在常规治疗的基础上 ,加用黄芪注射液和灯盏花素注射液 ,静脉点滴每日 1次 ,两组连续治疗 15d。结果 :有效率治疗组 92 .5 % ,对照组 70 % ,两组有明显差异 ,P值 <0 .0 5。结论 :在常规治疗的基础上加用黄芪和灯盏花素治疗可提高疗效。