Aim： To investigate the spatial and temporal expression of the cystatin-related epididymal spermatogenic （Cres） gene in mouse testis and epididymis during postnatal development. Methods： The QuantiGene assay and i...Aim： To investigate the spatial and temporal expression of the cystatin-related epididymal spermatogenic （Cres） gene in mouse testis and epididymis during postnatal development. Methods： The QuantiGene assay and indirect immunofluorescence technique were used to examine the Cres mRNA and Cres protein level in mouse testis and epididymis on postnatal days 14, 20, 22, 28, 35, 49, 70 and 420. Results： （1） In both the testis and epididymis, Cres mRNA was fast detected on day 20, then it increased gradually from day 20 to day 70, and the high expression level maintained till day 420. （2） In the testis, the Cres protein was exclusively localized to the elongating spermatids and was first detected on day 22. The number of Cres-positive spermatids increased progressively till day 49. From day 49 to day 420, the number of Cres-positive cells was almost stable. （3） The Cres protein was first detected on day 20 in the proximal caput epididymal epithelium. By day 35, the expression level of the Cres protein increased dramatically and the high level was maintained till day 420. Moreover, the luminal fluid of the midcaput epididymis was also stained Cres-positive from day 35 on. No Cres-positive staining was observed in distal caput, corpus and cauda epididymis throughout. Conclusion： The Cres gene displays a specific age-dependent expression pattern in mouse testis and epididymis on both the mRNA and protein level.展开更多
背景与目的探讨cystatin M基因在人乳腺癌组织中的表达及生物学意义。材料与方法采用半定量RT-PCR方法和免疫组织化学法检测50例乳腺癌组织和配对癌旁乳腺组织半胱氨酸蛋白酶抑制剂cystatin M mRNA和蛋白表达及其与临床病理特征的关系...背景与目的探讨cystatin M基因在人乳腺癌组织中的表达及生物学意义。材料与方法采用半定量RT-PCR方法和免疫组织化学法检测50例乳腺癌组织和配对癌旁乳腺组织半胱氨酸蛋白酶抑制剂cystatin M mRNA和蛋白表达及其与临床病理特征的关系。结果乳腺癌组织cystatin M mRNA表达阳性率为62%(31/50),其配对癌旁乳腺组织为94%(47/50),在乳腺癌细胞系MDA-MB-435S中未见其表达。乳腺癌组织cystatin M mRNA的表达指数为0.412±0.021,明显低于配对的癌旁乳腺组织(0.541±0.020),二者差异有统计学意义(P<0.01)。cystatin M蛋白在乳腺癌及癌旁组织表达阳性率分别为58%(29/50)和92%(46/50);其平均光密度值为0.318±0.058,显著低于癌旁乳腺组织(0.428±0.064,P<0.01);cystatin M表达下调与乳腺癌淋巴结转移和临床TNM分期显著相关(P<0.05),而与肿瘤的大小、病理分级、ER和PR状况无关(P>0.05)。结论人乳腺癌组织存在cystatin M基因表达显著下调且与肿瘤的浸润和转移有关。展开更多
目的:探讨同型半胱氨酸代谢酶蛋氨酸合成酶还原酶基因(MTRR)A66G及半胱氨酸蛋白酶抑制剂C(cystatin C)基因G73A多态性与缺血性脑血管病(ICVD)的关系。方法:采用聚合酶链反应-限制性片段长度多态性(PCR-RFLP)分析技术检测187例ICVD患者和...目的:探讨同型半胱氨酸代谢酶蛋氨酸合成酶还原酶基因(MTRR)A66G及半胱氨酸蛋白酶抑制剂C(cystatin C)基因G73A多态性与缺血性脑血管病(ICVD)的关系。方法:采用聚合酶链反应-限制性片段长度多态性(PCR-RFLP)分析技术检测187例ICVD患者和122例正常对照者MTRRA66G和cystatin C G73A基因多态性。结果:MTRRA66G3种基因型及等位基因频率在病例组和对照组中的分布差异均有统计学意义(χ2=7.008,P=0.030;χ2=6.045,P=0.014),其中GG型在病例组的分布频率35.3%高于对照组的23.0%(χ2=5.314,P=0.021);cystatinC3种基因型及等位基因频率在病例组和对照组中的分布差异均无统计学意义(χ2=2.859,P=0.239;χ2=2.886,P=0.089)。结论:MTRRA66G基因多态性可能与ICVD相关;GG基因型可能是ICVD的易感基因型;cystatin C G73A基因多态性与ICVD发病无关。展开更多
目的探讨蛇毒cystatin在Bac to Bac杆状病毒表达系统中的表达。方法PCR扩增蛇毒cystatin基因,将其克隆到pFastBacHTc中,通过转化E.coliDH10Bac筛选克隆,抽提重组Bacmid/cystatin,后者经Cell-fectin介导转染Sf9细胞,获取重组病毒,扩增病...目的探讨蛇毒cystatin在Bac to Bac杆状病毒表达系统中的表达。方法PCR扩增蛇毒cystatin基因,将其克隆到pFastBacHTc中,通过转化E.coliDH10Bac筛选克隆,抽提重组Bacmid/cystatin,后者经Cell-fectin介导转染Sf9细胞,获取重组病毒,扩增病毒并感染Sf9细胞进行表达,SDS-PAGE、Western-blot分析鉴定表达蛋白66。结果获得重组cystatin的杆状病毒,Sf9细胞能表达出与蛇毒cystatin单抗、5×His单抗结合的蛋白,相对分子质量约15 kD。结论蛇毒cystatin在Bac to Bac杆状病毒表达系统中成功表达。展开更多
文摘Aim： To investigate the spatial and temporal expression of the cystatin-related epididymal spermatogenic （Cres） gene in mouse testis and epididymis during postnatal development. Methods： The QuantiGene assay and indirect immunofluorescence technique were used to examine the Cres mRNA and Cres protein level in mouse testis and epididymis on postnatal days 14, 20, 22, 28, 35, 49, 70 and 420. Results： （1） In both the testis and epididymis, Cres mRNA was fast detected on day 20, then it increased gradually from day 20 to day 70, and the high expression level maintained till day 420. （2） In the testis, the Cres protein was exclusively localized to the elongating spermatids and was first detected on day 22. The number of Cres-positive spermatids increased progressively till day 49. From day 49 to day 420, the number of Cres-positive cells was almost stable. （3） The Cres protein was first detected on day 20 in the proximal caput epididymal epithelium. By day 35, the expression level of the Cres protein increased dramatically and the high level was maintained till day 420. Moreover, the luminal fluid of the midcaput epididymis was also stained Cres-positive from day 35 on. No Cres-positive staining was observed in distal caput, corpus and cauda epididymis throughout. Conclusion： The Cres gene displays a specific age-dependent expression pattern in mouse testis and epididymis on both the mRNA and protein level.
文摘背景与目的探讨cystatin M基因在人乳腺癌组织中的表达及生物学意义。材料与方法采用半定量RT-PCR方法和免疫组织化学法检测50例乳腺癌组织和配对癌旁乳腺组织半胱氨酸蛋白酶抑制剂cystatin M mRNA和蛋白表达及其与临床病理特征的关系。结果乳腺癌组织cystatin M mRNA表达阳性率为62%(31/50),其配对癌旁乳腺组织为94%(47/50),在乳腺癌细胞系MDA-MB-435S中未见其表达。乳腺癌组织cystatin M mRNA的表达指数为0.412±0.021,明显低于配对的癌旁乳腺组织(0.541±0.020),二者差异有统计学意义(P<0.01)。cystatin M蛋白在乳腺癌及癌旁组织表达阳性率分别为58%(29/50)和92%(46/50);其平均光密度值为0.318±0.058,显著低于癌旁乳腺组织(0.428±0.064,P<0.01);cystatin M表达下调与乳腺癌淋巴结转移和临床TNM分期显著相关(P<0.05),而与肿瘤的大小、病理分级、ER和PR状况无关(P>0.05)。结论人乳腺癌组织存在cystatin M基因表达显著下调且与肿瘤的浸润和转移有关。
文摘目的:探讨同型半胱氨酸代谢酶蛋氨酸合成酶还原酶基因(MTRR)A66G及半胱氨酸蛋白酶抑制剂C(cystatin C)基因G73A多态性与缺血性脑血管病(ICVD)的关系。方法:采用聚合酶链反应-限制性片段长度多态性(PCR-RFLP)分析技术检测187例ICVD患者和122例正常对照者MTRRA66G和cystatin C G73A基因多态性。结果:MTRRA66G3种基因型及等位基因频率在病例组和对照组中的分布差异均有统计学意义(χ2=7.008,P=0.030;χ2=6.045,P=0.014),其中GG型在病例组的分布频率35.3%高于对照组的23.0%(χ2=5.314,P=0.021);cystatinC3种基因型及等位基因频率在病例组和对照组中的分布差异均无统计学意义(χ2=2.859,P=0.239;χ2=2.886,P=0.089)。结论:MTRRA66G基因多态性可能与ICVD相关;GG基因型可能是ICVD的易感基因型;cystatin C G73A基因多态性与ICVD发病无关。
文摘目的探讨蛇毒cystatin在Bac to Bac杆状病毒表达系统中的表达。方法PCR扩增蛇毒cystatin基因,将其克隆到pFastBacHTc中,通过转化E.coliDH10Bac筛选克隆,抽提重组Bacmid/cystatin,后者经Cell-fectin介导转染Sf9细胞,获取重组病毒,扩增病毒并感染Sf9细胞进行表达,SDS-PAGE、Western-blot分析鉴定表达蛋白66。结果获得重组cystatin的杆状病毒,Sf9细胞能表达出与蛇毒cystatin单抗、5×His单抗结合的蛋白,相对分子质量约15 kD。结论蛇毒cystatin在Bac to Bac杆状病毒表达系统中成功表达。
