Objective:To investigate the value of serum human epididymis protein 4(HE4) in differential diagnosis of patients with low-grade serous(LGSC) and high-grade serous carcinoma(HGSC) serous ovarian cancer.Methods:LGSC an...Objective:To investigate the value of serum human epididymis protein 4(HE4) in differential diagnosis of patients with low-grade serous(LGSC) and high-grade serous carcinoma(HGSC) serous ovarian cancer.Methods:LGSC and HGSC serous ovarian cancer were diagnosed by the two-tier grade system,serum levels of HE4 and carbohydrate antigen 12S(CA125) were measured by ELBA and radioisotope method,respectively in 60 serous ovarian cancer patients. HE4 and TPS3 protein in cancer tissue were measured by immunohistochemical method. Results:The difference in density of HE4 and TP53 protein was significant between LGSC and HGSC tissue,while serum CA12S did not show significant difference between different serum samples.There was significant difference in serum HE4 levels between LGSC and HGSC and the result was different within FIGO(Ⅰ+Ⅱ) stage,suggesting HE4 was not a reliable biomarker for the discrimination between LGSC and HCSC.HE4 had potential as a biomarker for the discrimination between LGSC and HGSC but the role in early diagnosis was limited.Conclusions:HE4 may be a reliable marker for differential diagnosis of LGSC and HGSC.But its role in early diagnosis of LGSC and HGSC need to be confirmed from the perspective of two-tier grade system.展开更多
Vasectomy is a simple and reliable method of male contraception. A growing number of men after vasectomy request vasectomy reversal due to various reasons. The pregnancy rate is lower than the patency rate after vasov...Vasectomy is a simple and reliable method of male contraception. A growing number of men after vasectomy request vasectomy reversal due to various reasons. The pregnancy rate is lower than the patency rate after vasovasostomy and the pregnancy rate is time dependent. In this study, we evaluated the influence of reproductive tract obstruction on expression of epididymal proteins and their restoration after patency. Adult male Wistar rats were studied 30, 60 and 120 days after vasectomy, 30 days after vasovasostomy or after sham operations. Two-dimensional gel electrophoresis, mass-spectrometric technique, multidatabase search, Western blotting and real-time PCR were used to analyze the expression regulation of epididymal proteins. Total integrated intensity and total spot area of autoradiograms showed a consistent downward trend with time after obstruction, and this trend remained after patency. The intensity of the autoradiographic spots in three patency groups showed three trends： a downward trend, similar intensity and an upward trend compared with the correspondent obstruction group, respectively. Further verified experiments on human epididymis 2 （HE2）, fertilization antigen-1 （FA-1）, clusterin and PH20 demonstrated that compared with the correspondent obstruction group, the translation levels of HE2 and the mRNA transcription levels of HE2 showed an upward trend in patency groups, especially in the groups of obstruction for 60 days where the expression levels of HE2 were significantly upregulated after patency （P〈O.05）. Reproductive tract obstruction provokes a disregulation of gene expression in the epididymis and this disregulation remained after patency. Successful reversal may recover some proteins and the recovery is time dependent, Obstruction differentially alters mRNA transcription of different proteins and the content of proteins seemed to be easier to be influenced than the gene transcription.展开更多
To explore the functions of human ribonuclease 9(RNase 9),we constructed a mammalian fusion expression vector pcDNA-hRNase9,prepared recombinant human RNase 9-His fusion protein from HEK293T cells and determined its N...To explore the functions of human ribonuclease 9(RNase 9),we constructed a mammalian fusion expression vector pcDNA-hRNase9,prepared recombinant human RNase 9-His fusion protein from HEK293T cells and determined its N-terminal amino acid sequences.According to the determined mature protein,recombinant human RNase 9 was prepared in E.coli.Ribonucleolytic activity and antibacterial activity of recombinant human RNase 9 were detected,and the distribution of human RNase 9 on tissues and ejaculated spermatozoa and in vitro capacitated spermatozoa were analyzed via indirect immunofluorescence assay.The results showed that recombinant human RNase 9 did not exhibit detectable ribonucleolytic activity against yeast tRNA,but exhibited antibacterial activity,in a concentration/time dependent manner,against E.coli.Immunofluorescent analyses showed that the predicted human RNase 9 was present throughout the epididymis,but not present in other tissues examined,and human RNase 9 was also present on the entire head and neck regions of human ejaculated spermatozoa and in vitro capacitated spermatozoa.These results suggest that human RNase 9 may play roles in host defense of male reproductive tract.展开更多
目的:分析血清人附睾分泌蛋白4(Human epididymis protein 4,HE4)水平和子宫内膜癌患者临床病理特征的相关性.方法:回顾性分析2019年1月至2022年6月医院收治的120例子宫内膜癌患者临床资料.所有患者均实施子宫全切术治疗,收集患者的临...目的:分析血清人附睾分泌蛋白4(Human epididymis protein 4,HE4)水平和子宫内膜癌患者临床病理特征的相关性.方法:回顾性分析2019年1月至2022年6月医院收治的120例子宫内膜癌患者临床资料.所有患者均实施子宫全切术治疗,收集患者的临床病理特征资料,检测所有患者HE4水平,分析血清HE4水平与子宫内膜癌患者临床病理特征的相关性.结果:120例子宫内膜癌患者中,病理类型:子宫内膜样腺癌46例、子宫内膜样透明细胞癌36例、子宫内膜样鳞癌38例;淋巴结转移:发生35例、未发生85例;肌层浸润深度:>1/2肌层40例、≤1/2肌层80例;TNM分期:Ⅰ期22例、Ⅱ期43例、Ⅲ期55例.病灶所处部位:子宫底55例、子宫角37例、宫颈管28例.不同病理类型、TNM分期、病灶所处部位患者的血清HE4水平比较,差异无统计学意义;肌层浸润深度>1/2肌层患者的血清HE4水平高于肌层浸润深度≤1/2肌层的患者,发生淋巴结转移患者的血清HE4水平较未发生淋巴结转移的患者明显增加(P<0.05).经Logistic回归显示,血清HE4水平升高是子宫内膜癌患者肌层浸润深度>1/2肌层的风险因子(OR>1,P<0.05);血清HE4水平升高是子宫内膜癌患者发生淋巴结转移的风险因子(OR>1,P<0.05).结论:血清HE4水平升高增加可加剧肌层浸润深度,促使子宫内膜癌患者发生淋巴结转移.展开更多
Previous studies have shown that rat epididymis-specific gene HongrES1 plays important roles in sperm capacitation and fertility. In this study, we cloned the mouse homologue gene by sequence alignment and RT-PCR meth...Previous studies have shown that rat epididymis-specific gene HongrES1 plays important roles in sperm capacitation and fertility. In this study, we cloned the mouse homologue gene by sequence alignment and RT-PCR methods and designated it as mHongl. The mHongl gene is located on chromosome 12p14, spanning five exons. The cDNA sequence consists of 1257 nucleotides and encodes a 419 amino-acid protein with a predicted N-terminal signal peptide of 20 amino acids. The mHongl mRNA shows similarity with HongrES1 in the expression patterns： （i） specific expression in epididymal tissue, especially in the cauda region; and （ii） androgen-dependence but testicular fluid factor independence. Its protein product shows 71% similarity with HongrES 1 and contains a classical serpin domain as does HongrES1. A polyclonal antibody against mHongl with high specificity and sensitivity was raised. Like HongrES1, the mHongl protein shows a checker-board expression pattern in the epididymal epithelium and is secreted into the epididymal lumen. The mHongl protein shows higher glycosylation than HongrES1. Although both of them are deposited onto the sperm head surface, mHongl is localized to the equatorial segment, which is different from that of HongrES 1. The mHongl protein can be removed from the sperm membrane by high ionic strength and therefore can be classed as an extrinsic membrane protein. Collectively, we conclude that mHongl is the homologue of HongrES1 and the present work paves the way for establishing animal models to elucidate the precise functions of HongrES1 and mHongl.展开更多
基金suuported by Young Researcher Foundation from Education Department of Jiangxi Province(Grand No.GJJ12161)
文摘Objective:To investigate the value of serum human epididymis protein 4(HE4) in differential diagnosis of patients with low-grade serous(LGSC) and high-grade serous carcinoma(HGSC) serous ovarian cancer.Methods:LGSC and HGSC serous ovarian cancer were diagnosed by the two-tier grade system,serum levels of HE4 and carbohydrate antigen 12S(CA125) were measured by ELBA and radioisotope method,respectively in 60 serous ovarian cancer patients. HE4 and TPS3 protein in cancer tissue were measured by immunohistochemical method. Results:The difference in density of HE4 and TP53 protein was significant between LGSC and HGSC tissue,while serum CA12S did not show significant difference between different serum samples.There was significant difference in serum HE4 levels between LGSC and HGSC and the result was different within FIGO(Ⅰ+Ⅱ) stage,suggesting HE4 was not a reliable biomarker for the discrimination between LGSC and HCSC.HE4 had potential as a biomarker for the discrimination between LGSC and HGSC but the role in early diagnosis was limited.Conclusions:HE4 may be a reliable marker for differential diagnosis of LGSC and HGSC.But its role in early diagnosis of LGSC and HGSC need to be confirmed from the perspective of two-tier grade system.
文摘Vasectomy is a simple and reliable method of male contraception. A growing number of men after vasectomy request vasectomy reversal due to various reasons. The pregnancy rate is lower than the patency rate after vasovasostomy and the pregnancy rate is time dependent. In this study, we evaluated the influence of reproductive tract obstruction on expression of epididymal proteins and their restoration after patency. Adult male Wistar rats were studied 30, 60 and 120 days after vasectomy, 30 days after vasovasostomy or after sham operations. Two-dimensional gel electrophoresis, mass-spectrometric technique, multidatabase search, Western blotting and real-time PCR were used to analyze the expression regulation of epididymal proteins. Total integrated intensity and total spot area of autoradiograms showed a consistent downward trend with time after obstruction, and this trend remained after patency. The intensity of the autoradiographic spots in three patency groups showed three trends： a downward trend, similar intensity and an upward trend compared with the correspondent obstruction group, respectively. Further verified experiments on human epididymis 2 （HE2）, fertilization antigen-1 （FA-1）, clusterin and PH20 demonstrated that compared with the correspondent obstruction group, the translation levels of HE2 and the mRNA transcription levels of HE2 showed an upward trend in patency groups, especially in the groups of obstruction for 60 days where the expression levels of HE2 were significantly upregulated after patency （P〈O.05）. Reproductive tract obstruction provokes a disregulation of gene expression in the epididymis and this disregulation remained after patency. Successful reversal may recover some proteins and the recovery is time dependent, Obstruction differentially alters mRNA transcription of different proteins and the content of proteins seemed to be easier to be influenced than the gene transcription.
基金The authors would like to thank Mr Shou-Xin Zhang and other members of the Research Center,Yuhuangding Hospital(Yantai,China)for technical assistance.
文摘To explore the functions of human ribonuclease 9(RNase 9),we constructed a mammalian fusion expression vector pcDNA-hRNase9,prepared recombinant human RNase 9-His fusion protein from HEK293T cells and determined its N-terminal amino acid sequences.According to the determined mature protein,recombinant human RNase 9 was prepared in E.coli.Ribonucleolytic activity and antibacterial activity of recombinant human RNase 9 were detected,and the distribution of human RNase 9 on tissues and ejaculated spermatozoa and in vitro capacitated spermatozoa were analyzed via indirect immunofluorescence assay.The results showed that recombinant human RNase 9 did not exhibit detectable ribonucleolytic activity against yeast tRNA,but exhibited antibacterial activity,in a concentration/time dependent manner,against E.coli.Immunofluorescent analyses showed that the predicted human RNase 9 was present throughout the epididymis,but not present in other tissues examined,and human RNase 9 was also present on the entire head and neck regions of human ejaculated spermatozoa and in vitro capacitated spermatozoa.These results suggest that human RNase 9 may play roles in host defense of male reproductive tract.
文摘目的:分析血清人附睾分泌蛋白4(Human epididymis protein 4,HE4)水平和子宫内膜癌患者临床病理特征的相关性.方法:回顾性分析2019年1月至2022年6月医院收治的120例子宫内膜癌患者临床资料.所有患者均实施子宫全切术治疗,收集患者的临床病理特征资料,检测所有患者HE4水平,分析血清HE4水平与子宫内膜癌患者临床病理特征的相关性.结果:120例子宫内膜癌患者中,病理类型:子宫内膜样腺癌46例、子宫内膜样透明细胞癌36例、子宫内膜样鳞癌38例;淋巴结转移:发生35例、未发生85例;肌层浸润深度:>1/2肌层40例、≤1/2肌层80例;TNM分期:Ⅰ期22例、Ⅱ期43例、Ⅲ期55例.病灶所处部位:子宫底55例、子宫角37例、宫颈管28例.不同病理类型、TNM分期、病灶所处部位患者的血清HE4水平比较,差异无统计学意义;肌层浸润深度>1/2肌层患者的血清HE4水平高于肌层浸润深度≤1/2肌层的患者,发生淋巴结转移患者的血清HE4水平较未发生淋巴结转移的患者明显增加(P<0.05).经Logistic回归显示,血清HE4水平升高是子宫内膜癌患者肌层浸润深度>1/2肌层的风险因子(OR>1,P<0.05);血清HE4水平升高是子宫内膜癌患者发生淋巴结转移的风险因子(OR>1,P<0.05).结论:血清HE4水平升高增加可加剧肌层浸润深度,促使子宫内膜癌患者发生淋巴结转移.
基金We thank Dr Qiang Liu for giving some advice and polishing the writing and thank Dr Yu-Chuan Zhou for fruitful discussions. We are also grateful to Ai- H ua Liu fur technical assistance in immunohistochemistry assay. This work was supported by grants from the National Natural Science Foundation of China （No. 30930053） and the Chinese Academy of Sciences Knowledge Innovation Program （No. KSCX2-EW-R-07）.
文摘Previous studies have shown that rat epididymis-specific gene HongrES1 plays important roles in sperm capacitation and fertility. In this study, we cloned the mouse homologue gene by sequence alignment and RT-PCR methods and designated it as mHongl. The mHongl gene is located on chromosome 12p14, spanning five exons. The cDNA sequence consists of 1257 nucleotides and encodes a 419 amino-acid protein with a predicted N-terminal signal peptide of 20 amino acids. The mHongl mRNA shows similarity with HongrES1 in the expression patterns： （i） specific expression in epididymal tissue, especially in the cauda region; and （ii） androgen-dependence but testicular fluid factor independence. Its protein product shows 71% similarity with HongrES 1 and contains a classical serpin domain as does HongrES1. A polyclonal antibody against mHongl with high specificity and sensitivity was raised. Like HongrES1, the mHongl protein shows a checker-board expression pattern in the epididymal epithelium and is secreted into the epididymal lumen. The mHongl protein shows higher glycosylation than HongrES1. Although both of them are deposited onto the sperm head surface, mHongl is localized to the equatorial segment, which is different from that of HongrES 1. The mHongl protein can be removed from the sperm membrane by high ionic strength and therefore can be classed as an extrinsic membrane protein. Collectively, we conclude that mHongl is the homologue of HongrES1 and the present work paves the way for establishing animal models to elucidate the precise functions of HongrES1 and mHongl.
