子宫内膜癌患者血清及组织YKL-40、HE4及IGF的表达及意义

目的:研究子宫内膜癌患者血清及组织甲壳质酶蛋白40(YKL-40)、人附睾蛋白4(HE4)以及胰岛素样生长因子受体(IGF)的表达及其意义。方法:选取2013年6月到2014年6月我院收治的子宫内膜癌患者为研究组(30例),另外选取正常子宫增生者为对照组(30例),采用蛋白质印迹法(western-blot)测量两组血清中YKL-40、HE4,应用PV-600Pic Ture TM免疫组化测量两组子宫内膜组织中IGF,比较研究组手术前后和对照组血清及组织中的YKL-40、HE4及IGF的水平。结果:研究组手术前血清中YKL-40、HE4分别为(102.4±0.9)μg/L和(105.7±1.8)pmol/L显著高于对照组的(41.7±1.3)μg/L和(31.5±1.1)pmol/L,两组比较差异具有统计学意义(t=11.321,12.973,P=0.021,0.019);研究组内膜组织中IGF阳性表达率为93.3%(28/30)显著高于对照组的66.7%(20/30),两组比较差异具有统计学意义(x2=11.967,P=0.013);研究组内膜组织中过度表达者43.3%(13/30)显著高于对照组的0.0%,两组比较差异具有统计学意义(x2=14.168,P=0.008);手术后研究组血清中YKL-40、HE4分别为(30.5±1.1)μg/L和(24.7±1.5)pmol/L,与手术前比较差异具有统计学意义(t=11.938,11.027,P=0.011,0.009);子宫内膜癌血清及组织YKL-40、HE4及IGF与子宫内膜癌的分期呈正相关(r=0.381,0.423,0.261,P=0.018,0.021,0.028)。结论:子宫内膜癌患者血清中以及组织中YKL-40、HE4及IGF的表达显著增高,可以辅助诊断早期子宫内膜癌。     

血清ProGRP、SCCAg及HE4与非小细胞肺癌患者病理特征的关系及其诊断价值分析

目的:探讨血清胃泌素释放肽前体(ProGRP)、鳞状上皮细胞癌抗原(SCCAg)、人附睾蛋白4(HE4)水平与非小细胞肺癌(NSCLC)患者病理特征的关系,分析其对NSCLC的临床诊断价值。方法:选择2015年9月至2020年2月我院接诊的110例NSCLC患者(观察组)和100例健康志愿者(对照组)为研究对象,检测血清ProGRP、SCCAg、HE4水平。分析血清ProGRP、SCCAg、HE4水平与NSCLC患者临床病理特征的关系。采用受试者工作特征(ROC)曲线分析血清ProGRP、SCCAg、HE4诊断NSCLC的价值。结果:观察组血清ProGRP、SCCAg、HE4水平均高于对照组(P0.05),低中分化、TNMⅢ～Ⅳ期、淋巴结转移患者血清ProGRP、SCCAg水平分别高于高分化、TNMⅠ～Ⅱ期、无淋巴结转移患者(P0.05);TNM分期Ⅲ～Ⅳ期、淋巴结转移患者血清HE4水平分别高于TNMⅠ～Ⅱ期、无淋巴结转移患者(P0.05)。ROC曲线分析结果显示ProGRP、SCCAg、HE4、ProGRP+SCCAg+HE4诊断NSCLC的曲线下面积(AUC)分别为0.834(95%CI:0.779～0.888)、0.584(95%CI:0.507～0.662)、0.743(95%CI:0.675～0.811)、0.947(95%CI:0.910～0.984)。结论:NSCLC患者血清ProGRP、SCCAg、HE4水平明显升高,血清ProGRP、SCCAg水平与NSCLC患者分化程度、TNM分期和淋巴结转移有关,血清HE4水平与TNM分期和淋巴结转移有关。联合检测ProGRP、SCCAg、HE4对NSCLC诊断具有较高价值,可提高早期诊断准确性。     

不同强度电针对肥胖大鼠脂肪组织炎症相关因子的影响

探讨不同强度电针对肥胖大鼠脂肪组织核因子-κBp65(NF-κBp65)、单核细胞趋化蛋白-1(MCP-1)和肿瘤坏死因子-α(TNF-α)的作用差异.将SD大鼠随机分为普通饮食组、高脂饮食组、5 V电针组、2.5 V电针组,除普通饮食组外其余各组大鼠均饲以高脂饲料.取"足三里"、"三阴交"穴,不同强度电针治疗14 d后,用蛋白质印迹技术(Western blot)检测肥胖大鼠附睾脂肪组织NF-κBp65的表达,酶联免疫吸附法(ELISA)检测肥胖大鼠附睾脂肪组织MCP-1、TNF-α的含量.研究发现两电针组肥胖大鼠体重、Lee’s指数、脂肪组织中NF-κBp65表达、MCP-1和TNF-α含量较高脂饮食组显著降低(P<0.01),5 V电针组较2.5 V电针组下降效果更为明显(P<0.01,P<0.05).结果表明电针可改善肥胖脂肪组织炎症反应状态,减轻肥胖大鼠体重,且5 V电针组效果优于2.5 V电针组.     

人附睾蛋白4在卵巢癌中的研究进展

卵巢癌是严重威胁女性健康的恶性疾病之一。糖类抗原125（carbohydrate antigen125,CA125）是目前临床上广泛用于诊断卵巢上皮性癌的肿瘤标记物,然而其缺乏敏感性和特异性,因此,迫切需要寻找一种新的标志物来提高卵巢癌的诊断。人附睾蛋白4（human epididymis protein4,HE4）是近年发现的肿瘤标志物,其在良性肿瘤和正常组织中含量极低,但在卵巢癌中高表达。本文就HE4在卵巢癌中的诊断,动态监测等方面的进展做一综述。     

Epididymal G protein-coupled receptor (GPCR): two hats and a two-piece suit tailored at the GPS motif

G protein-coupled receptors (GPCRs) are involved in cell recognition and signaling and their function has been experimentally determined by ligand activation and site-directed mutagenesis. Structurally, GPCRs consist of an extracellular N-terminus and an intracellular C-terminus separated by seven helical transmembrane domains (TM7). The extracellular region is highly glycosylated. The intracellular region binds to G proteins. An epididymal GPCR, designated HE6 (for human epididymis-specific protein 6), is present in the stereocilia projecting from the apical domain of principal cells into the epididymal lumen. In conceptual terms, HE6 wears two hats: an unusually long extracellular region characteristic of cell adhesion proteins, and an intracellular region with binding affinity to G protein. The binding partner to the long extracellular region has not been identified. HE6 has another remarkable feature comparable to the GPCR calcium-independent receptor of alpha-latrotoxin, designated CIRL. Both HE6 and CIRL are endogenously cleaved into two pieces at the GPCR proteolytic site (GPS) located adjacent to TM1, the first of the seven transmembrane helices. One fragment of the heterodimer wears the cell adhesion hat; the other retains the typical characteristics of GPCRs. This proteolytic processing may be regarded as a mechanism of molecular compartmentalization of cell adhesion and G protein activation functions. The latter may engage a beta-arrestin-driven endocytic trafficking mechanism independent from the adhesive properties of the mucin extracellular domain. It is also conceivable that events taking place in the epididymal lumen can be surveyed by the long adhesive rod and subsequently coupled inside principal cells to a signaling cascade.     

Identification and characterization of P31m, a novel sperm protein in Cynomolgus monkey (Macaca fascicularis)

We have previously described a hamster sperm glycoprotein, P26h, which is implicated in the cascade of events occurring during the interaction between mature spermatozoa and the oocyte's zona pellucida. The P26h is acquired on the acrosomal cap of the spermatozoon during its maturation arising within the epididymis. Lately, using a polyclonal antiserum raised against P26h, a 34 kDa protein, P34H, has been identified on the acrosomal cap of the human spermatozoon. The cloning and sequencing of the cDNA encoding P34H has revealed a 65% similarity between the P34H and P26h amino acid sequences. Considering that P26h shows total immunocontraceptive properties in the hamster, it is of relevant importance to have an animal model phylogenetically closer to the human. Using the Cynomolgus monkey, we searched for a protein autologous to the human P34H. A 31 kDa protein, the P31m, localized on the acrosomal cap of the monkey spermatozoon has been identified by a Western blot analysis and by immunohistochemical techniques using an anti-hamster P26h antiserum. Northern blot analysis showed increasing high levels of the P31m mRNA through the epididymis and at lower levels in the testis. In situ hybridization showed the presence of the P31m mRNA in the principal cells of the epididymis. The cloning and sequencing of the cDNA encoding the P31m showed a high homology of 97% identity between the P31m and P34H nucleotidic sequences. This study clearly demonstrates that the monkey P31m is the homologous protein of the hamster P26h and of the human P34H. Mol. Reprod. Dev. 59: 431-441, 2001.     

Biogenesis of lysosomes in marshall cells and in cells of the male reproductive system

The mechanism of plasma membrane trafficking and degradation is still poorly understood. This investigation deals with the biogenesis of lysosomes during endocytic flow in Marshall cells and in various cell types of the male reproductive system. Marshall cells were exposed to ammonium chloride (NH4Cl) and leupeptin after labeling with cationic ferritin. In some experiments, the treated cells were immunogold labeled with anti-prosaposin antibody. NH4Cl and leupeptin are lysosomotropic agents that affect the endosomal-lysosomal progression. Testes, efferent ducts and epididymis from mouse mutants with defects affecting plasma membrane degradation were also used to analyze this process. NH4Cl produced a retention of cationic ferritin in endosomes and hindered the endosomal/lysosomal progression. Leupeptin did not affect this process. NH4Cl decreased the labeling of prosaposin in endosomes and lysosomes, while leupeptin increased the labeling of prosaposin in lysosomes. The number of lysosomes per cytoplasmic area was higher in treated cells than in controls. These findings suggest that leupeptin affected lysosomes whereas NH4Cl affected both endosomes and lysosomes. The endosomal and lysosomal accumulation of prosaposin induced by the treatment with NH4Cl and leupeptin indicated that the site of entry of prosaposinwas both the lysosome and endosome. Electron microscopy (EM) of tissues from mouse mutants with defects affecting plasma membrane degradation substantiated these observations. The EM analysis revealed a selective accumulation of multivesicular bodies (MVBs) and the disappearance of lysosomes, in testicular fibroblasts, nonciliated cells of the efferent ducts and principal cells of the epididymis, suggesting that MVBs are precursors of lysosomes. In conclusion: (1) endosomes and MVBs are a required steps for degradation of membranes; (2) endosomes and MVBs are precursors of lysosomes; and (3) endosomes, MVBs, and lysosomes appear to be transient organelles.     

CASK is in the mammalian sperm head and is processed during epididymal maturation

Upon adhesion to the zona pellucida or egg extracellular matrix, sperm undergo regulated exocytosis of the acrosomal vesicle. CASK is an adaptor protein that has been implicated in coupling neuronal cell adhesion to regulated exocytosis. In neurons, this scaffolding molecule is associated with several types of transmembrane receptor complexes and connects cell adhesion molecules with ion channels, the actin cytoskeleton, and the cell's exocytotic machinery. We hypothesized CASK might also be an important link between zona pellucida binding and the sperm acrosome reaction. RT-PCR experiments indicated CASK is transcribed in mouse testis. The full size (120 kDa) CASK protein was present in testis from mouse and pig. Immunoblots of mature porcine and murine sperm revealed that the 120 kDa molecule was much less abundant than in testis but the antibody also recognized a group of smaller proteins migrating at 55-65 kDa. Immunofluorescence experiments indicated both the full length and smaller CASK immunoreactive products were found only in the acrosomal region of spermatids and mature sperm and not in other testicular cell types. CASK immunofluorescence was lost following the acrosome reaction. During epididymal maturation, the abundance of the full size CASK decreased and the CASK fragments increased. These results suggest that CASK may be proteolytically processed during epididymal maturation. Because sperm acquire the ability to bind the zona pellucida, acrosome react, and fertilize eggs during epididymal maturation, CASK processing may play a role in the acquisition of these functions.     

Effects of FSH receptor deletion on epididymal tubules and sperm morphology, numbers, and motility

Follicle stimulating hormone (FSH) interacts with its cognate receptor (R) on Sertoli cells within the testis and plays an important role in the maintenance of spermatogenesis. Male FSH-R knockout (FORKO) mice show fewer Sertoli cells and many that are structurally abnormal and as a consequence fewer germ cells. Lower levels of serum testosterone (T) and androgen binding protein (ABP) also occur, along with reduced fertility. To assess the effects of FSH-R depletion as an outcome of testicular abnormalities, sperm from the cauda epididymidis were counted and examined ultrastructurally. As reduced fertility may also reflect changes to the epididymis, the secondary responses of the epididymis to lower T and ABP levels were also examined by comparing differences in sizes of epididymal tubules in various regions of FORKO and wild type (WT) mice. Sperm motility was evaluated in FORKO mice and compared to that of WT mice by computer assisted sperm analysis (CASA). Quantitatively, the data revealed that epithelial areas of the caput and corpus epididymidis were significantly smaller in FORKO mice compared to WT mice. Cauda epididymal sperm counts in FORKO mice were also much lower than in WT mice. This resulted in changes to 9 out of 14 sperm motility parameters, related mostly to velocity measures, which were significantly lower in the FORKO mice. The greatest change was observed relative to the percent static sperm, which was elevated by 20% in FORKO mice compared to controls. EM analyses revealed major changes to the structure of the heads and tails of cauda luminal sperm in FORKO mice. Taken together these data suggest a key role for the FSH receptor in maintaining Sertoli cells to sustain normal sperm numbers and proper shapes of their heads and tails. In addition, the shrinkage in epididymal epithelial areas observed in FORKO mice likely reflect direct and/or indirect changes in the functions of these cells and their role in promoting sperm motility, which is noticeably altered in FORKO mice.