子宫脱垂患者子宫主韧带和阴道前壁组织神经丝蛋白的表达及意义

目的探讨子宫脱垂(Uterine prolapse,UP)患者子宫主韧带和阴道前壁中神经丝蛋白(Neurofilament protein,NFP)组织的表达及其与子宫脱垂临床因素的关系。方法应用免疫组织化学S-P法对50例子宫脱垂患者的子宫主韧带和阴道前壁组织中神经丝蛋白表达进行测定,并与10例对照组组织对比,结合子宫脱垂的临床参数进行分析。结果免疫组化结果显示子宫脱垂患者阴道壁、主韧带组织中神经丝蛋白表达的阳性率为28.0%、32.0%,明显低于对照70.0%、80.0%,Ⅱ度子宫脱垂患者阴道壁、主韧带组织中神经丝蛋白表达阳性表达率明显低于Ⅲ+Ⅳ度子宫脱垂组(45.45%vs14.29%、50.0%vs17.86%)。结论子宫脱垂患者子宫主韧带和阴道前壁组织中神经丝蛋白表达下降,并与脱垂程度相关,提示神经丝蛋白可能在子宫脱垂的发生发展中起重要的作用。     

血清miR-21表达水平与子宫脱垂和阴道壁脱垂的相关性

摘要 目的：探讨血清miR-21表达水平与子宫脱垂和阴道壁脱垂的相关性。方法：选取我院2017年12月到2020年12月共收治的60例子宫脱垂和阴道壁脱垂的患者作为观察组,另取60例来我院体检的健康女性作为对照组,分析两组患者胶原蛋白Ⅰ、Ⅲ和miR-21表达水平,并分析血清miR-21表达水平与子宫脱垂和阴道壁脱垂的相关性。对所有患者经盆底重建术和相关治疗之后,将术后1年子宫和阴道壁基本复位未见脱垂,且POP-Q分度＜Ⅱ度的40例患者分为预后良好组,将术后1年内再次发生子宫脱垂和阴道壁脱垂,且POP-Q分度≥Ⅱ度的20例患者分为预后不良组,对比两组患者的临床情况,并分期血清miR-21表达水平对于子宫脱垂和阴道壁脱垂的预后预测价值。结果：两组患者胶原蛋白Ⅰ、Ⅲ和血清miR-21表达水平对比差异显著,观察组患者胶原蛋白Ⅰ、Ⅲ水平低于对照组,观察组患者血清miR-21水平高于对照组（P＜0.05）;Spearman相关分析结果显示：胶原蛋白Ⅰ、Ⅲ水平与子宫脱垂和阴道壁脱垂呈负相关,血清miR-21与子宫脱垂和阴道壁脱垂呈正相关（P＜0.05）;预后良好组与预后不良组患者孕次、脱垂部位、胶原蛋白Ⅰ、Ⅲ情况对比无差异（P＞0.05）,预后良好组与预后不良组患者年龄、BIM、产次、miR-21情况对比有差异（P＜0.05）;logistic回归分析结果得出,只有血清miR-21水平和BMI是子宫脱垂和阴道壁脱垂的预后独立因素（P＜0.05）。结论：胶原蛋白Ⅰ、Ⅲ和血清miR-21表达水平与子宫脱垂和阴道壁脱垂疾病具有明显相关性,因此可以考虑应用三种指标对患者进行诊断和治疗参考。而仅有血清miR-21水平和BIM水平是子宫脱垂和阴道壁脱垂的预后独立因素,可以用两者的水平来判断患者的预后情况,及时采取措施提升治疗方法。     

DcR3在甲状腺乳头状癌中的表达及临床意义

应用RT-PCR、Westem blot、免疫组化分别检测甲状腺乳头状癌组织与癌旁正常甲状腺组织标本中DcR3mRNA及蛋白的表达情况,探讨DcR3在甲状腺乳头状癌组织中的表达及,临床意义。RT-PCR检测显示,甲状腺乳头状癌中DcR3 mRNA的表达明显高于正常甲状腺组织（P〈0．05）：Western blot提示,DcR3蛋白在甲状腺乳头状癌中表达比正常甲状腺组织高（P〈0．05）;免疫组化显示,DcR3蛋白在甲状腺乳头状癌中高表达（P〈0．05）。DcR3mRNA及蛋白质在甲状腺乳头状癌及正常甲状腺组织间的表达差异有统计学意义（P〈0．05）。DcR3基因及蛋白在甲状腺乳头状癌中高表达,提示DcR3可能促进了甲状腺乳头状癌的发生发展。     

Fibulin-5在主动脉夹层血管壁中表达异常

目的:研究Stanford A型主动脉夹层(aortic dissection,AD)升主动脉和正常升主动脉血管组织中Fibulin-5表达的差异。方法:收集Stanford A型AD患者手术中切除的升主动脉血管组织标本12例(AD组),多器官捐献患者升主动脉血管12例(对照组)。采用EVG染色观察主动脉中膜弹性纤维形态结构;应用SP免疫组织化学法及Western blot法对标本组织中的Fibulin-5进行检测分析。结果:AD组主动脉中膜弹性纤维形态和排列不规则、破碎、丢失,结构紊乱。免疫组织化学显示Fibulin-5阳性表达见于主动脉壁平滑肌细胞胞质中,AD组与对照组比较,Fibulin-5表达明显较少。Western blot蛋白印迹示AD组Fibulin-5表达明显减少,差异有统计学意义(P0.05)。结论:Fibulin-5在AD主动脉中膜中表达下调,可能在AD的发生中发挥作用。     

感觉神经损伤性盐敏感性高血压大鼠心、肾AT1R的表达

目的对感觉神经损伤性盐敏感性高血压大鼠的心肌、肾脏组织中的血管紧张素Ⅱ1型受体（AT1R）在mRNA和受体水平的表达进行检测,探讨AT1R与盐敏感性高血压的关系。方法用乳鼠皮下注射辣椒辣素法建立模型。哺乳期后,大鼠被随机分成4组：对照＋正常盐饮食组（CON-NS）;对照＋高盐饮食组（CON-HS）;辣椒辣素＋正常盐饮食组（CAP-NS）;辣椒辣素＋高盐饮食组（CAP-HS）。至7周龄（分组饲养后第4周）处死大鼠,免疫组织化学方法和反转录-聚合酶链式反应（RT-PCR）分别检测大鼠心肌和肾脏AT1R蛋白,以及AT1 R mRNA的表达。结果①Wistar大鼠在给予不同程度的感觉神经损伤和饲料干预后,各组大鼠尾部收缩压均有明显增加,最终CAP-HS组的尾收缩压显著高于其他三组（P〈0.01）。②免疫组织化学结果显示,CAP-HS组组织有显著的AT1R蛋白表达（P〈0.01）;CON-HS组肾脏、心肌组织中AT1R蛋白表达高于CON-NS组（P〈0.05）。③RT-PCR检测基因表达,与对照组CON-NS相比,实验组CAP-HS的AT1R mRNA表达显著升高（P〈0.01）;CON-HS组肾脏、心肌组织中AT1 R mRNA表达有显著性（P〈0.05）。结论感觉神经损伤性盐敏感性高血压大鼠心、肾AT1R表达升高,AT1R表达水平的差异可能与盐敏感性高血压的形成有关。     

基质金属蛋白酶及其抑制剂mRNA在正常及异位子宫内膜中的表达

目的：观察正常及异位子宫内膜中基质金属蛋白酶－1,－2（MMP－1,－2）和基质金属蛋白酶组织抑制剂－1（TIMP－1）基因表达的变化,以探讨其与子宫内膜异位症的关系。方法：应用原位技术,采用地高辛生物素标记的cDNA探针（MMP－1,MMP－2,TIMP－1）对子宫内膜异位症患者（EM组）的子宫内膜（14例）、异位病灶组织（20例）及非子宫内膜异位症患者（对照组）的子宫内膜（12例）,分3批进行分子杂交检测,结果：3批结果相似。MMP－1,－2,TIMP－1mRNA在腺上皮细胞和间质细胞均有表达,EM组的子宫内膜MMP－1,－2及TIMP－1mRNA表达量与对照组无显著性差异（P＞0．05）,异位病灶组织的MMP－1,－2mRNA表达量明显高于对照组（P＜0．05）,而TIMP－1mRNA表达量则低于对照组（P＜0．05）。结论：子宫内膜异位症患者的异位病灶中,存在MMP－1,－2和TIMP－1明显的平衡失调,这可能与子宫内膜异位症的发生、发展和不孕有关。     

PKC、PKA信号通路在调控体外培养人牙囊细胞VEGF表达中的作用

该文主要探讨PKC、PKA信号通路在调控体外培养人牙囊细胞VEGF表达中的作用。选取生长状态良好的第4代人牙囊细胞,采用Real-time PCR和Western blot分别检测PKC激动剂（PMA）、PKC非特异性抑制剂（G 6983）、PKC-α和γ特异性抑制剂（HBDDE）、PKC-β特异性抑制剂（LY333531）、PKA激动剂（dbcAMP）和抑制剂（KT5720）对体外培养人牙囊细胞VEGF mRNA和蛋白表达的影响。结果显示,PMA组和PMA＋HBDDE组VEGF mRNA和蛋白的表达水平明显高于对照组,差异有统计学意义（P〈0.05）;而PMA＋G 6983组和PMA＋LY333531组VEGF mRNA和蛋白的表达水平与对照组之间无明显差异（P〉0.05）。dbcAMP组VEGF mRNA和蛋白的表达水平明显高于对照组,差异有统计学意义（P〈0.05）;而dbcAMP＋KT5720组VEGF mRNA和蛋白的表达水平与对照组之间无明显差异（P〉0.05）。这表明,PKC、PKA信号通路均参与了体外培养人牙囊细胞VEGF表达的调控,其中PKC信号通路中参与调控的亚型是PKC-β。     

糖尿病性膀胱病逼尿肌中SCFmRNA和SCF蛋白的表达

目的：检测DCP逼尿肌中SCF表达水平,探讨SCF基因表达与DCP关系及其发病机制。方法：按1：2病例对照研究,采用链脲佐菌素（STZ）及尿动力学检测成功建立DCP豚鼠20只为实验组,并以同质豚鼠40为对照组,应用RT-PCR和Western-blotting方法分别检测各组膀胱逼尿肌中SCF mRNA、SCF蛋白的表达。结果：DCP豚鼠组织中SCF mRNA表达与正常对照组比较无明显显著差异（P〉0.05）,DCP豚鼠组织中SCF蛋白表达明显低于正常对照组（P〈0.01）。结论：DCP组织中SCF蛋白表达减少与SCF基因翻译水平异常有关,因此高血糖环境下SCF基因表达异常可能是DCP的发病机制之一。     

盆底超声参数预测剖宫产术后压力性尿失禁的效能及其与盆底组织E-cadherin和Vimentin表达的关系研究

摘要 目的：研究盆底超声参数预测剖宫产术后压力性尿失禁（SUI）的效能及其与盆底组织钙粘附蛋白E（E-cadherin）和波形蛋白（Vimentin）表达的关系。方法：选取2020年1月～2021年1月湖南省妇幼保健院收治的124例剖宫产患者。将其按照是否发生SUI分为SUI组18例以及无SUI组106例。对所有患者均进行盆底超声检查,比较两组静息期（R）、张力期Valsalva动作（V）的膀胱尿道后角（PUA）以及膀胱颈到耻骨联合下缘水平垂直距离（BNS）,膀胱颈移动度（BND）、尿道旋转角度（UR）。通过受试者工作特征（ROC）曲线分析盆底超声参数预测剖宫产术后SUI的效能。另外,对比两组盆底组织E-cadherin和Vimentin mRNA水平,采用Pearson相关性分析各项盆底超声参数和盆底组织E-cadherin、Vimentin mRNA表达的相关性。结果：SUI组R-PUA、V-PUA、BND以及UR均高于无SUI组,而R-BNS以及V-BNS均低于无SUI组（均P＜0.05）。ROC曲线分析结果：各项盆底超声参数联合预测剖宫产术后SUI的效能均优于各项参数单独预测。SUI组盆底组织E-cadherin mRNA水平低于无SUI组,而Vimentin mRNA水平高于无SUI组（均P＜0.05）。经Pearson相关性分析发现,R-PUA、V-PUA、BND、UR均和盆底组织E-cadherin mRNA水平呈负相关关系,与Vimentin mRNA水平呈正相关关系;而R-BNS、V-BNS均和盆底组织E-cadherin mRNA水平呈正相关关系,与Vimentin mRNA水平呈负相关关系（均P＜0.05）。结论：盆底超声参数预测剖宫产术后SUI的效能较高,且和盆底组织E-cadherin、Vimentin表达有关。     

乌司他丁对机械通气致肺损伤大鼠肺组织CC16表达的影响

目的通过观察乌司他丁（UTI）对大潮气量机械通气大鼠肺组织Clara细胞分泌蛋白（CC16）表达的影响,探讨CC16在机械通气所致肺损伤（VILI）发病中作用以及UTI对VILI的干预作用。方法 24只雄性Wistar大鼠随机分为对照组、大潮气量组和UTI干预组,观察其肺组织病理学改变,测定肺组织中丙二醛（MDA）和BALF中总蛋白（TP）含量,采用逆转录聚合酶链反应（RT-PCR）法检测肺组织CC16 mRNA的表达,采用免疫组织化学染色法检测肺组织中CC16蛋白表达及Clara细胞计数。结果与对照组比较,大潮气量肺组织MDA的含量及BALF总蛋白含量明显升高（P〈0.01）,而肺细支气管上皮细胞CC16mRNA及其蛋白表达水平明显降低（P〈0.01）;与大潮气量组比较,UTI干预组大鼠肺组织中MDA的含量及BALF总蛋白含量明显降低（P〈0.01）,而肺细支气管上皮细胞CC16mRNA及其蛋白表达水平明显升高（P〈0.01）。结论大潮气量机械通气导致肺组织CC16 mRNA及其蛋白表达水平降低在VILI发病中起重要作用,乌司他丁不但能促进Clara细胞分泌CC16而抑制肺组织炎症反应,还能抑制脂质过氧化物的产生,对VILI有一定保护作用。     

Coordinate expression of fibulin-5/DANCE and elastin during lung injury repair

Fibulin-5, previously known as DANCE and EVEC, is a secreted extracellular matrix protein that functions as a scaffold for elastin fiber assembly and as a ligand for integrins alphavbeta3, alphavbeta5, and alpha9beta1. Fibulin-5 is developmentally regulated in the lung, and lung air space enlargement develops in mice deficient in fibulin-5. Fibulin-5 is also induced in adult lung following lung injury by hyperoxia. To further examine the role of fibulin-5 during repair of lung injury, we assessed fibulin-5 expression during elastase-induced emphysema in C57/b mice. Mice were treated with either saline or elastase via the trachea, and the lung was examined 20 days after treatment. Fibulin-5 mRNA was induced almost fourfold, whereas elastin mRNA was minimally elevated. Immunohistochemistry studies showed that fibulin-5 was induced in cells within the alveolar wall following elastase treatment. Western analysis demonstrates that fibulin-5 was strongly expressed in isolated primary lung interstitial fibroblasts. Fibulin-5 protein was localized to the fibroblast cell layer in culture, and brief elastase treatment degraded the protein. Intact fibulin-5 did not accumulate in the culture media. Treatment of fibroblasts with the proinflammatory cytokine interleukin-1beta abolished fibulin-5 mRNA expression. Our results indicate that fibulin-5 is coordinately expressed and regulated with elastin in lung fibroblasts and may serve a key role during lung injury and repair.     

Decrease of Fibulin-3 in Hepatocellular Carcinoma Indicates Poor Prognosis

Fibulin-3, originally identified in senescent and Werner syndrome fibroblasts, has been implicated in cell morphology, growth, adhesion and motility. Fibulin-3 exhibits both antitumor and oncogenic activities towards human cancers; however, the role of Fibulin-3 in hepatocellular carcinoma (HCC) remains elusive. In this study, we showed that both the mRNA and protein levels of Fibulin-3 were remarkably downregulated in HCC cell lines and fresh tissues. Immunohistochemical data revealed that Fibulin-3 was decreased in tumorous tissues in 67.1% (171/255) of cases compared to the corresponding adjacent nontumorous tissues. The results of statistical analysis indicated that low Fibulin-3 expression, defined by the receiver operating characteristic curve (ROC), was significantly associated with tumor differentiation (P = 0.008), clinical stage (P = 0.014) and serum AFP levels (P<0.01). Furthermore, Kaplan-Meier and multivariate analysis suggested that Fibulin-3 is an independent negative prognostic indicator for both overall (P<0.001) and recurrence-free (P = 0.036) survival. In addition, an in vitro study demonstrated that knockdown of Fibulin-3 by siRNA markedly increased cell viability and promoted cell invasion in HCC cells. Collectively, our data suggest that Fibulin-3 exhibits antitumor effects towards HCC and serves as a biomarker of unfavorable prognosis for this deadly disease.     

Fibulin-5, an integrin-binding matricellular protein: its function in development and disease

Interactions between the extracellular matrix (ECM) and cells are critical in embryonic development, tissue homeostasis, physiological remodeling, and tumorigenesis. Matricellular proteins, a group of ECM components, mediate cell-ECM interactions. One such molecule, Fibulin-5 is a 66-kDa glycoprotein secreted by various cell types, including vascular smooth muscle cells (SMCs), fibroblasts, and endothelial cells. Fibulin-5 contributes to the formation of elastic fibers by binding to structural components including tropoelastin and fibrillin-1, and to cross-linking enzymes, aiding elastic fiber assembly. Mice deficient in the fibulin-5 gene (Fbln5) exhibit systemic elastic fiber defects with manifestations of loose skin, tortuous aorta, emphysematous lung and genital prolapse. Although Fbln5 expression is down-regulated after birth, following the completion of elastic fiber formation, expression is reactivated upon tissue injury, affecting diverse cellular functions independent of its elastogenic function. Fibulin-5 contains an evolutionally conserved arginine-glycine-aspartic acid (RGD) motif in the N-terminal region, which mediates binding to a subset of integrins, including α5β1, αvβ3, and αvβ5. Fibulin-5 enhances substrate attachment of endothelial cells, while inhibiting migration and proliferation in a cell type- and context-dependent manner. The antagonistic function of fibulin-5 in angiogenesis has been demonstrated in vitro and in vivo; fibulin-5 may block angiogenesis by inducing the anti-angiogenic molecule thrompospondin-1, by antagonizing VEGF₁₆₅-mediated signaling, and/or by antagonizing fibronectin-mediated signaling through directly binding and blocking the α5β1 fibronectin receptor. The overall effect of fibulin-5 on tumor growth depends on the balance between the inhibitory property of fibulin-5 on angiogenesis and the direct effect of fibulin-5 on proliferation and migration of tumor cells. However, the effect of tumor-derived versus host microenvironment-derived fibulin-5 remains to be evaluated.     

Packed Cell Volume (PCV) in Connection with Uterine Prolapse and in Parturient Paresis in Cows

Packed cell volume was determined in 317 cows with uterine prolapse, the condition being in more than half of these cases associated with varying degrees of hypocalcaemia, in 133 cows with parturient paresis and in 73 clinically healthy newly calved cows. Independently of each other uterine prolapse and fall in plasma calcium were associated with a significant rise in PCV. The present investigation demonstrated a marked fall in circulating blood volume in a considerable number of cows suffering from uterine prolapse. The risk of irreversible hypovolemic shock developing in connection with uterine prolapse is not to be neglected, and may be aggravated by the simultaneous occurrence of hypocalcaemia, as several of the compensatory mechanisms are calcium dependent.     

Steroidal regulation of uterine edema and tissue inhibitors of metalloproteinase (TIMP)-3 messenger RNA expression is altered in TIMP-1-deficient mice

Tissue inhibitors of metalloproteinases (TIMPs) are expressed within the uteri of virtually all species where they are postulated to control extracellular matrix turnover, cellular apoptosis, and proliferation. The objective of the current study was to examine the steroidal regulation of uterine TIMP expression and to determine the potential role of the TIMP-1 gene product in this regulation. To accomplish these goals, ovariectomized female TIMP-1 wild-type and null mice were treated with estradiol, progesterone, or estradiol and progesterone and killed at various times after steroid administration. Estradiol induced a significant reduction in uterine TIMP-3 expression in wild-type mice at 8 and 24 h post-steroid administration, but the ability of this steroid to decrease TIMP-3 expression was impaired in the uteri of TIMP-1 null mice. Further, estrogen-induced uterine wet-weight gain/edema was enhanced in the TIMP-1 null mice, and the antiestrogen compound ICI 182780 or progesterone could only partially block this estrogenic effect. It is concluded from this study that steroidal modulation of uterine TIMP-3 expression and regulation of wet-weight gain/edema are altered in TIMP-1 null mice. These observations suggest that steroids induce uterine TIMP-1 expression and, in turn, that TIMP-1 influences TIMP-3 mRNA expression and uterine edema.     

Incidence of Some Diseases in Connection with Parturition in Dairy Cows

The incidence of dystocia, uterine, prolapse, retained placenta, puerperal metritis and vaginal prolapse was investigated in 283 herds over a period of eight years (about 2,590 cows per year). The incidence of dystocia (0.012 cases per cow and year), of uterine prolapse (0.002) and puerperal metritis (0.002) varied with the monthly variation in parturition. Retained placenta (incidence 0.025) occurred more frequently in March-June relative to parturition. The cows then calving were older than average. Vaginal prolapse (incidence 0.002) was also commoner in the spring months. Herds with a high incidence of retained placenta also showed more mastitis, ketosis and parturient paresis.     

Clinical implications of steroid receptor coactivator (SRC)-3 in uterine endometrial cancers

Estrogen is recognized as a significant modifier in the development, growth and invasion of uterine endometrial cancer. Steroid receptor coactivator-3 (SRC-3; AIB1, ACTR, RAC3, TRAM-1, and pCIP) is a member of the p160 family of coactivator for nuclear hormone receptors including estrogen receptor (ER). It is reported that SRC-3 is overexpressed in various cancers. However, SRC-3 expression manner in uterine endometrial cancer is not fully understood. In this study, we showed SRC-3 mRNA expression correlates with clinical stage, depth of myometrial invasion and dedifferentiation. The prognosis of the 25 patients with higher expression of SRC-3 mRNA in uterine endometrial cancers was extremely poor (36%), whereas the 24-month survival rate of the 15 patients with lower expression of SRC-3 mRNA was 96%. These data indicate that SRC-3 might be an important indicator of uterine endometrial cancer advancement and survival.