睾丸精子冻融后行卵胞浆内单精子注射术在无精子症患者中的临床应用

目的探讨对无精子症患者实施睾丸精子经冷冻复苏后行卵胞浆内单精子注射术（ICSI）的临床效果。方法回顾性分析了既往在我中心有冷冻睾丸精子的34个ICSI周期,评估其冷冻精子复苏、卵子受精、卵裂、可移植胚胎、优质胚胎、临床妊娠及其分娩情况。结果 34个拟定复苏冷冻睾丸精子的周期精子复苏均获得成功,受精率为76.9%（249/324）,2PN受精率69.7%（226/324）,卵裂率98.8%（246/249）,可利用胚胎率84.2%（207/246）,优质胚胎率46.3%（114/246）;所有周期均有胚胎移植;34个周期中行新鲜胚胎移植30个,种植率为34.4%（21/61）,临床妊娠17例（临床妊娠率56.7%）包括13例单胎和4例双胎,其中1例单胎妊娠流产（流产率为5.9%）。目前,有14个周期已经出生了18个健康婴儿（10个男婴,8个女婴）未发现先天缺陷儿,另2例单胎继续妊娠中（32＋w,29＋w）。结论睾丸精子经冷冻后有很好的复苏率,结合ICSI受精可以得到较好的临床结局。     

经皮睾丸精子抽吸术治疗无精子症的研究

目的探讨经皮睾丸精子抽吸术(PTSA)获取睾丸精子结合卵胞浆内单精子注射术(ICSI)治疗梗阻性和非梗阻性无精子症,使之获得亲生子女.方法对121例因男性梗阻性及非梗阻性无精子症患者进行诊断性穿刺,均证实有精子后进行119个周期PTSA ICSI治疗.结果共获卵子1514个,成熟卵985个,胚胎741个,平均每例6.23个胚胎,总受精率74.4%,卵裂率97.6%;共移植114个周期和冷冻胚胎移植5个周期,平均移植2.86个胚胎,B超证实临床妊娠48例,临床妊娠率40.3%.结论采用PTSA技术获取的睾丸精子进行ICSI是治疗梗阻性及非梗阻性无精子症的一种安全、简单、有效的方法.     

睾丸精子抽吸后行卵细胞单精子显微注射治疗梗阻性无精子症

无精子症约占男性不育患者的7％～14％。目前尚无确切的治疗方法。自Palerm等于1992年首次报道卵细胞浆内单精子注射（ICSI）获得妊娠成功，为男性不育症治疗提供了新方法，同时附睾或睾丸取精术彻底改变了无精子症不可治疗的局面。采用此方法只要在男性生殖道或睾丸内发现并分离到精子，利用ICSI技术就能获得一定的妊娠率，     

无精子症患者睾丸生精功能与血清性激素水平的相关性分析

目的通过对广东佛山地区104例无精子症患者的血清性激素水平进行检测并分析,探讨其与睾丸生精功能的相关性。方法 104例研究对象均于我院确诊为无精子症患者,测定其血清中总睾酮(T,nmol/L)、卵泡刺激素(FSH,IU/L)、黄体生成素(LH,IU/L)、泌乳素(PRL,ng/m L)、雌二醇(E2,ng/L)水平,根据睾丸活检病理结果分为生精功能正常组(1组)、生精功能低下组(2组)、唯支持细胞综合征组(3组)。结果各组患者的年龄均无统计学差异,1组与2组患者各性激素水平无统计学差异;3组患者血清FSH、LH、PRL水平高于2组,E2水平低于2组,有显著的统计学差异,虽然3组T水平低于2组,但两组间没有统计学差异;3组患者血清FSH、LH水平高于1组,T水平低于1组,有显著的统计学差异,但两组之间的PRL、E2水平并无统计学差异。相关分析显示睾丸生精功能与T水平呈正相关,与FSH、LH水平呈负相关,与PRL、E2水平没有相关性。结论血清性激素水平测定对于预测无精子症患者睾丸生精功能有重要意义,并可用于指导治疗及判断预后情况。     

经皮附睾或睾丸抽吸取精结合ICSI治疗无精子症

目的评价经皮附睾精子抽吸术（percutaneus epididymal sperm aspiration,PESA）或睾丸精子抽吸术（testicu1ar sperm aspiration,TESA）结合卵胞浆内单精子注射（intracytoplasmic sperm injection,ICSI）治疗无精子症的临床效果。方法对290例因男性梗阻性及非梗阻性无精子症（non-obstructive azoospermia,NOA）采用PESA或TESA穿刺获取精子,女方采用长方案超排卵,然后对处于细胞分裂中期的成熟卵母细胞进行单精予注射。结果梗阻性无精子症组203例,受精率77．5％,临床妊娠率46．1％;非梗阻性无精子症组87例,受精率73．O％,临床妊娠率41．4％,两组比较其受精率及临床妊娠率均无显著性差异（P〉0．05）。结论采用PESA或TESA获取精子结合ICSI是治疗梗阻性及非梗阻性无精子症等严重的男性不育症的一种有效的方法。     

附睾精子和睾丸精子妊娠结局比较

目的比较经皮附睾穿刺抽吸术和经皮睾丸精子抽吸术两种方法获得的精子妊娠结局。方法83例无精子症患者经皮附睾穿刺抽吸术取得附睾精子；35例无精子症患者经皮睾丸精子抽吸术（TESA）获得睾丸精子。女方进行常规超排卵。采用卵胞浆内单精子注射技术获得妊娠，比较两者的受精率、种植率和临床妊娠率。结果附睾精子组和睾丸精子组的受精率分别为75．20％和74．61％，比较其差异无显著性（P〉0．05）；两者的种植率和临床妊娠率分别为29．18％VS23．89％和52．43％VS40．21％，差异具有显著性（P〈0．05）。结论附睾是精子获能、成熟的重要部位，附睾精子优于睾丸精子，对无精子症患者行ICSI之前尽可能首先选取附睾精子。     

无精子症患者附睾穿刺及睾丸活检结果分析

无精子症占男性不育病例的10％左右,其病因多不确定,治疗效果也不好。本文通过对无精子症患者经皮附睾精子抽吸术（PESA）及睾丸活检（TESE）,分析睾丸病变,为其病因分析、临床诊疗、辅助生殖等提供实验室依据。     

无精子症患者血清中FSH的测定及临床意义

目的通过检测无精子症患者血清FSH水平,分析鉴别梗阻性无精子症和非梗阻性无精子症两种类型;通过比较血清FSH水平与睾丸体积,分析血清FSH水平与睾丸体积的关系,探讨其判断无精子症分型的意义。方法对2007年10月-2008年4月间76例来自大连市妇产医院白山路生殖中心的无精子症患者的血液样本,采用ROCHE2010全自动免疫分析仪及其配套试剂盒检测血清FSH水平,采用Prader睾丸模型比拟法测定睾丸体积。结果与对照组相比,非梗阻性无精子症组血清FSH水平显著升高（P〈0.05）,而对照组与梗阻性无精子症组血清FSH水平差异无显著性（P〉0.05）。非梗阻性无精子症组与梗阻性无精子症组血清FSH水平差异显著（P〈0.05）。与睾丸体积≥12ml组相比,睾丸体积〈12ml组血清FSH水平显著升高（P〈0.05）。以睾丸体积12ml为分界标准,≥12ml为梗阻性无精子症,〈12ml为非梗阻性无精子症。该标准对梗阻性和非梗阻性无精子症诊断的符合率分别为89.3%（25/28）、75%（36/48）。以生殖激素中FSH12.4mIU/ml为分界标准,小于该值为梗阻性无精子症,大于该值为非梗阻性无精子症,该标准对梗阻性无精子症诊断的符合率为92.9%（26/28）,对非梗阻性无精子症诊断的符合率为89.6%（43/48）。结论血清FSH水平为分析睾丸功能衰竭原因提供依据,对判定精道梗阻具有重要意义,是鉴别非梗阻性与梗阻性无精子症的首要临床指标;睾丸体积在判定无精子症分型中有一定意义;血清FSH水平与睾丸体积联合检测更有助于鉴别梗阻性无精子症和非梗阻性无精子症。     

两种TESA方法在腮腺炎致无精子症患者取精中的比较

目的探讨腮腺炎致无精症患者两种睾丸活检取精（TESA）即开放性睾丸活检和显微切割睾丸活检取精的优劣。方法每组24例患者,分别用开放性睾丸活检和显微切割睾丸活检取精,观察取精成功率、术后并发症发生率,术后并发症包括术后血肿的形成,术后3月睾丸萎缩、睾酮下降。结果开放性睾丸活检取精组,能找到活动的精子14例,取精成功率58．3％,术后并发症10例,约41．7％,分别为血肿2例、三月后睾丸萎缩3例、睾酮下降5例;显微切割睾丸活检取精组,能找到活动的精子18例,取精成功率75％,术后并发症5例,约20．8％,分别为血肿0例、三月后睾丸萎缩1例、睾酮下降4例。取精成功率和开放性睾丸活检取精组比较（P〈0．05）,术后并发症和开放性睾丸活检取精组此较（P〈0．01）。结论显微切割睾丸活检取精较开放手术睾丸活检取精成功率高、术后并发症少,值得推广。     

少弱精子症、死精子症、无精子症患者细胞遗传学分析

目的了解少弱精子症、死精子症、无精子症与染色体之间的联系。方法对2011年上半年生殖门诊101例少弱精子症、死精子症、无精子症患者,进行染色体检测,并对结果进行比较分析。结果染色体异常总数占23例,异常发生率达22.8%。性染色体异常9例,占异常总数的39%,常染色体异常14例,占异常总数61%。结论男性不育少弱精子症、死精子症、无精子症患者常规做染色体检查是必要的,对确定其是否有治疗价值提供重要依据。     

9-顺式维甲酸对甲状腺鳞癌细胞周期及CyclinD1、CDK4表达的影响

 摘 要：目的 探讨9-顺式维甲酸（9-cisRA）对甲状腺鳞癌细胞株SW579细胞周期及周期因子表达的影响。方法以终浓度分别为为10-7 mol/L、10-6 mol/L、5×10-6 mol/L、10-5 mol/L 的9-cisRA处理SW579细胞株,采用MTT比色法测定细胞增殖活性;流式细胞仪检测细胞周期;用RT-PCR方法检测SW579细胞CDK4、CyclinD1mRNA的表达水平;用Western blotting检测SW579细胞CDK4、CyclinD1的蛋白表达。结果 9-cisRA作用后,MTT结果显示,各组细胞均出现显著的生长抑制作用,生长抑制率明显升高,并呈剂量依赖性。流式细胞仪分析,各组细胞均随着药物浓度的升高,S期细胞在细胞周期中所占比例逐渐减少,G1期细胞则明显增加（P<0.01）,细胞滞留在G1期。RT-PCR检测结果表明,经不同浓度9-cisRA处理的细胞CDK4 mRNA表达水平没有明显变化;CyclinD1的表达水平明显下调;Western blotting检测结果表明经不同浓度9-cisRA处理的细胞CDK4蛋白表达水平没有明显变化,CyclinD1蛋白表达水平明显下降。结论9-cisRA对SW579细胞有显著的生长抑制作用,可能与抑制CyclinD1的表达,从而将细胞阻滞于G1期有关。     

Zinc finger of the cerebellum 5 promotes colorectal cancer cell proliferation and cell cycle progression through enhanced CDK1/CDC25c signaling

IntroductionColorectal cancer (CRC), mostly caused by external or environmental factors, is the third most common and lethal cancer worldwide. Although a large number of investigations have been carried out to reveal the evolution of CRC, the underlying mechanisms of CRC remain unclear.Material and methodsExpression of zinc finger of the cerebellum 5 (ZIC5) in CRC tissues and cell models was measured by qRT-PCR and IHC. Cell transfection was carried out for ZIC5 overexpression or knockdown. The MTT assay was applied to examine the capacity of glioma cell proliferation. Wound healing assay and tumor invasion assay were used to test the capacity of glioma cell migration and invasion respectively. Cell cycle analysis and western blot were used to verify the apoptosis rates of CRC cells upon ZIC5 overexpression or downregulation. A further tumor Xenograft study was used to examine the effects of ZIC5 on tumor malignancy in vivo.ResultsCell models using HCT116 and SW620 cells were established to study the ZIC5 function upon ZIC5 overexpression of knockdown. Consistently, we discovered that ZIC5 also significantly increased in Chinese CRC patients. In addition, ZIC5 promoted CRC cell proliferation through increasing the proportion of cells maintained in the S phase. ZIC5 overexpression facilitated the capacity of CRC cell migration and invasion. Inhibition of ZIC5 mitigated such malignant effects.ConclusionsCollectively, investigations of the ZIC5 in CRC provided a new insight into CRC diagnosis, treatment, prognosis and next-step translational therapeutic developments from bench to clinic.     

Expression of adhesion molecules in Langerhans' cell histiocytosis

Expression of adhesion molecules was investigated in six biopsy specimens of Langerhans' cell histiocytosis using immunocytochemistry. Cells with Langerhans' cell histiocytosis morphology were stained for ICAM-1, for the beta-1 integrins alpha-4 (VLA-4) and alpha-5 (VLA-5), and for the beta-2 integrins LFA-1, MAC-1 and p150,95. This pattern of reactivity was different from that of epidermal Langerhans' cells of the normal skin which were not immunostained. A variable number of CD68+ multinucleated giant cells was present in five biopsies. They were less reactive than the cells of Langerhans' cell histiocytosis for alpha-4 (VLA-4) and LFA-1, were positive for MAC-1 and p150,95 and were characterized by prominent expression of the beta-1 integrins alpha-2 (VLA-2), alpha-3 (VLA-3) and of VnR (alpha-v/ beta-3). The repertoire of adhesion molecules expressed by giant cells is indicative of profound cell-matrix interactions, whereas that of Langerhans' histiocytosis cells suggests particularly active cell–cell interactions. Blood vessels of the lesions were stained for beta-1 integrins, for vitronectin receptor and for molecules involved in adhesion and trans-endothelial migration of circulating leukocytes, such as ICAM-1, VCAM-1 and E-selectin. Additional findings were the observation of CD1a+ multinucleated giant cells in a single case, suggesting a possible lineage relationship with the histiocytosis cells, and the demonstration of some Ki-67+ Langerhans' cell histiocytosis cells and CD1a+ mitotic figures in four of six cases, indicating local proliferation of Langerhans' histiocytosis cells.     

CD40L、可溶性黏附分子及MMP9在川崎病中的表达及意义

目的：研究T细胞CD40配体（CD40L）、血浆中可溶性E选择素（sE-selectin）、可溶性细胞间黏附分子1（sICAM-1）和基质金属蛋白酶9（MMP9）在川崎病中的表达及意义。方法：用流式细胞双色荧光标记技术检测20例川崎病急性期及静脉注射免疫球蛋白（IVIG）治疗后的患儿，19例正常对照，外周血T细胞表达CD40L阳性细胞率及平均荧光强度；用ELISA方法检测上述3组血浆中sE—selectin、sICAM-1和MMP9水平。结果：急性期川崎病组T细胞CD40L表达明显高于IVIG治疗后组（P〈0．05），川崎病患儿T细胞CD40L表达较正常对照持久；2例伴冠状动脉损害（CAL）的川崎病患儿急性期T细胞CD40L表达高于无CAL者。急性期川崎病组血浆sICAM-1、MMP9明显高于IVIG治疗后组，急性期川崎病患儿血浆sE—selectin、sICAM—1明显高于正常对照（P〈0.05）。川崎病急性期T细胞表达CD40L与血浆sE—selectin、sICAM-1、MMP9水平无明确相关性（P〉0．05）。结论：T细胞表达CD40L升高在川崎病血管炎及冠状动脉病变中可能起一定作用。血浆sE-selectin、sLCAM-1和MMP9水平可作为反映川崎病血管炎的指标。     

Expression of IL-1Rrp2 by human myelomonocytic cells is unique to DCs and facilitates DC maturation by IL-1F8 and IL-1F9

We report for the first time that expression of the novel IL-1 cytokine receptor IL-1Rrp2 (IL-1R6) is unique to DCs within the human myelomonocytic lineage. IL-1Rrp2 was expressed by monocyte-derived dendritic cells (MDDCs) which was dose-dependently increased by IL-4 and correlated with increased numbers of differentiated MDDCs. Human plasmacytoid DCs also express IL-1Rrp2 but the receptor is not expressed by either myeloid DC type 1 (mDC1) or mDC2 cells. We also show that IL-1F8 or IL-1F9 cytokines, which signal through IL-1Rrp2, induce maturation of MDDCs, as measured by increased expression of HLA-DR and CD83 and decreased expression of CD1a. Furthermore, IL-1F8 stimulated increased CD40 and CD80 expression and IL-18 and IL-12 p70 production by MDDCs, which induced proliferation of IFN-γ-producing CD3(+) lymphocytes (indicative of inflammatory Th1 subsets). IL-1F8 and IL-1F2 were equipotent in their ability to stimulate IL-18 secretion from MDDCs but IL-1F8 was not as potent as IL-1F2 in stimulating secretion of IL-12p70 from MDDCs or inducing lymphocyte proliferation Therefore, IL-1Rrp2 expression by some DC subsets may have an important function in the human immune response in vivo via its role in differentiation of inflammatory Th1 lymphocytes.     

结直肠肿瘤中CXCR7、NDRG1的表达

目的 探讨CXCR7、NDRG1蛋白在结直肠肿瘤中的表达及其对结直肠肿瘤发生、发展的影响.方法 采用免疫组化染色技术观察人结直肠正常黏膜组织、结直肠腺瘤、无淋巴结转移(无LNM)结直肠腺癌组织及伴有淋巴结转移(有LNM)结直肠腺癌组织中CXCR7、NDRG1蛋白的表达水平.结果 (1)结直肠正常黏膜组织、腺瘤组织、无LNM结直肠腺癌组织及有LNM结直肠腺癌组织中CXCR7蛋白表达逐渐增高,其中后者CXCR7的表达与其他组别相比,差异有统计学意义(P＜0.05);CXCR7的表达与淋巴结转移呈正相关关系(rs=0.341),与肿瘤TNM分期有关(P＜0.05).(2)NDRG1蛋白表达逐渐增高,差异均有统计学意义(P＜0.05);NDRG1的表达与淋巴结转移呈正相关关系(rs＝0.402),与肿瘤TNM分期有关(P＜0.05).(3)CXCR7与NDRG1在结直肠肿瘤中的表达呈正相关关系(P=0.003,rs=0.334),2者均具有诊断准确性.结论 CXCR7、NDRG1在结直肠腺癌发生发展整个过程的不同阶段起着重要的作用,两者在促进肿瘤转移方面具有正相关性.     

Expression of mucosa-related integrin alphaEbeta7 on alveolar T cells in interstitial lung diseases.

The expression of alphaEbeta7 integrin has been related to the selective retention of lymphocytes in mucosal tissues of gut, urogenital tract and lung. To identify potential disease-associated alphaEbeta7 expression patterns on cells accounting for lymphocytic alveolitis in interstitial lung disease (ILD), alphaE expression on CD4+ and CD8+ T cell subsets was evaluated by dual-colour flow cytometry in peripheral blood and bronchoalveolar lavage fluid (BALF) of patients with idiopathic pulmonary fibrosis (IPF; n = 18), hypersensitivity pneumonitis (HP; n = 20) and sarcoidosis (n = 44) in comparison with healthy controls (n = 15). In both healthy individuals and all patient groups the proportion of alphaE-bearing T cells in peripheral blood was < 2%, whereas the vast majority of alveolar CD8+ T cells consistently co-expressed alphaE. Absolute alveolar CD8+alphaE+ cell numbers/ml were up to 30-fold increased in HP patients. Proportions of alphaE-bearing CD4+ cells in BALF were significantly elevated in IPF (74.0 +/- 2.7%) and HP (70.0 +/- 2.4%) compared with normals (30.0 +/- 1.8%) (mean +/- s.e.m.; P < 0.01). In sarcoidosis, the alphaE expression on BALF CD4+ cells displayed subgroup dependency: proportions significantly lower than normal were noted in chest radiographic stage I (14.3 +/- 1.5%), but increased proportions in stages II (50.0 +/- 3.8%) and III (64.0 +/- 4.8%). Correlations between common markers of T cell activation or BALF transforming growth factor-beta (TGF-beta ) bioactivity and alphaE expression were not noted. We conclude that the vast majority of alveolar CD8+ T cells consistently express alphaEbeta7 and that distinct patterns of alphaEbeta7 expression on alveolar CD4+ lymphocytes in sarcoidosis are related to the diverse manifestations of the sarcoid inflammatory process in the lung.     

非小细胞肺癌组织中MMP-9、TIMP-1表达及意义

目的 探讨非小细胞肺癌(non-small cell lung cancer,NSCLC)组织中基质金属蛋白酶9(MMP-9)、组织金属蛋白酶抑制剂-1(TIMP-1)的表达及其生物学行为的关系.方法 应用免疫组织化学SP法检测76例NSCLC和癌旁正常组织中MMP-9、TIMP-1的表达,并分析其表达与肺癌组织类型、肿瘤大小、TNM分期、分化程度、淋巴结转移的相关性.结果 MMP-9、TIMP-1在肺癌组织中的阳性表达率明显高于癌旁正常组织(P<0.05).NSCLC中MMP-9、TIMP-1表达与肿瘤的分化程度、临床分期、淋巴结转移有相关性(P<0.05),与肿瘤病理分型无关(P>0.05).结论 检测NSCLC中组织的MMP-9、TIMP-1的表达对判断肿瘤的恶性程度和预后评估有一定的意义.