联合检测E-cadherin, VEGF表达水平在卵巢良恶性肿瘤诊断中的临床价值

目的:探讨联合检测E-钙粘素(E-cadherin)、血管内皮生长因子(VEGF)表达水平在卵巢良恶性肿瘤诊断中的临床价值。方法:收取2012年1月至2016年1月间我院收治的卵巢良恶性肿瘤患者共83例,使用免疫组化方法检测对E-cadherin及VEGF表达情况进行分析与比较。结果:E-cadherin在卵巢良性肿瘤、卵巢交界性肿瘤及卵巢恶性肿瘤中的阳性表达率分别为64.71%、44.44%及25.00%,VEGF在卵巢良性肿瘤、卵巢交界性肿瘤及卵巢恶性肿瘤中的阳性表达率分别为11.76%、33.33%及83.33%,上述差异均具有统计学意义(P0.05)。E-cadherin及VEGF在卵巢恶性肿瘤中的表达与分化程度、FIGO分期有关(P0.05)。VEGF表达与淋巴结转移有关(P0.05),但E-cadherin与淋巴结转移关系不大(P0.05)。E-cadherin及VEGF表达呈负相关(r=-0.472,P0.05)。结论:E-cadherin低表达及VEGF高表达与卵巢恶性肿瘤发生、发展及侵袭有密切关系。     

剪切波弹性成像在乳腺实性病变良恶性鉴别中的诊断价值

目的:探讨剪切波弹性成像在乳腺实性病变良恶性鉴别中的诊断价值。方法:收集2012年3月-2013年6月于我院收治的乳腺实性病变患者54例,共65个病灶,先后给予乳腺二维超声检查与剪切波弹性成像检查,采用弹性模量值与钼靶BI-RADS分级方法诊断,比较两种方法诊断的准确性。结果:良性病灶组弹性最小、最大值以及平均值、标准差与恶性病灶组比较差异具有统计学意义(P0.05);ROC曲线分析显示,在乳腺实性病变良恶性的鉴别中,弹性最大值明显优于平弹性均值;剪切波弹性成像对乳腺实性病变良恶性鉴别的敏感度、特异度、准确度、阳性预测值、阴性预测值高于二维超声技术(P0.05)。结论:剪切波弹性成像在乳腺实性病变良恶性鉴别中具有良好的诊断价值,能够提高诊断的准确性。     

Nestin 和SOX2 在皮肤恶性黑色素瘤中的表达及临床意义

目的:研究巢蛋白(Nestin)和SOX2在皮肤恶性黑色素瘤中的表达及临床意义。方法:选取2010年1月-2013年1月我院收治的恶性黑色素瘤21例患者(研究组),另选黑色素痣患者21例(对照组),应用免疫组织化学法检测Nestin和SOX2,并比较两组Nestin和SOX2表达,分析其与患者生存期和转移的关系。结果:研究组Nestin和SOX2阳性表达率显著高于对照组,两组比较差异具有统计学意义(P0.05);Nestin、SOX2阳性的恶性黑色素瘤患者生存期显著低于Nestin、SOX2阴性恶性黑色素瘤患者,两组比较差异有统计学意义(P0.05),Nestin、SOX2阳性的恶性黑色素瘤患者与Nestin、SOX2阴性的恶性黑色素瘤患者转移率比较无统计学意义(P0.05)。结论:Nestin和SOX2表达阳性与恶性黑色素有关,且与恶性黑色素瘤的生存期有关。。     

可溶性CD40配体的表达及其对树突状细胞和恶性B细胞的影响(英文)

CD4 0配体 (CD4 0L)是TNF超家族成员 ,表达于人活化的CD4 T细胞表面 ,与存在于B细胞、抗原递呈细胞表面的相应受体CD4 0分子相互作用 ,促发并调节机体的体液免疫和细胞免疫应答。用PCR法从CD4 0L全长cDNA质粒中扩增CD4 0L胞外段编码cDNA ,即可溶性CD4 0L(sCD4 0L)编码cDNA ,并克隆入 pSK质粒 ;cDNA序列经测序证实后与 pPICZαA质粒重组构建成 pPICZαA sCD4 0L分泌型表达载体并在GS1 1 5毕氏酵母菌株中成功地表达出人可溶性CD4 0配体(rhsCD4 0L) ,最高表达量为 35mg/L(培养上清 )。结果显示 :在无TNFα存在时 ,重组人可溶性CD4 0L(rhsCD4 0L)可有效地诱导人外周单核细胞向树突状细胞 (DC)的分化 ,具有典型的DC形态特征和特殊表面标志 (CD1a、CD80、CD83等 ) ,为DC在体外的大量扩增提供新手段 ;进一步研究发现 ,rhsCD4 0L能直接并显著地抑制恶性淋巴瘤细胞株Daudi和多发性骨髓瘤细胞株XG 2的体外增殖 ;并证实rhsCD4 0L与CD4 0L基因转染细胞 (L/CD4 0L TC)一样 ,在体外均诱导XG2细胞的凋亡 ;rhsCD4 0L有希望成为一种重要的免疫调节剂和新的抗肿瘤药物     

Evaluation of the Potential In Vitro Antiproliferative Effects of Millimeter Waves at Some Therapeutic Frequencies on RPMI 7932 Human Skin Malignant Melanoma Cells

The potential antiproliferative effects of low power millimeter waves (MMWs) at 42.20 and 53.57 GHz on RPMI 7932 human skin melanoma cells were evaluated in vitro in order to ascertain if these two frequencies, comprised in the range of frequency used in millimeter wave therapy, would have a similar effect when applied in vivo to malignant melanoma tumours. Cells were exposed for 1 h exposure/day and to repeated exposure up to a total of four treatments. Plane wave incident power densities <1 mW/cm² were used in the MMWs-exposure experiments so that the radiations did not cause significant thermal effects. Numerical simulations of Petri dish reflectivity were made using the equations for the reflection coefficient of a multilayered system. Such analysis showed that the power densities transmitted into the aqueous samples were ≤0.3 mW/cm². Two very important and general biological endpoints were evaluated in order to study the response of melanoma cells to these radiations, i.e. cell proliferation and cell cycle. Herein, we show that neither cell doubling time nor the cell cycle of RPMI 7932 cells was affected by the frequency of the GHz radiation and duration of the exposure, in the conditions above reported.     

靶向Ang2的RNAi慢病毒载体的构建及其对恶性黑色素瘤细胞中Ang2基因表达的影响

目的 构建携带促血管生成素2-小干扰RNA（Ang2-siRNA）慢病毒载体,观察其对恶性黑色素瘤细胞中Ang2基因表达的干扰作用.方法 将经XbaⅠ酶切电泳鉴定的带有加强绿色荧光蛋白的转移质粒（pNL-EGFP）载体与pSilencer 1.0-U6启动子-促血管生成素2-小干扰RNA（pSilencer 1.0-U6-Ang2-siRNA）重组质粒连接,产生加强绿色荧光蛋白的转移质粒-U6启动子-促血管生成素2-Ⅰ（pNL-EGFP-U6-Ang2-Ⅰ）、加强绿色荧光蛋白的转移质粒-U6启动子-促血管生成素2-Ⅱ（pNL-EGFP-U6-Ang2-Ⅱ）慢病毒转移质粒,电泳筛选阳性克隆,测序鉴定.用连接成功的慢病毒转移质粒、水疱性口炎病毒G蛋白（pVSVG）包膜质粒和pHelper包装质粒共转染293T细胞,产生pNL-EGFP-U6-Ang2-Ⅰ、pNL-EGFP-U6-Ang2-Ⅱ慢病毒.收集病毒上清,测定病毒滴度.将收集的病毒上清感染恶性黑色素瘤细胞,通过实时荧光定量RT-PCR测定抑制Ang2基因表达的效率.结果 酶切电泳与测序鉴定证实成功构建了Ang2-SiRNA慢病毒载体,293T细胞测定病毒原液滴度为8.0×103/ml.实时荧光定量RT-PCR结果显示：Ang2-siRNA慢病毒载体感染恶性黑色素瘤细胞,抑制了恶性黑色素瘤细胞中Ang2基因的表达（P＜0.05）.结论 成功构建了Ang2-SiRNA慢病毒载体,体外研究显示Ang2-SiRNA慢病毒载体能抑制恶性黑色素瘤细胞中Ang2 mRNA的表达,为下一步进行裸鼠恶性黑色素瘤移植瘤生长的干预实验奠定基础,为肿瘤的基因治疗提供实验依据.     

甲状腺结节数对钙化甲状腺病变良恶性的鉴别价值

目的:通过甲状腺病变术前超声声像图与术后病理间的对照,分析结节单发或多发在鉴别伴钙化甲状腺病变良恶性中的意义。方法:回顾性分析我院2010年1月至2013年12月收治的218例甲状腺病变患者的临床资料,将患者术前超声声像图与术后病理结果进行对比。结果:术前超声探及甲状腺单发结节伴钙化其术后病理恶性比例显著高于多发结节伴钙化,差异具有统计学意义(P0.05);多发结节伴钙化其术后病理恶性比例显著高于多发结节不伴钙化,差异具有统计学意义(P0.05)。结论:术前超声探及甲状腺单发结节伴钙化其术后病理恶性比例较高,应积极手术治疗,多发结节伴钙化因其术后病理恶性病理较低,不宜作为手术治疗甲状腺病变的绝对指征。     

The reaction of Acridine Orange with proteoglycans in the articular cartilage of the rat

Summary Acridine Orange in concentrations from 0.01% to 0.2% was added to the first fixative solution in order to stain vibratome sections and small blocks of the articular cartilage of 2 month old rats. The interterritorial matrix of the radial or deep zone (zone 3) was examined. It contained reaction products with different morphology depending on the specimens used. In vibratome sections filaments were seen arranged in a homogenous pattern and changing in size with the concentration of the dye: diluted solutions produced finer filaments than concentrated ones. In contrast, in tissue blocks the staining pattern was not altered by different concentrations of Acridine Orange. However, with increase of the distance from the surface of the specimens the size of the filaments gradually decreased and formed a finer network. Since after preincubation with chondroitin ABC lyase only minute reaction products remained, an interaction of the dye with the sulphated glycosaminoglycans of the proteoglycans in the articular cartilage is suggested.The experiments show that by using mainly monocationic monomers of Acridine Orange the proteoglycans can be stained in a more expanded state than with polycationic dye polymers.     

Sensitive Immunofluorescent Staining of Cells via Generation of Fluorescent Nanoscale Polymer Films in Response to Biorecognition

Immunofluorescent staining is central to nearly all cell-based research, yet only a few fluorescent signal amplification approaches for cell staining exist, each with distinct limitations. Here, the authors present a novel, fluorescent polymerization-based amplification (FPBA) method that is shown to enable similar signal intensities as the highly sensitive, enzyme-based tyramide signal amplification (TSA) approach. Being non-enzymatic, FPBA is not expected to suffer from nonspecific staining of endogenous enzymes, as occurs with enzyme-based approaches. FPBA employs probes labeled with photopolymerization initiators, which lead to the controlled formation of fluorescent polymer films only at targeted biorecognition sites. Nuclear pore complex proteins (NPCs; in membranes), vimentin (in filaments), and von Willebrand factor (in granules) were all successfully immunostained by FPBA. Also, FPBA was demonstrated to be capable of multicolor immunostaining of multiple antigens. To assess relative sensitivity, decreasing concentrations of anti-NPC antibody were used, indicating that both FPBA and TSA stained NPC down to a 1:100,000 dilution. Nonspecific, cytoplasmic signal resulting from NPC staining was found to be reduced up to 5.5-fold in FPBA as compared to TSA, demonstrating better signal localization with FPBA. FPBA’s unique approach affords a combination of preferred attributes, including high sensitivity and specificity not otherwise available with current techniques.     

线粒体核糖体蛋白质与人类恶性肿瘤

线粒体拥有自身独特的核糖体--线粒体核糖体,用于翻译线粒体DNA（mitochondrial DNA, mtDNA）编码的基因。线粒体核糖体由核基因编码的线粒体核糖体蛋白质(mitochondrial ribosomal protein, MRPs)和线粒体自身编码的rRNA组装而成。MRPs表达失调会引发代谢紊乱、呼吸链受损,导致细胞发生功能障碍和异常增殖,甚至发生癌变等恶性转化。大量研究证明,MRPs在不同的肿瘤细胞中表达异常,提示着MRPs在肿瘤发生发展过程中发挥着重要作用。本文就线粒体核糖体蛋白质与人类恶性肿瘤发生的关系作一综述,为进一步阐明其在恶性肿瘤发生过程中的作用机制奠定基础。     

Ad-GFP-nm23-H1裸鼠体内抑制人恶性黑色素瘤A375作用的研究

目的探讨Ad—GFP—nm23-H1对人恶性黑素瘤裸鼠皮下移植瘤的抑制作用,从而为后期nm23-H1基因和腺病毒载体用于人恶性黑色素瘤及其它肿瘤的基因治疗提供一定的理论和方法。方法在裸鼠真皮下建立人A375细胞黑色素瘤动物模型后,设对照组及10^9 PFU／ml、10^10 PFU／ml的Ad—GFP—nm23-H1干预组。经Ad—GFP—nm23-H1干预治疗后,取瘤体称重,计算抑瘤率,通过光镜进行瘤组织病理形态学观察。结果对照组及10^9 PFU／ml、10^10 PFU／ml的Ad—GFP-nm23-H1干预组的平均肿瘤体积和瘤重分别为：1．4129&#177;0．4832mm^3、1．1914&#177;0．3304mm^3、0．75&#177;0．2548mm^3和1．924&#177;0．539g、1．655&#177;0．5754g、1．195&#177;0．2639g。与另两组比较,10^10 PFU／ml的Ad．GFP—nm23-H1干预组对A375具有明显的抑制作用。结论10^10 PFU／ml的Ad—GFP—nm23-H1干预组对裸鼠移植A375实体瘤有明显的抑制作用。     

The Non-Coding RNA Llme23 Drives the Malignant Property of Human Melanoma Cells

Several lines of evidence support the notion that increased RNA-binding ability of polypyrimidine tract-binding (PTB) protein-associated splicing factor (PSF) and aberrant expression of long non-coding RNAs (lncRNAs) are associated with mouse and human tumors. To identify the PSF-binding lncRNA involved in human oncogenesis, we screened a nuclear RNA repertoire of human melanoma cell line, YUSAC, through RNA-SELEX affinity chromatography. A previously unreported lncRNA, termed as Llme23, was found to bind immobilized PSF resin. The specific binding of Llme23 to both recombinant and native PSF protein was confirmed in vitro and in vivo. The expression of PSF-binding Llme23 is exclusively detected in human melanoma lines. Knocking down Llme23 remarkably suppressed the malignant property of YUSAC cells, accompanied by the repressed expression of proto-oncogene Rab23. These results may indicate that Llme23 can function as an oncogenic RNA and directly associate the PSF-binding lncRNA with human melanoma.     

Pro-MMP-9 activation by the MT1-MMP/MMP-2 axis and MMP-3: role of TIMP-2 and plasma membranes

MMP-9 (gelatinase B) is produced in a latent form (pro-MMP-9) that requires activation to achieve catalytic activity. Previously, we showed that MMP-2 (gelatinase A) is an activator of pro-MMP-9 in solution. However, in cultured cells pro-MMP-9 remains in a latent form even in the presence of MMP-2. Since pro-MMP-2 is activated on the cell surface by MT1-MMP in a process that requires TIMP-2, we investigated the role of the MT1-MMP/MMP-2 axis and TIMPs in mediating pro-MMP-9 activation. Full pro-MMP-9 activation was accomplished via a cascade of zymogen activation initiated by MT1-MMP and mediated by MMP-2 in a process that is tightly regulated by TIMPs. We show that TIMP-2 by regulating pro-MMP-2 activation can also act as a positive regulator of pro-MMP-9 activation. Also, activation of pro-MMP-9 by MMP-2 or MMP-3 was more efficient in the presence of purified plasma membrane fractions than activation in a soluble phase or in live cells, suggesting that concentration of pro-MMP-9 in the pericellular space may favor activation and catalytic competence.     

Multiplication of Legionella pneumophila in HeLa Cells in the Presence of Cytoskeleton and Metabolic Inhibitors

A study has been carried out on the action of cytoskeleton and metabolic inhibitors on intracellular multiplication in HeLa cells of a virulent strain of Legionella pneumophila serogroup 6. The effects of the substances were separately tested on both penetration and intracellular multiplication of L. pneumophila. Only cytochalasin A and 2-deoxy-d -glucose (2dG) affected bacterial internalisation, whereas intracellular multiplication was inhibited by cytochalasins A, B, C, D and J (D being the most active) and by 2dG with a dose-response effect. The action of 2dG was counteracted by 50 mM glucose. Experiments carried out with cytochalasin D and a rhodamine-phalloidin conjugate showed the involvement of cytoskeletal elements in intracellular multiplication of Legionella; compounds acting on microtubules had no effect.     

经直肠实时组织超声弹性成像诊断前列腺良恶性病灶的价值

目的:探索经直肠实时组织超声弹性成像技术在前列腺良恶性病灶诊断中的应用价值。方法:选取2013年12月至2014年5月我科疑似前列腺癌(PCa)并拟行穿刺活检的患者49例,以病例活检结果作为金标准,对比经直肠实时组织超声弹性成像技术、经直肠超声(TRUS)及直肠指诊(DRE)在疑似PCa患者中的诊出结果,并对直肠超声进行弹性图像评分及应变指数分析。结果:弹性图像评分≥4分时,其对PCa的敏感性、特异性及准确性分别为92.3%、91.3%和93.9%;良性病灶的应变指数为2.84±4.72,恶性病灶的应变指数为32.12±15.05,差异有统计学意义(P0.05)。结论:经直肠实时组织超声弹性成像技术可提高PCa的诊出率,在前列腺良恶性病灶的鉴别及指导治疗与预后方面有重要价值。     

常规高频超声联合超声弹性成像在甲状腺良恶性结节鉴别诊断中的应用价值

目的:研究常规高频超声与超声弹性成像在甲状腺结节良恶性鉴别诊断中的应用价值。方法:选择住院的甲状腺结节性疾病患者120例(146个甲状腺结节)。对患者进行常规高频超声和超声弹性成像检查,以术后病理检查为金标准,计算不同方法对甲状腺良恶性结节鉴别诊断的灵敏度、特异度、准确度。结果:120例患者,经手术病理检查证实有146个结节,其中良性结节98个,恶性结节48个。常规高频超声显示在形态、边界、包膜、内部回声、微钙化及RI值上,甲状腺良、恶性结节之间的差异有统计学意义(P0.05)。常规高频超声与弹性成像相比,诊断效能之间的差异无统计学意义(P0.05);常规高频超声与联合超声诊断的灵敏度(X~2=12.22,P0.01)、特异度(X~2=10.21,P0.01)、准确度(X~2=9.31,P0.01)相比较,差异具有统计学显著性;超声弹性成像与联合超声诊断灵敏度(X~2=6.51,P0.01)、特异度(X~2=5.82,P0.05)、准确度(X~2=4.56,P0.05)相比,差异具有统计学显著性。结论:常规高频超声联合超声弹性成像对甲状腺良恶性结节的诊断灵敏度和准确度高,值得进一步推广临床应用。     

The Role of Alpha-Synuclein in Melanin Synthesis in Melanoma and Dopaminergic Neuronal Cells

The relatively high co-occurrence of Parkinson’s disease (PD) and melanoma has been established by a large number of epidemiological studies. However, a clear biological explanation for this finding is still lacking. Ultra-violet radiation (UVR)-induced skin melanin synthesis is a defense mechanism against UVR-induced damage relevant to the initiation of melanoma, whereas, increased neuromelanin (NM), the melanin synthesized in dopaminergic neurons, may enhance the susceptibility to oxidative stress-induced neuronal injury relevant to PD. SNCA is a PD-causing gene coding for alpha-Synuclein (α-Syn) that expresses not only in brain, but also in skin as well as in tumors, such as melanoma. The findings that α-Syn can interact with tyrosinase (TYR) and inhibit tyrosine hydroxylase (TH), both of which are enzymes involved in the biosynthesis of melanin and dopamine (DA), led us to propose that α-Syn may participate in the regulation of melanin synthesis. In this study, by applying ultraviolet B (UVB) light, a physiologically relevant stimulus of melanogenesis, we detected melanin synthesis in A375 and SK-MEL-28 melanoma cells and in SH-SY5Y and PC12 dopaminergic neuronal cells and determined effects of α-Syn on melanin synthesis. Our results showed that UVB light exposure increased melanin synthesis in all 4 cell lines. However, we found that α-Syn expression reduced UVB light-induced increase of melanin synthesis and that melanin content was lower when melanoma cells were expressed with α-Syn, indicating that α-Syn may have inhibitory effects on melanin synthesis in melanoma cells. Different from melanoma cells, the melanin content was higher in α-Syn-over-expressed dopaminergic neuronal SH-SY5Y and PC12 cells, cellular models of PD, than that in non-α-Syn-expressed control cells. We concluded that α-Syn could be one of the points responsible for the positive association between PD and melanoma via its differential roles in melanin synthesis in melanoma cells and in dopaminergic neuronal cells.     

ADNEX模型联合ROMA指数、CA199鉴别卵巢肿瘤良恶性的临床价值及卵巢恶性肿瘤的影响因素分析

摘要 目的：探讨ADNEX模型联合ROMA指数、糖类抗原199（CA199）鉴别卵巢肿瘤良恶性的临床价值,并分析卵巢恶性肿瘤的影响因素。方法：选取2019年4月～2021年4月我院收治的150例卵巢肿瘤患者,以病理结果为金标准,其中良性肿瘤111例（良性组）,恶性肿瘤39例（恶性组）。所有患者均进行ADNEX模型分析,开展卵巢恶性肿瘤风险预测模型（ROMA）指数分析,并检测血清CA199水平。通过受试者工作特征（ROC）曲线分析ADNEX模型联合ROMA指数、CA199鉴别诊断卵巢肿瘤良恶性的效能。此外,以单因素、多因素Logistic回归分析卵巢恶性肿瘤的影响因素。结果：ROC曲线分析结果显示：ADNEX模型联合ROMA指数、CA199鉴别诊断卵巢肿瘤良恶性的曲线下面积为0.974,明显高于三项单独应用时的0.845、0.772、0.763。单因素分析结果显示：恶性组年龄、病灶最大径大于良性组,流产次数多于良性组,CA199、人附睾蛋白4（HE4）、糖类抗原125（CA125）水平以及腹水、乳头数＞4个比例高于良性组,差异均有统计学意义（P＜0.05）。多因素Logistic回归分析结果显示：年龄、病灶最大径较大,CA199水平较高以及乳头数＞4个是卵巢恶性肿瘤的危险因素（P＜0.05）。结论：ADNEX模型联合ROMA指数、CA199鉴别卵巢肿瘤良恶性的临床价值较高,年龄、CA199水平、病灶最大径以及乳头数是卵巢恶性肿瘤的影响因素。     

Double Fluorescent Staining for the Separate Demonstration of Chromosomes and Microtubules in Mitotic Cells In Vitro

A simple fluorescent method for double staining of mitotic cells using a rhodamine B indirect immunofluorescent method for tubulin and the DNA-specific fluorescent dye Hoechst 33258 for nuclei and chromosomes is described. This procedure enables one through the use of appropriate excitation filters to view at will either chromosomes and nuclei or tubulin within the same cell.