甘油二酯激酶ζ在发育大鼠脊髓和背根神经节的表达和亚细胞定位

目的:探讨甘油二酯激酶ζ(DGKζ)的表达模式和亚细胞定位及其在大鼠脊髓和背根神经节(DRG)发育中的作用。方法:取不同胎龄和出生时间的Wistar大鼠,用DGKζ-c、DGKζ-n抗体对其脊髓及DRG冰冻切片进行免疫组织化学染色。结果:DGKζ-c和DGKζ-n免疫组化结果显示,在脊髓,E15.5时DGKζ呈较强阳性广泛表达,神经上皮、基板腹侧、基板和翼板交界处的细胞胞浆中等阳性、胞核强阳性; E18.5时表达阴性;出生后P0.5-P3,DGKζ再次广泛表达在灰质和白质,灰质内细胞多为胞核表达强于胞浆; P5-P8,DGKζ的表达再次明显降低,在前角和后角细胞的胞核呈弱阳性; P14时,DGKζ在后角细胞胞核的表达增强。在DRG,E15.5时DGKζ在神经元胞浆和胞核表达,多数细胞胞核表达强于胞浆; E18.5时,仅少量神经元胞浆呈阳性表达;出生后P0.5-P3,DGKζ的表达多样,阴性细胞、胞核弱阳性细胞、胞浆或胞核阳性细胞、胞核和胞浆强阳性细胞共存; P5时,DGKζ主要表达在中型神经元胞浆、或胞浆与胞核; P8-P14,DGKζ主要表达在小型神经元胞浆、或胞浆与胞核。结论:DGKζ在大鼠发育中脊髓和DRG的表达不完全同步,核内和胞浆内定位的DGKζ可能在DRG内神经元分化和脊髓内细胞增殖时发挥作用,核内定位的DGKζ可能参与脊髓神经元分化。     

急性淋巴细胞白血病患者GLUT3、TSLC1的表达及临床意义

目的:探讨急性淋巴细胞白血病患者葡萄糖转运蛋白-3(Glucose transporter 3,GLUT3)、肿瘤抑制物1(Tumor suppressor in lung cancer 1,TSLC1)的表达及临床意义.方法:选取2018年4月至2020年4月期间我院诊治的59例急性淋巴细胞白血病患者作为研究对象的研究组,另选取53例同期非急性淋巴细胞白血病患者作为研究对象的对照组.比较两组患者骨髓单个核细胞中GLUT3、TSLC1的表达情况,以及患者的不同临床病理特征对骨髓单个核细胞中GLUT3、TSLC1表达的影响.结果:研究组GLUT3阳性表达率高于对照组、TSLC1阳性表达率低于对照组(P<0.05);血红蛋白(异常)、白细胞计数(异常)者GLUT3阳性率明显高于血红蛋白(正常)、白细胞计数(正常)者(P<0.05).白细胞计数(异常)者TSLC1阳性率明显高于白细胞计数(正常)者(P<0.05).结论:在急性淋巴细胞白血病患者骨髓单个核细胞中GLUT3阳性表达率高、TSLC1阴性表达率高.     

用胞质轻链免疫荧光原位杂交技术检测多发性骨髓瘤细胞遗传学异常

目的 建立骨髓单个核细胞涂片胞浆轻链免疫荧光原位杂交技术(immunofluorescence of the cytoplasmic light chain in situ hybridization,cIg-FISH),检测多发性骨髓瘤(multipte myeloma,MM)患者的染色体异常,为MM患者的分子细胞遗传学检测提供新方法.方法 以骨髓单个核细胞涂片为载体,采用8号染色体着丝粒探针进行间期荧光原位杂交技术,检测MM浆细胞(plasma cells,PCs)的8号染色体异常情况,并且通过联结7-氨基-4-甲基香豆素-3-乙酸(7-amino-4-methylcoumarin-3-acetic acid,AMCA)的抗人κ或λ轻链抗体标记MM标本中PCs,然后通过联结AMCA的二抗增强荧光效果.结果 单个核细胞涂片上胞浆荧光抗体筛选的PCs有明显的蓝色荧光,18例患者中7例(38.9%)有8号染色体异常,其中8号染色体单体(-8)5例(27.8%),8号染色体三体(+8)2例(11.1%).结论 骨髓单个核细胞涂片cIg-FISH具有简便、特异、准确的特点,可用于MM分子遗传学异常的检测.     

乳腺癌中高迁移率族蛋白B1蛋白表达及其临床病理意义

目的探讨高迁移率族蛋白B1(HMGB1)蛋白在乳腺癌中的表达及临床病理意义。方法收集2006—2018年浙江省东阳市人民医院病理科浸润性乳腺癌石蜡标本417例和同时切除的正常乳腺组织石蜡标本26例。采用免疫组织化学EnVision法检测和比较浸润性乳腺癌组织和正常乳腺组织中HMGB1蛋白的表达。分析HMGB1蛋白胞核高表达及胞质阳性表达与乳腺癌患者临床病理特征的关系。结果(1)乳腺癌中HMGB1蛋白胞核高表达率及胞质阳性率分别为80.8%(337/417)和16.8%(70/417),而其在正常乳腺组织中分别为46.2%(12/26)和0。乳腺癌中HMGB1蛋白胞核高表达率和胞质阳性率均明显高于正常乳腺组织(分别P<0.001,P=0.046)。(2)组织学级别高、雌激素受体(ER)阴性、孕激素受体(PR)阴性的乳腺癌患者,其HMGB1蛋白胞核高表达率明显更高(分别P=0.006,P=0.004,P<0.001),胞质阳性率也明显更高(均P<0.001)。Logistic回归模型多因素分析显示乳腺癌患者HMGB1蛋白胞核高表达的独立相关因素为肿瘤组织学分级(OR=2.188,95%CI=1.078~4.443,P=0.030),而HMGB1蛋白胞质阳性表达的独立相关因素包括肿瘤组织学分级(OR=3.031,95%CI=1.600~5.742,P=0.001)、ER(OR=0.129,95%CI=0.034~0.494,P=0.003)及TNM分期(OR=3.820,95%CI=1.042~14.001,P=0.043)。(3)Cox比例风险模型多因素分析显示HMGB1蛋白胞核高表达是影响乳腺癌患者总生存的独立危险因素(HR=0.366,95%CI=0.138~0.972,P=0.044)。结论HMGB1蛋白胞核高表达及胞质阳性表达与乳腺癌患者多项预后不良因素密切相关,有望成为抗乳腺癌治疗的一个潜在生物学标志。     

MCUR1在多发性骨髓瘤中的作用

目的探讨线粒体钙单向转运体调节分子1(MCUR1)在多发性骨髓瘤(MM)患者中的表达变化,观察其对MM细胞周期、凋亡的作用,初步分析其内在分子机制。方法设瞬时转染MCUR1干涉片段1和2的人MM细胞系RPMI 8226细胞作为实验组(命名为si MCUR1-1和si MCUR1-2细胞),阴性干涉片段转染的RPMI 8226细胞(命名为si Ctrl细胞)作为对照组;实时定量PCR测定MCUR1在MM患者及正常对照者骨髓单个核细胞中的表达水平;实时定量PCR和Western blot检测MCUR1沉默效果;用CCK-8法和流式细胞计量术分别测定沉默MCUR1后细胞增殖、细胞周期和凋亡,Western blot检测MCUR1沉默后p53及其下游分子表达变化。结果与对照者相比,MCUR1在MM患者中的表达显著升高(P0. 05); MM细胞系RPMI 8226中干扰MCUR1后,与对照组相比,细胞增殖显著减慢(P0. 05),G1期细胞比例增加,S期细胞比例减少,细胞凋亡比例增加;与对照组相比,干扰MCUR1组细胞内p53、BAX蛋白表达增加,BCL2、cyclin D1和cyclin E蛋白表达减少。结论 MCUR1在MM患者中表达上调,MCUR1通过抑制p53通路进而促进MM细胞增殖。     

香烟烟雾提取物通过蛋白激酶C-核因子相关因子2调节大鼠气道上皮细胞血红素加氧酶1表达

 目的：探讨蛋白激酶C（PKC）-红系衍生的核因子相关因子2 (Nrf2)对香烟烟雾提取物（CSE）诱导的大鼠气道上皮细胞血红素加氧酶1（HO-1）表达的影响。方法：通过CSE刺激雄性SD大鼠气道上皮细胞,使用PKC抑制剂RO318220和Nrf2 siRNA,将细胞分为对照组、CSE 3 h组、RO318220组、Nrf2 siRNA组和RO318220+Nrf2 siRNA组,用Western blotting法分别检测HO-1、Nrf2和p-PKC蛋白表达,免疫细胞化学法观察HO-1蛋白表达,逆转录－聚合酶链反应（RT-PCR）法检测HO-1 mRNA表达,免疫荧光法检测Nrf2蛋白定位,测定HO-1活性。结果：暴露CSE 3 h后,Nrf2蛋白主要表达在胞核, 胞核蛋白表达增强,p-PKC蛋白、HO-1的mRNA和蛋白高表达,HO-1活性增强。预先给予RO318220,PKC蛋白、Nrf2胞浆和胞核蛋白、HO-1的mRNA和蛋白表达均明显减弱,HO-1活性显著降低。预先用siRNA沉默Nrf2,胞浆和胞核的 Nrf2蛋白表达均减弱,HO-1活性、mRNA和蛋白水平明显降低。RO318220联合Nrf2 siRNA处理后,PKC蛋白、Nrf2胞浆和胞核蛋白、HO-1 mRNA和蛋白表达均明显降低,HO-1活性明显降低。结论：CSE通过PKC激活Nrf2,诱导Nrf2核转位,从而上调HO-1的表达水平。     

ClC-3在人胶质瘤中的表达及分布

目的探讨ClC-3在人脑胶质瘤中的表达及其生物学意义。方法应用免疫组织化学染色技术检测24例人胶质瘤以及4例脑转移癌手术标本中ClC-3蛋白的表达;RT-PCR检测ClC-3蛋白表达阳性的手术标本中其mRNA表达。结果4例正常脑组织ClC-3蛋白的表达为阴性;而蛋白表达阳性的19例胶质瘤及4例脑转移癌中,ClC-3主要位于瘤细胞和微血管内皮细胞的胞浆或胞膜上。蛋白表达阳性的手术标本中16例胶质瘤和4例脑转移癌检测到ClC-3mRNA的表达。少突胶质细胞瘤Ⅲ级中ClC-3的mRNA和蛋白表达明显高于其Ⅱ级。结论ClC-3在人胶质瘤及脑转移癌组织中普遍表达,并且可能与少突胶质细胞瘤病理分级相关。     

信号转导子与转录激活子3和细胞因子信号转导抑制因子-3蛋白在成年大鼠脊髓内的表达与定位

目的分析JAK-STAT信号转导途径重要成员STAT3蛋白和该途径抑制蛋白SOCS-3在大鼠脊髓内的表达和细胞内定位分布。方法免疫细胞化学技术和形态计量学方法。结果STAT3免疫阳性产物主要分布于脊髓前角运动神经元的胞浆内；SOCS-3免疫阳性产物广泛分布于脊髓前角及后角神经元内、部分胶质细胞及神经纤维内。在前角运动神经元内，SOCS-3免疫阳性产物主要分布于核内，有时也可分布于胞浆中。结论STAT3和SOCS-3广泛表达于正常大鼠脊髓内，其中SOCS-3具有核蛋白或胞浆蛋白两种存在形式。     

Neuropilin-1基因在CML骨髓基质细胞中的表达及其意义

目的了解Neruophlin-1基因在慢性粒细胞白血病(CML)骨髓基质细胞中的表达情况.方法收集14例CML和20例正常对照骨髓标本,分离单个核细胞.进行体外长期培养,收集贴壁细胞(骨髓基质细胞).利用RT-PCR方法分析两组骨髓基质细胞中的cDNA,了解Neuropilin-1 基因表达情况.结果建立了CML和正常人骨髓基质细胞培养方法,CML骨髓基质细胞中的Neuropilin-1基因表达率(50%)明显低于正常对照(85%)(p<0.05).结论参与调控骨髓基质细胞的Neuropilin-1基因可表达于大部分正常人和部分CML患者骨髓基质细胞中,Neuropilin-1基因在部分CML骨髓基质细胞中的表达缺失可能与其调节造血功能异常有关.     

丙型肝炎病毒核心蛋白在患者外周血单个核细胞内的核表达检测

目的　检测和研究丙型肝炎病毒 (hepatitisCvirus,HCV)核心蛋白在患者外周血单个核细胞 (peripheralbloodmononuclearcells ,PBMC)内核表达的意义 ,并探讨其与临床的关系。方法　对 6 6例慢性丙型肝炎患者PBMC标本进行免疫组化检测 ,并将HCV蛋白抗原定位分布情况与患者临床状况进行比较分析 ,对其中 2 7例患者PBMC进行HCVRNA和HCVAg的平行检测和分析。结果　免疫组化结果显示 ,慢性丙型肝炎患者PBMCHCVAg(core +NS3)阳性检出率为 77 2 7% (5 1 6 6 )。结果还证实 ,HCV核心蛋白均定位于胞核内 ,且呈强表达 ;NS3蛋白主要定位于胞质内 ,呈弱表达。当进行HCVAg在PBMC内定位情况与患者临床状况比较分析时显示 ,病情较重患者PBMC内核心蛋白表达阳性率 (35 2 9% )明显高于病情较轻者 (5 88% ) (P <0 0 0 1)。结论　HCV核心蛋白在PBMC内核表达与患者临床状况相关 ,提示其可能是丙型肝炎慢性化的一个指标 ,并可能在肝硬化和肝癌发生上起一定作用     

Nuclear Localization of Annexin A1 Correlates With Advanced Disease and Peritoneal Dissemination in Patients with Gastric Carcinoma

Annexin A1 (ANXA1) is a multifunctional molecule, which mediates various important physiologic processes depending on its subcelluar localization. The purpose of this study was to investigate the expression of ANXA1 level and its subcellular localization in paired clinical samples of gastric adenocarcinoma and adjacent normal counterpart. The study also assesses the clinical significance of ANXA1 subcelluar localization in gastric adenocarcinoma. A total of 104 paired resected gastric adenocarcinoma and corresponding normal specimens were collected in this study. Expression of ANXA1 was examined by immunohistochemical staining. Both cytoplasmic and nuclear ANXA1 expression levels and their correlation with clinicopathological parameters were assessed. ANXA1 protein expression was positive in 72 of 104 (69.2%) normal tissues and 47 of 104 (45.2%) gastric adenocarcinoma tissues. ANXA1 staining was predominantly localized in the cytoplasm in all 72 ANXA1‐positive normal specimens, whereas 12 ANXA1‐positive gastric adenocarcinoma specimens showed positive nuclear staining. The positive nuclear staining correlated well with serosal invasion, peritoneal dissemination and TNM stage. Cases with positive nuclear staining presented more peritoneal dissemination (41.7%, 5/12) than those with negative nuclear staining (8.7%, 8/92; P = 0.007). A logistic regression model revealed that positive ANXA1 nuclear staining had an independent association with peritoneal dissemination (P = 0.039; hazards ratio, 9.499; 95% confidence interval, 1.159–77.815). These results indicated that ANXA1 is expressed in both gastric adenocarcinoma and normal tissues. In gastric adenocarcinoma tissues ANXA1 is expressed both in cytoplasm and nucleus and its nuclear localization correlates with advanced disease stage and peritoneal dissemination. Anat Rec 293:1310–1314, 2010. © 2010 Wiley‐Liss, Inc.     

Diagnostic and prognostic value of the MB2 monoclonal antibody in paraffin-embedded bone marrow sections of patients with multiple myeloma and monoclonal gammopathy of undetermined significance.

Routinely processed bone marrow biopsies of 59 patients with untreated multiple myeloma (MM) and of 41 patients with monoclonal gammopathies of undetermined significance (MGUS) were immunocytochemically studied with the MB2 monoclonal antibody. In 54 of 59 biopsies of patients with MM, most neoplastic plasma cells showed strong cytoplasmic positivity to MB2. In contrast, only three biopsies of patients with MGUS contained highly MB2-positive plasma cells, whereas the plasma cells in the remaining biopsies were either negative (18 of 41) or revealed a weak dot-like staining of the cytoplasm (20 of 41). Plasma cells in tonsillar tissue, gastric and duodenal mucosae, and bone marrow with reactive plasmacytosis were not stained with MB2. These findings suggest that MB2 helps to distinguish between MM and MGUS. Because the five MB2-negative patients with MM were all in stage III and had very short survival time, neoplastic plasma cell staining with MB2 could also have a prognostic significance.     

The expression profile and prognostic value of APE/Ref‐1 and NPM1 in high‐grade serous ovarian adenocarcinoma

To analyze the expression trends and clinical significance of Apurinic/Apyrimidinic Endodeoxyribonuclease 1 (APE1/Ref‐1) and Nucleophosmin (NPM1) proteins in high‐grade serous ovarian adenocarcinoma (HGSC). The expressions of APE1/Ref‐1 and NPM1 proteins in 94 patients with HGSC were determined using the immunohistochemical (IHC) method, and their relationships with clinicopathological features were analyzed by the χ² test or Fisher's exact test. The follow‐up data, Cox proportional hazards univariate and multivariate survival analyses were integrated to evaluate the prognostic factors affecting patients with HGSC. In the normal fallopian tubes, APE1/Ref‐1 and NPM1 protein were mainly distributed in the nuclear. The HGSC experienced changes in the cellular localization of APE1/Ref‐1 and NPM1 protein expressions, which were abnormally expressed in the cytoplasm. The rates of abnormal cytoplasmic expression of APE1/Ref‐1 and NPM1 proteins in 94 patients with HGSC were 69.1% and 73.4%, respectively, which were significantly higher than the normal fallopian tube tissues (p < 0.05). The abnormal cytoplasmic APE1/Ref‐1 and NPM1 are significantly correlated with the lymph node metastasis, chemosensitivity, FIGO staging, and prognosis. The COX multivariate survival analysis showed that the abnormal expression of APE1/Ref‐1 protein, FIGO staging, and lymph node metastasis are independent prognostic factors. Collectively, the abnormal cytoplasmic APE1/Ref‐1 and NPM1 proteins are associated with the oncogenic progression and chemoresistance of HGSC, and predict a poor prognosis.     

Effect of aging on intracellular distribution of abasic (AP) endonuclease 1 in the mouse liver

The abasic (AP) endonuclease (APE1) plays a central role in the base excision repair (BER) pathway for repairing oxidatively damaged bases and abasic sites in mammalian genomes. We have investigated age-dependent changes in APE activity, contributed primarily by APE1, in total extracts as well as in nuclear, mitochondrial, and cytoplasmic compartments of mouse hepatocytes. The APE1 protein and mRNA levels did not differ significantly between the livers of 4-mo (young), 10-mo (middle-aged), and 20-mo (old) mice, and corresponds with similar APE activity. However, we observed a 2-fold increase in specific activity of APE1 in the nucleus, a 2-fold decrease in the cytoplasm, and a 6-fold increase in the mitochondrial matrix of hepatocytes of the old relative to the young animals. Surprisingly, in the middle-age animals we observed 30% increase in APE activity in the nucleus but 6-fold in the mitochondrial matrix. These results indicate age-dependent accumulation of APE1 in the nucleus and mitochondria. Such redistribution occurred early in the mitochondria during the aging process and preferential accumulation of APE in the nucleus was more gradual which may reflect distinct levels of oxidative stress in these organelles.     

A dual regulatory role of apurinic/apyrimidinic endonuclease 1/redox factor-1 in HMGB1-induced inflammatory responses

Apurinic/apyrimidinic endonuclease 1/Redox factor-1 (APE1) is a multifunctional protein involved in reduction-oxidation regulation. High-mobility group box 1 (HMGB1) is released by necrotic cells and various inflammatory stimuli, acting as an inflammatory marker in sepsis and autoimmune diseases. Here, we report the dual regulatory role of APE1 in inflammatory signaling to extracellular HMGB1 or in the release of endogenous HMGB1 in human monocytes/macrophages. Forced cytoplasmic overexpression of APE1 profoundly attenuated the upregulation of HMGB1-mediated reactive oxygen species generation, cytokine secretion, and cyclooxygenase-2 expression by primary monocytes and macrophage-like THP-1 cell lines. In addition, HMGB1-induced activation of p38 and c-Jun N-terminal kinase (JNK), but not extracellular signal-regulated kinase 1/2, was strongly abrogated by the overexpression of APE1. The activation of apoptosis signal-regulating kinase 1 was required for both the p38 and JNK activation challenge with HMGB1. The extracellular release of HMGB1 by activated macrophages was inhibited by APE1 transfection. Small interfering RNA (siRNA) knockdown of endogenous APE1 impaired HMGB1-mediated cytokine expression and MAPK activation in THP-1 cells. HMGB1 stimulation induced the translocation of APE1 to the nucleus of the cell. In addition, APE1 silencing via siRNA transfection inhibited both the nuclear and cytoplasmic expression of APE1. These data identify APE1 as a novel dual regulator of inflammatory signaling to HMGB1 by human monocytes/macrophages. The modulation of cytosolic APE1 expression might be useful as a potential therapeutic modality for the treatment of inflammatory or autoimmune diseases.     

硫氧还蛋白和氧化/还原因子的融合荧光蛋白载体的构建及在293T细胞中的分布

目的构建硫氧还蛋白和氧化/还原因子的融合荧光蛋白真核表达载体,并在293T细胞中得到表达和定位。方法以RT-PCR方法从PC12细胞中克隆氧化/还原因子(APE/ref-1)的cDNA,然后亚克隆构建APE/ref-1-GFP融合真核表达载体。以PCR方法将质粒pQE30-TRX上的硫氧还蛋白cDNA亚克隆到pDsred1-1质粒上,然后将硫氧还蛋白-DsRed融合基因序列亚克隆到pCMV5质粒上,构建硫氧还蛋白-DsRed融合真核表达载体.通过磷酸钙转染293T细胞,以荧光显微镜分析融合蛋白的表达及其亚细胞定位。结果成功构建了硫氧还蛋白-DsRed和APE/ref-1-GFP融合表达载体,并在293T细胞中得到表达,APE/ref-1-GFP融合蛋白定位在293T细胞核内;硫氧还蛋白-DsRed融合蛋白定位在293T细胞质和细胞核内。结论为进一步研究硫氧还蛋白和APE/ref-1之间的动态相互作用奠定基础。     

Megakaryocytes in myelodysplasia: an immunohistochemical study on bone marrow trephines

Megakaryocytes in 63 bone marrow trephine biopsies were examined for their staining characteristics, location and size distribution using the monoclonal antibody Y2/51 directed against platelet glycoprotein IIIa (CD61). Megakaryocytes in normal bone marrow were evenly distributed and demonstrated homogeneous staining with Y2/51. In addition, there was little variation in their size or shape. In contrast, myelodysplastic and myeloproliferative bone marrow trephines showed considerable dysmegakaryopoiesis demonstrated by heterogeneity of staining, an altered architectural distribution with a predominantly paratrabecular location and considerable variation in size and shape. Furthermore, in myelodysplasia 25% of the CD61 positive cells were micromegakaryocytes as opposed to less than 10% in normal or reactive marrows. Such dysmegakaryopoiesis is believed to be a clinically important feature of myelodysplasia, although until now it has only been possible to assess it subjectively. The availability of the monoclonal antibody Y2/51 provides a rapid and reproducible means of studying megakaryocyte size, shape and distribution in routine trephine specimens and may help to overcome some of the diagnostic problems currently associated with myelodysplasia and other intrinsic bone marrow neoplasias.     

Differentiation of plasma and myeloma cells of man combined planimetric and cytophotometric studies

Summary Plasmacytoid cells in the bone marrow of 3 patients with myeloma and plasma cells in the bone marrow of a 6-year-old boy with an infectious disease were assessed cytophotometrically, first after Giemsa-and second after Feulgen staining. The cell and nuclear surface and the nuclear/cytoplasmic ratio were determined from the number of measuring points. The nuclear DNA content of individual cells was registered and the distribution of DNA within the nucleus was assessed by the distributional error. Both the mean nuclear/cytoplasmic ratio and the distributional error of myeloma cells varied from patient to patient but could not be used to differentiate between normal plasma cells and myeloma cells. It was not possible either to differentiate these cell types by multiplying the mean nuclear/cytoplasmic ratio with the mean distributional error of the nuclear DNA. A strong correlation between cell and cytoplasmic surface area was observed both in normal plasma cells and in myeloma cells.Supported by the Swiss National Foundation for Scientific Research and by the Swiss Cancer League     

Cyclin D1 overexpression in multiple myeloma. A morphologic, immunohistochemical, and in situ hybridization study of 71 paraffin-embedded bone marrow biopsy specimens

Cyclin D1 expression was evaluated by immunohistochemical analysis and biotin-labeled in situ hybridization (ISH) in a series of 71 decalcified, paraffin-embedded bone marrow biopsy specimens from patients with multiple myeloma (MM). Cyclin D1 messenger RNA (mRNA) overexpression was detected by ISH in 23 (32%) of 71 cases, whereas cyclin D1 protein was identified by immunohistochemical analysis in 17 (24%) of 71 specimens. All cases that were positive by immunohistochemical analysis also were positive by ISH. Statistically significant associations were found between cyclin D1 overexpression and grade of plasma cell differentiation and between cyclin D1 overexpression and extent of bone marrow infiltration. Our findings demonstrate the following: (1) ISH for cyclin D1 mRNA is a sensitive method for the evaluation of cyclin D1 overexpression in paraffin-embedded bone marrow biopsy specimens with MM. (2) ISH is more sensitive than immunohistochemical analysis in the assessment of cyclin D1 expression. (3) Cyclin D1 overexpression in MM is correlated positively with higher histologic grade and stage.     

骨髓基质干细胞成骨分化过程中Wnt、骨形态发生蛋白2信号通路相关因子与辛伐他汀的作用

背景：辛伐他汀可促进体外培养的人或鼠骨髓基质干细胞向成骨细胞分化,但作用机制尚不清楚。 目的：观察辛伐他汀对大鼠骨髓基质干细胞向成骨细胞分化过程中Wnt与骨形态发生蛋白2信号途径中相关因子表达的影响。 方法：取6周龄雌性SD大鼠双侧股骨、胫骨全骨髓进行体外成骨细胞诱导培养。实验分为对照组及SIM组。SIM组加入浓度为10-7 mol/L辛伐他汀,对照组加入等量无水乙醇和PBS。培养14 d,行碱性磷酸酶染色,28 d时,行von Kossa染色观察细胞外基质矿化情况;培养14,21 d,免疫荧光细胞化学染色观察成骨细胞中β-catenin,Smad1/5,Cbfa1的表达及分布。  结果与结论：大鼠骨髓基质干细胞经体外诱导后可分化为具有碱性磷酸酶活性和矿化细胞外基质能力的成熟成骨细胞。辛伐他汀可显著上调骨髓基质干细胞成骨分化过程中碱性磷酸酶的表达。同时,与对照组比较,SIM组β-catenin,Smad1/5,Cbfa1表达明显增多(P < 0.05),且呈现明显的核内聚集趋势。说明辛伐他汀促进骨髓基质干细胞向成骨细胞分化的作用可能与调控Wnt与骨形态发生蛋白2信号通路中相关因子的表达及细胞内分布有关。