补肾方药对骨质疏松大鼠骨中LIM矿化蛋白1表达的影响

目的：探讨补肾方药预防糖皮质激素诱导大鼠骨质疏松发生的机理。方法：通过肌肉注射地塞米松造成骨质疏松模型,观察补肾中药对骨密度的影响,从骨骼中提取mRNA,采用RT-PCR方法对LIM矿化蛋白（LMP-1）进行半定量。结果：病理组大鼠的骨密度及骨骼中LMP-1的表达明显降低（P＜0.05),而补肾方药组的骨密度及LMP-1的表达明显高于地塞米松组（P＜0.05)。结论：地塞米松抑制LMP-1在骨骼的表达,而补肾方药可通过提高LMP-1在骨的表达而预防骨质疏松的发生。     

肝细胞癌组织中CD15 mRNA与CD44v6 mRNA的表达

目的: 研究CD15mRNA表达与肝细胞癌(HCC)侵袭转移和预后的关系,以及与CD44v6mRNA表达的相关性 。方法: 应用原位杂交和免疫组织化学方法,检测分析HCC组织中CD15mRNA及其蛋白和CD44v6mRNA的表达情况 。结果: 99例HCC中,CD15mRNA及其蛋白和CD44v6阳性率表达分别为38.4%、36.4%和41.4%。CD15mRNA表达与其蛋白表达一致,与CD44v6mRNA表达关联性密切。CD15mRNA及其蛋白和CD44v6mRNA表达,均与HCC侵袭转移倾向和病人预后相关 。结论: 检测CD15表达有可能成为HCC侵袭转移和预后判断的一项新的病理生物学指标。     

c-met mRNA在乳腺癌术后腋窝引流液中表达的意义

目的探讨c-met mRNA在乳腺癌患者术后腋窝引流液中表达的意义。方法用RT—PCR法检测乳腺癌患者术后腋窝引流液中c-met mRNA的表达，分析其表达与肿瘤大小、腋窝淋巴结转移、雌激素受体（estrogen receptor，ER）、孕激素受体（progesterone receptor，PR）及生长因子受体2（c—erbB-2）的关系，以及5-FU加蒸馏水冲洗手术野对c—met mRNA表达的影响。结果①c—met mRNA能在乳腺癌术后腋窝引流液中表达，其阳性率高于病理检查腋窝淋巴结转移的阳性率（P〈0．05）。②腋窝淋巴结转移、肿瘤大小与c—met mRNA的表达有关。③肿瘤ER、PR及c—erbB-2的表达与c—met mRNA的表达无关。④5-FU加蒸馏水冲洗手术创面能明显降低c-met mRNA的表达。结论c—met mRNA是乳腺癌微转移研究的理想特异性标志物，与常规病理检查腋窝淋巴结相比，RT—PCR检测腋窝引流液中c—met mRNA的表达能更早检测出肿瘤细胞在胸壁淋巴管中的转移，并具有更高的准确性。     

尿Cox-2蛋白与CK20 mRNA诊断膀胱移行细胞癌的临床价值

目的：评价尿环氧化酶-2（Cox-2）蛋白、CK20 mRNA诊断膀胱移行细胞癌的临床价值。方法：选择78例膀胱移行细胞癌和30例非膀胱肿瘤患者,同时行尿Cox-2蛋白、CK20 mRNA和尿脱落细胞学检查,比较各种方法的敏感性、特异性和Youden指数。结果：尿Cox-2蛋白、CK20 mRNA和尿脱落细胞学的敏感性分别为89.7％．66.7％．26.9％;特异性分别为93.3％、86.7％和100％;Youden指数分别为83.1％、53.4％和26.9％。尿Cox-2蛋白和CK20 mRNA的敏感性和Youden指数均高于尿脱落细胞学（P＜0．05）,同时尿Cox-2蛋自的敏感性和Youden指数高于CK20 mRNA（P＜0．05）。结论：尿Cox-2蛋白和CK20 mRNA检测的高敏感性、高特异性为膀胱癌提供了一个简单无创的检测方法,非常适合于复发率很高的膀胱癌的诊断与随访。     

T-cell activation Rho GTPase–activating protein expression varies with inflammation location and severity in Crohn's disease

Background

The T-cell activation Rho GTPase–activating protein (TAGAP) gene has a regulatory role in T cell activation. We have previously suggested a correlation between the TAGAP-associated single nucleotide polymorphism rs212388 and protection from anal sepsis in Crohn's disease (CD) patients. The present study sought to evaluate TAGAP's expression in colonic tissue of CD patients with varying disease severity and location.

Materials and methods

Five transverse, 17 left, and five sigmoid colectomy specimens from 27 CD patients with varying disease severity (16 male, mean age at diagnosis 26.4 ± 2.2 y) were evaluated for TAGAP messenger RNA expression. Fisher exact, Mann–Whitney, and Welch two-sample t-tests were used for statistical evaluation. Immunohistochemistry confirmed results.

Results

Patients with tissue demonstrating lower TAGAP messenger RNA expression (less than the overall mean) were younger at diagnosis (mean age 21.1 ± 6.3 versus 32.5 ± 13 y, P = 0.009). Increased TAGAP expression was seen in moderate or severely diseased tissue versus tissue with no or mild disease (RQ = 1.3 ± 0.34 versus 0.53 ± 0.09, P = 0.050). This was the most dramatic in the sigmoid colon (P = 0.041). TAGAP expression was increased in more distal tissue with a significant difference seen when comparing transverse versus sigmoid colon with moderate or severe disease (0.51 ± 0.14 versus 1.9 ± 0.37, P = 0.049).

Conclusions

Colonic expression of TAGAP in CD patients varied according to disease severity and location, being the most elevated in patients with severe disease in the sigmoid colon. Whether changes in TAGAP expression are a result of disease response or inherent to the disease pathophysiology itself remains to be determined. This gene warrants further investigation for its role in CD.     

Tumoral load quantification of positive sentinel lymph nodes in breast cancer to predict more than two involved nodes

AimOne-Step Nucleic Acid Amplification (OSNA) can detect isolated tumour loads in axillary lymph nodes of breast cancer patients. We investigated the predictability of the non-sentinel lymph node (SLN) metastatic involvement (MI) based on the OSNA SLN assessment in surgical invasive breast cancer.MethodsWe studied surgical breast invasive carcinoma patients, not taking neoadjuvant chemotherapy, having SLN positive by OSNA and having received axillary lymphadenectomy. Age, basic histopathological, immunohistochemical, SLN biopsy and lymphadenectomy data were compared between patients with or without MI of more than 2 non-SLN in both univariate and multivariate analyses. The discriminating capacity of the multivariate model was characterized by the ROC AUC.Results726 patients from 23 centers in Spain aged 55.3 ± 12.2 years were analysed. The univariate analysis comparing patients with or without MI of more than 2 non-SLN detected statistically significant differences in primary tumour size, multifocality, presence of lymphovascular infiltration, positive proliferation index with ki67, immunophenotype and logTTL (Tumour Total Load). The multivariate logistic analyses (OR (95% CI)) confirmed multifocality (2.16 (1.13–4.13), p = 0.019), lymphovascular infiltration (4.36 (2.43–7.82), p < 0.001) and logTTL (1.22 (1.10–1.35), p < 0.001) as independent predictors, and exhibit an AUC (95% CI) of 0.78 (0.72–0.83) with an overall fit (Hosmer–Lemeshow test) of 0.359. A change in the slope of both sensitivity and specificity is observed at about 10,000 copies/μL, without relevant changes in the Negative Predictive Values.ConclusionsUsing OSNA technique, the MI of more than 2 non-SLN can be reliably predicted.     

Is OSNA better than imprint cytology for intraoperative diagnosis of cancer involvement of axillary sentinel node in breast cancer?

ObjectiveThe study aim was to establish Sensitivity, Specificity, Positive Predictive Value, Negative Predictive Value (NPV), and Accuracy Values of both imprint cytology (IC) and the OSNA assay for intraoperative assessment of axillary sentinel node (SN) cancer involvement in breast cancer. Specifically, we wished to find out if true positive and false negative results of IC were associated to axillary lymphadenectomy (ALND). Also, we addressed a comparative cost analysis between techniques.Methods244 patients treated for breast cancer in the Breast Unit of Hospital Germans Trias i Pujol from 2011 to 2015 were prospectively included. A transversal, consecutive design was applied to assess IC compared to the reference test (OSNA). Inclusion criteria were: T1 and T2 tumors with negative nodes, both clinically and on ultrasound.ResultsSensitivity of IC for macrometastases was 70%. The NPV of IC for macrometastases was 95,75%. Accuracy of IC was 96,12%. In the comparative cost analysis, the release time of results for OSNA doubled that of IC and was associated with an increased cost of € 370.ConclusionsIC has been stated as a good technique for intraoperative cancer involvement SN with high sensitivity and NPV compared to the OSNA assay. It allows keeping the whole node tissue and thus the possibility of improved histopathological evaluation, which can be useful for adjuvant, and offers the advantage of being less time consuming. Cost analysis shows a higher cost for OSNA, which may exceed the benefit of sorting out false negatives from IC.     

AML1/ETO阳性急性髓系白血病患者galectin-3的表达及临床意义

目的 探讨AML1/ETO阳性急性髓系白血病（AML1/ETO+ AML）患者半乳凝集素3（galectin-3,gal-3）表达及其临床意义。方法 采用实时定量RT-PCR（RQ-PCR）法检测53例AML1/ETO+ AML患者骨髓单个核细胞中gal-3 mRNA表达水平,ELISA法检测患者外周血gal-3蛋白水平,分析gal-3 mRNA表达与疾病预后的相关性。结果 ①gal-3 mRNA及蛋白在初诊AML1/ETO+ AML患者中呈高表达（ P<0.001）,初诊患者gal-3 mRNA与蛋白表达呈正相关（r=0.732, P<0.001）,疾病完全缓解期外周血gal-3蛋白表达量下降（ P<0.001）;②初诊时gal-3 mRNA表达水平越高,患者白细胞数量越低（ P=0.014）,CD34表达和附加染色体异常的比例越高（ P=0.001, P=0.026）;③gal-3 mRNA高表达组与低表达组在完全缓解（CR）、部分缓解（PR）、诱导期死亡（即早期死亡）率上无明显差异（ P＞0.05）,但两组患者在复发率上差异有统计学意义（ P=0.029）;④gal-3 mRNA高表达组患者中位总生存率（OS）及无病生存率（DFS）均较低表达组缩短（ P=0.007, P=0.015）,累积复发率增高（ P=0.045）,但累积死亡率无明显差异（ P＞0.05）。Cox风险比例模型提示gal-3 mRNA是影响AML1/ETO+ AML患者OS及DFS的独立指标。结论 骨髓gal-3 mRNA有可能成为AML1/ETO+ AML患者评估预后及指导治疗的重要参考指标。     

以肿瘤-睾丸抗原mRNA为特异性标记评价肝癌患者肝移植的预后

目的探讨肝细胞癌 (hepatocellularcarcinoma ,HCC)患者原位肝移植 (orthotopiclivertrans plantation ,OLT)手术前后外周血中MAGE 1、SSX1及CTp11基因mRNA表达与预后的关系。方法用巢式RT PCR方法检测OLT手术前后外周血单个核细胞 (PBMC)中上述 3种基因的表达 ,并对 2 6例患者进行随访。结果HCC患者术前PBMC中MAGE 1、SSX1和CTp11基因mRNA的表达率分别为4 0 0 % (14 / 35 )、37 1% (13/ 35 )和 31 4 % (11/ 35 ) ,6 0 0 % (2 1/ 35 )患者的PBMC中至少可以检测到其中一种基因表达。 16例术前PBMC中检测到上述基因表达者 ,术后肿瘤复发 /转移率 (75 0 % )显著高于10例不表达这 3种基因者 (30 0 % ) (P =0 0 4 3) ;在术前及术后这 3种基因mRNA持续呈阳性或者由阴性转为阳性的 2 0例患者中 ,15例 (75 0 % )术后发生肿瘤复发 /转移 ,而 3种基因mRNA持续阴性或者由阳性转为阴性的 6例患者无 1例复发 /转移 (P =0 0 0 2 )。结论MAGE 1、SSX1及CTp11基因mR NA可以联合作为肿瘤特异性标志物用于检测HCC患者血循环中的肿瘤细胞 ,并有助于选择OLT受体及判断预后。