肝癌患者肝移植术后肿瘤复发相关的血清蛋白质标记物筛选

Objective To appraise and compare protein expression profiles in sera of patients without or with recurrence following liver transplantation for hepatocellular carcinoma (HCC) using SELDI-TOF-MS technique,and establish the diagnostic and predictive model. Methods A total of 76 sera (41 from disease free survival patients and 35 from recurrence individuals) were collected pretransplantation and differentially expressed proteins were identified by SELDI-TOF-MS. The intensity values for each peak were analyzed by Biomarker Wizard Software to screen serum proteome biomarkers related to the recurrence post-transplantation. By using Biomarker Patterns Software, the classification trees were generate. from randomly selected samples (30 fingerprints obtained from each group). The sensitivity and specificity of best decision tree were then chosen for blind test with 16 samples (5 from recurrence individuals and 11 from recurrence-free survival patients). Results There were significant differences only in tumor size and the presence of vascular invasion between recurrence group and recurrence-free survival group (P＜0.05). According to serum protein fingerprints, a total of 368 protein peaks were identified at the mass-to-charge ratio (M/Z) value ranging from 2000 to 300 00. There were 22 significant differential proteins between two groups. Among them, 9 proteins were up-regulated and 13 proteins were down-regulated -espectively in recurrence group. The intensity values of differential proteins were input into BPS for classification tree analysis and the best performing tree could distinguish two groups successfully. As a result of blind assessment for this model,a sensitivity of 80.0 % (4/5) and specificity of 72.7 % (8/11) were obtained. Conclusion Some of differential proteins screened by SELDI-TOF-MS technique in the serum may be correlated with the prognoses of liver transplantation patients with HCC. The decision tree may be useful for the clinical application of formulating the indication for liver transplantation, detecting extrahepatic micrometastasis and setting up the diagnostic and treatment strategies.     

肝癌患者肝移植术后肿瘤复发相关的血清蛋白质标记物筛选

Objective To appraise and compare protein expression profiles in sera of patients without or with recurrence following liver transplantation for hepatocellular carcinoma (HCC) using SELDI-TOF-MS technique,and establish the diagnostic and predictive model. Methods A total of 76 sera (41 from disease free survival patients and 35 from recurrence individuals) were collected pretransplantation and differentially expressed proteins were identified by SELDI-TOF-MS. The intensity values for each peak were analyzed by Biomarker Wizard Software to screen serum proteome biomarkers related to the recurrence post-transplantation. By using Biomarker Patterns Software, the classification trees were generate. from randomly selected samples (30 fingerprints obtained from each group). The sensitivity and specificity of best decision tree were then chosen for blind test with 16 samples (5 from recurrence individuals and 11 from recurrence-free survival patients). Results There were significant differences only in tumor size and the presence of vascular invasion between recurrence group and recurrence-free survival group (P＜0.05). According to serum protein fingerprints, a total of 368 protein peaks were identified at the mass-to-charge ratio (M/Z) value ranging from 2000 to 300 00. There were 22 significant differential proteins between two groups. Among them, 9 proteins were up-regulated and 13 proteins were down-regulated -espectively in recurrence group. The intensity values of differential proteins were input into BPS for classification tree analysis and the best performing tree could distinguish two groups successfully. As a result of blind assessment for this model,a sensitivity of 80.0 % (4/5) and specificity of 72.7 % (8/11) were obtained. Conclusion Some of differential proteins screened by SELDI-TOF-MS technique in the serum may be correlated with the prognoses of liver transplantation patients with HCC. The decision tree may be useful for the clinical application of formulating the indication for liver transplantation, detecting extrahepatic micrometastasis and setting up the diagnostic and treatment strategies.     

乳腺癌和乳腺良性肿物体表傅里叶变换红外光谱研究

目的 探讨利用傅里叶变换红外(FTIR)光谱技术鉴别乳腺癌和乳腺良性肿物的可行性.方法 联合运用衰减全反射(ATR)探头与中红外光纤、FTIR光谱分析仪,测定26例乳腺癌患者和34例乳腺良性肿物患者体表的FTIR光谱,比较两组光谱各个谱带的峰位(P)、相对峰强(I)及半高宽指标(F),总结乳腺癌体表光谱的特征.结果 与乳腺良性肿物体表光谱比较,乳腺癌体表光谱中(1)蛋白质酰胺Ⅱ带P1546发生明显地红移(向低波数移动,P＜0.05);(2)与核酸相关的相对峰强11256/I1460(P＜0.05)明显升高;(3)与C-H或C-O-H的变角振动的谱带峰位P1303发生明显地红移(向低波数移动)(P＜0.05).结论 乳腺癌体表FTIR光谱与乳腺良性肿物体表的光谱比较,在蛋白质、核酸和脂类相关谱带均有明显差异.

Abstract：

Objective To explore the feasibility of superficially differentiating breast cancer from the benign fourier transform infrared ( FTIR) spectroscopy. Methods Twenty-six cases of breast cancer and 34 cases of the benign were examined superficially by FTIR spectroscopy. For each FTIR spectrum,the peak positions (P), relative intensity (I) and full width at half maximum ( F) of the band were recorded and the relative intensity ratios were calculated. Results In the group of breast cancer, both the peaks of 1546 cm-1 and 1303 cm-1 shifted toward lower wave number (P ＜0. 05), as compared to those of the benign group. The relative intensity ratio of I1256/I1460) was increased significantly (P ＜ 0. 05). Conclusion There existed many significant differences in FTIR spectra between malignant and benign breast masses.     

腹腔镜胃肠手术治疗2型糖尿病的血清蛋白质组学初步研究

目的 探讨腹腔镜胃肠手术前后2型糖尿病患者血清蛋白质表达的差异.方法 收集2008年6月至2010年9月第二军医大学附属长海医院行腹腔镜胃肠手术治疗的12例2型糖尿病患者的血清标本,双向凝胶电泳分离2型糖尿病患者术前、术后1周及术后1个月的血清总蛋白,对手术前后差异表达明显的蛋白点行质谱鉴定及生物信息学分析.结果 建立了稳定、重复性好的血清凝胶蛋白图谱.筛选出2型糖尿病患者胃肠手术前后血清中差异表达明显的20个蛋白点,并成功鉴定出8个蛋白点,其中术后表达增高的有5个,分别是Rho GDP-dissociation inhibitor 1、Prohibitin、Alpha-1-antitrypsin precursor、Serotransferrin precursor和Fibrinogen gamma chain precursor;术后表达降低的有3个,分别是MAP3K12-binding inhibitory protein 1、Coronin-1A和Isovaleryl-CoA dehydrogenase.结论 2型糖尿病患者行腹腔镜胃肠手术后的血清蛋白质与术前比较,存在着明显的差异.共成功鉴定出8个蛋白点.

Abstract：

Objective To investigate the impact of laparoscopic gastrointestinal surgery on serum protein expression in patients with type 2 diabetes mellitus(T2DM).Methods Twelve patients with T2DM received gastrointestinal surgery at Changhai Hospital of the Second Medical University from June 2008 to September 2010.Their serum samples were collected at different time points(before surgery,1 week and 1 month after surgery).Total proteins were seperated by two-dimensional(2D)gel electrophoresis.The differentially expressed proteins were analyzed by mass spectrometry and bioinformatics.Results Protein extracts of the serum samples were separated on 2D gels successfully.Twenty differentially expressed proteins in the serum after surgery were screened out.Eight proteins were successfully identified,in which the expression of 5 proteins(Rho GDP-dissociation inhibitor 1,Prohibitin,Alpha-1-anfitrypsin precursor,Serotransferrin precursor and Fibrinogen gamma chain precursor)was increased after operation,and the expression of 3 proteins(MAP3K12-binding inhibitory protein 1,Coronin-1A and Isovalery1-COA dehydrogenase) was decreased.Conclusions The expression of 20 proteins have been changed significantly in serum samples after laparoscopic gastrointestinal surgery in patients with T2DM,and 8 proteins were successfully identified.     

Expression and clinical significance of FPARγ and β-catenin in breast cancer

Objective To study the expression of peroxisome proliferator-activated receptor γ (PPARγ) and β-catenin in breast cancer, and their correlations with clinicopathological parameters and prognosis. Methods Tissue samples obtained from 70 patients with breast cancer and 20 patients with breast benign mass were immunohistochemically examined for the expression of PPARγ and p-catenin. Results Overexpression rate of PPARγ protein was 34. 3% in breast cancer, significantly lower than that in breast benign mass. Abnormal expression rate of β-catenin in breast cancer was 67. 1%. A significant negative-correlation was found between the expression of PPARγ and β-catenin (r=-0. 398,P<0.05 ). PPARγ expression was inversely associated with histologic grade, tumor size, axillary lymph node metastasis,TNM stage and Ki-67 expression (P<0. 05), while positively correlated with ER status and overall survival rate (P<0. 05). Abnormal β-catenin expression was positively associated with histologic grade, axillary lymph node metastasis and TNM stage (P<0. 05), while inversely correlated with overall survival rate (P<0.05). Conclusion PPARγ and β-catenin are correlated with development of breast carcinoma,suggesting that detection of PPARγ and β-catenin may be of value in evaluating the biological behaviors and the prognosis of breast cancer.     

Expression and clinical significance of FPARγ and β-catenin in breast cancer

Objective To study the expression of peroxisome proliferator-activated receptor γ (PPARγ) and β-catenin in breast cancer, and their correlations with clinicopathological parameters and prognosis. Methods Tissue samples obtained from 70 patients with breast cancer and 20 patients with breast benign mass were immunohistochemically examined for the expression of PPARγ and p-catenin. Results Overexpression rate of PPARγ protein was 34. 3% in breast cancer, significantly lower than that in breast benign mass. Abnormal expression rate of β-catenin in breast cancer was 67. 1%. A significant negative-correlation was found between the expression of PPARγ and β-catenin (r=-0. 398,P<0.05 ). PPARγ expression was inversely associated with histologic grade, tumor size, axillary lymph node metastasis,TNM stage and Ki-67 expression (P<0. 05), while positively correlated with ER status and overall survival rate (P<0. 05). Abnormal β-catenin expression was positively associated with histologic grade, axillary lymph node metastasis and TNM stage (P<0. 05), while inversely correlated with overall survival rate (P<0.05). Conclusion PPARγ and β-catenin are correlated with development of breast carcinoma,suggesting that detection of PPARγ and β-catenin may be of value in evaluating the biological behaviors and the prognosis of breast cancer.     

原发性腹膜后神经鞘瘤的外科治疗

目的 探讨原发性腹膜后神经鞘瘤的诊治与预后.方法 回顾性分析1995年1月至2009年12月北京大学第一医院诊治的47例原发性腹膜后良、恶性神经鞘瘤患者的临床资料.结果 良性神经鞘瘤36例,中位年龄41岁,体检发现25例,有症状者11例;恶性神经鞘瘤11例,中位年龄38岁,体检发现5例,有症状者6例.术前CT和MRI诊断阳性率分别为36.2%(17/47)和58.3%(7/12).免疫组化染色良性组S-100阳性率100%;恶性组S-100阳性者率81.8%(9/11).47例患者均行手术治疗,良性组手术切除率100%,恶性组手术切除率90.9%(10/11),两组均无围手术期死亡患者,术后并发症5例(10.6%).良、恶性神经鞘瘤术后5年生存率分别为100%与45.5%.良性组术后复发2例;恶性组术后复发4例,远处转移3例.结论 腹膜后神经鞘瘤确诊依靠病理及免疫组化检查,手术完整切除是治疗主要方法.良性神经鞘瘤预后良好,恶性神经鞘瘤易转移和复发.

Abstract：

Objective To explore the clinical diagnosis and surgical treatment of primary retroperitoneal neurilemoma (schwannoma). Methods Clinical data of 47 patients of primary retroperitoneal schwannoma admitted and surgically treated from January 1995 to December 2009 were retrospectivelly reviewed. Results As diagnosed by pathology there were 36 cases of Benign schwannoma,with a median age at onset of 41 years, among those 11 patients were symptomatic, and 25 were asymptomatic. There were 11 malignant 11 cases, the median age was 38 years, among those 6 patients were symptomatic, and 5 were asymptomatic. The positive diagnostic rate of preoperative CT and MRI were 36. 2％ ( 17/47 ) and 58. 3％ ( 7/12 ) respectively. Immunohistochemically positive rates of S-100 were 100％ and 81.8％ (9/11) in benign and malignant group respectively. All cases underwent surgical treatment. Surgical resection rates for benign and malignant groups were 100％ and 90. 9％ (10/11)respectively. There was no perioperative death, Overall 5-year survival rates were 100％ and 45.5％ for benign and malignant tumors groups respectively. In benign group 2 cases recurred, in malignant group 4 cases recurred, and 3 had distant metastasis. Conclusions Primary retroperitoneal schwannomas are less common. It is difficult to make an accurate preoperative diagnosis. Surgery is the most effective therapy.Prognosis is good for benign and poor for malignant retroperitoneal neurilemomas.     

临床检查阴性的乳腺X线照片簇状钙化对乳腺癌诊断的意义

目的 探讨乳腺X线片发现乳腺内簇状钙化在乳腺癌诊断的临床意义。方法 对临床检查阴性,而乳腺X线摄片发现乳腺内簇状钙化共91例病人102个病灶,并进行手术活检。根据病理组织学结果,对其形态、数量及分布作进行回顾性分析及对照研究。结果 91例病例发现102个乳腺内簇状钙化灶,其中43个恶性病变（42．15％）,59个良性病变（57．42％）。结论根据乳腺内钙化的形态、数目、分布的X线特征,是有助于对乳腺癌早期诊断。     

超声探测甲状腺结节钙化的临床诊断价值

目的：探讨超声探测甲状腺结节钙化对甲状腺结节良恶性的诊断价值。 方法：回顾性分析2005年1月—2010年1月经手术治疗的4 011例甲状腺疾病患者的超声资料和临床病理结果,分析钙化与否及不同钙化类型与甲状腺结节良恶性的关系,比较单发和多发钙化结节的恶性率以及不同年龄组甲状腺结节伴钙化的恶性率。 结果：甲状腺结节伴钙化的恶性率明显高于非钙化结节,微小钙化结节的恶性率明显高于非微小钙化,单发钙化结节恶性率均明显高于多发钙化结节,年龄<45岁钙化结节者的恶性率明显高于年龄≥45岁者,组间差异均有统计学意义（均P<0.05）。钙化、微小钙化、粗大钙化和环状钙化对诊断甲状腺恶性肿瘤的敏感度分别为80.42%、51.65%、26.52%、2.25%,特异度分别为67.53%、93.19%、76.79%、97.55%。39例颈部淋巴结钙化者中,37例为甲状腺乳头状癌淋巴结转移。 结论：甲状腺结节钙化和微小钙化对甲状腺癌的诊断具有重要意义。单发结节伴钙化、微小钙化、年龄<45岁钙化结节患者以及淋巴结钙化者具有更高的恶性率。

     

利用二维胶内差示凝胶电泳技术检测胃癌的血清学肿瘤标记物

目的 分离胃癌、胃溃疡、正常人之间的差异蛋白质,检测胃癌的血清学肿瘤标记物.方法 从定量比较蛋白质组分析入手,利用二维胶内差示凝胶电泳技术(2D-DIGE)分离包括胃癌、胃溃疡和正常人共27例血清样本,以基质辅助激光解吸附电离串联质谱对差异蛋白点进行检测,检索Mascot数据库.结果 共检测出14个差异表达蛋白.正常人和胃溃疡比较有3种差异蛋白质,2种被鉴定出;胃溃疡和胃癌比较发现1个下调的蛋白质;胃癌与正常人中发现10个差异表达蛋白质,6个点在胃癌患者血清中上调.结论 2D-DIGE技术是一项有效的筛选胃癌患者血清中差异蛋白的技术手段,检测出的蛋白质有可能成为胃癌诊断的分子标记物.     

乳腺钙化组织中骨桥蛋白的表达及其临床意义

目的 检测骨桥蛋白（osteopontin protein,OPN）mRNA在含钙化灶乳腺组织中的表达,及其在乳腺癌转移发生发展中的意义.方法 采用RT-PCR法检测128例乳腺病变组织及相应癌旁乳腺组织和9例乳癌转移淋巴结中骨桥蛋白mRNA的表达.结果 骨桥蛋白mRNA表达在含钙化灶乳腺癌组织及转移淋巴结组织中表达最高,良性钙化灶乳腺组织次之,而在癌旁乳腺组织及良性无钙化病变组织中表达最低,差异有显著意义（P＜0.05）.结论 骨桥蛋白的表达与乳腺癌组织钙化有关,并可能与乳腺癌的发生有关,检测其表达水平,可望能指导临床治疗,评判患者预后.     

胆囊癌血清标志物的蛋白质组学研究

目的 分析胆囊癌患者和健康人血清蛋白质组的差异,筛选胆囊癌血清分子标志物.方法 以6例健康人和6例早胆囊癌患者的血清为样本,图像分析两组血清图谱寻找差异蛋白质点,基质辅助激光解吸电离飞行时间质谱对差异蛋白点进行鉴定.Western blot和免疫组织化学方法检测差异蛋白质S100A10和结合珠蛋白在胆囊癌患者和健康人血清中的表达.结果 建立了健康人和胆囊癌患者血清样品的二维凝胶电泳图谱,质谱鉴定出24种非冗余的血清蛋白质.Westernblot证实了S100A10蛋白和结合珠蛋白在胆囊癌患者和健康人血清中的差异表达.免疫组织化学证实胆囊癌组织中S100A10及结合珠蛋白高表达.结论 24种血清差异蛋白质为筛选胆囊癌的血清分子标志物提供了实验依据.     

Breast biopsy for calcifications in nonpalpable breast lesions. A prospective study

We prospectively studied 239 consecutive patients who underwent breast biopsy for 277 nonpalpable lesions characterized by mammographic microcalcifications. Clinical and mammographic characteristics were correlated with histologic findings in an attempt to identify patients more likely to have early breast cancer. The distribution of clinical risk factors was equal between patients with benign or malignant outcomes. The predominant Wolfe pattern on mammography was P2 (38%); however, no relationship was observed between the Wolfe pattern and malignancy. A marked correlation was observed between malignancy and small lesions, more than 15 calcifications, and calcifications in a linear or branching pattern. Twenty-four percent (n = 67) of the biopsy specimens contained either ductal or lobular breast cancer. This study highlights the necessity of an aggressive approach toward suspicious calcifications found by mammography.     

Breast cancer screening in women with chronic kidney disease: the unrecognized effects of metastatic soft-tissue calcification

Patients with chronic kidney disease (CKD) or end-stage renal disease (ESRD) are known to develop metastatic soft-tissue calcification, secondary to hyperparathyroidism, in tissues including the breast. Such calcifications in women could pose a problem for interpretation of mammograms, since they are thought to mimic malignant lesions and interfere with differentiation of benign from malignant disease. Investigation of this issue is important to provide high-quality, accurate breast care to women with CKD or ESRD, but little evidence is so far available. In a systematic review of the literature on the types and patterns of breast calcifications, we found only three studies that examined metastatic soft-tissue calcifications of the breast. The studies did, however, confirm that women with CKD or ESRD have a higher frequency of breast calcification than women with normal kidney function. The two older studies reported that these breast calcifications are not associated with malignancy, but the later study reported a raised rate of suspicious breast calcification among women with ESRD receiving hemodialysis, leading to an increased biopsy referral rate. In this Review we discuss the strengths and limitations of the available data and whether mammography is recommended in women with CKD or ESRD.     

Microcalcifications Associated with Breast Cancer: An Epiphenomenon or Biologically Significant Feature of Selected Tumors?

Radiographic mammary calcifications occur in 30–50% of breast cancers and constitute one of the most important diagnostic markers of both benign and malignant lesions of the breast. The presence of oxalate-type microcalcification appears to be a reliable criterion in favor of the benign nature of the lesion or, at most, of a lobular carcinoma in situ. In contrast, calcium hydroxyapatite (HA) crystals are associated with both benign and malignant breast tumors. Although the diagnostic value of microcalcifications in breast cancer is of great importance, the genesis of these calcifications is unclear. Despite numerous histological ultrastructure studies of HA deposits in breast carcinomas, to date there have been limited investigations of the potential role of these crystals in breast cancer. We review the literature examining the biological effects of HA crystals in breast cancer cell lines, specifically the mechanism of HA-induced mitogenesis and upregulation of gene expression.     

乳腺癌组织中EG-1 mRNA的表达与临床病理特征及预后的关系

目的 探讨EG-1基因在乳腺癌组织中的表达及其与乳腺癌临床特征、预后的相关性.方法 采用RT-PCR技术,检测EG-1基因在72例乳腺癌及18例良性乳腺组织中的表达,分析其表达与临床特征及预后的相关性.结果 71%(51/72)患者的乳腺癌石蜡标本中EG-1mRNA阳性,而良性乳腺组织中的表达率为24%(4/18).EG-1基因在乳腺癌组织中的表达与淋巴结转移、VEGF蛋白有关(P＜0.05),与年龄、月经状况、肿瘤大小、组织学分级、Her-2蛋白、p53蛋白、激素受体状况无关,EG-1mRNA阳性患者远处转移率和复发率高于阴性患者.多因素回归分析发现EG-1mRNA阳性是乳腺癌的独立预后因素之一. 结论 EG-1基因在乳腺癌组织中异常高表达,且与淋巴结转移及VEGF蛋白相关,是乳腺癌潜在的预后标记物.     

乳腺癌患者血清和肿瘤组织VEGF表达与临床预后的关系

目的探讨乳腺癌患者血清和肿瘤组织中血管内皮细胞生长因子（VEGF）的表达以及与临床预后的关系。方法以44例乳腺癌患者、13例乳腺良性疾病患者和40例正常人为研究对象，分别采用酶联免疫吸附法（ELISA）检测其血清VEGF水平，采用免疫组化LSAB法检测其组织中VEGF、雌激素受体（ER）和原癌基因C-erbB-2蛋白的表达，并分析其与临床预后因素如淋巴结转移情况及临床分期的关系。结果乳腺癌组血清和组织中VEGF表达水平（113．88pg／ml，20723．99）均明显高于乳腺良性疾病组（55．79pg／ml，3594．74），P〈0．001，而乳腺良性疾病组与正常对照组（41．30pg／ml，-）比较差异无统计学意义（P〉0．05）；且血清和组织中VEGF的表达呈正相关（r=0．48，P〈0．01）。有淋巴结转移者血清和组织中VEGF的表达水平（129．60pg／ml，28506．82）明显高于无淋巴结转移者（80．80pg／ml，14656．73），P〈0．01；血清和组织中VEGF的表达与肿瘤的临床分期有关（P〈0．01），但与患者年龄和肿瘤大小无关（P〉0．05）。乳腺癌患者血清和组织中VEGF表达水平与组织中ER表达呈负相关（r=-0．45，P〈0．05），与C-erbB-2表达呈正相关（r=0．48，P〈0．01）。结论乳腺癌患者血清和肿瘤组织中VEGF表达呈正相关，且与其临床预后有关，可作为乳腺癌患者预后判断的重要参考指标之一。