PCR-based assays for the detection of monoclonality in non-Hodgkin's lymphoma: application to formalin-fixed,paraffin-embedded tissue and decalcified bone marrow samples

In crucial cases the diagnosis of non-Hodgkin's lymphoma (NHL) still represents a challenge to the pathologist since morphological criteria do not always help to distinguish between reactive and malignant lymphoproliferations. Clonality assays are a useful supplement since monoclonal cell proliferation is strong evidence for malignancy. The polymerase chain reaction (PCR) can be utilized to establish the clonal origin of B-or T-cell lymphocyte populations by amplification of rearranged immunoglobulin and T-cell receptor (TCR) genes. In the present study DNA was isolated from a variety of neoplastic and nonneoplastic formalin-fixed, paraffin-embedded lymph nodes (n=62), cutaneous tissue (n=9), samples of miscellaneous origin (n=11), and, reported here for the first time, decalcified bone marrow samples (n=35). These samples were submitted to PCR-based assays directed against the immunoglobulin heavy-chain (IgH), immunoglobulin light-chain (IgL), and TCR chain genes. The impact of various decalcifying agents on the ability to amplify DNA was investigated by PCR-based amplification of a single copy gene. Buffered and nonbuffered EDTA was found not to impede amplification of DNA fragments up to 300 bp in length. In lymph node and cutaneous specimens monoclonality was detected in 83% of B-NHL cases using a seminested PCR approach for the amplification of IgH, whereas the same approach gave rise to monoclonal bands in 80% of bone marrow samples. The subsequent amplification of IgL helped to raise the sensitivity of detection to 94%. Monoclonality was detected in seven of nine T-cell NHLs by amplification of TCR. Most of the false-negative results were related to DNA extracted from centroblastic-centrocytic lymphoma and lymphoplasmocytic immunocytoma (37% negative each). PCR-based rearrangement analysis of immunoglobulin and TCR chain genes should be used in diagnostic pathology for cases which are histopathologically and immunohistochemically questionable. The application of clonality assays to bone marrow samples previously decalcified with EDTA provides a new tool for the detection of minimal residual disease.Abbreviations BALT bronchus-associated lymphoid tissue - dNTP deoxynucleoside triphosphate - Ig immunoglobulin - IgH immunoglobulin heavy chain - IgL immunoglobulin light chain - MALT mucosa-associated lymphoid tissue - NHL non-Hodgkin's lymphoma - PCR polymerase chain reaction - TCR T-cell receptor     

乙肝病毒感染患者胆囊胆汁成分改变及其临床意义

目的：探讨乙肝病毒感染与胆石形成的关系。方法：收集38例乙肝病毒感染患者和35例非乙肝病毒感染者的胆囊内胆汁,对其胆红素、胆汁脂类、钙离子进行检测。结果：乙肝病毒感染患者胆囊胆汁的非结合性胆红素、钙离子明显高于对照组（P＜0.01,P＜0.05）,总胆汁酸、胆固醇明显低于对照组（P＜0.01）。结论：乙肝病毒感染患者胆囊胆汁成分改变与胆石症发病率增高有密切关系。     

Factores de riesgo de enfermedad residual en la re-RTU en una gran cohorte de pacientes con enfermedad T1G3

Introduction and objectivesThe goals of transurethral resection of a bladder tumor (TUR) are to completely resect the lesions and to make a correct diagnosis in order to adequately stage the patient. It is well known that the presence of detrusor muscle in the specimen is a prerequisite to minimize the risk of under staging.Persistent disease after resection of bladder tumors is not uncommon and is the reason why the European Guidelines recommended a re-TUR for all T1 tumors. It was recently published that when there is muscle in the specimen, re-TUR does not influence progression or cancer specific survival.We present here the patient and tumor factors that may influence the presence of residual disease at re-TUR.Material and methodsIn our retrospective cohort of 2451 primary T1G3 patients initially treated with BCG, pathology results for 934 patients (38.1%) who underwent re-TUR are available. 74% had multifocal tumors, 20% of tumors were more than 3 cm in diameter and 26% had concomitant CIS.In this subgroup of patients who underwent re-TUR, there was no residual disease in 267 patients (29%) and residual disease in 667 patients (71%): Ta in 378 (40%) and T1 in 289 (31%) patients. Age, gender, tumor status (primary/recurrent), previous intravesical therapy, tumor size, tumor multi-focality, presence of concomitant CIS, and muscle in the specimen were analyzed in order to evaluate risk factors of residual disease at re-TUR, both in univariate analyses and multivariate logistic regressions.ResultsThe following were not risk factors for residual disease: age, gender, tumor status and previous intravesical chemotherapy. The following were univariate risk factors for presence of residual disease: no muscle in TUR, multiple tumors, tumors > 3 cm, and presence of concomitant CIS. Due to the correlation between tumor multi-focality and tumor size, the multivariate model retained either the number of tumors or the tumor diameter (but not both), p < 0.001. The presence of muscle in the specimen was no longer significant, while the presence of CIS only remained significant in the model with tumor size, p < 0.001.ConclusionsThe most significant factors for a higher risk of residual disease at re-TUR in T1G3 patients are multifocal tumors and tumors more than 3 cm. Patients with concomitant CIS and those without muscle in the specimen also have a higher risk of residual disease.     

恶性梗阻性胆管炎29例临床分析

我院在1978～1988年间共收治胆管及壶腹周围肿瘤162例,其中29例有恶性梗阻性胆管炎,占同期收治的胆管及壶腹周围肿瘤的17.9％,其中表现为急性梗阻性化脓性胆管炎10例。病变依次为乳头部癌、胰头癌、胆管癌和肝门部转移癌。部分伴有胆石与胆道手术史。通过B超、经皮肝穿刺胆道造影术、胆道引流术,以及逆行胰胆管造影术而确诊。对发病情况、临床特点、诊治进行了讨论。     

脂多糖对胆汁分泌的影响

[目的 ]通过脂多糖对胆汁分泌的影响实验 ,初步探讨引起肝功能障碍及肝内结石形成的机理 .[方法 ]在麻醉状态下 ,直接往实验动物肝内注入脂多糖等实验药物 ,测定相关数据 .[结果 ]脂多糖对胆汁分泌有抑制作用 ,并对胆汁酸、磷脂浓度有降低作用 ,而对胆红素、钙的浓度无显著作用 .[结论 ]脂多糖对胆汁分泌有明显作用 ,对诱发肝功能障碍及形成肝内胆结石有一定的影响     

人体胆结石高能震波碎石的动物实验

目的　研究人体 3种胆结石在一定能量下 ,冲击次数和能量的关系 ,确定不损伤胆囊和小肠内膜的能量安全阈值 .方法　犬胆囊和小肠内放入固醇石、混合石和色素石 ,在不同的输出电压下 ,放电碎石 .结果　固醇石、混合石和色素石破碎所需的能量不同 ,在同样电压下 ,固醇石所需的次数最少 ,混合石次之 ,色素石最多 ;病理检查表明 ,在电压 <15 k V时 ,胆囊和小肠粘膜处 ,细胞膜轻度充血 ,出现小出血点 .当电压 >15 k V后 ,粘膜有脱落 ,膜下组织水肿 .结论　电压具有依赖性 ;该仪器临床应用的冲击电压应小于 15 k V     

Acid maltase deficiency in non-identical adult twins

Summary Acid maltase deficiency is described in non-identical adult twins. The onset of the disease can be traced into late infancy; the clinical picture is one of severe muscular dystrophy; respiratory insuficiency was the cause of death in one case. The autopsy showed the central nervous system, heart and liver to be spared. Glycogen filled vacuoles are found in skin, mesenchymal cells, small nerves and skeletal muscles. The light microscopic study of 9 different muscles showed extremely variable involvement ranging from normal appearance to overt vacuolization. A 6–20% residual acid -glucosidase activity was found in visceral organs, cultured fibroblasts and in some skeletal muscles. No satisfactory explanation can be given why this generalized acid -glucosidase deficiency produces a selective involvement of skeletal muscles. If compared with infantile AMD (Pompe's disease) our cases have a much higher residual acid -glucosidase activity and show the presence of an antigenically detectable protein.From our study and from a similar report in the literature (de Barsy et al., 1975), it appears that a combined approach of light microscopy, electron microscopy and biochemical analysis (determination of acid -glucosidase) is necessary to make a diagnosis of AMD in adults.Dr. Th. de Barsy is a Research Fellow of the Fonds National de la Recherche Scientifique.     

The endoscopic evaluation of gastritis,gastric remnant residue,and the incidence of secondary cancer after pylorus-preserving and transverse gastrectomies

Background Pylorus-preserving gastrectomy (PPG) and transverse gastrectomy (TrG) have been accepted as function-preserving procedures for node-negative early gastric cancer. It is believed that a better quality of life is guaranteed after PPG or TrG compared to that after distal subtotal gastrectomy (DSG) with Billroth type-I reconstruction. However, objective evaluations of the gastric remnant following gastrectomy have not been widely reported, and the real advantages and disadvantages of PPG or TrG over DSG remain unclear. Moreover, the risk of secondary cancer after PPG or TrG is uncertain.Methods Between 1991 and 2000, 834 DSGs were carried out in our institute for preoperatively diagnosed patients with early gastric cancer. The degree of residual gastritis and the amount of diet residue in the gastric remnant were evaluated by annual gastrointestinal endoscopic investigations prospectively for 72 patients after PPG, 95 patients after TrG, and 60 patients after DSG. These analyses were performed using the RGB classification (residue, gastritis, bile). The incidence of disease greater than or equal to grade 2 was calculated, and the time trends of the incidence for each procedure were also studied for 3 years after gastrectomy. In addition, secondary cancer cases in the gastric remnant mucosa were checked for each procedure during this period, and the incidence of secondary cancer after each operation was calculated.Results The incidence of gastritis, of grade 2 or more, found in the gastric remnant was significantly lower after PPG (1.4%) and TrG (2.1%) than after DSG (43.3%). However, the incidence of moderate or greater residue in the gastric remnant, grade 2 or more, was significantly higher after PPG (45.8%) and TrG (40.0%) than after DSG (11.7%). The analysis of time trends of gastritis and diet residue reflected the significant advantage or disadvantage for each procedure 1 year after surgery. The analysis also included these factors without consideration of elapsed time following surgery. Two patients after PPG (2.8%) and three patients after TrG (3.2%) developed secondary cancer in the gastric remnant. No DSG-treated patient showed new cancer genesis in the remaining stomach.Conclusion PPG and TrG have the advantage over DSG in preventing postoperative gastritis in the gastric remnant. On the other hand, moderate or greater diet residue in the gastric remnant is more common after PPG or TrG than after DSG. For the risk of carcinogenesis in the remnant gastric mucosa, we could not conclude that there was any apparent difference between these range-limited gastrectomies and conventional DSG. Further study is necessary to determine the significant advantages and disadvantages of using PPG or TrG.     

肺癌术后支气管切缘癌发生的相关因素分析

目的　探讨肺癌术后支气管切缘癌发生的相关因素。方法　对 2 43例原发性非小细胞肺癌患者的病例资料进行回顾性分析 ,采用Logistic回归分析判断术后支气管切缘癌发生的相关因素。 结果　 2 43例肺癌患者中 ,支气管切缘癌发生 3 1例 ,发生率为 12 .8% ,多因素分析显示 ,中央型肺癌、肺叶支气管淋巴结转移及组织学类型为鳞癌是支气管切缘癌发生的危险因素。结论　中央型肺癌、肺叶支气管淋巴结转移及组织学类型为鳞癌的肺癌病例应考虑发生术后支气管切缘癌的可能