Villous adenoma of the urinary tract: a clinicopathological study / 中华病理学杂志

<p><b>OBJECTIVE</b>To explore the clinicopathological features, immunophenotype, differential diagnosis, pathogenesis and prognosis of villous adenoma with poorly differentiated adenocarcinoma of the urinary tract.</p><p><b>METHODS</b>Clinical and pathologic findings of 3 cases of villous adenoma with poorly differentiated adenocarcinoma of the urinary tract were analyzed by gross examination, microscopic investigation and immunohistochemical staining. The related literatures were reviewed.</p><p><b>RESULTS</b>All of the three cases were middle-aged or elderly patients. Three cases all presented with hematuria and mucusuria. Endoscopic examination identified that case 1 had a polyp with broad attachment in the dome of bladder, case 2 had a solid mass in the ureter, and case 3 had a exophytic fungating tumor in the renal pelvis. Microscopically, case 1 revealed a papillary lesion with finger-like processes lined by pseudostratified columnar epithelium with abundant goblet cells. The cells demonstrated moderate degree dysplasia. In case 2 and case 3, both villous adenomas and poorly differentiated adenocarcinoma were observed, the adenoma cells arranged in a cribriform pattern, and the tumor cells showed severe atypia, mitotic activity, and transition with invasive poorly differentiated adenocarcinoma. Immunohistochemically, the tumor cells in three cases were positive for CK20, CEA,EMA and MUC-1; none of them expressed cdx-2 and PSA; In case 2 and 3, the same immunophenotype of villous adenomas and their associated adenocarcinomas was observed, but the number of the positive cells of p53 and Ki-67 staining were significantly increased in the area of adenocarcinomas than in that of the villous adenomas.</p><p><b>CONCLUSIONS</b>Villous adenoma of the urinary tract is rare. It can occur in the urinary bladder, urachus, renal pelvis, ureter and urethra. These lesions may have malignant potential and frequently coexist with other malignant tumors. So, villous adenoma of the urinary tract should be removed completely and sampled thoroughly to avoid missing a more aggressive component.</p>

Forward genetic screening for zebrafish mutants defective in erythropoiesis / 南方医科大学学报

<p><b>OBJECTIVE</b>To screen and identify zebrafish mutants with erythropoiesis defects by N-ethyl-N-nitrosourea (ENU) mutagenesis and large-scale forward genetic screening using beta e 1 as the marker.</p><p><b>METHODS</b>The chemical mutagen ENU was used to treat healthy wild-type male fish (AB strain, F0). The surviving ENU-treated fish were mated with wild-type female fish to generate F1, and further F2 family was generated by F1 family intercross. The adult F2 fish were intercrossed within each F2 family and the resulting F3 embryos from each crossing were subjected to whole mount in situ hybridization (WISH) with the beta e 1 probe. Mutagenesis was performed by treating the male zebrafish with ENU to induce mutations in pre-meiotic germ cells to generate the founders, which were outcrossed to obtained the F1 fish. The F1 fish from different founders were mated to generate the F2 families. F3 embryos from the sibling cross in the F2 family were examined by whole mount in situ hybridization using beta e 1-globin probe. The putative mutants were then characterized with different hematopoiesis markers.</p><p><b>RESULTS AND CONCLUSION</b>We identified 4 beta e 1-deficient mutants with erythropoiesis defects, including two with specific erythiod lineage defects and two with concurrent lymphopoiesis defects.</p>

Preparation of RNA probe for cd99l2 gene of zebrafish labeled with digoxingenin-UTP / 南方医科大学学报

<p><b>OBJECTIVE</b>To study the expression pattern of cd99l2 gene during zebrafish development, the RNA probes for whole-mount in situ hybridization were prepared in this study.</p><p><b>METHODS</b>The cd99l2 fragment obtained by RT-PCR was cloned into pGM-T Easy, then the plasmids were linearized with the restriction enzymes SacII or SalI. Using Sp6 or T(7) RNA polymerase, the digoxingenin-labeled antisense and sense probes were synthesized and confirmed by whole-mount in situ hybridization.</p><p><b>RESULTS</b>The plasmid cd99l2/pGM-T was constructed. cd99l2 gene expression pattern during embryogenesis of zebrafish was examined using the antisense probe, and intense expression was detected in the central nervous system during zebrafish development.</p><p><b>CONCLUSION</b>The antisense probe can be used for study of the spatial and temporal distribution of cd99l2 during zebrafish development using the sense probe as control.</p>

Forward genetic screening for zebrafish mutants defective in myelopoiesis / 南方医科大学学报

<p><b>OBJECTIVE</b>To identify zebrafish mutants with myelopoiesis defects by ENU mutagenesis and large-scale forward genetic screening.</p><p><b>METHODS</b>Male zebrafish were mutagenized with N-ethyl N-nitrosourea to induce mutations in the spermatogonial cells to generate the founders, which were outcrossed with AB to raise F1 fish. The F1 fish from different founders were mated to generate the F2 families. The F3 embryos from F2 sibling crosses were screened by Sudan black B staining and neutral red staining.</p><p><b>RESULTS</b>A total of 350 F2 families from F1 sibling crosses were screened, and 1424 F2 crosses were analyzed. Six mutations were identified resulting in abnormal Sudan black B staining and neutral red staining, indicating the involvement of neutrophil deficiency or macrophage abnormalities.</p><p><b>CONCLUSION</b>It is simple and cheap to induce and screen myelopoiesis deficiency in zebrafish by ENU chemical mutagenesis and Sudan black B staining and neutral red staining. These mutants shed light on the identification of the genes important to myelopoiesis in zebrafish.</p>

Relationship between lymphocyte immunophenotypes and histologic subtypes of Hodgkin's lymphoma and its significance / 中国实验血液学杂志

This study was aimed to investigate the relationship between immunophenotype of the background lymphocytes and histological subtype of Hodgkin's lymphoma (HL), and its significance. The relative protein expressions of background lymphocytes were detected in 37 HL specimens on the basis of instant-rapid MaxVision(TM) immunohistochemical method and assessed quantitatively with image analysis software IPP6.0. The adoptive antibody included anti-CD3/CD45RO, anti-CD20/CD79a, anti-CD4, anti-CD8, anti-GrB, anti-TIA-1. The results indicated that out of 37 cases 4 were NLPHL, 33 were CHL including 6 of MCHL, 14 of NSHL, 13 of LRHL. In addition, 10 cases (1 was NLPHL, 4 were NSHL, 5 were LRHL) were involved in the analysis of T/B ratios. The ratio of T/B in NLPHL was 0.28 +/- 0.07, in CHL 4.34 +/- 2.45 (p = 0.001), in CHL the ratio was LRHL > NSHL > MCHL (p = 0.649); CD4(+)/CD8(+) ratio in NLPHL was 4.55 +/- 1.28, in CHL 4.10 +/- 1.50 (p = 0.574), in CHL it was MCHL > NSHL > LRHL (p = 0.037); GrB(+)/TIA-1(+) ratio in NLPHL was 0.71 +/- 0.57, in CHL 0.74 +/- 0.39, it was MCHL > NSHL > LRHL (p > 0.05). It is conduced that immune cell composition of the diagnostic HL lymph node represents the immune microenvironment. It is different between NLPHL and CHL in terms of the T- and B-lymphocyte distribution. NLPHL is of unique feature. The subtypes of CHL are of peculiar. The T/B ratio is in order as LRHL > NSHL > MCHL, but CD4(+)/CD8(+) and GrB(+)/TIA-1(+) ratios are in the opposite order. Combining with prognosis of subtypes of CHL i.e, LRHL > NSHL > MCHL, these data suggest that low ratio of T/B with high ratios of CD4(+)/CD8(+) and GrB(+)/TIA-1(+) may represent biological markers predicting an unfavorable outcome of CHL subtypes.

Construction of mic2/CD99 gene vector and its transfection in Hodgkin lymphoma L428 cell line / 南方医科大学学报

<p><b>OBJECTIVE</b>To construct a eukaryotic expression vector of CD99 gene for transfection into Hodgkin lymphoma L428 cells.</p><p><b>METHODS</b>The full-length cDNA of CD99 gene was amplified from Jurkat cells by RT-PCR and cloned into the pcDNA3.1(+) vector and transfected into L428 cell line using Lipofextamine 2000. The sequence of CD99 mRNA in the transfected cells was confirmed by restriction endonuclease digestion and DNA sequencing, and the expression of CD99 protein was identified using immunocytochemistry.</p><p><b>RESULTS</b>A gene fragment of 558 bp was amplified from the transfected cells and the sequence was verified by DNA sequencing. Immunocytochemistry identified the presence of CD99 expression in the transfected cells.</p><p><b>CONCLUSION</b>A eukaryotic expression vector pcDNA3.1(+)-CD99 is successfully constructed and stably expressed in L428 cell line.</p>