Cytoplasmic FOXP1 expression is correlated with ER and calpain II expression and predicts a poor outcome in breast cancer.

BACKGROUND: Nuclear forkhead box protein P1 (N-FOXP1) expression in invasive breast cancer has been documented in the literature. However, the FOXP1 expression patterns at different stages of breast cancer progression are largely unknown, and the significance of cytoplasmic FOXP1 (C-FOXP1) expression in breast cancer has not been well illustrated. The aims of this study were to investigate FOXP1 expression patterns in invasive ductal carcinoma (IDC), ductal carcinoma in situ (DCIS), atypical ductal hyperplasia (ADH) and usual ductal hyperplasia (UDH), and to analyze the clinicopathological relevance of C-FOXP1 and its prognostic value in IDC. METHODS: N-FOXP1 and C-FOXP1 expression in cases of IDC, DCIS, ADH and UDH was determined using immunohistochemistry. The correlation between C-FOXP1 expression and clinicopathological parameters as well as the overall survival (OS) and disease-free survival (DFS) rates of patients with IDC were analyzed. RESULTS: Exclusive N-FOXP1 expression was found in 85.0% (17/20), 40.0% (8/20), 12.2% (5/41) and 10.8% (9/83) of UDH, ADH, DCIS, and IDC cases, respectively, and exclusive C-FOXP1 expression was observed in 0% (0/20), 0% (0/20), 4.9% (2/41), and 31.3% (26/83) of the cases, respectively. Both N- and C-FOXP1 staining were observed in 15.0% (3/20), 60.0% (12/20), 82.9% (34/41) and 48.2% (40/83) of the above cases, respectively, while complete loss of FOXP1 expression was observed in only 9.6% (8/83) of IDC cases. Estrogen receptor (ER) expression in C-FOXP1-positive IDC cases (31/66, 47.0%) was significantly lower than that in C-FOXP1-negative cases (13/17, 76.5%) (p = 0.030). Calpain II expression was observed in 83.3% (55/66) of C-FOXP1-positive IDC cases, which was significantly higher than that in C-FOXP1-negative cases (9/17, 52.9%) (p = 0.007). Calpain II was significantly associated with pAKT (p = 0.029), pmTOR (p = 0.011), p4E-BP1 (p < 0.001) and p-p70S6K (p = 0.003) expression levels. The 10-year OS and DFS rates of the C-FOXP1-positive patients were 60.5% and 48.7%, respectively, both of which were lower than those of the C-FOXP1-negative patients (93.3, 75.3%). The OS curve showed a dramatic impact of C-FOXP1 status on OS (p = 0.045). CONCLUSIONS: Cytoplasmic relocalization of FOXP1 protein was a frequent event in breast IDC. Calpain II might play an important role in nucleocytoplasmic trafficking of FOXP1 and the AKT pathway might be involved in this process. C-FOXP1 expression was inversely associated with ER expression and might be a predictor of poor OS in patients with IDC.

Rhabdomyosarcoma of the breast: a clinicopathologic study and review of the literature.

BACKGROUND: Rhabdomyosarcoma (RMS) is an uncommon malignancy of the breast. The aim of this study was to summarize its clinicopathologic features and biological behavior. METHODS: Five primary or secondary breast RMSs were collected. Their clinicopathological characteristics and all published literature about breast RMS were reviewed. Immunohistochemical study of desmin, myogenic differentiation 1 (MyoD1), myogenin, leukocyte common antigen (LCA), vimentin, cytokeratin (AE1/AE3), E-cadherin, neuron specific enolase (NSE), CD99, chorioallantoic membrane 5.2 (CAM5.2) and epithelial membrane antigen (EMA) expression were performed. RESULTS: The five patients were all female with ages ranging from 16 to 46 years old (mean, 30 years). Three were metastatic breast RMSs, two embryonal and one solid variant alveolar, with the primary tumor sites the right labium majus, left nasal meatus and nasopharynx, respectively. The other two, one embryonal and one alveolar, were primaries. Grossly, the surgical specimens revealed round or oval, well-demarcated but nonencapsulated masses. Their cut surfaces consisted of homogeneous grayish yellow or white tissue. Microscopically, most tumor cells were poorly differentiated small round, oval or small polygons with eosinophilic cytoplasm. All cases were positive for vimentin, desmin, MyoD1 and myogenin. One embryonal RMS also had a few cells with perinuclear staining of AE1/AE3. The other markers were negative. CONCLUSIONS: Although primary or metastatic RMS in breast was almost confined to young adolescent females, our cases suggested that it can also happen to the middle-aged women. Embryonal RMS has a certain metastatic potential. MyoD1 and myogenin are two useful markers when making differential diagnosis. Axillary lymph node status and age may play a role in the prognosis of primary breast RMS patients.

[Clinical implications of positive peritoneal cytology in endometrial cancer].

OBJECTIVE: To evaluate the clinical significance of positive peritoneal cytology in patients with endometrial cancer. METHODS: The records of 315 patients with endometrial cancer who were operated at Cancer Hospital, Fudan University between January 1996 and December 2008 were reviewed. Peritoneal cytology were performed and diagnosed in all patients. Factors related with peritoneal cytology were analyzed by correlation analysis. Log-rank test and Cox regression test was used for the analysis of prognosis, respectively. RESULTS: (1) Peritoneal cytology were positive in 30 (9.5%) patients. Positive peritoneal cytology was associated with pathological subtype (P = 0.013), stage (P = 0.000), myometrial invasion (P = 0.012), lymph-vascular space invasion (P = 0.012), serosal involvement (P = 0.004), cervical involvement (P = 0.016), adnexal involvement (P = 0.000), and omental involvement (P = 0.000), with no association with grade (P = 0.152) and lymph node metastasis (P = 0.066). (2) Three-year overall survival (OS) and progression-free survival (PFS) were 93.0% and 85.5%, respectively. Positive peritoneal cytology, surgical stage, pathological subtype, myometrial invasion, grade, and lymph-vascular space invasion were significantly associated with worse prognosis by univariate analysis (P < 0.05), while only surgical-pathology stage and myometrial invasion were independent prognostic factors by multivariate analysis (P < 0.05). For 30 cases with positive peritoneal cytology, the patients with no high risk factors shown significantly prognoses better than those with any risk factors. The results shown that for patients with late stage (stage III-IV) endometrial cancer with positive peritoneal cytology was significantly associated with the worse OS and PFS by multivariate analysis (P = 0.006). CONCLUSIONS: Positive peritoneal cytology was associated with serosal involvement, cervical involvement, adnexal involvement, omental involvement, and late stage. Therefore, peritoneal cytology should be performed and reported separately as a part of full surgical staging procedure.

Separation of normal and premalignant cervical epithelial cells using confocal light absorption and scattering spectroscopic microscopy ex vivo.

Confocal light absorption and scattering spectroscopic (CLASS) microscopy can detect changes in biochemicals and the morphology of cells. It is therefore used to detect high-grade cervical squamous intraepithelial lesion (HSIL) cells in the diagnosis of premalignant cervical lesions. Forty cervical samples from women with abnormal Pap smear test results were collected, and twenty cases were diagnosed as HSIL; the rest were normal or low-grade cervical squamous intraepithelial lesion (LSIL). The enlarged and condensed nuclei of HSIL cells as viewed under CLASS microscopy were much brighter and bigger than those of non-HSIL cells. Cytological elastic scattered light data was then collected at wavelengths between 400 and 1000 nm. Between 600 nm to 800 nm, the relative elastic scattered light intensity of HSIL cells was higher than that of the non-HSIL. Relative intensity peaks occurred at 700 nm and 800 nm. CLASS sensitivity and specificity results for HSIL and non-HSIL compared to cytology diagnoses were 80% and 90%, respectively. This study demonstrated that CLASS microscopy could effectively detect cervical precancerous lesions. Further study will verify this conclusion before the method is used in clinic for early detection of cervical cancer.

[Establishment and characterization of a nude mice model of human diffuse large B-cell lymphoma].

OBJECTIVE: To establish a diffuse large B-cell lymphoma (DLBCL)-mice model using human DLBCL cell line LY8, to investigate its characteristics of growth and to provide a model for in vivo study of DLBCL pathogenesis and treatment. METHODS: LY8 cells were injected subcutaneously into the right flank of nude mice. Harvested tumor tissues were cut into small pieces of 1.5 mm × 1.5 mm × 1.5 mm and implanted subcutaneously into nude mice. Tumor growth was visualized and the histologic characteristics were documented. Expression of LCA, CD20, CD79α, Ki-67, CD3, CD45RO, bcl-6, MUM-1, CD10 and bcl-2 were examined by using immunohistochemistry. IgH clonal rearrangement and status of three microsatellite loci (D14S68, D18S69, D20S199) in the xenografted tumor samples and the parental cell line LY8 were detected using PCR amplification followed by PAGE. RESULTS: The subcutaneous xenograft DLBCL model was successfully established by using cell line LY8, and a stable growth was achieved up to the 9th generation. The tumor in each generation showed similar growth characteristics and the rate of subcutaneous tumor formation was 91.9% (114/124). The tumor growth was observed from the 2nd week after implantation, reaching 1.3 cm in major diameter at the 3rd week and 2.0 cm at the 4th week. The tumor had identical morphological characteristics with those of human DLBCL, and expressed LCA, CD20, CD79α, bcl-6, MUM-1, CD10 and bcl-2. The tumor of xenograft mice and cell line LY8 showed identical IgH rearrangement and microsatellite length. CONCLUSIONS: A human DLBCL bearing mouse model was successfully established. The mice model is similar to human counterpart with high stability and repeatability. Therefore, it provides an ideal animal model for in vivo studies of the biological characteristics and treatment of DLBCL.

Germline mutation analysis of hPMS2 gene in Chinese families with hereditary nonpolyposis colorectal cancer.

AIM: To study the germline mutation of hPMS2 gene in 26 unrelated Chinese hereditary nonpolyposis colorectal cancer (HNPCC) probands and to fulfill the screening strategy for HNPCC in Chinese. METHODS: Genomic DNA was extracted from the peripheral blood. To avoid the interference of pseudogene in detection of the remaining 11 exons (exon 1-5, 9, 11-15), long-range polymerase chain reaction (PCR) was conducted to amplify the complete coding region of hPMS2 gene firstly. Then 1/8 of the PCR products were used as template to amplify the individual exon respectively and DNA sequencing was done. Direct DNA sequencing of the conventional PCR products of exon 6, 7, 8 and 10 of hPMS2 gene was performed. The same analysis was made in 130 healthy persons without family histories of HNPCC to further investigate the pathological effects of the detected missense mutation. RESULTS: One HNPCC proband fulfilled Bethesda guidelines and was found to carry the germline mutation of hPMS2 gene, which has not been reported in Chinese HNPCC families. It was a missense mutation at c.1532C>T of exon 11. It was detected in three controls as well with an occurrence rate of 2.3% (3/130). Since it could not be found in the PMS2-single nucleotide polymorphism (SNP) database, this missense mutation is a new SNP unreported up to date. Meanwhile, 260 reported SNPs of hPMS2 gene were detected in the 26 HNPCC probands. The 2nd and 5th exons were probably the hot SNP regions of hPMS2 gene in Chinese HNPCC families involving 53.1% of all reported SNP. CONCLUSION: The germline mutation of hPMS2 gene may be rare in Chinese HNPCC families. The 2nd and 5th exons are hot SNP regions of hPMS2 gene.

[Clinical significance of Her-2/neu status in patients with uterine papillary serous carcinoma].

OBJECTIVE: The purpose of this study was to evaluate gene amplification by chromogenic in situ hybridization (CISH) and the protein expression of Her-2/neu gene in patients with uterine papillary serous carcinoma (UPSC) and to determine its prognostic value. METHODS: Thirty-six patients with confirmed pathologic diagnosis of UPSC in Cancer Hospital of Fudan University from Jan. 1996 to Jan. 2006, were analysed retrospectively. CISH was performed to assess Her-2/neu gene amplification, and protein expression was evaluated by immunohistochemistry (IHC). The prognostic factors were analyzed by log-rank test or Cox proportional hazard model. RESULTS: Among 36 cases with UPSC, 13 patients (36.1%) showed moderate staining (++) to strong staining (+++) for Her-2/neu protein, while amplification of the Her-2/neu gene by CISH was observed in 4 of the 36 (11.1%) cases. Her-2/neu protein over-expression was significantly associated with advanced surgical stage and worse prognosis by univariate analysis (P=0.030 and P=0.002, respectively), while the multivariate analysis shown that only Her-2/neu protein over-expression and deep myometrial invasion were associated with a poor prognosis (P<0.05). In 13 patients with Her-2/neu protein over-expression, the mean survival period with chemotherapy was shorter than those without chemotherapy (20 vs. 42 months, P=0.370). CONCLUSION: Her-2/neu protein over-expression is significantly associated with advanced surgical stage UPSC and poor survival outcome, and might reduce the chemotherapy sensitivity.

[Cutaneous anaplastic large cell lymphoma: clinicopathologic, immunohistochemical and prognostic study of 44 cases].

OBJECTIVE: To study the clinicopathologic features, immunophenotype and prognosis of primary cutaneous anaplastic large cell lymphoma (CALCL). METHODS: Histopathologic evaluation and immunohistochemical study by Envision method were carried out in 44 archival cases of CALCL. The clinical information and follow-up data were analyzed. RESULTS: The patients presented with skin nodules, masses or plaques, sometimes associated with ulceration. The commonest sites of involvement were the extremities. Follow-up data were available in 39 patients. The overall survival rate was 87.2% (34/39). Disease relapses were detected in 46.2% (18/39) of the patients. Statistical analysis indicated that patients older than 50 years of age or with no less than two involved anatomic sites were more likely to have disease relapses (P < 0.05). Histologically, 31 cases were classified as common variant, 6 cases as small cell variant and 7 cases as neutrophil/eosinophil-rich variant. Immunohistochemical study showed that the rates of expression of CD30, CD45, CD45RO, CD43, CD3, cytotoxic protein and epithelial membrane antigen were 100% (44/44), 91.2% (31/34), 82.6% (19/23), 94.7% (18/19), 70.0% (28/40), 73.3% (22/30) and 31.8% (7/22), respectively. The CD4(+)/CD8(-), CD4(-)/CD8(+) and CD4(-)/CD8(-) immunophenotypes were found in 58.3% (21/36), 22.2% (8/36) and 19.4% (7/36) of the CALCL cases, respectively. Only one case (3.7%) expressed CD56. CONCLUSIONS: CALCL is a form of low-grade primary cutaneous T-cell lymphoma with a wide spectrum of clinicopathologic pattern. Special variants of CALCL should not be confused with other types of cutaneous lymphomas and inflammatory lesions. CALCL patients older than 50 years of age or with no less than two involved anatomic sites are more likely to have disease relapses.

[Sclerosing angiomatoid nodular transformation of spleen: a clinicopathologic study of 10 cases with review of literature].

OBJECTIVE: To study the clinicopathologic features, differential diagnosis and pathogenesis of sclerosing angiomatoid nodular transformation of spleen. METHODS: Ten cases of sclerosing angiomatoid nodular transformation of spleen were retrieved from the archival file. Histochemical and immunohistochemical (EnVision method) studies were performed. Ultrastructural findings were also available in one of them. RESULTS: Sclerosing angiomatoid nodular transformation was characterized by micronodular appearance of vascular spaces lined by plump endothelial cells with interspersed ovoid spindle cells. Immunohistochemical study showed that the endothelial cells of vessels in the angiomatoid nodules had various expressions of immunologic phenotypes and could be mainly classified into 3 types: CD34(+)/CD31(+)/CD8⁻ endothelial cells of the capillaries, CD8(+)/CD31(+)/CD34⁻ lining cells of the sinusoids and CD31(+)/CD8⁻/CD34⁻ endothelial cells of the small veins. Collagen network and dilated lymphatic sinuses were evident under transmission electron microscope. CONCLUSIONS: Sclerosing angiomatoid nodular transformation of spleen is a rare benign entity. It may represent a reactive condition and bears some relationship with splenic angioma. It needs to be distinguished from borderline or malignant vascular tumors of spleen.

[Study on PI3K inhibitor LY294002 for chemotherapeutic sensitization in diffuse large B cell lymphoma cell lines].

OBJECTIVE: To investigate the effects on chemotherapeutic sensitization of the PI3K inhibitor LY294002 in diffuse large B cell Lymphoma (DLBCL) cell lines ly1, ly8, ly10. METHODS: The three cell lines were treated with LY294002, or doxorubicin alone or combined or sequentially respectively. Western blotting was used to detect the level of phospho-AKT after the treatment. Flow cytometry combined with annexin V-FITC assay and Brdu incorporation assay were used to analyze the alterations of cell cycle, proliferation, and apoptosis, respectively. RESULTS: LY294002 decreased the level of phospha-AKT efficiently in the three DLBCL cell lines. The ratio of S phase cells was significantly decreased (P < 0.05). Sequential use of LY294002 and doxorubicin increased the ratio of apoptosis and there was significant difference between the sequential group and the other four groups (P < 0.05) at 24, 48, 72(ly1), 48, 72 (ly8) or 24 h (ly10). CONCLUSION: LY294002 can sensitize doxorubicin-induced apoptosis and may be a potential molecular therapeutic agent targeted at AKT signaling pathway in DLBCL.

[Melanotic epithelioid clear cell tumor of kidney: report of three cases].

OBJECTIVE: To study the pathologic features and immunophenotype of 3 cases of melanotic epithelioid clear cell tumor of kidney. METHODS: More than 2000 cases of renal tumors were retrospectively reviewed. Three cases of melanotic epithelioid clear cell tumor were identified. Immunohistochemical study was carried out using the paraffin-embedded tissue samples. Electron microscopy was also performed in 1 case. RESULTS: Amongst the 3 cases studied, the male-to-female ratio is 1:2. Histologically, 2 cases showed a clear cell carcinoma-like pattern. Papillary structures covered by clear cells and eosinophilic cells were observed in 1 case. Immunohistochemical study showed that the tumor cells in all cases expressed HMB 45. Two of them were also positive for Melan A. The staining for epithelial markers and S-100 protein was negative. Melanosomes were not identified by ultrastructural examination. CONCLUSIONS: Melanotic epithelioid clear cell tumor is a rarely seen neoplasm of kidney. There are some histologic overlaps with renal cell carcinoma, epithelioid angiomyolipoma and melanoma. Immunohistochemical study is useful in confirming the diagnosis. The tumor represents a morphologic variant of epithelioid angiomyolipoma.

[Evaluation of intraoperative molecular assay in sentinel lymph node biopsy from breast cancer patients].

OBJECTIVE: to evaluate the application of GeneSearch(TM) breast lymph node assay in intraoperative detection of metastases in sentinel lymph node (SLN) from breast cancer patients. METHODS: a total of 225 SLN from 88 patients was prospectively studied. Each SLN was cut into 2 mm slabs which were examined by intraoperative imprint cytology (IIC) first, followed by GeneSearch assay and post-operative serial sectioning. GeneSearch used real-time fluorescence quantitative RT-PCR technology to detect the expression of CK19 and mammaglobin in SLN. The results of GeneSearch assay were correlated with those of IIC and post-operative serial sectioning. RESULTS: amongst the 88 cases studied, 225 SLNs were found, and obvious metastatic carcinoma cells were identified in 27 SLNs and micrometastasis in 9 SLNs. One hundred and eight-nine SLNs were considered as "negative" (with "isolated tumor cells" present in 5 SLNs). The turn-around time of intraoperative GeneSearch assay ranged from 35 to 45 minutes (mean = 40 minutes). The concordance rate between GeneSearch assay and post-operative serial sectioning was 95.6% (215/225), with a sensitivity of 86.1% (31/36), compared with 94.7% (213/225) and 72.2% (26/36) respectively for IIC. The size of metastatic foci correlated with the Ct value of CK19 and mammaglobin (P < 0.01). CONCLUSIONS: GeneSearch assay for intraoperative detection of metastase in SLN has a satisfactory performance and demonstrates a relatively higher sensitivity than IIC. The potential clinical application still requires further evaluation of larger number of cases.

[Clinicopathologic characteristics and chromosomal abnormalities in salivary mucosa associated lymphoid tissue lymphomas].

OBJECTIVE: To study the morphological and genetic characteristics in salivary gland marginal zone B cell lymphoma of mucosa associated lymphoid tissue (MALT) lymphomas. METHODS: Twenty-eight cases of MALT lymphomas of salivary gland were collected from Department of Pathology, Cancer Hospital of Fudan University. Morphological review based on HE sections, and specific chromosomal abnormalities were detected by two-color interphase fluorescent in situ hybridization (FISH). Four different probes were available to detect for API2-MALT1 fusion gene, bcl-10, IgH and MALT1 gene, respectively. RESULTS: There were 16 females and 12 males, median age was 52. In those cases, 18 originated from parotid gland, 6 from submandibular and 4 from sublingual gland. Ten were localized mass and 18 were masses diffusely involved the glands. According to the clinical information, only 8 cases showed symptoms of dry mouth, dry nose or dry eye. Pathological findings showed that all cases had a dense lymphoid infiltration and obliteration and atrophy of acini and ducts. Twenty-two (78.6%) showed prominent monocytoid B cells and more often formed broad halos around epithelial islands. Eighteen (64.3%) showed clusters of lymphoblastic cells or plasma cells, Russel' and Dutcher' body were easily seen. Ten (35.7%) showed nerve or blood vessel infiltration. Interphase FISH showed that 3 cases harbored t(11;18) and 2 cases harbored trisomy 18, but none of all found IgH and bcl-10 translocations. After operation, 22 patients' follow-up information was available. One case died on 15 months later after operation, the rest of 21 cases were alive. Except surgical resection, patients did not get systematic radio-or chemotherapy. Eight to fifteen months after operation, 8 cases found recurred nodules on the original resected sites or cervical lymph nodes, but did not get repeated biopsy. All follow-up time was from 23 to 54 months. CONCLUSIONS: Most salivary MALT lymphomas are arising from parotid glands. Most patients do not have the symptoms of the Sjogren's syndrome. The final diagnosis depends on the pathological findings, the number and distribution of monocytoid B cells and clusters of plasmacytoid cells are hints for diagnosis of salivary MALT lymphomas, invasion of blood vessels or nerve also help for malignant diagnosis. t(11;18) and trisomy 18 may be the main chromosomal abnormalities in salivary gland MALT lymphomas, but with low morbidity. This genetic characteristic may connect with the low malignancy and slow progression in biological behavior.

Neutrophil/eosinophil-rich type of primary cutaneous anaplastic large cell lymphoma: a clinicopathological, immunophenotypic and molecular study of nine cases.

AIMS: To elucidate the clinicopathological, immunophenotypic and molecular features of neutrophil/eosinophil-rich primary cutaneous anaplastic large cell lymphoma (CALCL), and to emphasize the cutaneous manifestations, differential diagnosis and prognosis of this peculiar entity. METHODS AND RESULTS: We described the clinical presentations, histopathology, immunophenotype, molecular features and follow-up courses of nine neutrophil/eosinophil-rich CALCL cases. Various clinical lesions including multiple nodules, plaques and solitary exophytic masses with or without ulceration or crusting were noted in nine patients. Two patients died of disease progression, with one developing multiple lymph node involvement. Histologically, cohesive sheets or small clusters of neoplastic cells were admixed with large numbers of neutrophils and/or eosinophils, representing 10-40% of cells per high-power field. All nine cases showed T-cell phenotypes. The frequency of rearranged TCRB, TCRG and TCRD genes in six cases with available paraffin-embedded tissue was 100%, 83% and 33%, respectively. CONCLUSIONS: Neutrophil/eosinophil-rich CALCL should be differentiated from various infectious and non-infectious diseases, especially from non-neoplastic cutaneous CD30+ infiltrates rich in neutrophils and eosinophils. Precise correlation of clinical presentation, morphological features, phenotypic and molecular analysis can help to establish the correct diagnosis. Whether this rare variant has a significantly different prognosis from classical CALCL needs further investigation.

[Clinical significance in detection of immunoglobulin heavy chain clonal rearrangement in bone marrow of patients with B cell lymphoma].

OBJECTIVE: To explore the feasibility of semi-nested PCR technique for detection of immunoglobulin heavy chain (IgH) clonal rearrangement in bone marrow of B-cell lymphoma patient and to further evaluate its clinicopathological value. METHODS: Gene clonal rearrangement of IgH was detected by semi-nested PCR using primers of FR2 & FR3A in 105 bone marrow samples of patients with B-cell lymphoma. The PCR detection results were compared with the cytomorphology of bone marrow aspiration biopsy. The correlation between PCR detection results and clinicopathological factors were evaluated. RESULTS: Among 105 cases of B-cell lymphoma, bone marrow involvement was detected by PCR technique in 48 cases (45.7%), while only 22 cases (21.0%) were detected by bone marrow cytological analysis. There was a significant difference between two methods (P < 0.05), and the concordance rate was 71.4%. The incidence of bone marrow involvement at the time of initial diagnosis detected by PCR technique was 30.8% for diffuse large B cell lymphoma (DLBCL), 25.0% for follicular lymphoma (FL), and 100.0% for small lymphocytic lymphoma (SLL), respectively. Bone marrow involvement detected by PCR detection correlated with Ann Arbor stage. Rate of clonal IgH gene rearrangement by PCR in early B-cell lymphoma was lower than that in advanced stage B-cell lymphoma patients (P = 0.02). There was no statistically significant difference in efficacy between patients with positive and negative results detected by PCR (P > 0.05). But difference in complete response (CR) rate (23.3% and 46.3%) had significant difference (P = 0.019). CONCLUSION: Semi-nested PCR analysis may be an effective method for detection of abnormalities in bone marrow in patients with B-cell lymphoma and is superior to cytomorphology. The positive rate in patients with advanced Ann Arbor stage is higher than that in patients with early Ann Arbor stage, and patients with PCR negative result have more chances to achieved CR after treatment.

[Activation and clinicopathologic significance of AKT/mTOR signaling pathway in diffuse large B-cell lymphoma].

OBJECTIVE: (1) To investigate the activation of AKT/mTOR signaling transduction pathway in DLBCL and its association with the expression of bcl-6 and some other clinical pathologic factors. (2) To estimation of the signaling pathway function in diverse subtypes of DLBCL and its potential value in the targeted treatment of DLBCL. METHODS: Immunohistochemical (IHC) EnVision staining was used to detect the expressions of pAKT and pmTOR in 100 DLBCL and 10 reactive hyperplasia fresh lymph node samples; TaqMan real-time reverse transcription polymerase chain reaction (real-time RT-PCR) technique was used to explore the expression of bcl-6 mRNA in the DLBCL samples. IHC staining was used to detect the expressions of bcl-6, CD10 and MUM1 in 75 of the 100 corresponding paraffin-embedded samples and these 75 DLBCL samples were subdivided into GCB and non-GCB subgroups. RESULTS: (1) The expression of pAKT and pmTOR was 76% (76/100) and 75% (75/100), respectively, and the expression of the two proteins correlated with each other. (2) The expression of bcl-6 protein and mRNA significantly correlated with each other. The expression of bcl-6 protein and mRNA in pAKT and pmTOR high-expression group was significantly lower than that in low-expression group (both P < 0.01). (3) The expression of pAKT and pmTOR in non-GCB group was 82.5% (47/57) and 84.2% (48/57), respectively, which were significantly higher than that in GCB group, which showed an expression rate of 44.4% (8/18) and 44.4% (8/18), respectively (both P < 0.01). (4) The expression of pAKT and pmTOR in male was higher than that in female, and the percentage of patients with abnormal LDH in pAKT and pmTOR positive groups was higher than that in negative groups, although there were no statistical significance (both P > 0.05). There was no relationship between the expression of pAKT and pmTOR and age, sex, stage, KPS and B symptoms (P > 0.05). CONCLUSIONS: Activation of AKT/mTOR signaling pathway plays an important role in the development of DLBCL, and it is closely related to the low or non-expression of bcl-6 and non-GCB subgroup. Molecules in this pathway might serve as the new targets in the treatment of certain group of DLBCL.

[Clinicopathologic study of centrally necrotizing carcinoma of breast].

OBJECTIVE: To study the clinicopathologic features and immunophenotype of centrally necrotizing carcinoma (CNC) of breast; and to study its relationship with basal-like breast cancer. METHODS: The clinical and pathologic characteristics of 35 cases of CNC were analyzed. Immunohistochemical study for estrogen receptor, progesterone receptor, HER2, CK8/18, 34betaE12, CK5/6, CK14, CK17, smooth muscle actin, p63, vimentin and epidermal growth factor receptor was performed using EnVision method. The surival information of 10 case were obtained. RESULTS: The age of patients with CNC ranged from 30 to 82 years (mean = 54.2 years). Macroscopically, all tumors were relatively circumscribed, with a mean diameter of 2.4 cm. Histologically, there was a prominent central, necrotic or acellular zone surrounded by a narrow rim of viable tumor cells. The central necrotic foci had the following morphologic patterns: (1) coagulative tumor necrosis associated with various degree of fibrosis or hyaline degeneration (24 cases), (2) predominance of fibrous and scar tissue, with small amount of necrotic debris (8 cases), and (3) infarction (3 cases). The peripheral zone of tumor cells showed features of grade 3 invasive ductal carcinoma in 32 cases and grade 2 in 3 cases. Twenty cases of CNC were associated with ductal carcinoma in-situ. A component of invasive micropapillary carcinoma was identified in 5 cases. Peripheral lymphocytic infiltrates were seen in 17 cases. Immunohistochemical study of 31 cases showed that the expression rate of basal-like markers (83.9%, 26 cases) was higher than that of myoepithelial markers (38.7%, 12 cases). The percentage of basal-like subtype (64.5%, 20 cases) was higher than luminal-A (9.7%, 3 cases), luminal-B (9.7%, 3 cases), HER2 over-expression (12.9%, 4 cases) and null (3.2%, 1 case) subtypes. In 20 cases of basal-like carcinoma, the expression ratio of CK5/6 was highest amongst basal-like markers (18 cases), the other markers ratios of CK17, CK14 and epidermal growth factor receptor were 8/10, 14/19 and 8/16, respectively. Follow-up data were available in 10 patients. The follow-up duration ranged from 15 to 42 months (mean = 21.5 months). The median disease-free and overall survivals were 14.0 and 18.0 months, respectively. Disease progression (as defined by the presence of recurrence, metastasis or tumor-related death) occurred in 9 patients. The mean and median time to disease progression was 16.6 and 13.0 months, respectively. CONCLUSIONS: CNC is a rare subtype of breast carcinoma and has distinctive, easily discernible morphologic features. The majority of CNC exhibits basal-like immunophenotype and carries a poor prognosis. CNC is the typical representative of basal-like breast cancer.

[Prevalence of API2-MALT1 fusion gene in gastrointestinal mucosa-associated lymphoid tissue lymphomas and diffuse large B cell lymphomas].

OBJECTIVE: To investigate the difference of the prevalence of t(11;18)(q21;q21)/API2-MALT1 fusion gene between gastrointestinal mucosa-associated lymphoid tissue (MALT) lymphoma and diffuse large B cell lymphoma (DLBCL). METHODS: A total of 57 cases gastrointestinal MALT lymphomas (38 gastric and 19 intestinal lymphomas), 32 DLBCL (28 gastric and 4 intestinal lymphomas) and 7 cases gastric DLBCL accompanied MALT lymphoma were collected from the Cancer Hospital of Fudan University. API2-MALT1 fusion gene was detected by fluorescent in situ hybridization (FISH) using both dual fusion translocation and break apart probes. RESULTS: Among gastrointestinal MALT lymphomas, API2-MALT1 fusion gene was found in 12 of 57 cases (21.1%, 10 gastric and 2 intestinal lymphomas). In contrast, the fusion gene was not found in all 32 DLBCL and 7 gastric DLBCL with MALT lymphoma component. There was statistical significant difference between two groups (chi(2) = 9.383, P = 0.001). CONCLUSIONS: API2-MALT1 fusion gene is a distinctive genetic aberration in MALT lymphomas, and is not present in DLBCL. The findings suggest that gastrointestinal tract MALT lymphomas with API2-MALT1 fusion gene may not transform into DLBCL, which may represent primary lymphoma or transformed API2-MALT1 negative MALT lymphomas.

Intravascular large B-cell lymphoma with cutaneous manifestations: a clinicopathologic, immunophenotypic and molecular study of three cases.

BACKGROUND: Intravascular large B-cell lymphoma (IVLBCL) is an extremely rare type of malignant lymphoma characterized by exclusive or predominant growth of neoplastic cells within the lumen of blood vessels. Cases in the literature predominantly involve the skin and central nervous system, with special emphasis on the 'cutaneous variant'. METHODS: Three cases of IVLBCL with cutaneous manifestations, including two systemic IVLBCL and one cutaneous variant, were described in this study. In all cases, clinical presentation and follow-up data were meticulously evaluated and immunophenotypic and molecular studies were performed. RESULTS: All three cases displayed the B-cell phenotype and showed monoclonality with immunoglobulin heavy chain gene rearrangement. Bcl2 was expressed in the two systemic IVLBCL cases with fatal outcomes. The third patient with the 'cutaneous variant' achieved complete remission and a longer survival time of 15 months after chemotherapy. CONCLUSIONS: Skin manifestations and neurological findings, although to different degrees, are important clues to the diagnosis of IVLBCL. As most IVLBCL are grouped into the post-germinal center B-cell subtype of diffuse large B-cell lymphoma, Bcl2 expression may be correlated with a worse prognosis in IVLBCL. The cutaneous variant of IVLBCL has a significantly better outcome than that of systemic IVLBCL.