Gene Rearrangement ; Humans ; RNA-Binding Protein EWS ; genetics ; Soft Tissue Neoplasms ; genetics

Purpose To evaluate the application of fluorescence in situ hybridization ( FISH) in detection of EWSR1 gene translocation in formalin-fixed paraffin-embedded tissue samples of Ewing family tumor ( EFT) . Methods Four cases of EFT were collected and their clinical pathological features were obsered. Interphase FISH was carried out in paraffin-embedded tissue of EFT cases, using a EWSR1 dual colar break-apart probe. Fifteen cases of other soft tissue tumor were selected as negative control. Results Translocation of EWSR1 was detected in four EFT cases, which may be a adjuvantly diagnositic value for the diagnosis of atypical cases. Conclu-sions FISH may serve as an adjunctive diagnostic tool in problematic cases of EFT, however, FISH results should be interpreted con-cerning clinical pathological features, morphological patterns and immunohistological results.

Objective To establish a quantitative detection method for Mycobacterium tuberculosis by immunomagnetic capture combined with PCR-ELISA detection system with double internal standards(IMC-PCR-ELISA) .Methods The immunomagnetic (Dynabeads? ) which could specifically capture Mycobacterium tuberculosis were prepared .According to Mtp40 and IS6110 gene sequence of Mycobacterium Tuberculosis ,2 pairs of primers(upstream primer was modified with Biotin at 5′end) ,2 same-length mutant fragments with PCR amplified fragments ,and 3 capture probes(modified with digoxigenin at 3′end) were designed .Myco-bacterium tuberculosis were captured by immunomagnetic ,then detected by PCR-ELISA with double internal standards .Results The IMC-PCR-ELISA method could yield quantitative results in about 4 h with a detection limit at 5 × 103 copies/mL .There was a fine linear relationship between the copies of Mtp40(IS6110)in fact and in the calculation through formula when the concentrations of low internal standards were 30-70 copies/mL and the concentrations of high internal standards were 8 000-12 000 copies/mL (r2 =0 .998) .No nonspecific amplification was observed .Conclusion A rapid and quantitative method for the detection of Myco-bacterium tuberculosis was established successfully .The IMC-PCR-ELISA method was rapid ,sensitive ,secific and quantitative .

Objective: To study the clinicopathologic features, diagnosis and differential diagnosis of pulmonary microcystic fibromyxoma. Methods: In March 2014, at the First Affiliated Hospital to Nanjing Medical University a 58-year-old female patient of pulmonary microcystic fibromyxoma was collected. The clinicopathologic, immunohistochemical and genetic profile of a case of pulmonary microcystic fibromyxoma were studied, and the relevant literature reviewed. Results: The patient was a 58-year-old female who presented with cough and sputum for 1 month. CT scan disclosed a 15 mm nodule in her right middle lobe of lung. The patient underwent a wedge resection with negative margin. Grossly, a well-demarcated peripheral lung nodule was detected, measuring 1.5 cm×1.5 cm×1.0 cm, with myxoid tan-white cut surface containing microcysts. Microscopically, the tumor was composed of bland spindled to stellate-shaped cells widely spaced within prominent fibromyxoid stroma with prominent cystic change. No mitosis or necrosis was present. There were inconspicuous slim curvilinear capillaries and occasional collection of stromal lymphocytes and plasma cells. Immunohistochemically, the tumor cells were positive for vimentin, but negative for CD34, SMA, desmin, S-100 protein, ALK, CKpan, EMA, calretinin and TTF1. Fluorescence in situ hybridization did not show chromosomal translocation involving EWSR1, DDIT3 or FUS genes. The patient was recurrence or metastasis free after follow-up for 38 months. Conclusion Pulmonary microcystic fibromyxoma is a rare benign lesion that should be differentiated from other lung tumors with myxoid characteristics.

Objective: To study clinicopathologic features, diagnosis and differential diagnosis of myxoid variant of angiomatoid fibrous histiocytoma (AFH). Methods: Three cases of myxoid variant of AFHs were collected from First Affiliated Hospital of Nanjing Medical University during 2008 and 2017. EnVision method and fluorescence in situ hybridization(FISH) were used to detect immunophenotype and EWSR1 gene rearrangement, respectively. Results: There were 2 males and l female with age at 13, 31, and 42 years, respectively. The patients presented with a painless mass located superficially (subcutaneous or submucosal) in two cases or deep-seated (retroperitoneum) in one case. Grossly, the diameters of tumors were 1, 7, and 2 cm, respectively. The cut surface was solid and firm, tan to gray in colour. Histologically, the circumscribed tumor had fibrous pseudocapsule and peritumoal lymphoplasmacytic infiltrates. The tumor cells arranged in vaguely nodular growth pattern, with prominent myxoid stroma (present in 60% to 100% of the entire tumor). In hypocellular myxoid areas, the spindle to stellate tumor cells arranged in cords or reticular pattern, or in a haphazard manner. However, histiocytoid cells arranged in fascicular, sheet-like, or whorled growth pattern, as in classical AFH, were also identified in hypercelluar areas. Mild to moderate atypia was observed with low mitotic rate of (0-2)/10 HPF. Tumor necrosis was not seen. One case presented with slit-like hemorrhage and sclerosing collagen intermingled with myxoid matrix was identified in 1 case. Immunohistochemically, all cases were positive for CD68 and CD163. Two of three were positive for desmin, EMA, CD99 and one for Calponin, SMA. All cases were negative for S-100 protein, CD34, CD31, CD35, CD21 and CKpan. FISH detection was positive for EWSRl gene in all cases. Available clinical follow-up was obtained in 2 cases, revealing no evidence of disease in 6 and 89 months, respectively. Conclusions Myxoid variant of AFH is a histological subtype of AFH, with clinical features, immunophenotypes, genomic profiles and biological behavior similar to typical AFH. Their unusual morphology is easily confused with a variety of other myxoid mesenchymal neoplasms, including myoepithelioma and nerve sheath tumors.

Objective: To study the clinicopathological features of primary hepatic extranodal marginal zone lymphoma of mucosa associated lymphoid tissue (MALT lymphoma) and hepatic pseudolymphoma, and to discuss their differential diagnosis, treatment and prognosis. Methods: Three primary hepatic MALT lymphomas and two hepatic pseudolymphomas collected from January 2012 to March 2017 in the First Affiliated Hospital of Nanjing Medical University were evaluated by HE and immunohistochemistry(IHC), in-situ hybridization and immunoglobulin (Ig) gene rearrangement detection, and the relevant literature reviewed. Results: In the three MALT lymphomas, tumor cells infiltrated the portal areas with nodular pattern, and invaded the surrounding normal liver with serpiginous configuration and formation of confluent sheets. A number of bile ducts were entrapped within the lesions, and showed lymphoepithelial lesion. Reactive lymphoid follicles were present and surrounded by tumor cells, consisting of predominantly centrocyte-like cells and monocytoid B cells. There were clusters of epithelioid histiocytes in one case. The tumor cells were positive for CD20, PAX5 and negative for CD5, CD23, CD10, bcl-6, and cyclin D1. In the two hepatic pseudolymphomas, the lesions presented as solitary nodules well-demarcated from the surrounding liver tissue; one case was partially encapsulated with fibrous tissue. Entrapped bile ducts were only found at the edge of the lesions without lymphoepithelial lesion. The lesions comprised of massive lymphoid proliferation consisting predominantly of reactive lymphoid follicles, but not monocytoid B-cells or atypical cells. By IHC, a mixture of B- and T-cell population was identified. A monoclonal rearrangement of the Ig gene was detected in all three MALT lymphomas but not in two pseudolymphomas. Interphase fluorescence in situ hybridiazation test for MALT1 break-apart gene was positive in two cases of MALT lymphomas and EBER was negative in all studied cases. Conclusions Primary heptic MALT lymphoma and pseudolymphoma are both rare lymphoid proliferative lesions of liver. These two lesions have overlapping histological and IHC features and are top differential diagnosis to each other. A combination analysis of morphology, immunophenotype and Ig gene rearrangement is helpful to distinguish between them.

Objective To study the clinicopathologic features , diagnosis and differential diagnosis of succinate dehydrogenase ( SDH ) deficient gastrointestinal stromal tumors ( GISTs ) as a unique tumor subtype.Methods SDHB and SDHA immunohistochemistry was performed in 120 gastric GISTs, in addition to CD117, DOG-1, CD34, smooth muscle actin (SMA), desmin,S-100 protein, cytokeratin (CK) and Ki-67.Subset of the cases was further evaluated for the presence of mutations in CKIT exons 9, 11, 13 and 17 mutations and platelet derived growth factor receptor alpha ( PDGFRA) exons 12 and 18.Results Eight of 120 (6.6%) GIST cases were found SDH-deficient including 3 male and 5 female patients (median age of 36.2 years;ranging 16 to 65 years of age).The tumors involved antrum (6 cases), lesser curvature (1 case) and fundus (1 case).Macroscopically, the dominant tumor masses varied from 3 to 10 cm in diameter with a multinodular or plexiform pattern involving the gastric wall .Microscopically ,tumor cells had predominantly epithelioid morphology , with occasional mixed spindle cell nodules .Lymphovascular invasion was identified in 5 cases.Immunohistochemistry for SDHB was negative in all 8 cases, and SDHA was negative in 5 cases.All 8 SDHB negative cases also expressed CD 117, DOG-1 and CD34, but were negative for SMA, desmin, S-100 and CK.All 8 cases were found to have wild-type CKIT and PDGFRA genes. Available clinical follow-up were obtained in 7 cases, ranging from 2 to 60 months ( median follow-up 23.3 months) , and all patient were alive .Three cases were found to have liver metastases at their first diagnosis , and one developed omental and mesenteric metastases in 17 months.Conclusions SDH-deficient GIST is a distinct subtype of GIST , with a predilection to occur in young and female patients .Characteristic pathological findings include multinodular gastric wall involvement , epithelioid cell morphology , frequently lymphovascular invasion with occasional lymph node and liver metastases , but an overall indolent clinical behavior.Immunohistochemistry for SDHB is required for the diagnosis .

OBJECTIVETo analyze the survival time and its related factors among AIDS patients in Liangshan prefecture of Sichuan province from 1995 to 2012.

METHODSA retrospective cohort study was conducted to analyze the information of 5 263 AIDS patients. The data were collected from Chinese HIV/AIDS Comprehensive Information Management System. Life table method was applied to calculate the survival proportion, and Kaplan-Meier and Cox proportion hazard regression model were used to identify the factors related to survival time.

RESULTSAmong 5 273 AIDS patients, 819 (15.6%)died of AIDS related diseases; 2 782(52.9%) received antiretroviral therapy. The average survival time was 126.7 (117.1-136.2) months, and the survival rate in 1, 5, 10, 15 years were 95.4%, 78.8%, 54.2%, and 31.8% respectively. Univariate analysis showed a significant difference in survival time of age diagnosed as AIDS patients, nationality, transmission route, AIDS phase, CD4(+)T cell counts in the last testing, receiving antiretroviral therapy or not. Multivariate Cox regression showed age diagnosed AIDS below 50 years old ( < 15 years old:HR = 0.141, 95%CI:0.036-0.551;15-49 years old:HR = 0.343, 95%CI:0.241-0.489), HIV infection diagnosed phase (HR = 0.554, 95%CI:0.432-0.709), CD4(+)T cell counts last testing ≥ 350/µl (HR = 0.347, 95%CI:0.274-0.439) reduced the risk of dying of AIDS related diseases among AIDS patients. The patients having not received antiretroviral therapy had a higher risk of death(HR = 3.478, 95%CI:2.943-4.112) compared to those who received antiretroviral therapy.

CONCLUSIONSurvival time of AIDS patients was possibly mainly influenced by the age of diagnosed as AIDS patients, AIDS phase, CD4(+)T cell counts and whether or not received antiretroviral therapy. The early initiation of antiretroviral therapy could extend the survival time.

Acquired Immunodeficiency Syndrome ; mortality ; China ; epidemiology ; Cohort Studies ; HIV Infections ; Humans ; Proportional Hazards Models ; Retrospective Studies ; Survival Analysis ; Survival Rate

Objective:To investigate the clinicopathological characteristics, genetic features, diagnosis and differential diagnosis of pulmonary artery intimal sarcoma (PAIS).Methods:Three cases of PAIS were collected from Jiangsu Province People′s Hospital (from February 2016 to November 2019). The clinical data, imaging examination, morphology, immunostaining, and molecular changes were retrospectively analyzed.Results:There were 1 male and 2 females (age: 32, 50, 60 years), who had symptoms of cough, asthma or chest tightness. Imaging findings indicated low density filling defects which were suspected as thrombus, embolism or myxoma. Grossly, the main tumor was located in the elastic arteries and their lobar branches, also extended into the atrium and ventricle, with lung parenchymal infiltration focally. Microscopically, tumor cells were predominantly composed of abundant spindle cells with obvious atypia and myxoid background, resembling fibroblastic or myofibroblastic differentiation. Active mitotic figures and necrosis could be seen in some areas. Immunohistochemical staining of vimentin was strongly positive, while pan-cytokeratin, S-100, desmin, Fli-1, CD31, SMA and ERG etc were variably positive only in focal areas. FISH detection showed amplification of MDM2 gene in three cases and EGFR gene in two cases. Metastatic lesions were found in one case by 18, 32 and 42 months after surgery respectively. There was no recurrence or metastasis in the other two cases.Conclusions:PAIS is one of exceptionally poor differentiated mesenchymal tumor that arises from the arterial intima of elastic pulmonary arteries. There was no definite differention in morphology. Gene detection shows amplification of MDM2 and EGFR gene. This tumor often has poor prognosis with aggressive behavior. Complete resection is the only effective therapeutic option. There is disagreement as to whether chemotherapy and radiotherapy can improve survival.

Objective:To study the clinicopathological features of large B-cell lymphoma (LBCL) with IRF4 rearrangement.Methods:Seven cases of LBCL with IRF4 rearrangement collected at the First Affiliated Hospital of Nanjing Medical University from November 2018 to October 2019 were evaluated by hematoxylin and eosin staining, immunohistochemistry and fluorescence in situ hybridization detection. The relevant literature was reviewed.Results:Four tumors were located in the tonsils, 2 tumors in the lymphoid nodes and one tumor in the adenoid.The patients were 3 males and 4 females patients with a median age of 24 years (range, 6 to 39 years).Microscopically, entirely follicular pattern was present in one case, entirely diffuse pattern in 2 cases, and follicular and diffuse pattern in other 4 cases. The tumor cells were medium to large in size and showed the morphology of centroblasts or blastoid cells with irregular nuclei, brisk mitotic activity in 3 cases and starry sky in 2 cases. All of the cases were positive for CD20, PAX-5, bcl-6, and MUM1 and had a Ki-67 index>80%, while CD10 and bcl-2 were positive in 3 cases. IRF4 gene rearrangement was identified in all cases and bcl-6 gene rearrangement in 2 cases. All patients presented with localized disease with clinical stage Ⅰ or Ⅱ, except one with stage Ⅳ at presentation and a new lesion in the mediastinum developed 8 months later.Conclusions:LBCL with IRF4 rearrangement is a clinicopathologically distinct entity. The observations reveal a broader spectrum of morphology and biological behaviors. The relationship between clinical stage and prognosis needs to be determined in more cases.