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Objective@#To investigate the morphological features, diagnosis and differential diagnosis of extrapleural sarcomatoid malignant mesothelioma (SMM).@*Methods@#Six cases of extrapleural SMM were evaluated for their clinical, histological, immunohistochemical features, and prognosis.@*Results@#Patients included 3 men and 3 women, with a median age of 60 years (range 41-75 years). All patients had no asbestos exposure in history and no pleural lesions. The tumors involved peritoneum (3 cases), bone (2 cases), and neck soft tissue (1 case). Histologically, the tumors were mainly composed of slender to plump spindle cells with occasional polymorphic cells, arranged in fascicular to storiform pattern or haphazardly organized, closely resembling those of fibromatosis, fibrosarcoma or malignant fibrous histiocytoma. The tumor cells were imunohistochemically positive for cytokeratin (pan, 6/6), calretinin (5/6), podoplanin (6/6), D2-40 (4/6), vimentin (6/6), WT1 (4/6), CD10 (3/6), SMA (4/6), and variably positive for CK7, and CK8/18, but were negative for other linage-specific markers. The Ki-67 proliferation indexes ranged from 25% to 55%, consistent with the diagnosis of malignant mesothelioma of the sarcomatous type. Ultrastructurally, the tumor cells possessed discontinuous external lamina, cytoplasmic processes, microfilaments and desmosomal intercellular junctions. Local recurrence or metastasis was seen in 1 case and 4 cases, respectively, after surgery, and all the patients died of the disease within 9 months.@*Conclusions@#Extrapleural SMM, although rare, should be considered as a differential diagnosis among other benign or malignant sarcomatoid tumors and sarcomas. Along with clinical and radiological presentation, the combination of broad-spectrum cytokeratin, vimentin, and a series of mesothelial markers are useful for diagnosis of SMM.
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Objective@#To investigate the multidirectional differentiation potential in epithelioid sarcoma (ES), with special emphasis on its mesothelial and lymphatic endothelial markers expression.@*Methods@#Ten cases of distal-type ES were included. The clinical, histological, and immunohistochemical(including mesothelial and lymphatic endothelial markers expression)features and follow-up data were evaluated.@*Results@#The patients aged between 8 to 66 years. Five cases were male and five were female. The tumors were located at the palm (2 cases), wrist (3 cases), upper arm (2 cases), poplitealfossa (1 case), lower leg (1 case) and thigh (1 case), respectively. Clinically, most cases presented as painful, firm subcutaneous nodules. Histologically, the tumors were mainly composed of epithelioid, rhabdoid, spindle, or transitional cells, with abundant eosinophilic cytoplasm, oval and vesicular nucleus, and one or more prominent nucleoli. They were arranged in nodular, diffuse nodular or sheet like growth patterns, frequently with necrosis at the center with vague granulomatous configuration. Immunohistochemically, all tumors expressed cytokeratins, epithelial membrane antigen, CD34, desmin, mesothelial markers such as Calretinin, WT1, D2-40, M2A, vascular and lymphatic endothelial markers FLI-1, and VEGFR-3. The tumor cells did not express CD31, FⅧRAg, HHF35, HMB45, Melan A, MyoD1, myogenin, S-100 protein and SMA. All 10 patients underwent radical resection or extended excision, with additional radiotherapy or chemotherapy. During the follow-up from October 2012 to August 2016, seven cases showed recurrences and metastases within 2 months to 2 years. Five patients died of the disease due to widespread metastases.@*Conclusions@#ES may show a wide spectrum of morphology, and display a multidirectional differentiating capabilities including towards mesothelial and lymphatic endothelial cells. As such, its diagnosis and differential diagnosis are particularly important as it is easily confused with other tumors with similar morphology or immune phenotype.
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Objective@#To analyze the clinical and pathological features of Kaposiform hemangioendothelioma (KHE), and to investigate the role of master transcriptional factor Prox-1 in the regulation of lymphatic differentiation.@*Methods@#Nine cases of KHE (during the period from October 2009 to June 2016) were collected with clinical and pathological data. H&E stained section review and immunohistochemietry using the Dako EnVision method were performed.@*Results@#There were 6 female and 3 male patients with age ranging from 2 months to 8 years (median 3 years and 4 months). The patients presented with either single subcutaneous soft tissue mass, or bone tumors, with the duration of disease onset ranging from 1 month to 1 year. The sites of involvement included the skins of neck (2 cases), nose root (1 case), inguinal (1 case), thigh root (1 case), humerus (2 cases), lumbar vertebrae(1 case), and mesentery (1 case). These tumors were histologically composed of nodules of densely packed spindle or ovoid cells and deformed small blood vessels in an invasive growth pattern. The tumor cells were immunohistochemically positive for both blood vessels and lymphatic endothelial markers, including Prox-1, the master transcriptional factor, and VEGFR-3. With followed-up from 1 to 60 months (median 26 months), two patients died of the disease, while the remaining patients were alive without recurrence.@*Conclusions@#KHE is a rare vascular tumor with at least partial lymphatic endothelial differentiation, in which Prox-1 may act as a master regulator for such differentiation. KHE is an aggressive tumor of intermediate malignant potential, with local invasion and recurrence tendency, and long term follow-up is required.
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<p><b>OBJECTIVE</b>To study the clinicopathologic features, immunophenotype and differential diagnosis of extraskeletal myxoid chondrosarcoma (EMC).</p><p><b>METHODS</b>The clinicopathologic features of 5 cases of EMC (during the period from 2008 to 2013) were retrospectively analyzed. Immunohistochemical study (EnVision method) was carried out using the archival material. The literature was reviewed.</p><p><b>RESULTS</b>There were altogether 3 female patients and 2 male patients. Their age ranged from 38 to 63 years (average = 51 years). The patients primarily presented with a tender soft tissue mass. All the tumors studied were solitary and the duration of disease onset varied from 3 months to 1 year. The sites of involvement included toe (number = 2), intracranial (number = 1), thigh (number = 1) and shoulder (number = 1). Gross examination showed white nodular masses with a gelatinous cut surface. The average tumor size measured 5.2 cm in greatest dimension. Histologically, a multinodular architecture with fibrous or loose fibrovascular septa separating lobules of tumor cells was identified. The lobules contained abundant myxoid stroma, with peripheral accentuation of tumor cellularity. Two cases were diagnosed as cellular variant of EMC, with invasive growth pattern and hemorrhage. The tumor cells in cellular EMC were arranged in solid nodules, with rare myxoid matrix in between. The nuclei were relatively uniform, round to oval and contained prominent nucleoli. The mitotic figure ranged from 5 to 10 per 10 high-power fields. Immunohistochemical study showed that all of the 5 cases were positive for vimentin, mitochondria and CD56. Two cases expressed synaptophysin and NSE. Focal positivity for these neuroendocrine markers was detected in the other 2 cases. Chromogranin and S-100 protein expression was demonstrated in 2 cases. The staining for epithelial membrane antigen was positive in case 2 and negative in the other 4 cases. CD117 showed diffuse positivity in case 1, the other 4 cases were not expressed.</p><p><b>CONCLUSIONS</b>EMC is a rare soft tissue sarcoma characterized by distinctive histopathologic features and often shows neuroendocrine differentiation. Although EMC is a slow-growing tumor, it carries a high local recurrence rate and even metastases, warranting long-term follow up.</p>
Subject(s)
Adult , Female , Humans , Male , Middle Aged , CD56 Antigen , Metabolism , Chondrosarcoma , Metabolism , Pathology , General Surgery , Chordoma , Metabolism , Pathology , Chromogranins , Metabolism , Diagnosis, Differential , Neoplasms, Connective and Soft Tissue , Metabolism , Pathology , General Surgery , Phosphopyruvate Hydratase , Metabolism , Retrospective Studies , Rhabdomyosarcoma , Metabolism , Pathology , S100 Proteins , Metabolism , Shoulder , Synaptophysin , Metabolism , Thigh , Toes , Vimentin , MetabolismABSTRACT
Objective To confirm that astrocytes from cerebral cortex of newborn rat can be the target cells of Hantaan virus (HTNV)and Seoul virus (SEOV)infection and to observe changes of astrocytes after different infection time. Methods Astrocytes were prepared from cerebral cortex of newborn rat, and then infected with HTNV and SEOV. The established virus infections were confirmed by detection of virus nucleocapsid protein (NP) and S segment RNA in astrocytes using double-label immunofluoreseence, Western-blot and reverse transcriptase-polymerase chain reaction (RT-PCR). Results The astrocytes from cerebral cortex of newborn rat were cultured successfully in vitro, which could be infected by HTNV and SEOV. The number of infected astrocytes and the virus titer in the infected astrocytes kept on increasing along with the extended infection duration. Conclusions Astrocytes from cerebral cortex of newborn rat are the target cells for HTNV and SEOV infection. Then establishment of in vitro cultured astrocytes model for Hantaviruses infection will be helpful for the study on the pathogenesis of hemorrhagic fever with renal syndrome.
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Objective: To observe the targeting therapy of the hdsFv-RC-RNase recombinant single chain immunotoxin on the xenograft of the human hepatocellular carcinoma in nude mice and to explore its clinical potentiality. Methods: The prokaryotic expression vector TIG-hdsFv-RC-RNase was transformed into E.coli BL21(DE3)plys to largely express recombinant single chain immunotoxin hdsFv-RC-RNase against hepatocellular carcinoma induced by IPTG. The expressed product was purified via Ni-NTA affinity chromatography under native conditions and mildly refolded. The ELISA was used to analyze its immunological activity of antigen-binding capability. The xenogrft model of the human hepatocellular carcinoma in nude mice was established and the targeting therapy of hdsFv-RC-RNase was evaluated. Results:After induced by IPTG, a new protein band with M_r 41 000 was found in the supernatant of the bacteria and expressed in a soluble form. The expressed product was purified to homogeneity via Ni-NTA affinity chromatography under native conditions. The results of ELISA showed the refolded hdsFv-RC-RNase had the specific antigen-binding capability to the human hepatocellular carcinoma cell, but not to the normal hepatocyte (P
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Objective:To study heat shock proteins(HSP) and their association with Hantavirus nucleocapsid protein(HV-NP) in Vero-E6 cells infected with Hantaan76-118.Methods: The HV-NP was identified by immunocytochemical staining after infected with Hantaan76-118.The expression of HSP was detected by Western-blot and analyzed by double specific antibody sandwich ELISA.Results: Western-blot exhibited that the expressions of HSP27,HSP70 and Grp94 in the Vero-E6 cells infected with Hantaan 76-118 were significantly higher than those in the control cells(P
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Objective:To prepare polyclonal antibody against human immunoglobulin G(human IgG) and to use this antibody in the clinical diagnosis of nephrosis. Methods:New Zealand rabbit was immunized subcutaneously with human IgG.Enzyme linked immunosorbent assay(ELISA) was applied to reveal the titer of the prepared polyclonal antiserum against human IgG.Antiserum was purified with affinity chromatography,and the purified antibody was confirmed for its specificity by Western blot and immunohistochemical staining.The purified antbodies which have been indentified were fused with FITC(fluorescein isothiocyanate) and then analyzed by immunohistochemical staining.Finally the fused antibody was useed in the clinical diagnosis of nephrosis. Results: The titer of the obtained antiserum was up to(1∶128 000,) double agar diffusion test showed the titer of the antibody was 1∶32.By the method of affinity chromatography,we obtained purified antibody with the purity of 85%.ELISA,double agar diffusion and immunohistochemical staining tests showed that the specificity and titer of the antibody were not decreased sharply.The purified antibody fused with FITC also kept the specificity of the primitive antibody.When the FITC fused antibody was tried in nephrosis patients,it detected human IgG effectively. Conclusion:The polyclonal antibody can specifically recognize human IgG.This purified antibody fused with FITC can be used in the diagnosis of nephrosis.
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Aim To explore clinicalpathological features and the pathogenesis of lymphocytic mastitis, an uncommon breast benign disease. Methods The clinical pathological characteristics of 7 patients with lymphocytic mastitis were studied retrospectively. All cases were evaluated by using paraffin-embedded sections and by immunohistochemical staining with mouse anti-CD20, -CD45RO, and -CD68 antibodies. T and B lymphocytes infiltrated in the lobules of mammary gland were quantitatively analyzed according to stereoscope. The glucose regulation protein 94 (grp 94)/glycoprotein 96(gp 96), a member of heat shock protein family was also investigated in these cases by using immunohistochemical staining. Results It was showed that 4 cases were women suffering from insulin dependent diabetic mellitus (IDDM). One case was a woman without diabetic history. The history of the other two cases was not clear. The histopathological features of all 7 cases were lobulitis, perilobulitis and catheter ductitis with infiltration of lymphocytes accompanied with atrophy of lobule in mammary glands and homologous fibrosis of stroma. The result of immunohistochemical staining showed that most of infiltrated lymphocytes were B lymphocytes, while the small proportion of the cells were T lymphocytes, and the difference was significant(P〈 0.01). There was the expression of grp 94 in the cytoplasm of epithelium cells of lobules and ducts in normal mammary glands. A proportion of lymphocytes infiltrated in lobules and perilobules expressed grp 94. Some infiltrated cells expressed CD68. Conclusion A portion of lymphocytic mastitis is related closely to insulin dependent diabetic mellitus. Both humoral immunity and cellular immunity are probably involved in the pathogenesis of lymphocytic mastitis. Because of its unique pathologic and clinical features, lymphocytic mastitis should be defined as an independent mastitis and distinguished from other chronic inflammatory and fibrosing conditions in breast.
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Objective To study Heat shock proteins (HSPs) induction and their association with Hantaan virus (HTNV) structural proteins in the brain tissues of mice experimentally infected with HTNV 76～118. Methods Newborn mice less than 72 h of age were experimentally infected with HTNV by intracerebral inoculation. The brains of mice at the 8th day post infection were removed and prepared for tissue extracts. The possible interaction or association of Hantaan virus nucleocapsid protein (HTNV NP) and glycoprotein G2 (HTNV G2) with GRP94, HSP70 and HSP27 were analyzed by double specific antibo dies sandwich ELISA and immunoprecipitation methods. Results HTNV infection induced the expression a set of HSPs including GRP94 and HSP70 in the brain tissues of mice experimentally infected with HTNV. HTNV NP was associated with GRP94, HSP70 and HSP27, which led to formation of HTNV NP HSP70 GRP94 HSP27 complexes. It was also observed that HTNV G2 was associated with NP and HSP27, which led to formation of HTNV G2 NP HSP27 complexes. Conclusions HTNV infection induces expression of a set of HSPs, which were associate with viral structural proteins in the brain tissues of mice experimentally infected with HTNV. This association may demonstrate the chaperone roles of HSPs in the synthesis, transportation and maturity of viral proteins during viral replication in the host cells.
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Objective To study the impact of viral infection on stress protein gene expression and investigate the possibility that heat shock proteins interact with viral components during viral assembly in the brain of suckling mice experimentally infected with Hantaan virus (HV). Methods New born mice less than 72 h of age were inoculated into brain with the 100 LD50 of Hantaan virus (strain 76 118). At the 8th day mice died from brain disease. Brains were prepared for tissue sections and extracts for the experiments. The Hantaan virus infection, HSP70 induction and their co localization in brain of infected suckling mice were demonstrated by immunohistochemical staining and laser scan co focal microscopic double labling. The possible intereaction between HV structural proteins and HSP70 was further analyzed by immunoprecipitation of infected tissue extracts with antibodies to either HSP70 or to viral nuclear capsule proteins HV NP by using SDS PAGE and Western blotting. Results HV infection induced an increased levels of HSP70 expression at the cytoplasma of neurons in the brain of suckling mice HV NP protein proved to be co localized with HSP70. Monoclonal antibody (mAb) to HSP70 could co immunoprecipitate the HV NP, and similarly mAb to HV NP could co immunoprecipitate HSP70 from virus infected tissue extracts, but not from mock infected tissue extracts. Antibody, which recognized one of these two proteins, did not cross react with the other. Thus, the association between HSP70 and HV NP appeared to be specific.Conclusions Hantavirus infection can directly induce HSP70 expression, leading to the formation of HV HSP 70 viral antigen peptide complex which plays some roles in the course of viral infection and replication.