[Radiation-associated sarcomas of bone and soft tissue: a clinicopathological analysis of 46 cases].

Objective: To investigate the clinical, imaging, histological, and molecular features and the differential diagnosis of radiation-associated sarcomas of bone and soft tissue. Methods: Forty-six cases of radiation-associated sarcomas of the bone and soft tissue in Beijing Jishuitan Hospital from January 2010 to January 2022 were retrospectively analyzed; and the imaging, histological features and immunophenotype were examined. Results: There were 33 females and 13 males, aged from 18 to 74 years, with a mean of 52 years. The most common site of radiation-associated sarcomas were the limbs and spine (15 cases), followed by the chest (9 cases). The primary diseases included epithelial tumors (15 breast cancer, 6 cervical cancer, and 5 bowel cancer), hematolymphoid tumors, bone and soft tissue tumors and infectious lesions. The latent period of radiation-associated sarcomas ranged from 2-22 years, with an average of 11.6 years. Histopathologically, the morphology was divergent from the primary tumor. The most common malignant tumor type was undifferentiated sarcoma (22 cases), followed by osteosarcoma (16 cases). The immunophenotype of radiation-related sarcoma was almost the same as the corresponding soft tissue sarcoma. Conclusions: Radiation-induced sarcoma has a wide range of primary tumor types and its imaging, morphology and immunohistochemical features are similar to those of the primary sarcoma of bone and soft tissue. Clinical correlation is often recommended for the differential diagnosis.

[Pediatric myofibroma/myofibromatosis of the soft tissue and bone: a clinicopathological analysis of 28 cases].

Objective: To investigate the clinicopathological features, diagnosis and differential diagnosis of pediatric myofibroma/myofibromatosis of the soft tissue and bone. Methods: All cases of pediatric myofibroma/myofibromatosis of the soft tissue and bone diagnosed between January 2011 and December 2018 were retrieved from the surgical pathology records in the Department of Pathology, Beijing Jishuitan Hospital, Beijing, China. Clinical and radiological data were collected. H&E and immunohistochemistry were used to examine histological and immunophenotypic features and to make the diagnosis and differential diagnosis. The relevant literature was also reviewed. Results: Twenty-eight cases of pediatric myofibroma/myofibromatosis of the soft tissue and bone were respectively collected. The patients' ages ranged from 2 months to 14 years, with a mean age of 7 years. There were 7 females and 21 males. There were 12 cases located in soft tissue, including the finger (n=9), upper arm (n=1) and foot (n=2). There were 14 cases located in the bone of limb, including the femur (n=8), tibia (n=4), clavicle (n=2), fibula (n=2) and radius (n=1). There were 2 cases of myofibromatosis involving multiple bones. Radiology showed lytic lesions in the bone. The proliferation of spindle-shaped myofibroblasts arranged in fascicles with indistinct eosinophilic cytoplasm and bland nuclei, with no pleomorphism and cytological atypia. The characteristic histologic structure was the biphasic nodular growth pattern with cellular and paucicellular regions. The tumors might arrange in a hemangiopericytoma-like pattern. The stroma varied between dense fibrosis and myxoid changes. The reactive new bone formation and inflammatory cell infiltration also existed. Immunohistochemical study showed that the SMA was positive. The surgical resections were performed. One of the patients had tumor recurrence as a result of 11-month follow-up. Conclusions: The pediatric myofibroma/myofibromatosis of the soft tissue and bone is a very rare benign tumor and has a good prognosis. It has a characteristic morphology and its differential diagnosis from other spindle cell tumors could be made with the immunohistochemical analysis.

[The diagnostic value of CCNB3 and BCOR expression in BCOR-CCNB3 sarcoma].

Objective: To investigate the diagnostic value of expression of CCNB3 and BCOR in BCOR-CCNB3 sarcoma (BCS). Methods: Fifteen cases of BCS confirmed by fluorescence in situ hybridization (FISH) and/or reverse transcription-polymerase chain reaction (RT-PCR) from January 2014 to October 2021 at Beijing Jishuitan Hospital were collected. Immunohistochemical EnVision method was used to detect the expression of CCNB3 and BCOR in 15 cases of BCS and in 65 non-BCS tumors (54 cases of Ewing's sarcoma, 5 cases of CIC rearranged sarcoma, 4 cases of synovial sarcoma, 1 case of mesenchymal chondrosarcoma and 1 case of soft tissue clear cell sarcoma). Results: Immunohistochemical staining for CCNB3 revealed strongly diffuse nuclear staining in 14 of 15 (14/15) BCS cases, whereas none of the 65 non-BCS tumors showed any staining. Immunohistochemical staining for BCOR showed strongly diffuse nuclear staining in 11 (11/14) BCS cases; seven of the 65 (7/65, 10.8%) non-BCS tumors showed variable staining (five cases of Ewing sarcoma, one cases of synovial sarcoma, and one case of mesenchymal chondrosarcoma). The sensitivity and specificity of CCNB3 in diagnosing BCS were 93.3% and 100% and these of BCOR were 78.6% and 89.2%, respectively. Conclusions: CCNB3 is a highly sensitive and specific marker for BCS.The antibody may help screening BCS.

Histopathology-Based Diagnosis of Oral Squamous Cell Carcinoma Using Deep Learning.

Oral squamous cell carcinoma (OSCC) is prevalent around the world and is associated with poor prognosis. OSCC is typically diagnosed from tissue biopsy sections by pathologists who rely on their empirical experience. Deep learning models may improve the accuracy and speed of image classification, thus reducing human error and workload. Here we developed a custom-made deep learning model to assist pathologists in detecting OSCC from histopathology images. We collected and analyzed a total of 2,025 images, among which 1,925 images were included in the training set and 100 images were included in the testing set. Our model was able to automatically evaluate these images and arrive at a diagnosis with a sensitivity of 0.98, specificity of 0.92, positive predictive value of 0.924, negative predictive value of 0.978, and F1 score of 0.951. Using a subset of 100 images, we examined whether our model could improve the diagnostic performance of junior and senior pathologists. We found that junior pathologists were able to delineate OSCC in these images 6.26 min faster when assisted by the model than when working alone. When the clinicians were assisted by the model, their average F1 score improved from 0.9221 to 0.9566 in the case of junior pathologists and from 0.9361 to 0.9463 in the case of senior pathologists. Our findings indicate that deep learning can improve the accuracy and speed of OSCC diagnosis from histopathology images.

Lnc-AC145676.2.1-6-3 can influence STX3-induced abnormal autophagy by sponging hsa-miR-1292-3p in intestinal aGVHD.

OBJECTIVE: Intestinal acute graft-versus-host disease (aGVHD) is a serious complication of allogeneic hematopoietic stem cell transplantation (allo-HSCT). Abnormal autophagy levels in intestinal aGVHD have been confirmed in many studies. LncRNAs exert coregulatory functions and participate in a variety of intracellular regulatory processes. In this study, we investigated how lnc-AC145676.2.1-6-3 regulates dysregulated STX3-related autophagy in aGVHD. MATERIALS AND METHODS: First, we established a mouse model of aGVHD by transplanting a mononuclear cell suspension from Balb/c donor mice treated with 60Co X-rays into CB6F1 recipient mice. STX3-related indicators were analyzed by Western blotting (WB) and immunohistochemistry which confirmed that STX3 plays an important role in dysregulating autophagy in intestinal aGVHD. TNF-αinduced Caco-2 cells, which is an in vitro model of intestinal barrier dysfunction, were established to verify the effect of STX3. The direct interaction between the partners of lnc-AC145676.2.1-6-3-mediated hsa-miR-1292-3p and STX3 axis was evaluated by the Dual-Luciferase activity assay. We performed PCR, WB, and immunofluorescence in Caco-2 cells to determine whether the abnormal autophagy levels were influenced by lnc-AC145676.2.1-6-3. RESULTS: The results showed that lnc-AC145676.2.1-6-3 could significantly suppress the number of autophagic vacuoles, the LC3-II/I ratio, and beclin1 levels by increasing STX3 levels. CONCLUSIONS: Lnc-AC145676.2.1-6-3 may play an important role in intestinal aGVHD by targeting STX3.

[Reassessment of EWSR1 gene rearrangement-negative undifferentiated round cell sarcomas in bone and soft tissues by fluorescence in situ hybridization].

Objective: To unravel the CIC rearrangement sarcomas and BCOR-CCNB3 sarcomas from EWSR1 rearrangement-negative undifferentiated round cell sarcomas in the bone and soft tissues. Methods: Twenty-eight cases of EWSR1 rearrangement-negative undifferentiated round cell sarcomas of bone and soft tissues, tested for CIC rearrangement and BCOR rearrangement by fluorescence in situ hybridization and related immunostaining were analyzed, and some of the BCOR rearrangement cases were verified by reverse transcription-polymerase chain reaction. Results: Five of 28 (17.9%) tested cases were positive for CIC rearrangement and six (21.4%) for BCOR rearrangement. Histopathologically, CIC rearrangement sarcomas comprised nodular aggregates of round to polygonal cells, containing hyperchromatic nuclei, prominent nucleoli and moderate cytoplasm, with focal variable necrosis and myxoid stroma. BCOR-CCNB3 sarcomas mostly comprised diffusely arranged, round to oval to short spindly cells with angulated nuclei, vesicular chromatin, inconspicuous nucleoli and interspersed vessels. Immunohistochemically, five of six BCOR-CCNB3 sarcomas showed CCNB3 immunostaining, which could be helpful for diagnosis. Two patients with CIC rearrangement sarcoma died of the diseases in seven months and twenty-two months. One patient with BCOR-CCNB3 sarcoma died of the diseases in forty-six months. Conclusions: Overall, 39.3% of the EWSR1 rearrangement-negative undifferentiated round cell sarcomas are CIC rearrangement sarcomas and BCOR-CCNB3 sarcomas. Molecular testing is helpful for diagnosis.

[DNA sequencing of H3F3A mutations in H3.3 immunohistochemistry-negative giant cell tumors of bone].

Objective: To investigate the subtypes of H3F3A DNA mutation in H3.3 immunohistochemistry (IHC) negative giant cell tumors of bone (GCTB). Methods: IHC expression of G34W mutated protein was evaluated in 181 cases GCTB. In H3.3 IHC negative cases, Sanger DNA sequencing analysis was used to detect the subtypes H3F3A mutations. Results: Overall, 164 (90.61%) cases of GCTB showed nuclear expression of H3.3, and 17 cases were negative. These 17 H3.3 negative cases were subjected to Sanger DNA sequencing analysis; results showed that eight presented rare mutation subtypes occurring at glycine 34 to leucine (G34L, 3/181, 1.66%), glycine 34 to valine (G34V, 3/181, 1.66%) and glycine 34 to arginine (G34R, 2/181, 1.10%), and the other nine cases were wild type (glycine 34, 9/181, 4.97%). Sanger DNA sequencing analysis confirmed the absence of G34W mutation in the H3.3 negative cases. Combining IHC and DNA sequencing analysis increased the detection rate of H3F3A mutation in the GCTB to 95.03%. Conclusions: H3.3 IHC could detect H3F3A G34W mutation in GCTB, but not for other rare mutation and wild types loci.

APLNR stimulates the development of glioma via the NFAT5/AKT feedback loop.

OBJECTIVE: To detect the role of APLNR in influencing the proliferative ability and development of glioma. PATIENTS AND METHODS: APLNR levels in 42 matched glioma tissues and adjacent normal brain tissues were detected by quantitative Real Time-Polymerase Chain Reaction (qRT-PCR). The correlation between APLNR level and clinical features of glioma patients was assessed. Regulatory effects of APLNR on glioma cell functions were evaluated by cell counting kit-8 (CCK-8), colony formation, and 5-Ethynyl-2'-deoxyuridine (EdU) assay, respectively. At last, the involvement of NFAT5 in APLNR-regulated glioma cell phenotypes was examined. RESULTS: APLNR was upregulated in glioma tissues than the adjacent ones. Glioma patients expressing higher level of APLNR had more advanced stage and worse prognosis. Knockdown of APLNR inhibited proliferative ability of glioma. NFAT5 level was negatively regulated by APLNR. Notably, NFAT5 could partially abolish the regulatory effect of APLNR on glioma cell phenotypes. CONCLUSIONS: APLNR level is closely linked to tumor grading and prognosis of glioma patients. It stimulates proliferative ability in glioma cells by targeting NFAT5.

[Primary intraosseous Rosai-Dorfman disease: a clinicopathological analysis of fourteen cases].

Objective: To investigate the clinicopathological characteristics, histogenesis, immunophenotypes and molecular genetic features of primary intraosseous Rosai-Dorfman disease (RDD) for improving diagnostic accuracy and differential diagnosis. Methods: This retrospective study included 14 RDD cases diagnosed from January 2009 to January 2019 at Beijing Jishuitan Hospital, China. The immunohistochemical staining for S-100, cyclin D1, CD1a and CD207 expression was analyzed. The BRAF V600E and KRAS mutation analyses were performed using the Scorpions amplification refractory mutation system (ARMS) fluorescence quantitative PCR. Results: There were 6 female and 8 male patients, aged from 2 to 64 years (mean 31.4 years). All of the 14 cases occurred in the bone without lymph node disease, while one patient developed additional lesions within vertebra and nasal cavity. Radiographically, the lesions were lytic with sclerotic margins. Histologically, the lesions percolated through the medullary cavity in an infiltrative fashion and alternating hyper- and hypo-cellular regions of histiocytic clusters (seen as alternating dark and light zones at low magnification). Large histiocytes also showed emperipolesis. Some cases had areas of fibrosis and dense lymphoplasmacytic infiltrates. There were vasculitis and an increased number of plasma cells in the cases involving multiple sites. One case showed concurrence of RDD and Langerhans cell histiocytosis(LCH) with inconspicuous increase of Langerhans histiocytes. Immunohistochemical staining showed that the large histiocytes were positive for S-100, CD68 and CD163 in all cases. The nuclear immunoreactivity for cyclin D1 was observed in 13 of the 14 cases. S-100, CD1a and CD207 were positive in the case with concurrence of RDD and LCH. ARMS-PCR results showed that BRAF V600E mutation was observed in the cases with concurrence of RDD and LCH, while there were no KRAS mutations (7/7). Follow-up information was available for 12 patients and ranged from 9 to 49 months. Three of the 12 patients experienced recurrences after the first surgery. Conclusions: Primary intraosseous RDD is rare, and its concurrence with LCH is a very rare phenomenon. Its clinical symptoms, imaging, and pathological manifestations need to be distinguished from other bone lesions. The molecular detection of BRAF V600E and the nuclear expression of cyclin D1 mutations can be used for the diagnosis and differential diagnosis of RDD.

[Long-term observation of morphological changes of the inner retinal after internal limiting membrane peeling in macular hole surgery].

Objective: To investigate the characteristics of morphological changes of inner retinal layer after internal limiting membrane peeling in macular hole surgery. Methods: Retrospective case study. Patients with idiopathic macular hole from 2015 to 2018 underwent vitrectomy+internal limiting membrane peeling (inverting)+ gas tamponade in China-Japan Friendship Hospital were investigated. A total of 19 eyes (17 patients) were enrolled, including 4 males (4 eyes) and 13 females (15 eyes). The average age was 62.74±5.25 years. Optical coherence tomography (OCT) was used to obtain the topographic maps of retinal thickness, the thickness of retinal ganglion cell complex and probability maps, and the Angio/en-face maps of macular retina. The characteristics of the morphological changes of the inner retinal were comprehensively analyzed. Results: Among 19 eyes, 9 eyes had internal limiting membrane peeling, 8 eyes had lotus-like internal limiting membrane inverting, and 2 eyes had the uper180 degrees internal limiting membrane inverting. The minimum diameter of macular hole was (543.06+220.17) µm and the maximum diameter was (947.18+319.12) µm. The follow-up time was (21.05+9.66) months, and the visual acuity was 0.45+0.35 at the last follow-up. In the 19 eyes, all the macular holes were closed postoperatively and dissociated optic nerve fiber layer appearance (DONFL) and concentric macular dark spots (CMDS) all showed, as well as mGCC thinning. The changes of CMDS and mGCC in 2 eyes in the uper180 degrees internal limiting membrane inverting group were mainly seen in the upper retina and in the remaining 17 eyes were seen diffused around the macula, which roughly corresponded to the extent of internal limiting membrane peeling. Two eyes showed clear decrease of retinal capillary density on Angio/map. Conclusions: Long-term morphological changes of the inner retinal after internal limiting membrane peeling in macular hole surgery are obvious. In addition to the appearance changes like DONFL and CMDS, the macular ganglion cell complex (mGCC) is also involved. (Chin J Ophthalmol, 2019, 55:747-756).

[Dedifferentiated liposarcoma of extremities: a clinicopathologic analysis].

Objective: To study the clinicopathologic features of dedifferentiated liposarcoma of extremities. Methods: Nine cases of dedifferentiated liposarcoma of extremities diagnosed at Beijing Jishuitan Hospital from 2009 to 2017 were selected. The histological features of cases of dedifferentiated liposarcoma of extremities were evaluated by HE and immunohistochemistry, together with the clinical and radiological features. Flourescence in situ hybridization(FISH) was used to detect MDM2 amplification. Results: They were located in the thigh (6 cases), calf (2 cases) and buttock (1 case). There were six females and three males. Patients' age ranged from 61 to 79 years (mean 68 years). Histologically, there were two components, well-differentiated liposarcoma and dedifferentiated sarcoma with or without transitional lesions between them. The histology of dedifferentiated liposarcoma included undifferentiated sarcoma and fibrosarcoma. Heterologous elements such as bone and cartilage were present in two cases. Immunohistochemical study showed the tumor cells expressed vimentin, CDK4 and p16. MDM2 were positive in 6 cases (6/9) and p53 was positive in one case(1/9). CKpan was positive in the epithelioid differentiation area. S-100 protein was positive in the well-differentiated liposarcoma component. FISH showed the amplification of MDM2(6/9). Conclusions: Dedifferentiated liposarcoma of extremities is very rare. The diagnosis should be combined with the histological characteristics and immunohistochemical results and differentiated from the other tumors and tumor-like lesions.