[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.

[Development of an asthenopia survey questionnaire for general surveys].

Objective: To design a visual fatigue questionnaire that can be used for population surveys. Methods: This was a cross-sectional study that involved three stages of subjects' recruitment. In the first stage, by convenience sampling, 150 individuals who complained of visual fatigue were selected at public places in Wenzhou City in May 2016. The 19-Item Asthenopia Survey Questionnaire (ASQ-19) was used to conduct the survey, and the questionnaire was adjusted. In the second stage, 200 outpatient participants were recruited from Wenzhou Medical University Affiliated Eye and Optometry Hospital from June 2016 to May 2017 and were divided into a visual fatigue group and a control group based on clinical diagnosis. The adjusted visual fatigue questionnaire was used for validation. In the third stage, 64 outpatient participants who met the inclusion criteria were continuously recruited from the Wenzhou Medical University Affiliated Eye and Optometry Hospital in July 2022. They were tested using the adjusted visual fatigue questionnaire and retested one week later. During the questionnaire adjustment stage, factor analysis and feedback were used to adjust the scoring method and items of the ASQ-19 questionnaire. The adjusted questionnaire was then analyzed for reliability, validity, accuracy, and subject acceptance during the validation and retest stages. Results: A total of 403 participants were included, and 456 questionnaires were distributed. Eventually, 432 valid questionnaires were collected from 379 participants, resulting in a valid response rate of 94.7%. During the questionnaire adjustment phase, there were 140 valid questionnaires from 140 participants consisting of 56 males and 84 females with an average age of (35.2±12.4) years. In the questionnaire validation phase, there were 186 valid questionnaires from 186 participants. Sixty-two participants had visual fatigue and 124 were controls. During the questionnaire retesting phase, 53 participants yielded 106 valid questionnaires. The group consisted of 20 males and 33 females with an average age of (22.8±4.9) years. After factor analysis, the symptom severity graded as none, mild, moderate, severe, and very severe was scored as 0, 1, 2, 3, and 4 points, respectively. The total score was 44, and the final questionnaire consisted of 11 items (numbered 1, 2, 3, 5, 6, 8, 10, 15, 17, 18, and 19). The 11-Item Asthenopia Survey Questionnaire (ASQ-11) had a Cronbach's α coefficient of 0.89, a split-half reliability of 0.82, and a test-retest Pearson correlation coefficient of 0.90 (P<0.001). The structural validity was 51.26%, and the discriminative validity was a t-value of 9.19 (P<0.001). On average, it took (2.82±0.43) minutes for participants to complete the questionnaire. The receiver operating characteristic curve had a cutoff value of 8.5, with a sensitivity of 74.19% and a specificity of 80.65%. Conclusion: The ASQ-11, with fewer items and a shorter completion time, is easy for participants to use and is suitable for screening or self-assessment of visual fatigue in the general population. Additionally, it is convenient for clinical and epidemiological studies related to visual fatigue.

[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.

Comparing responses of dairy cows to short-term and long-term heat stress in climate-controlled chambers.

Heat stress (HS) in dairy cows can be classified into short-term heat stress (STHS) and long-term heat stress (LTHS) according to the number of consecutive days in HS. The comparative study of these 2 types of HS is limited in terms of their effects on the production and energy metabolism of cows. In this study, 4 lactating Holstein cows (102.5 ± 12 days in milk, 605 ± 22 kg of body weight, second parity) fitted with rumen fistulae were randomly assigned to 1 of 2 groups in a 2 × 2 crossover design and allocated to 1 of 2 climate-controlled chambers. This study contained 2 periods, each with a control phase and a HS phase. There was a recovery phase between 2 periods. The HS phase comprised either STHS (3 d) or LTHS (7 d) treatments. Data collected from the 3 d of STHS and the last 3 d of LTHS were compared. The chambers were set at thermal neutral conditions (20°C, 50% humidity) during the control and recovery phases or cyclical HS conditions (26-38°C, 50% humidity) during the HS phase. Compared with STHS, LTHS decreased milk yield by 17.2% and dry matter intake by 12.6%, indicating that LTHS caused a more severe decline in milk production and feed intake. In addition, LTHS decreased milk protein concentration by 6.8% and milk protein yield by 22.4%. In comparison with STHS, LTHS decreased rumen liquor volatile fatty acid (29.7%), blood glucose (11.6%), and nonesterified fatty acid (13.6%) concentrations, but increased milk urea nitrogen by 15.1%, blood urea nitrogen by 8.6%, and creatine concentrations by 15.4%. Our results suggest that although reduced feed intake may be mainly responsible for reduced milk production during STHS, impaired rumen metabolism and suppressed mobilization of adipose tissue could be the main reasons for further reduction in milk yield during LTHS.

Diagnostic value of miRNA-122 in Kawasaki disease.

OBJECTIVE: To explore the expression pattern and diagnostic value of microRNA-122 (miRNA-122) in childhood Kawasaki disease (KD). PATIENTS AND METHODS: A total of 150 children with KD were included in the KD group. During the same period, 150 children with respiratory infection complicated with fever and without myocardial involvement were included in the control group.  Serum level of miRNA-122 in children with acute phase of KD and those in the control group was detected. The relationship between serum level of miRNA-122 and clinical features of KD was analyzed by Pearson correlation test. ROC curves were depicted to assess the diagnostic value of miRNA-122 in KD. RESULTS: Serum level of miRNA-122 was higher in the KD group than controls. In the acute phase of KD, the serum level of miRNA-122 was positively correlated to CRP and NT-proBNP, while negatively correlated to the sodium level. The specificity and sensitivity of miRNA-122 in diagnosing KD was 78.67% and 84.67%, respectively (AUC=0.8861, cut-off value=2.905). CONCLUSIONS: Serum level of miRNA-122 is significantly enhanced in the acute phase of KD, and highly expressed miRNA-122 is related to systematic inflammation. MiRNA-122 may be used as a diagnostic hallmark of KD.

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.