Objective:To investigate the clinicopathological features, immunophenotype and molecule characteristics of EWSR1::CREM fusion malignant epithelioid neoplasm in soft tissue.Methods:The clinical and pathological data of 2 cases of EWSR1::CREM fusion malignant epithelioid neoplasm in soft tissue diagnosed at the Department of Pathology, Beijing Jishuitan Hospital, Beijing, China from May 2023 to May 2024 were analyzed. Immunohistochemical study, fluorescence in situ hybridization (FISH) and next generation sequencing (NGS) were performed. Relevant literature was reviewed.Results:There were one male and one female patients, aged 35 and 29 years, respectively. The tumors developed in the somatic soft tissue, including calf and chest wall, and were 6.0 and 6.2 cm in size, respectively. The imaging studies suggested space-occupying lesions in muscle tissue. Case 1 did not involve the bone, while Case 2 showed fracture of the 8th rib. Microscopically, a fibrous pseudocapsule surrounded by lymphocytes and plasma cells was identified. The tumors were composed of small to medium-sized round and short spindle-shaped cells, showing nodular or sheet-like pattern. The tumor cells showed round nuclear outline, coarse chromatin with prominent nucleoli. Immunohistochemically, tumor cells showed diffuse positivity of ALK (D5F3), MUM1 and Syn, focal or patchy positivity of CKpan, EMA, S-100, NSE, WT-1 and SMA, and a high Ki-67 index (20%-30%). FISH demonstrated break-apart signals of EWSR1 gene in the 2 cases. NGS revealed EWSR1::CREM gene fusion. Case 2 showed an ATRX gene mutation. The two patients were free of recurrence or metastasis at the 10-month and 1-month follow-up, respectively.Conclusions:EWSR1::CREM fusion malignant epithelioid neoplasm is rare and lacks distinctive morphological and immunohistochemical features. FISH and NGS can help make a definitive diagnosis.

Objective:To investigate the clinicopathological and genetic characteristics of malignant transformation of polyostotic fibrous dysplasia (FD) in long bone.Methods:A retrospective analysis of clinical characteristics and morphological features was conducted from 4 cases of malignant transformation of FD diagnosed at Beijing Jishuitan Hospital from January 2016 to December 2023. Hotspot mutations for GNAS gene were tested in 4 cases by Sanger sequencing, in which both FD and malignant tissues were detected in 3 cases respectively.Results:There were 2 female and 2 male patients, aged 46 to 53 years [mean (49±3.2) years], and the course of the disease spanned from 2 months to 36 years. The tumor involved the femur ( n=2), tibia ( n=1) and humerus ( n=1). Three of them were diagnosed with FD before surgery. Single photon emission computed tomography showed multiple increases in bone metabolism, CT showed poorly margin, cortical destruction and soft tissue mass with uneven enhancement. Three cases had both FD and sarcoma components, while the remaining case exhibited exclusively sarcoma. The sarcomas displayed significant morphological variation, with 1 case diagnosed as osteosarcoma and 3 cases classified as low to high grade spindle cell sarcoma. Immunohistochemical results did not provide any indications for clear classification. Sanger sequencing demonstrated GNAS mutations of p.R201H (c.CGT>CAT, n=2) and p.R201C (c.CGT>TGT, n=2). All 4 cases were followed-up for 18 to 76 months, and received chemotherapy after surgery; 2 cases maintained disease-free, one case was diagnosed with invasive breast cancer through a core needle biopsy 3 months after chemotherapy, and another one was found to relapse 18 months after surgery. Conclusions:Some cases of polyostotic FD occur in association with café-au-lait macules and/or endocrine hyperfunctioning in McCune-Albright syndrome (MAS); polyostotic FD and MAS have more malignant potential than monostotic FD, but they are not the risk factors for FD malignancy. GNAS mutations may be involved in the occurrence and development of FD. The histologic types of malignant transformation of polyostotic FD in long bone are diverse, the sarcoma components of FD also present the GNAS mutation, suggesting potential involvement in the pathogenesis of FD malignancy.

Objective:To investigate the clinicopathological and genetic characteristics of malignant transformation of polyostotic fibrous dysplasia (FD) in long bone.Methods:A retrospective analysis of clinical characteristics and morphological features was conducted from 4 cases of malignant transformation of FD diagnosed at Beijing Jishuitan Hospital from January 2016 to December 2023. Hotspot mutations for GNAS gene were tested in 4 cases by Sanger sequencing, in which both FD and malignant tissues were detected in 3 cases respectively.Results:There were 2 female and 2 male patients, aged 46 to 53 years [mean (49±3.2) years], and the course of the disease spanned from 2 months to 36 years. The tumor involved the femur ( n=2), tibia ( n=1) and humerus ( n=1). Three of them were diagnosed with FD before surgery. Single photon emission computed tomography showed multiple increases in bone metabolism, CT showed poorly margin, cortical destruction and soft tissue mass with uneven enhancement. Three cases had both FD and sarcoma components, while the remaining case exhibited exclusively sarcoma. The sarcomas displayed significant morphological variation, with 1 case diagnosed as osteosarcoma and 3 cases classified as low to high grade spindle cell sarcoma. Immunohistochemical results did not provide any indications for clear classification. Sanger sequencing demonstrated GNAS mutations of p.R201H (c.CGT>CAT, n=2) and p.R201C (c.CGT>TGT, n=2). All 4 cases were followed-up for 18 to 76 months, and received chemotherapy after surgery; 2 cases maintained disease-free, one case was diagnosed with invasive breast cancer through a core needle biopsy 3 months after chemotherapy, and another one was found to relapse 18 months after surgery. Conclusions:Some cases of polyostotic FD occur in association with café-au-lait macules and/or endocrine hyperfunctioning in McCune-Albright syndrome (MAS); polyostotic FD and MAS have more malignant potential than monostotic FD, but they are not the risk factors for FD malignancy. GNAS mutations may be involved in the occurrence and development of FD. The histologic types of malignant transformation of polyostotic FD in long bone are diverse, the sarcoma components of FD also present the GNAS mutation, suggesting potential involvement in the pathogenesis of FD malignancy.

Objective:To investigate the clinicopathological features, immunophenotype and molecule characteristics of EWSR1::CREM fusion malignant epithelioid neoplasm in soft tissue.Methods:The clinical and pathological data of 2 cases of EWSR1::CREM fusion malignant epithelioid neoplasm in soft tissue diagnosed at the Department of Pathology, Beijing Jishuitan Hospital, Beijing, China from May 2023 to May 2024 were analyzed. Immunohistochemical study, fluorescence in situ hybridization (FISH) and next generation sequencing (NGS) were performed. Relevant literature was reviewed.Results:There were one male and one female patients, aged 35 and 29 years, respectively. The tumors developed in the somatic soft tissue, including calf and chest wall, and were 6.0 and 6.2 cm in size, respectively. The imaging studies suggested space-occupying lesions in muscle tissue. Case 1 did not involve the bone, while Case 2 showed fracture of the 8th rib. Microscopically, a fibrous pseudocapsule surrounded by lymphocytes and plasma cells was identified. The tumors were composed of small to medium-sized round and short spindle-shaped cells, showing nodular or sheet-like pattern. The tumor cells showed round nuclear outline, coarse chromatin with prominent nucleoli. Immunohistochemically, tumor cells showed diffuse positivity of ALK (D5F3), MUM1 and Syn, focal or patchy positivity of CKpan, EMA, S-100, NSE, WT-1 and SMA, and a high Ki-67 index (20%-30%). FISH demonstrated break-apart signals of EWSR1 gene in the 2 cases. NGS revealed EWSR1::CREM gene fusion. Case 2 showed an ATRX gene mutation. The two patients were free of recurrence or metastasis at the 10-month and 1-month follow-up, respectively.Conclusions:EWSR1::CREM fusion malignant epithelioid neoplasm is rare and lacks distinctive morphological and immunohistochemical features. FISH and NGS can help make a definitive diagnosis.

Objective:To investigate the diagnostic value of detecting MDM2 gene amplification by fluorescence in situ hybridization (FISH) in low-grade osteosarcoma (LGOS).Methods:Thirty cases of parosteal osteosarcoma (POS) and 14 cases of low-grade central osteosarcoma (LGCOS) from April 2009 to August 2022 at Beijing Jishuitan Hospital, Capital Medical University were analyzed for the presence of MDM2 gene amplification by FISH. Fifty-eight additional cases were used as negative controls (including 28 cases of fibrous dysplasia, 5 cases of giant cell tumor, 4 cases of conventional osteosarcoma, 2 cases each of periosteal osteosarcoma, reparative changes after fracture, pleomorphic undifferentiated sarcoma, low grade myofibroblastic sarcoma, fibrous dysplasia with malignant transformation, one case each of leiomyosarcoma, sclerosing epithelioid fibrosarcoma, malignant peripheral nerve sheath tumor, desmoplastic fibroma of bone, solitary fibrous tumor, aneurysmal bone cyst, clear cell chondrosarcoma, osteofibrous dysplasia, and 3 cases of unclassified spindle cell tumor).Results:Among the 30 patients with POS, 15 were male and 15 were female, ranging in age from 10 to 59 years (mean 35 years, median 30.5 years). Among the 14 patients with LGCOS, four were male and 10 were female, ranging in age from 15 to 56 years (mean 37 years, median 36 years). All except one case were successfully detected by FISH. MDM2 gene amplification was detected in 27 cases of POS (27/29,91.3%) and 8 cases of LGCOS (8/14). All the negative controls were negative for MDM2 gene amplification. The positive rate of MDM2 gene amplification was significantly different between the case group and the control group ( P<0.05). The sensitivity and specificity of MDM2 gene amplification in diagnosing POS and LGCOS were 91.3% and 100.0%; and 57.1% and 100.0%, respectively. The sensitivity and specificity of MDM2 gene amplification in diagnosing LGOS (including POS and LGCOS) were 81.3% and 100.0%, respectively. In cases where MDM2 gene was amplified, the MDM2 amplified signal was clustered. Nine cases showed increased CEP12 signal different from polyploidy which was displayed as small and weak signal points or cloud flocculent and cluster signals. Conclusions:Detection of MDM2 gene amplification by FISH is a highly sensitive and specific marker for LGOS. The interpretation criteria for FISH detection of MDM2 amplification are currently not unified. The signal characteristics need more attention when interpreting.

Objective:To investigate the clinicopathological and genetic characteristics of neuromuscular choristoma-associated desmoid type fibromatosis (NMC-DF).Methods:The clinical morphological and immunohistochemical features of 7 NMC-DF cases diagnosed from January 2013 to January 2023 in Beijing Jishuitan Hospital were retrospectively analyzed. A series of neuromuscular choristoma and neuromuscular choristoma-associated desmoid type fibromatosis were evaluated for CTNNB1 mutations, and hotspot mutations for CTNNB1 were tested in 4 NMC-DF cases using Sanger sequencing.Results:The tumors were collected from 3 females and 4 males, aged 1 to 22 years (mean 7.1 years), involving the sciatic nerve ( n=4), brachial plexus ( n=2) or multiple nerves ( n=1). The course of the disease spanned from 3 months to 10 years. Two cases were recurrent tumors. All the 7 NMC cases showed endoneurial intercalation of mature skeletal muscle fibers among the peripheral nerve fascicles, and the histologic features of the NMC-DF were strikingly similar to the conventional desmoid-type fibromatosis. By immunohistochemistry, all NMC and NMC-DF cases showed aberrant nuclear staining of β-catenin (7/7), the muscle cells in NMC were intensely immunoreactive for desmin, and the admixed nerve fibers were highlighted by NF and S-100 (7/7). Four NMC and NMC-DF had CTNNB1 mutations, 3 c.121A>G (p.T41A) and 1 c.134C>T (p.S45F). Follow-up of the 7 cases, ranging from 22 to 78 months, showed tumor recurrence in 2 patients at 3 and 8 months respectively after the first surgical resection, of which 1 patient underwent above-knee amputation. No recurrence occurred in other cases with tumor excision and neurological reconstruction surgery. There was no metastasis occurred in the 7 cases. Conclusions:NMC is a rare congenital lesion with differentiated mature skeletal muscle tissue found in peripheral nerve fascicles, and approximately 80% of patients with NMC develop a soft tissue fibromatosis. CTNNB1 mutation in the Wnt signaling pathway may be involved in the pathogenesis of NMC and NMC-DF, and S45F mutations seems to have a higher risk of disease progression.

Objective To observe the effects of Xinkang Granules-contained serum on the proliferation of cardiac fibroblasts(CFs)induced by angiotensin Ⅱ(Ang Ⅱ);To explore its mechanism of inhibiting the proliferation of CFs.Methods Taking out the heart of SD suckling rats,extracting the CFs from the hearts by differential attachment,and then they were used to replicate cell proliferation model induced by Ang Ⅱ.After transfection with miR-21 overexpressing lentivirus and growth arrest specific transcript 5(GAS5)silencing lentivirus,Xinkang Granules-contained serum was intervened for 24 hours,respectively.Cell viability was detected by cell CCK-8 method,cell cycle was detected by flow cytometry,α-smooth muscle actin(α-SMA)was detected by immunofluorescence,RT-qPCR was used to detect the mRNA expression of GAS5,miR-2l,phosphatase and tensin homolog(PTEN),Western blot was used to detect the expressions of PTEN,protein kinase B(Akt),Cyclin D1 and cyclin-dependent kinase 4(CDK4)protein.Results Compared with the blank group,the activity of CFs in the model group increased,the proportion of G0/G1 phase cells decreased,the proportion of S phase cells increased,the positive expression of α-SMA increased,the expressions of GAS5,PTEN mRNA and PTEN protein decreased,the expressions of miR-21 mRNA and Akt,Cyclin D1,CDK4 protein increased,with statistical significance(P<0.05).Compared with the model group,the activity of CFs in the Xinkang Granules group decreased,the proportion of G0/G1 phase cells increased,the proportion of S phase cells decreased,the positive expression of α-SMA decreased,the expressions of GAS5,PTEN mRNA and PTEN protein increased,while the expressions of miR-21 mRNA and Akt,Cyclin D1 and CDK4 protein decreased;The activity of CFs increased in the miR-21 overexpression group and the GAS5 silencing group,and the proportion of G0/G1 phase cells decreased,the proportion of S phase cells increased,and the positive expression of α-SMA increased,the expressions of GAS5,PTEN mRNA and PTEN protein decreased,while the expressions of miR-21 mRNA and Akt,Cyclin D1 and CDK4 protein increased,with statistical significance(P<0.05).Compared with the miR-21 overexpression group and the GAS5 silencing group,the miR-21 overexpression+Xinkang Granules group and the GAS5 silencing+Xinkang Granules group showed a decrease in CFs activity and an increase in the proportion of G0/G1 phase cells,the proportion of S phase cells decreased,the positive expression of α-SMA decreased,the expressions of GAS5,PTEN mRNA and PTEN protein increased,and the expressions of miR-21 mRNA and Akt,Cyclin D1,CDK4 protein decreased,with statistical significance(P<0.05).Conclusion Xinkang Granules-contained serum can inhibit the increase of miR-21 and Akt expression caused by silence GAS5,resist the decrease of GAS5 and PTEN caused by overexpress miR-21,inhibit the proliferation cycle of CFs,promote GAS5 to play the role of"molecular sponge",and improve the excessive proliferation of CFs induced by Ang Ⅱ.

Humans ; Arthroplasty, Replacement, Knee/adverse effects* ; Arthroplasty, Replacement, Hip/adverse effects* ; Reoperation ; Prosthesis-Related Infections ; Retrospective Studies

Objective:To investigate the knowledge, attitudes, and practices (KAP) of pulse oximetry among pediatric healthcare providers in China and analyze the factor influencing the KAP.Methods:A self-developed questionnaire was used for an online research on the KAP of 11 849 pediatric healthcare providers from 31 provinces, autonomous regions, and municipalities of China from March 11 to 14, 2022.The factors influencing the KAP of pulse oximetry among pediatric healthcare providers were examined by Logistic regression. Results:The scores of KAP, of pulse oximetry were 5.57±0.96, 11.24±1.25 and 11.19±4.54, respectively.The corresponding scoring rates were 69.61%, 74.95%, and 55.99%, respectively. Logistic regression results showed that the gender and working years of pediatric healthcare providers, the region they were located, and whether their medical institution was equipped with pulse oximeters were the main factors affecting the knowledge score (all P<0.05). Main factors influencing the attitude score of pediatric healthcare providers included their knowledge score, gender, educational background, working years, region, medical institution level, and whether the medical institution was equipped with pulse oximeters (all P<0.05). For the practice score, the main influencing factors were the knowledge score, gender, age, and whether the medi-cal institution was equipped with pulse oximeters (all P<0.05). Conclusions:Chinese pediatric healthcare providers need to further improve their knowledge about and attitudes towards pulse oximetry.Pulse oximeters are evidently under-used.It is urgent to formulate policies or guidelines, strengthen education and training, improve knowledge and attitudes, equip more institutions with pulse oximeters, and popularize their application in medical institutions.

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.