Objective:To investigate the clinicopathological and molecular genetic characteristics of pan cytokeratin (CKpan)-positive EWSR1/FUS::CREB fusion malignant tumors in abdominopelvic cavity.Methods:Four cases of malignant tumor with CKpan-positive EWSR1/FUS::CREB fusion were selected from January 2019 to July 2024 in the Department of Pathology, Tianjin Medical University Cancer Hospital, Tianjin, China. Their clinical, pathological, and immunohistochemical characteristics were examined. Their molecular genetic characteristics were analyzed using fluorescence in situ hybridization (FISH) and next-generation sequencing (NGS).Results:Among the 4 patients, there were 2 males and 2 females, aged 44, 44, 48 and 66 years, respectively. The tumor sites included 1 case located between the stomach and transverse colon, 1 case on the serous surface of the gastric wall, 1 case in the transverse mesocolon, and 1 case in transverse mesocolon and small mesentery. The clinical manifestations were mostly abdominal distension and abdominal pain. The maximum diameter of the tumor in the surgical resection specimen was 3.5-8.5 cm. The tumor′s cut surface was grayish-white and gray-yellow in color, with medium consistency. Microscopically, the tumor cells were mainly composed of epithelioid tumor cells, and 2 of the tumors showed that tumor cells arranged in a solid sheet or multinodular pattern, and the cytoplasm of the tumor cells was abundant, lightly stained, and the boundaries were unclear, accompanied by the formation of capsules or microcapsules, and lymphocyte and plasma cell sleeves were seen. In one case, the pseudopapillary arrangement was present, and the tumor cells were radially distributed around the fibrovascular axis. In another case, it was arranged in a pseudoacinar pattern, and the nest was surrounded by slender reticular fibers. Immunohistochemistry showed that tumor cells expressed CKpan (4/4) and WT1 (4/4, including 1 focal positive). Vimentin, CK8/18, D2-40 and S-100 were expressed in various intensities, while Calretinin was locally positive or negative. FISH showed that 2 cases had EWSR1 break-apart and 2 cases had FUS break-apart. NGS confirmed the presence of EWSR1::CREM fusion (1 case) and FUS::CREM fusion (2 cases), respectively. Except for 1 recently diagnosed case, 3 cases were followed up: 1 patient died due to tumor recurrence and metastasis (overall survival was 33 months), and 2 patients survived (1 case had recurrence 58 months after surgery, and 1 case had no recurrence or metastasis after surgery).Conclusions:CKpan-positive EWSR1/FUS::CREB fusion malignant tumor is a rare malignancy tumor with undetermined classification that tends to occur in the abdominopelvic cavity and often involves the gastrointestinal tract. Molecular testing such as FISH and NGS is helpful for a definitive diagnosis.

Objective:To investigate the clinicopathological and molecular genetic characteristics of pan cytokeratin (CKpan)-positive EWSR1/FUS::CREB fusion malignant tumors in abdominopelvic cavity.Methods:Four cases of malignant tumor with CKpan-positive EWSR1/FUS::CREB fusion were selected from January 2019 to July 2024 in the Department of Pathology, Tianjin Medical University Cancer Hospital, Tianjin, China. Their clinical, pathological, and immunohistochemical characteristics were examined. Their molecular genetic characteristics were analyzed using fluorescence in situ hybridization (FISH) and next-generation sequencing (NGS).Results:Among the 4 patients, there were 2 males and 2 females, aged 44, 44, 48 and 66 years, respectively. The tumor sites included 1 case located between the stomach and transverse colon, 1 case on the serous surface of the gastric wall, 1 case in the transverse mesocolon, and 1 case in transverse mesocolon and small mesentery. The clinical manifestations were mostly abdominal distension and abdominal pain. The maximum diameter of the tumor in the surgical resection specimen was 3.5-8.5 cm. The tumor′s cut surface was grayish-white and gray-yellow in color, with medium consistency. Microscopically, the tumor cells were mainly composed of epithelioid tumor cells, and 2 of the tumors showed that tumor cells arranged in a solid sheet or multinodular pattern, and the cytoplasm of the tumor cells was abundant, lightly stained, and the boundaries were unclear, accompanied by the formation of capsules or microcapsules, and lymphocyte and plasma cell sleeves were seen. In one case, the pseudopapillary arrangement was present, and the tumor cells were radially distributed around the fibrovascular axis. In another case, it was arranged in a pseudoacinar pattern, and the nest was surrounded by slender reticular fibers. Immunohistochemistry showed that tumor cells expressed CKpan (4/4) and WT1 (4/4, including 1 focal positive). Vimentin, CK8/18, D2-40 and S-100 were expressed in various intensities, while Calretinin was locally positive or negative. FISH showed that 2 cases had EWSR1 break-apart and 2 cases had FUS break-apart. NGS confirmed the presence of EWSR1::CREM fusion (1 case) and FUS::CREM fusion (2 cases), respectively. Except for 1 recently diagnosed case, 3 cases were followed up: 1 patient died due to tumor recurrence and metastasis (overall survival was 33 months), and 2 patients survived (1 case had recurrence 58 months after surgery, and 1 case had no recurrence or metastasis after surgery).Conclusions:CKpan-positive EWSR1/FUS::CREB fusion malignant tumor is a rare malignancy tumor with undetermined classification that tends to occur in the abdominopelvic cavity and often involves the gastrointestinal tract. Molecular testing such as FISH and NGS is helpful for a definitive diagnosis.

Purpose To investigate the diagnosis and the clinicopathological characteristics of gastric adenocarcinoma with enteroblastic differentiation(GAED)in biopsy,and to provide data for the pathological evaluation of endoscopic resection of early gastric cancer.Methods 26 GAED biopsy specimens were collected,and the clinicopathological features were ana-lyzed by re-reading slides,immunohistochemistry and with paired radical surgery specimen.Results Serum AFP was de-tected in 16 patients before operation,and 11 patients(68.75％)were elevated.The initial diagnosis rate and follow-up rate of GAED in biopsy were 42.31％and 92.31％.Among the biopsy specimens,5 cases of GAED were accompanied by conventional gastric adenocarcinoma,and the positive rates of SALL4 and Glypican-3 were high(both 78.57％).The accom-panying cancers in the radical resection specimens include con-ventional gastric adenocarcinoma,hepatoid adenocarcinoma,and low adhesion adenocarcinoma,with the positive rate of SALL4(81.25％),Glypican-3(75％),and AFP(62.5％).GAED was more prone to deep invasion of the gastric wall and lymph node metastasis than conventional gastric adenocarcinoma.A-mong the 6 cases of early GAED,3 cases(50％)were lymph node-positive.Conclusion GAED is easy to be missed in biop-sy,and more attention should be paid to its tissue morphology and immunophenotype.GAED needs to be excluded in the histo-logical type of endoscopic curative resection of early gastric canc-er.

Purpose To investigate the diagnosis and the clinicopathological characteristics of gastric adenocarcinoma with enteroblastic differentiation(GAED)in biopsy,and to provide data for the pathological evaluation of endoscopic resection of early gastric cancer.Methods 26 GAED biopsy specimens were collected,and the clinicopathological features were ana-lyzed by re-reading slides,immunohistochemistry and with paired radical surgery specimen.Results Serum AFP was de-tected in 16 patients before operation,and 11 patients(68.75％)were elevated.The initial diagnosis rate and follow-up rate of GAED in biopsy were 42.31％and 92.31％.Among the biopsy specimens,5 cases of GAED were accompanied by conventional gastric adenocarcinoma,and the positive rates of SALL4 and Glypican-3 were high(both 78.57％).The accom-panying cancers in the radical resection specimens include con-ventional gastric adenocarcinoma,hepatoid adenocarcinoma,and low adhesion adenocarcinoma,with the positive rate of SALL4(81.25％),Glypican-3(75％),and AFP(62.5％).GAED was more prone to deep invasion of the gastric wall and lymph node metastasis than conventional gastric adenocarcinoma.A-mong the 6 cases of early GAED,3 cases(50％)were lymph node-positive.Conclusion GAED is easy to be missed in biop-sy,and more attention should be paid to its tissue morphology and immunophenotype.GAED needs to be excluded in the histo-logical type of endoscopic curative resection of early gastric canc-er.

Objective: To verify the safety and efficacy of IONTRIS particle therapy system (IONTRIS) in clinical implementation. Methods: Between 6.2014 and 8.2014, a total of 35 patients were enrolled into this trial: 31 males and 4 females with a median age of 69 yrs (range 39-80). Ten patients had locally recurrent head and neck tumors after surgery, 4 cases with thoracic malignancies, 1 case with hepatocellular carcinoma, 1 case with retroperitoneal sarcoma, and 19 cases with non-metastatic prostate carcinomas. Phantom dose verification was mandatory for each field before the start of radiation. Results: Twenty-two patients received carbon ion and 13 had proton irradiation. With a median follow-up time of 1 year, all patients were alive. Among the 16 patients with head and neck, thoracic, and abdominal/pelvic tumors, 2, 1, 12, and 1 cases developed complete response, partial response, stable disease, or disease progression, respectively. Progression-free survival rate was 93.8% (15/16). Among the 19 patients with prostate cancer, biological-recurrence free survival was 100%. Particle therapy was well tolerated in all 35 patients. Twenty-five patients (71.4%) experienced 33 grade 1 acute adverse effects, which subsided at 1 year follow-up. Six (17.1%) patients developed grade 1 late adverse effects. No significant change in ECOG or body weight was observed. Conclusions IONTRIS is safe and effective for clinical use. However, long term follow-up is needed to observe the late toxicity and long term result.

Objective To verify the safety and efficacy of IONTRIS particle therapy system ( IONTRIS) in clinical implementation. Methods Between 6.2014 and 8.2014, a total of 35 patients were enrolled into this trial:31 males and 4 females with a median age of 69 yrs ( range 39?80) . Ten patients had locally recurrent head and neck tumors after surgery, 4 cases with thoracic malignancies, 1 case with hepatocellular carcinoma, 1 case with retroperitoneal sarcoma, and 19 cases with non?metastatic prostate carcinomas. Phantom dose verification was mandatory for each field before the start of radiation. Results Twenty?two patients received carbon ion and 13 had proton irradiation. With a median follow?up time of 1 year, all patients were alive. Among the 16 patients with head and neck, thoracic, and abdominal/pelvic tumors, 2, 1, 12, and 1 cases developed complete response, partial response, stable disease, or disease progression, respectively. Progression?free survival rate was 93.8％ (15/16). Among the 19 patients with prostate cancer, biological?recurrence free survival was 100％. Particle therapy was well tolerated in all 35 patients. Twenty?five patients (71.4％) experienced 33 grade 1 acute adverse effects, which subsided at 1 year follow?up. Six ( 17.1％) patients developed grade 1 late adverse effects. No significant change in ECOG or body weight was observed. Conclusions IONTRIS is safe and effective for clinical use. However, long term follow?up is needed to observe the late toxicity and long term result.

Objective To verify the safety and efficacy of IONTRIS particle therapy system ( IONTRIS) in clinical implementation. Methods Between 6.2014 and 8.2014, a total of 35 patients were enrolled into this trial:31 males and 4 females with a median age of 69 yrs ( range 39?80) . Ten patients had locally recurrent head and neck tumors after surgery, 4 cases with thoracic malignancies, 1 case with hepatocellular carcinoma, 1 case with retroperitoneal sarcoma, and 19 cases with non?metastatic prostate carcinomas. Phantom dose verification was mandatory for each field before the start of radiation. Results Twenty?two patients received carbon ion and 13 had proton irradiation. With a median follow?up time of 1 year, all patients were alive. Among the 16 patients with head and neck, thoracic, and abdominal/pelvic tumors, 2, 1, 12, and 1 cases developed complete response, partial response, stable disease, or disease progression, respectively. Progression?free survival rate was 93.8％ (15/16). Among the 19 patients with prostate cancer, biological?recurrence free survival was 100％. Particle therapy was well tolerated in all 35 patients. Twenty?five patients (71.4％) experienced 33 grade 1 acute adverse effects, which subsided at 1 year follow?up. Six ( 17.1％) patients developed grade 1 late adverse effects. No significant change in ECOG or body weight was observed. Conclusions IONTRIS is safe and effective for clinical use. However, long term follow?up is needed to observe the late toxicity and long term result.

Objective:To highlight the developmental process of 3D cell culture technology system, which is more suitable for isolating and identifying lung cancer stem-like cells than 2D cell culture technology system, and to explore the application of 3D cell cultures in the evaluation of proliferation, apoptosis, invasion, and drug resistance of lung cancer. Methods:Cells (104/well) from the human lung adenocarcinoma cell lines A549 and RPMI 1640 were cultured in complete medium containing 10%fetal bovine serum. Cell suspension was cultured in a BME basal medium. A growth curve was drawn after 7 d of culture. The stem-like cell was identified through a mammosphere culture, drug resistance and invasion assay, and flow cytometry. Data of A549 cells cultured in 3D and 2D tra-ditional cell culture technologies were compared. Results: Cells from the 3D cell culture had higher tumor formation rates [(20.75 ± 0.85) d vs. (60.25 ± 1.49) d, P<0.01)] and tumor sphere formation (28.50%± 1.17%vs. 8.67%± 0.80%, P<0.01) than those from the 2D cell culture. Moreover, cells from 3D cell culture were more invasive and resistant to therapy (58.17%± 2.19%vs. 41.70%±5.81%in 48 h, P<0.01;33.27%±5.76%vs. 27.30%±4.25%in 72 h, P<0.01). Phenotype experimental results demonstrated that the CD44 and CD326 cells were double-positive, whereas the CD24 cell was negative. Conclusion:The proportion of stem-like cells in A549 cell line after 3D cell culture significantly increased compared with 2D cell culture. The 3D cell culture can promote the proliferation of lung cancer stem cells.

Objective:To observe the clinicopathological features of gastrointestinal stromal tumor (GIST) cases with concurrent carcinoma. Methods:Patient data of 24 GIST cases with concurrent carcinoma were collected from the 157 GIST cases reported be-tween 2002 and 2012. The clinicopathological features of the GIST cases with concomitant carcinoma were studied. The expression of CD117, CD34, and SMA by the tumors was assayed using the immunohistochemical EliVision method. In particular, the expression of the proliferation marker Ki-67 was studied. Results:GIST cases with concurrent carcinoma accounted for 15.3%of the total GIST cas-es studied. The GIST patients with concurrent carcinoma included 14 males and 10 females. The male-female ratio of these patients was 1.4∶1. The age of the patients ranged from 41 years to 66 years, with a median age of 55 years. Lesions at the inferior segment of the esophagus were found in 7 of the 24 selected GIST cases;lesions at the gastric wall and in the intestines were observed in 15 and 2 cas-es, respectively. The diameter of the GIST cases with concurrent carcinoma ranged between 0.6 and 3.8 cm, with an average of 1.50 ± 0.85 cm. Slight dysplasia was observed in 4 of the 24 cases; no heteromorphism was present in the remaining 20 cases. The mitotic counts of GIST cases with concurrent carcinoma ranged from 0/50 HPF to 5/50 HPF, with an average of (0.79±1.83)/50 HPF. The pro-liferative index of Ki-67 in the GIST cases with concurrent carcinoma ranged between 0 and 7.72, with an average of 2.16 ± 3.26. The concurrent carcinoma cases included 5 cases with esophageal carcinoma, 2 with cardiac carcinoma, 15 with gastric cancer, and 2 with intestinal cancer. In contrast to the GIST cases with concurrent carcinoma, the GIST cases without carcinoma complications included 74 males and 59 females. The male-female ratio was 1.25∶1. The age of the patients without concurrent carcinoma ranged from 43 years to 71 years, with a median age of 54 years. Among the 133 GIST cases without cancer complications, gastric, intestinal, and esophageal lesions were found in 114, 13, and 6 cases, respectively. The diameter of GISTs without cancerous complications ranged from 2.4 cm to 15.5 cm, with an average of 6.11 ± 7.09 cm. Different degrees of dysplasia were seen in 82 of the 133 cases. The mitotic counts in the GIST cases without cancer complications ranged from 0/50 HPF to 53/50 HPF, with an average of (3.81±23.67)/50 HPF. The prolifera-tive index of Ki-67 for these cases ranged from 0 to 39.21 and averaged at 6.22 ± 16.96. The male-female ratio of the GIST cases with cancer complications was higher compared with the GIST cases without. The average diameter of GISTs with complications was small-er compared with that of GISTs without complications. The mitotic counts and the proliferative index of Ki-67 were significantly lower in the GIST cases with cancer complications than in those without (t=1.981, P<0.05 vs. t=1.993 5, P<0.05). Conclusion:Concurrent car-cinomas were found in 15.3% of the total GIST cases. No special clinical symptoms were observed in most GIST cases with cancer complications, as revealed when the carcinomas were examined. The proliferative index of Ki-67 in the GIST cases with concurrent car-cinoma is significantly lower compared with that of the GIST cases without complications.

Tissue factor pathway inhibitor-2 (TFPI-2), a member of the Kunitz-type family, is a broad-spectrum serine proteinase inhibitor. The expression of TFPI-2 is inversely related to increasing degree of malignancy, suggesting a role of TFPI-2 in the mainte-nance of tumor stability and inhibition of the growth of neoplasma. Aberrant methylation of TFPI-2 promoter cytosine-phosphorothio-ate-guanine (CpG) islands has been widely documented to be responsible for diminished expression of TFPI-2 mRNA and protein dur-ing cancer progression. TFPI-2 expression is significantly up-regulated by the ERK1/2 and JNK signaling pathways and modestly in-creased by VEGF, TNF-alpha, and fibroblast growth factor in time-and dose-dependent manners. TFPI-2 can maintain the stability of the tumor environment and inhibit invasiveness and growth of neoplasms. TFPI-2 has also been shown to regulate proliferation, apopto-sis, and vasculogenic mimicry of tumor cells, which may contribute significantly to tumor growth inhibition. Restoration of TFPI-2 ex-pression in tumor tissue inhibits tumor growth and metastasis, which creates a novel possibility of cancer patient treatment. This review focuses on the expression and the molecular regulation mechanisms of TFPI-2 in malignant tumors that control the functions of TFPI-2 in proliferation, apoptosis, and angiogenesis. Insight into these processes will improve our understanding of TFPI-2 and provide new ap-proaches for rational treatment strategies.