Improving fine needle aspiration to predict the tumor biological aggressiveness in pancreatic neuroendocrine tumors using Ki-67 proliferation index, phosphorylated histone H3 (PHH3), and BCL-2.

INTRODUCTION: Surgery is the only known cure for sporadic pancreatic neuroendocrine tumors (PNETs). Therefore, the prediction of the PNETs biological aggressiveness evaluated on endoscopic ultrasound-guided fine-needle aspiration (EUS-FNA) has a significant impact on clinical management. The proliferation rate of Ki-67 in PNETs can help to predict the biological aggressiveness of the tumor. In addition, there is a relatively new proliferation marker called phosphorylated histone H3 (PHH3) that can identify and quantify dividing cells in tissue samples, which is a marker highly specific to mitotic figures. Other markers such as BCL-2 also contribute to tumorigenesis and may be involved in the differentiation of neuroendocrine cells. MATERIALS AND METHODS: A retrospective observational study was performed on patients undergoing surveillance for PNETs from January 2010 to May 2021. Data collection included the patients' age, sex, tumor location, tumor size in the surgical specimen, and tumor grade in FNA. The 2019 World Health Organization (WHO) classification guideline was followed to diagnose PNETs, including grade and stage. Immunohistochemical stainings for Ki-67, PHH3 and BCL-2 in PNETs were performed. RESULTS: After excluding cell blocks containing fewer than 100 tumor cells, 44 patients with EUS-FNA and surgical resection specimens were included in this study. There were 19 cases of G1 PNETs, 20 cases of G2 PNETs, and 5 cases of G3 PNETs. The grade assigned based on the Ki-67 index was higher and more sensitive than that based on the mitotic count using H&E slides in some cases of G2 and G3 PNETs. However, there was no significant difference between the mitotic count using PHH3-positive tumor cells and the Ki-67 index to grade PNETs. All grade 1 tumors (19 cases) on surgical resection specimens were correctly graded on FNA (100 % concordance rate). Within the 20 G2 PNETs, 15 cases of grade 2 on surgical resection specimens were graded correctly on FNA based on the Ki-67 index only. Five cases of grade 2 PNETs on surgical resection specimens were graded as grade 1 on FNA when using only the Ki-67 index. Three of five grade 3 tumors on surgical resection specimens were graded as grade 2 on FNA based on the Ki-67 index only. Using only FNA Ki-67 to predict PNET tumor grade, the concordance (accuracy) rate was 81.8 % in total. However, all these eight cases (5 cases of G2 PNETs and 3 cases of G3 PNETs) were graded correctly by using the Ki-67 index plus mitotic rate (using PHH3 IHC stains). Four of 18 (22.2 %) patients with PNETs were positive for BCL-2 stain. In these 4 cases positive for BCL-2 stains, 3 cases were G2 PNETs and one case was G3 PNETs. CONCLUSION: Grade and the proliferative rate in EUS-FNA can be used to predict the tumor grade in surgical resection specimens. However, when using only FNA Ki-67 to predict PNET tumor grade, about 18 % of cases were downgraded by one level. To solve the problem, immunohistochemical staining for BCL-2 and especially PHH3 would be helpful. Our results demonstrated that the mitotic count using PHH3 IHC stains not only improved the accuracy and precision of PNET grading in the surgical resection specimens, but also could reliably be used in routine scoring of mitotic figures of FNA specimens.

Analysis of levels and Clinical value of CA19-9, NLR and SIRI in patients with Pancreatic Cancer with different Clinical Features.

This study aimed to investigate the levels of cancer antigen 19-9 (CA19-9), neutrophil/lymphocyte ratio (NLR) and systemic immune-inflammatory index (SIRI) in patients with pancreatic cancer with different clinical features and their clinical value. For this purpose, 78 patients with pancreatic cancer treated in our hospital from January 2018 to January 2021 were divided into two groups according to the NLR and SIRI calculated by blood routine and biochemical examination before operation. To observe and analyze the relationship between serum CA19-19 levels, clinical features and NLR and SIRI values in patients with different clinical stages and tumor diameter, and the diagnostic value of CA19-9, NLR and SIRI in pancreatic cancer. The results showed that the level of serum CA19-9 in patients with stage IV pancreatic cancer was significantly higher than that in patients with stage I, II and III, and the level of CA19-9 in patients with stage III was higher than that in patients with stage I and II, and the level of CA19-9 in patients with stage II was higher than that in patients with stage I. The level of CA19-9 in patients with pancreatic cancer with tumor diameter ≥ 5cm was significantly higher than that in patients with tumor diameter < 5cm. The percentage of patients with age ≤ 60 years old in high NLR group (83.78%) was significantly higher than that in low NLR group (63.41%), and the rate of patients with tumor diameter ≥ 5cm in the high SIRI group (54.05%) was significantly higher than that in low NLR group (26.83%). The rate of patients with tumor diameter ≥ 5cm in the high SIRI group (61.92%) was significantly higher than that in the low SIRI group (31.58%). Among the single indexes, the sensitivity of CA19-9 in the diagnosis of pancreatic cancer was the highest (93.10%). The specificity of Siri in the diagnosis of pancreatic cancer was 89.70%. The sensitivity and specificity of combined detection were 89.7% and 70.00%, respectively. In general, the level of serum CA19-9 in patients with pancreatic cancer increases gradually with the increase of clinical stage and tumor diameter. NLR value is related to patient age and tumor diameter, and SIRI value is related to tumor diameter. Combined detection of CA19-9, NLR and SIRI are of higher value in the early diagnosis of pancreatic cancer.

Pomiferin targeting SLC9A9 based on histone acetylation modification pattern is a potential therapeutical option for gastric cancer with high malignancy.

Changes in histone acetylation status are associated with gastric cancer (GC) progression. Pomiferin is a natural flavonoid, however, the specific role of pomiferin in the treatment of GC is still unclear, and its targets are not well clarified. In this work, the prognostic genes related with histone acetylation in GC were screened by univariate Cox analysis. Next, a risk model of was constructed using least absolute shrinkage and selection operator-Cox regression analyses, and multivariate Cox analysis was used for identifying the independent risk factor. Molecular docking was performed using AutoDock Vina to validate the interaction between solute carrier family 9 member A9 (SLC9A9) and pomiferin. In vitro and in vivo models were applied to investigate the tumor-suppressive role of pomiferin against GC. The inhibitory effects of pomiferin on EGFR/PI3K/AKT signaling were valdiated by Western blotting, immunofluorescence staining and qPCR. Here, a prognostic risk model based on histone acetylation regulators was established, and SLC9A9 was identified as a risk factor associated with histone acetylation status in GC. SLC9A9 expression was associated with abnormal immune microenvironment of tumor. Pomiferin had a high binding affinity with SLC9A9, and both pomiferin treatment and depletion of SLC9A9 repressed the malignant phenotypes of GC cells. Mechanistically, pomiferin inactivates EGFR/PI3K/AKT signaling in GC cells. In summary, SLC9A9, as a indicator of abnormal histone acetylation status of GC, functions as an oncogenic factor. Pomiferin binds with SLC9A9 to inactivate EGFR/PI3K/AKT pathway, to block GC progression, suggesting it is a promising drug for the patients with highly malignant GC.

Simultaneous osseous metaplasia nodules of the submucosa and mesosalpinx after first trimester abortion: a case report.

OBJECTIVES: Here, we report a case of simultaneous osseous metaplasia nodules of the submucosa and mesosalpinx after a first trimester abortion. CASE PRESENTATION: A 36-year-old woman presented to the Women's Hospital, School of Medicine, Zhejiang University with vaginal bleeding and infertility resulting from osseous metaplasia nodules of the submucosa and mesosalpinx after a first trimester abortion. Diagnostic and operative hysteroscopy and laparoscope procedures were performed. The osseous metaplasia nodules disappeared after hysteroscopy and laparoscope interventions; 2 weeks postoperatively, the patient underwent a transvaginal ultrasound examination and the abnormal ultrasound appearance had resolved. CONCLUSIONS: Osseous metaplasia nodules in the submucosa and mesosalpinx can be a rare cause of vaginal bleeding and infertility. Autologous tissue, not persistent heterologous tissue, may be the true reason for metaplasia. Treatment is by ultrasound-guided curettage or by hysteroscopic and laparoscope removal.

Co-administration of inactivated avian influenza virus with CpG or rIL-2 strongly enhances the local immune response after intranasal immunization in chicken.

Intranasal delivery of vaccines is the most effective means of inducing effective immunity in the upper respiratory tract as well as other mucosal lymphoid tissues. To evaluate the effects of the H5N2 inactivated virus with adjuvant, 120 one-day-old chicks were intranasal immunized with the H5N2 inactivated virus respectively mixed with adjuvant CpG or recombinant IL-2 (rIL-2). The local immunocompetent cells on the respiratory tract were detected. The results showed that the number of intraepithelial lymphocytes (IELs), CD3(+) T lymphocytes and mast cells in respiratory tract increased significantly respectively and the number of IgA and IgG secreting cells increased significantly after immunization. However, there was no significant change in the immunocompetent cells of the animals administrated H5N2 inactivated virus alone compared to the control group. Our results indicated that intranasal administration of H5N2 inactivated virus with adjuvant CpG or rIL-2 could be beneficial to the local immune response in the respiratory tract.