To investigate the effect of VD3 on the concentration of Ca2+in neutrophils of subclinical hypocalcemia dairy cows and the autophagy of LC3,peripheral blood of dairy cows was collected and neutrophils were isolated and extracted.Western blot was used to detect the abundance of LC3 autophagy pathway-related proteins(LC3-Ⅱ,P62,ATG5)in cells.The intracellular Ca2+concen-tration of neutrophils was detected by flow cytometry.The LC3 autophagosome was detected by la-ser confocal technology.By adding VD3 to affect intracellular Ca2+concentration levels,the effects of vitamin D3 on calcium absorption and LC3 autophagy function were investigated.Western blot result showed that the protein abundances of LC3 and ATG5 in neutrophils of subclinical hypocal-cemia dairy cows decreased,while the protein abundance of p62 increased,indicating that the auto-phagy function of LC3 in subclinical hypocalcemia dairy cows was weakened.After VD3 treatment,the flow cytometry results showed that VD3 promoted the increase of Ca2+concentration levels in neutrophils.The Western blot results showed that the LC3 autophagy function of neutrophils from healthy cows and subclinical hypocalcemia cows was enhanced under the action of VD3,indicating that VD3 promoted the concentration of Ca2+in neutrophils and then affected the LC3 autophagy function.

Purpose To investigate the clinicopathological and molecular genetic characteristics of Warthin-like mucoepidermoid carcinoma(WLMEC).Methods Eight cases of WLMEC were collected.HE staining,immunohisto-chemistry,and fluorescence in situ hybridization were performed to observe their histological morphology,immunophe-notype,and molecular genetic characteristics.Clinical information was analyzed,and follow-up was conducted.Re-sults Among the eight cases of WLMEC,three were male and five were female,aged from 28 to 65 years(median age:47 years),all occurring in the parotid gland.All eight cases had clear boundaries and appeared as polycystic structures.Besides the homogeneously eosinophilic material,blue-stained mucoid material was visible in the cyst cavi-ty.The epithelium was arranged in various ways,including single-layer,double-layer,or multi-layer.The cells had a bland-looking morphology with round or oval nuclei.Compared to Warthin tumor,the cytoplasm was significantly less eosinophilic and flatter.Mitotic figures were rare.A prominent lymphoid stroma with abundant plasma cell infiltration was observed,especially in areas adjacent to the epithelium.The tumor cells expressed CK(AE1/AE3),CK5/6,p63,and CK7.The epithelial arrangement was disordered,unlike the typical double-layered structure seen in Warthin tumor.The Ki67 index ranged from 1％to 5％.MAML2 gene rearrangement was detected in all eight cases.No recur-rence was observed during the follow-up period of 1 to 54 months after surgical resection.Conclusion WLMEC is a rare low-grade malignant tumor originating from salivary glands.Compared with Warthin tumor,patients with WLMEC are younger and more commonly female.The presence of epithelial cells that are not typically arranged in a regular double layer with strong eosinophilia,as well as the abundance of plasma cells beneath the epithelium,serve as impor-tant diagnostic clues.Performing MAML2 gene testing on suspicious cases can aid in the accurate diagnosis of this dis-ease.

To investigate the effect of VD3 on the concentration of Ca2+in neutrophils of subclinical hypocalcemia dairy cows and the autophagy of LC3,peripheral blood of dairy cows was collected and neutrophils were isolated and extracted.Western blot was used to detect the abundance of LC3 autophagy pathway-related proteins(LC3-Ⅱ,P62,ATG5)in cells.The intracellular Ca2+concen-tration of neutrophils was detected by flow cytometry.The LC3 autophagosome was detected by la-ser confocal technology.By adding VD3 to affect intracellular Ca2+concentration levels,the effects of vitamin D3 on calcium absorption and LC3 autophagy function were investigated.Western blot result showed that the protein abundances of LC3 and ATG5 in neutrophils of subclinical hypocal-cemia dairy cows decreased,while the protein abundance of p62 increased,indicating that the auto-phagy function of LC3 in subclinical hypocalcemia dairy cows was weakened.After VD3 treatment,the flow cytometry results showed that VD3 promoted the increase of Ca2+concentration levels in neutrophils.The Western blot results showed that the LC3 autophagy function of neutrophils from healthy cows and subclinical hypocalcemia cows was enhanced under the action of VD3,indicating that VD3 promoted the concentration of Ca2+in neutrophils and then affected the LC3 autophagy function.

Purpose To investigate the clinicopathological and molecular genetic characteristics of Warthin-like mucoepidermoid carcinoma(WLMEC).Methods Eight cases of WLMEC were collected.HE staining,immunohisto-chemistry,and fluorescence in situ hybridization were performed to observe their histological morphology,immunophe-notype,and molecular genetic characteristics.Clinical information was analyzed,and follow-up was conducted.Re-sults Among the eight cases of WLMEC,three were male and five were female,aged from 28 to 65 years(median age:47 years),all occurring in the parotid gland.All eight cases had clear boundaries and appeared as polycystic structures.Besides the homogeneously eosinophilic material,blue-stained mucoid material was visible in the cyst cavi-ty.The epithelium was arranged in various ways,including single-layer,double-layer,or multi-layer.The cells had a bland-looking morphology with round or oval nuclei.Compared to Warthin tumor,the cytoplasm was significantly less eosinophilic and flatter.Mitotic figures were rare.A prominent lymphoid stroma with abundant plasma cell infiltration was observed,especially in areas adjacent to the epithelium.The tumor cells expressed CK(AE1/AE3),CK5/6,p63,and CK7.The epithelial arrangement was disordered,unlike the typical double-layered structure seen in Warthin tumor.The Ki67 index ranged from 1％to 5％.MAML2 gene rearrangement was detected in all eight cases.No recur-rence was observed during the follow-up period of 1 to 54 months after surgical resection.Conclusion WLMEC is a rare low-grade malignant tumor originating from salivary glands.Compared with Warthin tumor,patients with WLMEC are younger and more commonly female.The presence of epithelial cells that are not typically arranged in a regular double layer with strong eosinophilia,as well as the abundance of plasma cells beneath the epithelium,serve as impor-tant diagnostic clues.Performing MAML2 gene testing on suspicious cases can aid in the accurate diagnosis of this dis-ease.

Objective:To investigate the pathological subtypes and clinicopathological characteristics of the non-squamous immunophenotype nasopharyngeal carcinoma (NSNPC).Methods:The clinicopathological features of the non-squamous immunophenotype nasopharyngeal carcinoma diagnosed between 2011 and 2019 at the First Affiliated Hospital of Zhengzhou University were analyzed using hematoxylin and eosin staining, immunohistochemistry, in situ hybridization, transmission electron microscopy and PCR gene rearrangement. Follow-up data were also collected.Results:There were 14 males and 9 females with a median age of 46 years (ranging from 16 to 76 years) with an average age of 45 years. Microscopically, patterns were similar to the classic nasopharyngeal carcinoma. Immunohistochemistry showed that most NSNPC cases expressed low molecular weight keratin (CK8/18, CK8 and CKL) and expressed pathway proteins in a low level (EGFR, PI3K, p-AKT and p-mTOR), which had significant difference from classic nasopharyngeal carcinoma group ( P<0.05). Other proteins including CK5/6, CKpan, CK7, Syn, CD56, CgA, SOX-10, AKT, mTOR, Notch, STAT3 and p-STAT3 showed no statistical difference between the two groups. Pathogen detection showed that EBER was positive (18/23, 78.3%) and HPV positive(2/23, 8.7%)which were HPV35 and HPV38. The cancer suppressor gene BLU was highly expressed in NSNPC; RASSF1 and Rbms3 were less expressed in NSNPC, in line with classic NPC. As a whole, NSNPC was characterized by ultrastructures of low-differentiated squamous cell carcinoma. Compared with classic nasopharyngeal carcinoma, NSNPC had a lower recurrence rate and earlier clinical stage( P<0.05),but there was no significant correlation with age, sex, distant metastasis and death ( P>0.05). Conclusions:The histological morphology, etiology and gene changes of NSNPC are similar to those of classical nasopharyngeal carcinoma and ultrastructural findings show that NSNPC still belongs to undifferentiated type in non-keratinized squamous cell carcinoma. The malignant degree of NSNPC is low and the prognosis is good.

Objective:To study the clinical manifestations, pathological features, molecular features, differential diagnosis and prognosis of secretory carcinoma of salivary gland (SCSG).Methods:Twelve cases of SCSG diagnosed in the Department of Pathology, the First Affiliated Hospital of Zhengzhou University from January 2017 to December 2019 were collected and analyzed in terms of histological morphology, immunohistochemistry (Envision method) and molecular detection.Results:Among the 12 patients, there were 6 males and 6 females, aged 12-67 years old, with a median age of 41.5 years. The lesions in 11 patients were located in the parotid gland. The maximum diameter of the tumors ranged from 0.8 to 5.5 cm. Clinically, slow-growing painless or tender mass was the first symptom, and all patients underwent surgical resection, among which 5 patients received postoperative radiotherapy and chemotherapy, and 1 patient developed local recurrence. Histopathologically, the tumor is characterized by microcystic, solid or tubular structures with unique intravitary homogeneous secretions. Immunohistochemistry showed diffuse expression of CK7, mammaglobin, GATA-3 and S-100 in all cases, and pan-Trk in 10 of the 12 cases. Sox-10 was expressed focally in 9 cases, and Ki-67 index was 5%-20%. In molecular detection, 11 cases had ETV6 gene break/fusion.Conclusions:SCSG is a relatively rare low-grade malignant salivary gland tumor, with typical histological morphology and immunophenotype, Pan-Trk immunohistochemistry may be related to NTRK fusion, ETV6-NTRK3 gene rearrangement is not only of diagnostic significance, but also Trk-targeted therapy is expected to play a greater role in clinical treatment.

Objective To explore the application value of telepathological consultation in helping grassroots hospitals.Methods 578 cases of intraoperative telepathology consultation were reviewed,and the accuracy and the timely rate of diagnosis were calculated.The systematic distribution,benign and malignant distribution,and the distribution difference in different primary hospitals were analyzed,so as to evaluate the popularization value of the intraoperative telepathology consultation.Results The accuracy rate of 578 cases of intraoperative telepathology consultation was 99.83％.The timely rate of consultation in 30min was 96.02％,and most reports could be diagnosed in 2 to 5 mins.The source of tissues involved in consultation were thyroid,breast,ovary/fallopian tube and lung.In all cases,24.39％ of the malignant tumors were found.Among the diseases of different systems,the proportion of malignant tumors is the highest in breast diseases,followed by lung,thyroid and ovary.Among the four hospitals with most of the consultations,the rate of malignant tumor in Renmin Hospital of Jianli County was the highest,followed by Renmin Hospital of Yingshan County,Renmin Hospital of Xiaochang County,and Fifth Division Hospital of Xinjiang.Conclusion Intraoperative telepathology consultation can provide accurate and timely expert consultation for grassmots hospitals,avoid the "second operations" of the patients,improve the access of medical treatment for people living in relatively remote areas,solve the shortage of pathologists at the grassroots hospitals,and improve the level of doctors' diagnosis and treatment at the grassroots hospitals,which is worth popularizing and applying in Pathology Department of the grassroots hospitals.