Objective To investigate the mechanism of hippocampal neuron death,and to explore the relationship between necroptosis of hippocampal neurons and impaired learning and memory in vascular dementia(VaD)mice model.Methods Except the sham group,the mouse model of chronic cerebral hypoperfusion vascular dementia was constructed by bilateral occlusion of the common carotid arteries(2VO).Morris water maze was used to evaluate the cognitive of mice.The mRNA expressions of RIPK1,RIPK3 and MLKL in the hippocampal region of mice was detected by real-time fluorescence quantitative PCR(qPCR).The protein expressions of RIPK1,RIPK3,pRIPK3 and MLKL were detected by Western blotting,and their correlation with the cognitive ability was analyzed.The co-localization of RIPK1,RIPK3 and MLKL was detected by immunofluorescence double labeling.Results qPCR showed that compared with sham group,mRNA expressions of RIPK1,RIPK3 and MLKL in hipp-ocampal neurons in VaD group were significantly increased.Western blot results showed that the protein levels of RIPK1,pRIPK3 and MLKL in hippocampal neuron of VaD group were also significantly increased.The co-expression localization of RIPK1,RIPK3 and MLKL was significantly enhanced in the hippocampus of the mouse model by immunofluorescence double labeling.Further study showed that the expressions of RIPK1,RIPK3 and MLKL were negatively correlated with the target quadrant residence time and the number of platform crossing in the mouse model.Conclusion In the VaD mouse model,the loss of hippocampal neurons and the development of learning and memory dysfunction may be caused by the activation of RIPK1/RIPK3/MLKL signaling pathway,which leads to the programmed necrosis of neurons.

Objective To investigate the mechanism of hippocampal neuron death,and to explore the relationship between necroptosis of hippocampal neurons and impaired learning and memory in vascular dementia(VaD)mice model.Methods Except the sham group,the mouse model of chronic cerebral hypoperfusion vascular dementia was constructed by bilateral occlusion of the common carotid arteries(2VO).Morris water maze was used to evaluate the cognitive of mice.The mRNA expressions of RIPK1,RIPK3 and MLKL in the hippocampal region of mice was detected by real-time fluorescence quantitative PCR(qPCR).The protein expressions of RIPK1,RIPK3,pRIPK3 and MLKL were detected by Western blotting,and their correlation with the cognitive ability was analyzed.The co-localization of RIPK1,RIPK3 and MLKL was detected by immunofluorescence double labeling.Results qPCR showed that compared with sham group,mRNA expressions of RIPK1,RIPK3 and MLKL in hipp-ocampal neurons in VaD group were significantly increased.Western blot results showed that the protein levels of RIPK1,pRIPK3 and MLKL in hippocampal neuron of VaD group were also significantly increased.The co-expression localization of RIPK1,RIPK3 and MLKL was significantly enhanced in the hippocampus of the mouse model by immunofluorescence double labeling.Further study showed that the expressions of RIPK1,RIPK3 and MLKL were negatively correlated with the target quadrant residence time and the number of platform crossing in the mouse model.Conclusion In the VaD mouse model,the loss of hippocampal neurons and the development of learning and memory dysfunction may be caused by the activation of RIPK1/RIPK3/MLKL signaling pathway,which leads to the programmed necrosis of neurons.

Objective:To study the diagnostic clues and significance in serous effusion cytology associated with lymphoblatic lymphoma/acute lymphoblastic leukemia (LBL/ALL).Methods:Forty-five serous effusion specimens with final diagnosis of LBL/ALL were collected from August 2011 to December 2019 at the First Affiliated Hospital of Zhengzhou University. All cases were reviewed for their clinical profiles, cytomorphologic features and ancillary studies. Cell blocks and immunocytochemistry were prepared in 22 cases; flow cytometric immunophenotyping was performed in three cases and gene rearrangement analysis (T-cell recepter, TCR and immunoglobulin, Ig) was performed in five cases.Results:Among the 45 cases, there were 35 males and 10 females with male to female ratio of 3.5∶1.0. The median age was 15 years. Mediastinal mass was the initial presentation in 39 patients (86.7%) and high LDL level were observed in 34 patients (75.6%). Microscopically, the majority of the specimens (86.7%) were hypercellular. The smears demonstrated dispersed lymphoblasts that were predominantly small to intermediate in size with scanty basophilic cytoplasm and irregular or convoluted nuclei with fine chromatin condensation and inconspicuous nucleoli. Mitoses were frequently observed. Karyorrhexis and apoptosis were seen in all cases. By immunophenotyping, TdT was expressed in 19 cases (86.4%) and CD99 in 20 cases (90.9%). Ki-67 expression varied from 65% to 95%. Flow cytometry in three cases demonstrated positivity for TdT, CD2, CD3 and CD7. Monoclonal TCR gene rearrangement was found in 4 of 5 cases, and both monoclonal TCR and Igκ gene were found in 1 case.Conclusions:In LBL/ALL, primary diagnosis could be made basing on clinical features (younger male patients with a mediastinum mass) and cytomorphology (monotonous, small to medium sized lymphoid cells with prominent irregular nuclei, fine chromatin and frequent mitoses, karyorrhexis and apoptosis). If immunocytochemistry and other ancillary studies are performed, the accuracy and reliability of the results could be improved.