Objective:To investigate the effect of nuclear receptor subfamily 2 group F member 2(NR2F2)on the biological behaviors of human ovarian cancer SKOV3 cells,and to clarify its molecular mechauism and provide the new idea for treatment of ovarian cancer.Methods:Gene Expression Profiling Interactive Analysis(GEPIA)Database analyse the expression level of NR2F2 gene in ovarian tissue,and analyse its correlation with clinical prognosis of ovarian cancer patients.The human ovarian cancer SKOV3 cells were divided into control group and NR2F2 over-expression(NR2F2 OE)group,which were transfected with mCherry control virus and NR2F2 OE over-expression virus,respectively,when the cell deusity reached 70％,and the stable transfection SKOV3 cell lines were screened with puromycin(puro)48h lafter.Real-time fluorescence quantitative PCR(RT-qPCR)and Western blotting methods were used to detect the transfection efficiencies of the cells;RT-qPCR method was used to detect the expression levels of NR2F2 and sex-determining region Y-box 2(SOX2)mRNA in the cells in two groups;Western blotting method was used to detect the expression levels of NR2F2,ATP-binding cassette superfamily G member 2(ABCG2),and programmed cell death 1-ligand 1(PD-L1)protcins in the cells in two groups.CCK-8 assay was used to detect the proliferation activities of the cells in two groups;Wound assay was used to detect the migration rates of the cells in two groups;Transwell chamber assay was used to detect the number of transmembrane cells;Spheroidization assay was used to detect the numbers of spheroids in the cells;peripheral blood mononuclear cells(PBMCs)-mediated tumor cell killing assay was used to detect the relative densities of surviving tumor cells;CCK-8 assay was used to detect the half maximal inhibitory concentration(IC50)of paclitaxel(PTX)and carboplatin(CBP).Results:Compared with normal ovarian tissue,the expression level of NR2F2 gene in ovarian tumor tissue was decreased(P＜0.05),and decreased with the improvement of clinical pathological grading of ovarian tumor.The patients with higher expression level of NR2F2 gene had better clincal prognosis.The SKOV3 cells with NR2F2 over-expresson were successfully constructed,and the expression levels of NR2F2 mRNA and protein in the cells in NR2F2 OE group were increased compared with control group(P＜0.001).The CCK-8 assay results showed that compared with control group,the proliferation activities of the cells in NR2F2 OE group were decreased at different time points(1,2,3,and 4 d)(P＜0.05 or P＜0.01).The cell wound assay results showed that compared with control group,the migration rate of the cells in NR2F2 OE group was decreased(P＜0.001).The Transwell assay results showed that compared with control group,the number of transmembrane cells in NR2F2 OE group was decreased(P＜0.01).Compared with control group,the number of the spheroids in NR2F2 OE group was decreased(P＜0.05),and the expression levels of SOX2 mRNA(P＜0.01)and protein(P＜0.001)were increased.Compared with control group,the relative density of surviving tumor cells in NR2F2 OE group was decreased,but the difference was not significant(P＜0.05),and the expression level of PD-L1 protein was decreased(P＜0.05).Compared with control group,the proliferation activities of cells in NR2F2 OE group were decreased(P＜0.05),and the drug sensitivities of the cells to PTX and CBP were enhanced(P＜0.05);the IC50 of PTX was significantly reduced,while the IC50of CBP could not be calculated due to excessively high drug concentration;the expression level of ABCG2 protein was decreased(P＜0.05).Conclusion:The over-expression of NR2F2 may inhibit the proliferation,migration,and invasion of the human ovarian cancer SKOV3 cells,decrease the expression levels of SOX2,PD-L1 and ABCG2 proteins,suppress the stemness and immune evasion ability of the SKOV3 cells,and enhance the sensitivities of the SKOV3 cells to PTX and CBP.

Objective To analyze the influencing factors of diabetic cardiovascular autonomic neuropathy(DCAN)secondary to type 2 diabetes mellitus(T2DM).Methods A total of 63 T2DM patients admitted to Department of Endocrinology in our hospital from Nov.1,2022 to Nov.30,2023 were enrolled.All patients were examined by cardiovascular autonomic nervous system.According to the results,they were assigned to DCAN group or non-DCAN group.Baseline data and laboratory parameters were recorded for each group.Logistic regression analysis was used to identify the influencing factors of DCAN secondary to T2DM,and receiver operating characteristic(ROC)curve analysis was used to study the predictive value of the influencing factors on DCAN.Results There were 32(50.79％)patients in the DCAN group and 31(49.21％)patients in the non-DCAN group.The mean age of the patients was significantly older in the DCAN group than in the non-DCAN group(P=0.002).Logistic regression analysis showed that age was a risk factor for DCAN secondary to T2DM(odds ratio=1.095,95％confidence interval 1.029-1.166,P=0.04).The ROC curve showed that the area under curve value of age predicting DCAN was 0.718,and the best diagnostic value was 58.5 years old,with a sensitivity of 0.719 and a specificity of 0.645.Conclusion Age is a risk factor for DCAN secondary to T2DM,and an increase in age leads to a higher positive rate of DCAN.

AIM:This study aims to investigate the mechanism by which LINC00973 regulates multidrug re-sistance of non-small-cell lung cancer(NSCLC).METHODS:The GEPIA database was employed to analyze the expres-sion levels of LINC00973 in NSCLC and its correlation with patient prognosis in clinical settings.RT-qPCR was utilized to assess LINC00973 expression in various NSCLC cell lines and chemoresistant cells.The migration,invasion,stemness ca-pacity,and drug sensitivity of NSCLC cells with either overexpression or knockdown of LINC00973 were evaluated using wound healing assay,Transwell assay,sphere formation assay,and CCK-8 assay.RNA sequencing was conducted on LINC00973-overexpressing and parental NSCLC cells to identify dysregulated pathways and targets.The LncBase Pre-dicted v.2 and TargetScan databases were used to predict the microRNAs(miRNAs)co-bound by LINC00973 and their target genes.Mimics and inhibitors of candidate miRNAs were synthesized and transfected into NSCLC cells subjected to LINC00973 overexpression or knockdown.Changes in the expression level of LINC00973,and the mRNA and protein ex-pression levels of its target genes were assessed using RT-qPCR and Western blot.RESULTS:Analysis of the GEPIA da-tabase revealed that LINC00973 was significantly up-regulated in lung adenocarcinoma and lung squamous cell carcino-ma,correlating with poor prognosis of NSCLC patients.The LINC00973 expression was elevated in various NSCLC and chemoresistant cell lines(A549/DDP and A549/5-FU).Overexpression of LINC00973 in A549 and H520 cells markedly enhanced their migration,invasion,and stemness capabilities,while concurrently reducing sensitivity to chemotherapy and targeted therapies.RNA sequencing,along with RT-qPCR results,indicated that ATP-binding cassette transporter G5(ABCG5)was activated in LINC00973-overexpressing A549 and H520 cells.Further database analyses and dual trans-fection experiments confirmed that LINC00973 regulated ABCG5 expression through competitive binding with hsa-miR-150-5p.CONCLUSION:LINC00973,which is aberrantly up-regulated in NSCLC specimens and associated with poor clinical prognosis,promotes the expression of ABCG5 through competitive binding with hsa-miR-150-5p.This interaction leads to en-hanced invasion,stemness,and multidrug resistance in NSCLC cells.

AIM:This study aims to investigate the mechanism by which LINC00973 regulates multidrug re-sistance of non-small-cell lung cancer(NSCLC).METHODS:The GEPIA database was employed to analyze the expres-sion levels of LINC00973 in NSCLC and its correlation with patient prognosis in clinical settings.RT-qPCR was utilized to assess LINC00973 expression in various NSCLC cell lines and chemoresistant cells.The migration,invasion,stemness ca-pacity,and drug sensitivity of NSCLC cells with either overexpression or knockdown of LINC00973 were evaluated using wound healing assay,Transwell assay,sphere formation assay,and CCK-8 assay.RNA sequencing was conducted on LINC00973-overexpressing and parental NSCLC cells to identify dysregulated pathways and targets.The LncBase Pre-dicted v.2 and TargetScan databases were used to predict the microRNAs(miRNAs)co-bound by LINC00973 and their target genes.Mimics and inhibitors of candidate miRNAs were synthesized and transfected into NSCLC cells subjected to LINC00973 overexpression or knockdown.Changes in the expression level of LINC00973,and the mRNA and protein ex-pression levels of its target genes were assessed using RT-qPCR and Western blot.RESULTS:Analysis of the GEPIA da-tabase revealed that LINC00973 was significantly up-regulated in lung adenocarcinoma and lung squamous cell carcino-ma,correlating with poor prognosis of NSCLC patients.The LINC00973 expression was elevated in various NSCLC and chemoresistant cell lines(A549/DDP and A549/5-FU).Overexpression of LINC00973 in A549 and H520 cells markedly enhanced their migration,invasion,and stemness capabilities,while concurrently reducing sensitivity to chemotherapy and targeted therapies.RNA sequencing,along with RT-qPCR results,indicated that ATP-binding cassette transporter G5(ABCG5)was activated in LINC00973-overexpressing A549 and H520 cells.Further database analyses and dual trans-fection experiments confirmed that LINC00973 regulated ABCG5 expression through competitive binding with hsa-miR-150-5p.CONCLUSION:LINC00973,which is aberrantly up-regulated in NSCLC specimens and associated with poor clinical prognosis,promotes the expression of ABCG5 through competitive binding with hsa-miR-150-5p.This interaction leads to en-hanced invasion,stemness,and multidrug resistance in NSCLC cells.

Objective:To investigate the clinicopathological features and imaging differential diagnosis of intravascular large B-cell lymphoma (IVLBCL) in the central nervous system (CNS).Methods:A case of CNS IVLBCL with multiple intracerebral microbleeds (CMBs) diagnosed in the Department of Neurology, Qilu Hospital of Shandong University in 2017 was reported. The clinical and imaging data, histological and immunohistochemical markers were retrospectively analyzed, and the relevant literature was reviewed.Results:The patient was a 31-year-old woman presented with headache and seizures. Cranial magnetic resonance imaging (MRI) showed multifocal lesions involving mainly the cortical and subcortical white matter (bilateral cerebral hemisphere and right cerebellar hemisphere), hyperintense signal on T 2-weighted and fluid-attenuated inversion recovery images, with hypointense signal on T 1-weigthed and diffusion-weighted images and contrast enhancement in some lesions. The susceptibility weighted imaging revealed multifocal cortical or subcortical hypointense lesions, involving mainly the subcortical white matter. Brain magnetic resonance angiography was normal. Brain magnetic resonance venography showed left side transverse sinus was hypoplastic. Cranial magnetic resonance spectroscopy showed decreased N-acetylaspartylglutamate peak, elevated choline peak and inverted lipid double peak. Her symptoms and the lesions once improved after starting steroid treatment. However, CNS recurrence occurred after 1 week of steroid withdrawal. She underwent the biopsy of the right frontotemporal lobe. The pathological examination showed multiple microscopic hemorrhages and edema scattered in the brain tissue. A large number of heterologous mononuclear cells were aggregated in small blood vessels in the parenchyma and meninges. Immunohistochemical analysis revealed that the tumor was negative for Epstein-Barr virus encoded small RNAs, CD 3, CD 10, cytokeratin and CD 138, and positive for CD 20, CD 79α, B-cell lymphoma (BCL)-2, BCL-6, myelocytomatosis oncogene (C-myc) and multiple myeloma oncogene-1 (MUM-1). The Ki67 proliferation index was about 70%. The diagnosis of IVLBCL was confirmed. Conclusions:IVLBCL in CNS is a rare and swiftly progressive disease with poor prognosis. Its clinical symptoms and imaging are nonspecific. Early diagnosis and treatment is critical. Biopsy is the gold standard for diagnosis. Random skin biopsy may be helpful for the early diagnosis. Furthermore, regarding the cause of multiple CMBs, the possibility of IVLBCL should be considered in the differential diagnosis, in addition to the common causes, such as primary angiitis of the CNS and cerebral amyloid angiopathy.

Objective : To investigate the effect and mechanism of Urolithin B ( UB) on proliferation , migration , invasion , apoptosis and cell cycle of glioblastoma cells. Methods : The effects of UB on the proliferation , migration and invasion of glioblastoma U251 cells were detected by CCK⁃8 , clone formation assay , scratch healing assay , Transwell invasion assay respectively. The regulation effect of UB on cell cycle and apoptosis was detected by flow cytometry. Western blot was used to detect the effects of UB on the phosphorylation levels of downstream signaling pathway proteins ERK , AKT , p38 and JNK. Results : Compared with the control group , at 24 h and 48 h , the absorbance value of U251 cells was decreased by UB (P < 0. 01) in a dose dependent manner. UB reduced the percentage of cell clone formation (P < 0. 01) . The percentage of the scratch healing area was reduced by UB (P < 0. 05 or P < 0. 01) . The percentage of the number of invaded cells was reduced by UB (P < 0. 01) . UB could induce apoptosis (P < 0. 01) and cause the cells to stagnate in G2/M phase (P < 0. 01) . UB could significantly inhibit the phosphorylation of ERK (P < 0. 05 or P < 0. 01) and AKT (P < 0. 05 or P < 0. 01) . Conclusion UB can inhibit proliferation , migration , invasion , apoptosis and cell cycle of glioblastoma cells , and regulate the phosphorylation levels of ERK and AKT.

Objective:To explore the application effect of online and offline mixed teaching based on DingTalk platform in clinical practice teaching of respiratory department.Methods:The Batch 2016 ( n= 40) and Batch 2017 ( n=47) undergraduates of clinical medicine in Qiqihar Medical College were selected as the research objects, and they were divided into a control group and a research group. The control group adopted traditional offline theoretical teaching, while the research group adopted online-offline mixed teaching, focusing on teaching design and teaching objectives, teaching subjects, teaching resources, network platforms and teaching activities (classroom learning and evaluation). The theoretical and practical test scores of the two groups of interns were compared, and the evaluations of the students in the research group on the effect of online and offline mixed teaching were collected. Excel 2013 was used to sort out the data, and R software was used for data processing and statistical analysis. T-test was used for measurement data and Chi-square test was used for counting data. Results:The operational test scores [(28.16±1.70) points] and theoretical scores [(82.07±6.40) points] of the students in the study group were higher than those in the control group [(23.35±2.88) points and (76.20±5.67) points], with significant differences ( P<0.001). Compared with the students in the control group, the students in the research group were more satisfied with the mixed teaching method. Conclusion:The online and offline mixed teaching is conducive to improving students' comprehensive performance, learning effect, and promoting students' clinical diagnosis and treatment skills, self-study, innovation abilities and clinical competence.

Objective:To investigate the clinical manifestations and imaging features of spastic paraplegia caused by spinal cord Wallerian degeneration after pontine infarction, and to analyze its occurrence process and mechanism.Methods:The clinical manifestations and imaging features of two patients with spastic paraplegia caused by spinal cord Wallerian degeneration after pontine infarction were reported for the first time in China. Combined with anatomy and review of the literature, the mechanism, imaging manifestations and clinical features of Wallerian degeneration of lateral funiculus of spinal cord after pontine infarction were analyzed systematically.Results:Case 1 was a 65 years old female, and case 2 was a 58 years old male, who were treated in Qilu Hospital of Shandong University on December 7, 2018 and June 23, 2019 respectively. All the two patients presented with strength weakness of both limbs, hypertonia, symmetric hyperreflexia, and bilateral extensor plantar responses, which suggested spastic paraplegia secondary to upper motor neuron involvement. Spastic paraplegia appeared eight months after pontine infarction in case 1 and appeared six months after pontine infarction in case 2. Magnetic resonance imaging revealed continuous iso-T 1 and high-T 2 signals of bilateral pyramidal tracts below the pontine foci. Case 1 showed lesions of lateral cord of medulla oblongata, cervical spinal cord and thoracic spinal cord, and case 2 showed lesions of lateral cord of medulla oblongata and cervical spinal cord. At the same time, motor neuron disease and metabolic disease were excluded by electromyography and laboratory examination, inflammatory demyelinating disease was excluded by cerebrospinal fluid examination in one case. The syndromes, in combination with a continuous strip of abnormal signal revealed by magnetic resonance imaging which was consistent with the pyramidal tract and connected with the primary lesion suggested wallerian degeneration of spinal cord secondary to pontine infarction. The clinical symptoms of two cases were gradually aggravated in follow-up. Conclusions:Spinal cord Wallerian degeneration is a sequel after pontine infarction, which is related to the prognosis of the disease. A full understanding of its clinical manifestations and imaging features can avoid clinical misdiagnosis as other diseases.

Objective:To explore the etiology, clinical features and treatment of superficial siderosis of central nervous system (SSCNS) in China.Methods:The clinical data of four patients with SSCNS diagnosed by magnetic resonance imaging (MRI) and susceptibility weighted imaging (SWI) from Qilu Hospital of Shandong University during 2015—2019 were retrospectively reviewed. The etiology and clinical features of the four cases were summarized and analyzed.Results:All the four patients were male, with an average age of 60.5 years. Clinical symptoms included headache, hearing loss, and cerebellar ataxia. Brain MRI and SWI showed that hemosiderin mainly deposited (short linear T 2 signal, low SWI signal) on the surface of cerebellum, brainstem, temporal lobe, frontal lobe, and spinal cord. Potential bleeding sites were found in all four patients, including brain traumatic history, spinal intradural extramedullary cavernous hemangioma, brain metastasis and intracranial aneurysm. Patients were followed up for six months to four years. Headache symptoms improved in only one patient who received surgical treatment, while symptoms of the other three patients progressed. Conclusions:SSCNS is mainly characterized by hearing loss, progressive cerebellar ataxia and myelopathy. The diagnosis of this disease mainly depends on imaging examination. The linear low signal on the surface of T 2WI is the main basis for the diagnosis of SSCNS. Surgical treatment of bleeding sites and iron chelator are the main treatments of the disease.

Objective: To explore the clinical and radiological features of myelin oligodendrocyte glycoprotein (MOG) antibody associated disease. Methods: The clinical data of 22 MOG antibody associated disease cases treated in the Department of Neurology, Qilu Hospital of Shandong University from January 2017 to June 2019 were retrospectively analyzed. The clinical data of MOG antibody associated disease were summarized, including clinical and imaging features. Results: Of the 22 included patients with MOG antibody associated disease, the average age was 38.5 years, 13 were male and nine were female. Among them, 11 cases manifested as aquaporin-4 (AQP4)-negative neuromyelitis optica spectrum disorder (NMOSD), four cases optic neuritis, two cases transverse myelitis, one case acute disseminated encephalomyelitis (ADEM), two cases cortical encephalitis and two cases vestibular neuronitis. Magnetic resonance imaging (MRI) results showed that multiple anatomical areas were involved. Among the nine patients with optic nerve involvement, five patients had longitudinally extensive optic nerve lesions, which were longitudinally enhanced. In eight patients, MRI lesions in the spinal cord showed mostly long or short segments involvement, involving 2-5 spinal cord segments. Five cases involved the cervical spinal cord, six cases involved the thoracic spinal cord, and one case involved the lumbar spinal cord. Brain MRI abnormalities were found in 13 cases and the lesions were mostly patchy and point-shaped. MRI lesions demonstrated T₂ hyperintensity and some of them could be strengthened, which may involve the basal ganglia, thalamus, radiographic crown, frontal temporal lobe, brain stem and other parts. Among them, 16 patients were sensitive to high-dose intravenous/oral methylprednisolone in the acute phase. Seven patients had recurrence after two months to two years of follow-up. Conclusions MOG antibody associated disease include multiple manifestations. Among them, AQP4-negative NMOSD is the most common form. The clinical manifestations of patients showed diversity. Imaging is characterized by multiple parts involvement such as optic nerve, spinal cord, and brain. Most patients are sensitive to high-dose intravenous/oral methylprednisolone, and have a good prognosis in the acute phase, but some patients may relapse.