Objective:To discuss the effect of urolithin C(UC)on the proliferation,apoptosis,and autophagy of the acute myeloid leukemia(AML)HL-60 cells,and to clarify its mechanism.Methods:The HL-60 cells were divided into different concentrations(20,40,60,80,and 100 μmol·L-1)of urolithin A(UA)groups,urolithin B(UB)groups,and UC groups.CCK-8 assay was used to detect the proliferation activity of the cells in various groups;the morphology of the cells in different concentrations of UC groups was observed under optical microscope.The HL-60 cells were divided into different concentrations(0,20,40,and 80 μmol·L-1)of UC groups and 3-methyladenine(3-MA)combined with different concentrations(0,20,40,and 80 μmol·L-1)of UC groups.CCK-8 assay was used to detect the proliferation activities of the cells in various groups.The HL-60 cells were divided into control group(0 μmol·L-1)and different concentrations(20,40,and 80 μmol·L-1)of UC groups.The live/dead cell staining method was used to detect the dead rates of the cells in various groups;flow cytometry was used to detect the apoptotic rates of the cells in various groups;the autophagy of the cells was detected by autophagy staining kit(monodansylcadaverine,MDC)method;real-time fluorescence quantitative PCR(RT-qPCR)method was used to detect the expression levels of Beclin 1,autophagy related gene 9(ATG9),and autophagy related gene 7(ATG7)mRNA in the cells in various groups;Western blotting method was used to detect the expression levels of cysteinyl aspartate specific proteinase-3(Caspase-3),cleaved cysteinyl aspartate specific proteinase-3(Cleaved Caspase-3),microtubule-associated protein 1 light 3(LC-3),extracellular regulated protein kinases(ERK),phosphorylated ERK(p-ERK),AMP-activated protein kinase(AMPK),and phosphorylated AMPK(p-AMPK)in the cells in various groups.Results:The CCK-8 assay results showed that after cultured for 24,48,and 72 h,compared with 0 μmol·L-1 UA,UB,and UC groups,the proliferation activities of the cells in different concentrations of UA,UB,and UC groups were decreased(P＜0.01)with a concentration-and time-dependent manner;at 48 h,compared with UA and UB,the half-maximal inhibitory concentration(IC50)of UC was the lowest.The cell morphology observation results showed that compared with control group,the intercellular connection and the number of the cells were decreased with the increasing of UC concentration,and the cell fragment was increased.The CCK-8 assay results showed that compared with 40 and 80 μmol·L-1 UC groups,the proliferation activities of the cells in 3-MA combined with 40 and 80 μmol·L-1 UC groups were increased(P＜0.05 or P＜0.01).The live/dead cell staining results showed that compared with control group,the dead rates of the cells in 40 and 80 μmol·L-1 UC groups were increased(P＜0.01).The flow cytometry results showed that compared with control group,the apoptotic rate of the cells in 80 μmol·L-1 UC group was increased(P＜0.01).The MDC method results showed that with the increasing of UC concentration,the green fluorescence in the cells in different concentrations of UC groups was gradually intensified.The RT-qPCR results showed that compared with control group,the expression levels of Beclin 1,ATG9,and ATG7 mRNA in the cells in 80 μmol·L-1 UC group were increased(P＜0.01).The Western blotting results showed that compared with control group,the expression levels of Cleaved Caspase-3 protein in the cells in 20,40,and 80 μmol·L-1 UC groups were increased(P＜0.01),the ratio of membrane LC3/cytoplasmic LC3(LC3-Ⅱ/LC3-Ⅰ)in the cells in 80 μmol·L-1 UC group was increased(P＜0.05),and the ratios of p-AMPK/AMPK and p-ERK/ERK in the cells in 40 and 80 μmol·L-1 UC groups were increased(P＜0.01).Conclusion:UC can inhibit the proliferation of the AML HL-60 cells,induce the apoptosis and autophagy,and increase the phosphorylation levels of ERK and AMPK proteins in the cells.

Objective:To observe the changes of serum micro-inflammatory and nutritional status in elderly peritoneal dialysis patients after treatment with bifidobacterium triple viable bacteria, and the impact of Bifidobacterium triple viable bacteria treatment on the endpoint.Methods:Totally 180 elderly patients receiving peritoneal dialysis were divided into control group and observation group, with 90 cases in each.Both groups were treated on the basis of the routine treatment, the observation group was treated with oral Bifidobacterium triple viable bacteria for twelve months.Before treatment, 6 months and 12 months after treatment, fasting venous blood from the patients in the two groups were collected in the morning.The levels of serum micro-inflammatory indexes[tumor necrosis factor-α(TNF-α), interleukin-6(IL-6), interleukin-8(IL-8), C-reactive protein(CRP)]were detected by ELISA.The nutritional status and dialysis adequacy indexes[nutritional status: albumin(ALB), hemoglobin(Hb), transferrin(TRF), prealbumin(PA), calcium ion, phosphorus ion; indicators of dialysis adequacy: serum creatinine(Scr), blood uric acid(BUA), blood urea nitrogen(BUN), cystatin C(Cys-c)levels]were detected by automatic blood biochemical analyzer.After 24 months of follow-up, the occurrence of endpoint events(peritonitis, abdominal pain, malnutrition, abdominal infection, cardiovascular and cerebrovascular accidents)in the two groups was recorded.Results:After 24 months of treatment, the levels of TNF-α, IL-6, IL-8 and CRP in the two groups were lower than those before treatment, and the observation group was lower than control group[(25.7±4.0)μg/L vs.(33.6±6.0)μg/L, (2.9±0.7)ng/L vs.(4.9±1.2)ng/L, (17.0±7.2)ng/L vs.(22.8±7.9)ng/L, (4.6±0.7)mg/L vs.(6.9±1.2)mg/L]( t=10.272, 13.134, 5.040, 15.575, respectively, P<0.05 for all). After 24 months of treatment, the levels of ALB, Hb, TRF, PA and calcium ion of the two groups were higher than before treatment, and the levels of phosphorus ion were lower than before treatment, and the changes of the above indicators in observation group were more obvious than those in the control group[(45.7±5.2)g/L vs.(39.8±4.9)g/L, (72.7±8.0)g/L vs.(68.6±9.0)g/L, (4.3±1.0)g/L vs.(3.0±0.6)g/L, (321.5±29.0)mg/L vs.(297.6±25.1)mg/L, (4.9±1.3)mmol/L vs.(2.9±0.9)mmol/L, (1.3±0.9)/L vs.(1.8±0.3 mmol/L)]( t=7.737, 3.213, 9.880, 5.9 00, 11.937, 4.415, P<0.05 for all). There was no significant difference in intestinal flora between the two groups before treatment( P>0.05). After 24 months, an increase was observed in Lactobacilli and Bifidobacteria in both groups, while Enterobacteria and Enterococcus in both groups were decreased, and the changes of the above indicators in the observation group were also more obvious than those in the control group[(8.4±0.9)IgCFU/g ratio(6.4±0.9)IgCFU/g, (8.8±1.3)IgCFU/g ratio(7.9±1.3)IgCFU/g, (7.1±0.9)IgCFU/g ratio(8.0±1.1)IgCFU/g, (5.4±0.7)IgCFU/g ratio(6.9±0.9)IgCFU/g]( t=14.248, 4.339, 5.825, 12.753, P<0.05 for all). There was no significant difference in dialysis adequacy index between the two groups before treatment( P>0.05 for all). After 24 months of treatment, the levels of Scr, BUA, BUN and Cys-C in both groups decreased, and those of the observation group were lower than those of the control group[(471.5±50.5)μmol/L vs.(623.3±62.6)μmol/L, (17.5±0.5)mmol/L vs.(20.6±1.8)mmol/L, (16.4 ± 3.0)mmol/L vs.(22.5±2.0)mmol/L, (1.9±0.5)mg/L vs.(3.0±0.7)mg/L]( t=17.877, 14.976, 15.842, 11.749, P<0.05 for all). The incidence of endpoint events in the observation group was significantly lower than that in the control group(12.2% vs.2.2%, t=6.574, P<0.05 for all). Conclusions:After the treatment of Bifidobacterium triple viable bacteria in elderly peritoneal dialysis patients, the micro-inflammatory state of the patients was reduced, the nutritional status was improved, and the incidence of endpoint events was low, and has high clinical promotion and application value.

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 correlation between serum calcitonin gene-related peptide (CGRP) and the culture outcome of in vitro fertilization embryo in male patients with infertility.Methods:In this study, the randomized samples from 25 male patients who received in vitro fertilization-embryo transfer (IVF-ET) in the 1st Affiliated Hospital of Fujian Medical University from June 2019 to October 2019 were analyzed by multiple linear stepwise regression, with some important clinical outcomes, such as the logarithmic conversion index of serum CGRP, fertilization method, masturbation difficulty, age, infertility duration, and prolactin, as independent variables, while total fertilization rate, normal fertilization rate, high quality embryo rate at day 3, blastocyst formation rate as dependent variables.Results:The Pearson correlation analysis showed that the D3 high-quality embryo rate was related to the normal sperm morphology rate in the primary infertility group ( r=0.537, P=0.048), the blastocyst formation rate was correlated with sperm density ( r=0.760, P=0.002), the CGRP logarithm was correlated with the total fertilization rate ( r=0.693, P=0.006). The logarithmic conversion index of serum CGRP was related to the total fertilization rate and normal fertilization rate in the secondary infertility group ( r=0.614, P=0.042 and r=0.611, P=0.046). In the secondary infertility group, there was a linear relationship between normal fertilization rate and total sperm count, serum CGRP log conversion, and sperm normal morphology rate, with standardized regression coefficients of 0.2, -0.729, and 6.8, respectively. Conclusion:Serum CGRP level, together with total sperm count and normal sperm morphology rate may affects normal fertilization rate in male patients with infertility.

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 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 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.

Objective:To explore the effect of intervention of E-cadherin (E-cad) and B-lymphoma Moloney murine leukemia virus insertion region-1 (Bmi-1) mediated by transcription activator-like effector nuclease (TALEN) on the biological behaviors ofnasopharyngeal carcinoma cells.Methods:Multi-locus gene targeting vectors pUC-DS1-CMV-E-cad-2A-Neo-DS2 and pUC-DS1-Bmi-1 shRNA-Zeo-DS2 were constructed,and the E-cad and Bmi-1 targeting vectors were transferred with TALEN plasmids to CNE-2 cells individually or simultaneously.The integration of target genes were detected by PCR,the expressions of E-cad and Bmi-1 were detected by Western blot.The changes of cell proliferation were detected by cell counting kit-8 (CCK-8) assay.T-he cell cycle and apoptosis were detected by flow cytometry.The cell migration and invasion were detected by Transwell assay.Results:The E-cad and Bmi-1 shRNA expression elements were successfully integrated into the genome of CNE-2 cells,the protein expression level of E-cad was up-regulated,and the protein expression level of Bmi-1 was down-regulated.The intervention of E-cad and Bmi-1 didn't affect the proliferation,cell cycle and apoptosis of CNE-2 cells,but it significantly inhibited the migration and invasion ability of CNE-2 cells.Furthermore,the intervention of E-cad and Bmi-1 together significantly inhibited the migration ability of nasopharyngeal carcinoma cells compared with the intervention of E-cad or Bmi-1 alone (all P＜0.01).Conclusion:The joint intervention of E-cad and Bmi-1 mediated by TALEN can effectively inhibit the migration and invasion of nasopharyngeal carcinoma cells in vitro,which may lay the preliminary experimental basis for gene therapy of human cancer.