1.Ligustrazine induces rat bone morrow mesenchymal stem cells to differentiate into neuron-like cells: Screening of the optimal inductive concentration
Bing CHEN ; Yanqing YIN ; Junlong KE ; Xinhui ZOU ; Hao PENG ; Shanfeng TAN ; Zhien XU
Chinese Journal of Tissue Engineering Research 2010;14(6):1072-1077
BACKGROUND: There are numerous inducers used in inducing bone marrow mesenchymal stem cells (BMMSCs) differentiate into neuron-like cells, however, due to poisonous, most chemical inducers can not be used in human.OBJECTIVE: To investigate the effect of ligustrazine on differentiation of rat BMMSCs into neuron-like cells in vitro, and to search for the optimal inductive concentration.METHODS: After SD rats were anesthetized, bone marrow was obtained from the femoral and tibial bones, centrifuged, and the supernatant was discarded. The extracted cells were cultured in L-DMEM supplemented with 15% fetal bovine serum. The expression of CD44 and CD45 of the 5~(th) passage of BMMSCs were identified by immunocytochemical technique. Serum-free L-DMEM medium contains 1.00, 1.25 and 1.50 g/L ligustrazine concentrations were used to induce the 5~(th) passage of BMMSCs in vitro. Morphology changes of BMMSCs were observed under an inverted phase microscope. Expression of nestin, neuron-specific enolase and glial fibrillary acidic protein were identified by immunocytochemical technique, and the expression ratio of neuron-like cells' surface antigens induced by different concentrations of ligustrazine were compared.RESULTS AND CONCLUSION: ①Most primarily cultured BMMSCs adhered to the wall at 3 days after culture, which proliferated faster after passaged, and the 5~(th) passage of cells were mostly purified into BMMSCs, spread radially or vortex-likely. ②The 5~(th) passage of BMMSCs was positive expressed (98.02±0.81)% CD44, but negative for CD45. ③Neuron-like cells with prominence and bifurcation could be seen after induction. The immunocytochemical method showed that nestin and neuron-specific enolase in most induced cells were positive expressed, especially received a highest ration of neuron-specific enolase expressing in the induced group with 1.25 g/L concentration of ligustrazine. It revealed that ligustrazine can induce BMMSCs differentiated into neuron-like cells, and 1.25 g/L is the optimal inductive concentration.
2.Multiple acute cerebral infarcts as initial manifestation of occult systemic malignancy with 12 case report
Wei SUN ; Haiying XING ; Qing PENG ; Zhi ZHOU ; Lili SUN ; Junlong SHU ; Xi MEN ; Ran LIU ; Ke XU ; Yining HUANG
Chinese Journal of Nervous and Mental Diseases 2016;42(7):385-389
Objective To characterize the clinical manifestations, laboratory findings of patients with occult sys?temic malignant neoplasms, whose initial manifestation presented as multiple acute cerebral infarcts including coagula?tion function,radiological imaging and microembolic signals (MES) detection by transcranial Doppler sonography (TCD) and to explore the possible underlying mechanisms. Methods All clinical records, laboratory hematological tests includ?ing hypercoagulable states measured by D-dimer levels, brain MRI including DWI, and TCD monitoring MES, the treat?ment and prognosis were retrospectively reviewed in 12 patients with multiple acute cerebral infarcts as the first manifes?tation of occult systemic malignancy. Results The clinical manifestations presented as localized neurological dysfunction, e.g. hemiparesis, aphasia, hemiparesthesia, dysarthria, vertigo and seizures, etc. DWI revealed multiple disseminated acute cerebral infarcts in multiple arterial territories such as the bilateral anterior or anterior plus posterior cerebral circu?lation in all patients. Eleven of 12 patients tested had elevated D-dimer. TCD detected MES in 5 of 7 patients. There were 12 patients diagnosed with occult systemic malignancy including 5 lung cancer, 3 pancreatic cancer, 1 gastric can?cer, 1 colon cancer, 1 endometrial adenocarcinoma and 1 metastatic poorly differentiated mucinous adenocarcinoma with unknown primary. Ten patients already had remote metastasis at diagnosis. The prognosis was usually poor and there were 7 cases with ischemic stroke recurrence, 4 cases with acute myocardial infarction, 3 cases died during hospitaliza?tion. Conclusions When patients present with multiple disseminated acute cerebral infarcts involving multiple arterial territories as initial manifestation, the underlying occult systemic malignancy should be considered. Hypercoagulopathy and MES might provide the clues to the diagnosis.
3.Clinical manifestations and microemboli signals in patients with hypercoagulability related multiple acute cerebral infarcts within non-single arterial territories
Wei SUN ; Yajun YAO ; Haiying XING ; Qing PENG ; Junlong SHU ; Xi MEN ; Ran LIU ; Ke XU ; Yining HUANG
Chinese Journal of Nervous and Mental Diseases 2016;42(8):488-492
Objective To investigate the clinical features and TCD-detected microembolic signals in patients with hypercoagulability related multiple acute cerebral infarcts within non-single arterial territories, and to explore the possi?ble underlying mechanisms. Methods A retrospective review was conducted on all clinical, laboratory, radiological and TCD monitoring records from patients with hypercoagulability related multiple acute cerebral infarcts within non-single arterial territories, who admitted to the neurology department in our hospital. Results The data from twenty-two cases were finally included in this study. All patients presented with acute-onset localized neurological dysfunction, e.g. hemi?paresis, aphasia, hemiparesthesia, dysarthria, hemianopsia and cortical blindness. Their hypercoagulability related diseas?es included 10 cases of systemic malignancy, 5 moderate to severe hyperhomocystynemia (HCY>50μmol/L), 2 nephrot?ic syndrome, 2 antiphospholipid syndrome, 1 ulcerative colitis, 1 polycythemia vera,1 paroxysmal nocturnal hemoglobin?uria. In 18 cases, the hypercoagulability related diseases were diagnosed after their initial stroke onset. DWI showed mul?tiple disseminated acute cerebral infarcts in non-single arterial territories involving bilateral anterior or anterior plus pos?terior cerebral circulation simultaneously. Foci involved lobar cortex/subcortex of cerebral hemisphere in 22 cases, deep cerebral hemisphere in 12 cases, cerebellum foci in 10 cases,brainstem foci in 2 cases. TCD revealed microembolic sig? nals in ten of 22 patients monitored. Conclusions Patients with multiple acute cerebral infarcts involving non-single arte?rial territories, should be screened for hypercoagulability as in that hypercoagulability and microembolism might be in?volved in the etiology of cerebral infarction.
4.Effects of cardiac function and renal function on early neurological function recovery in patients with acute stroke
Ailing ZHANG ; Ke XU ; Haiying XING ; Wei SUN ; Qing PENG ; Fan LI ; Yining HUANG ; Weiping SUN ; Ran LIU ; Junlong SHU
Chinese Journal of Neurology 2019;52(6):463-471
Objective To investigate the effect of cardiac function and renal function on early neurological function recovery of acute stroke patients.Methods Hospitalized acute stroke patients from January 2010 to May 2018 were recruited in the Department of Neurology,Peking University First Hospital,who were divided into two groups according to the modified Rankin Scale (mRS) score at discharge,good recovery (mRS score ≤2) and poor recovery (mRS score >2).The clinical characteristics,laboratory and echocardiography data were collected respectively.All cases were classified according to Trial of Org 10 172 in Acute Stroke Treatment (TOAST) criteria.Estimated glomerular filtration rate (eGFR) was calculated by chronic kidney disease epidemiology collaboration equation in renal function assessment,and cardiac function was calculated by left ventricular ejection fraction (LVFF) and E-wave/A-wave ratio (E/A).Multivariate Logistic regression analysis and stratified analysis were performed to explore the effects of cardiac and renal function and interactions on stroke patients.Results A total of 517 patients with acute stroke were enrolled in this study,23.4% (121/517) of which presented with chronic kidney disease.Poor recovery was aggravated with the progression of eGFR stage in patients with acute stroke (x2=14.627,P=0.001).Compared with the good recovery group,LVEF and E/A were significantly lower in the poor recovery group,while left atrium diameter ((3.87±0.52) cm vs (3.77±0.49) cm,t=-2.139,P=0.033),interventricular septum thickness ((1.10±0.19) cm vs (1.04±0.16) cm,t=-4.056,P=0.000),left ventricular posterior wall thickness ((1.00±0.13) cm vs (0.98±0.13) cm,t=-2.190,P=0.029) and left ventricular mass index ((102.03±25.73) g/m2 vs (94.94±23.63) g/m2,t=-3.145,P=0.002) were significantly higher in the poor recovery group.Stratified analysis showed that the rate of poor recovery increased with the decrease of eGFR at different levels of LVEF and E/A.Compared with patients of normal renal function and mild impairment of renal function or with patients of high third tertile of LVEF,chronic kidney disease significantly increased the rate of poor recovery in patients with low third tertile of LVEF,while the rate of poor recovery was not significantly different in patients with chronic kidney disease among third tertile of E/A.The levels of eGFR and LVEF in different stroke subtypes were significantly different (F=7.433,P=0.000;F=2.617,P=0.034).The eGFR and LVEF levels of the cardioembolism (CE) group were the lowest compared with other subtypes of stroke.The eGFR levels in the CE group were significantly lower than that in other subtypes except the large artery atherosclerosis group,and the LVEF level was significantly lower in the CE group compared to the small artery occlusion group.Multivariate Logistic regression analysis revealed that eGFR (OR=0.866,95%CI 0.760-0.987,P=0.031),LVEF (OR=0.798,95%CI 0.688-0.925,P=0.003),E/A (OR=0.136,95%CI 0.034-0.548,P=0.005) and eGFR by LVEF (OR=1.002,95%CI 1.000-1.004,P=0.022) were significantly associated with early functional outcome in patients with acute stroke after adjusting for relevant clinical confounders (all P<0.05).Conclusions Renal function,cardiac systolic and diastolic function were associated with the early functional recovery in patients with acute stroke.Moreover,cardiac systolic dysfunction and renal dysfunction interacted significantly with the early neurological function recovery in patients with acute stroke.