1.Human albumin attenuates Mincle-associated early neuroinflammatory injury in subarachnoid hemorrhage rats
Yi XIE ; Qiushi LYU ; Hongquan GUO ; Nana ZHAO ; Ruidong YE ; Xinfeng LIU
Chinese Journal of Neurology 2017;50(7):531-537
Objective To investigate the protective role of human serum albumin in treatment of monocyte-inducible C-type lectin (Mincle)-associated neuroinflammation in subarachnoid hemorrhage (SAH) rats and its underlying mechanisms.Methods Vascular perforation model was used to induce SAH.Ninety-two male SD rats were randomly assigned to sham (n =23),vehicle (n =23),low-dose albumin (0.63 g/kg,n =23) and high-dose albumin (1.25 g/kg,n =23) groups.Saline and albumin were intravenously injected into rats two hours after surgery.Modified Garcia scale was employed to assess neurological functions.Iba-1 and myeloperoxidase (MPO) staining was used to examine the activation of microglial cells and infiltration of neutrophils.Real-time PCR was applied to determine the changes of IL-1β,inducible nitric oxide synthase,CD11b,monocyte chemoattractant protein-1,cytokine-induced neutrophil chemoattractant-1 and CXC motif chemokine ligand-2 mRNA levels.Co-immunoprecipitation was conducted to assess the binding ability of albumin with Mincle.Immunoblotting was carried out to evaluate protein levels of Minlce,Syk and p-Syk.SAH severity measurement was performed before conductions of all the experiments.Results SAH severity scores were 11.4 ± 1.6,12.8 ±2.5 and 11.2 ±3.2 in the vehicle,low-dose albumin and high-dose albumin groups,respectively,without statistically significant difference among groups (F =0.694,P =0.516).Neurological score was 7.5 ± 2.9 in the vehicle group,while the low-dose albumin (14.6 ± 2.2) and high-dose albumin groups (13.6 ± 2.7) exhibited better neurological perfomance (P < 0.01).Immunostaining showed that albumin significantly inhibited the activation of microglia,and reduced the percentage of MPO positive cells from 20.7% ± 1.9% in the vehicle group to 12.1% ±2.1% in the low-dose albumin group and 9.8% ±0.9% in the high-dose albunin group (F =32.216,P =0.001).mRNA levels of pro-inflammatory cytokines and chemotactic factors were also suppressed by albumin (P < 0.05).The results of co-immunoprecipitation displayed that albumin could directly bind Mincle and disrupt the association between Mincle and SAP130.Immunoblotting demonstrated that albumin depressed the protein levels of Mincle,Syk and p-Syk.Conclusion Human serum albumin can inhibit Mincle/Syk-induced neuroinflammation via directly binding Mincle receptor in SAH rats.
2.Clinical and neuroimaging features of hypoglycemia encephalopathy in the elderly
Xuewu LIU ; Wei WU ; Shuzhen WANG ; Lili CAO ; Xiuhe ZHAO ; Ruidong CHEN ; Lijun SU ; Guoqing DONG ; Zhaofu CHI
Chinese Journal of Neurology 2008;41(9):610-613
Objective To investigate the clinical and neuroimaging features of hypoglycemia encephalopathy in the elderly. Methods The history and clinical features of 36 patients who had undergone brain CT and MRI were analyzed retrospectively. Results Twenty-seven patients had infections and fevers as a trigger, presenting all kinds of symptoms. Eleven cases were found to have abnormal signals in bilateral caudate nucleus and lenticular nucleus in MRL But CT examination showed no new lesions in corresponding position. Hypoglycemia encephalopathy were commanly found in the elderly who had diabetes mellitus and treated with drugs. After being followed up for 6 months, their neuroimaging did not change. Conclusions Because the patients often present unconsciousness and weakness with a sudden onset, hypoglycemia is easily mistaken for other disorders, especially in the elderly. For those with consciousness, we should pay more attations to hypoglycemia. Brain CT has no value of diagnosing hypoglycemia encephalopathy, while MRI plays an impotant role in diagnosing the disease. The characteristic MRI features predicts a bad prognosis.
3.The expression of circulating microRNA-595 in patients with hepatitis B virus-related liver failure and its relationship with disease prognosis
Peng WANG ; Xiaogang XIANG ; Ruidong MO ; Rongtao LAI ; Weijing WANG ; Yuhan LIU ; Gangde ZHAO ; Simin GUO ; Hong YU ; Huijuan ZHOU ; Wei CAI ; Hui WANG ; Yan HAN ; Qing XIE
Chinese Journal of Infectious Diseases 2014;(9):545-549
Objective To measure the expression of circulating microRNA (miRNA)in patients with hepatitis B virus (HBV)-related liver failure and its relationship with disease prognosis.Methods The miRNA expressions in serum of 5 patients with HBV-related liver failure and 5 healthy control subjects were compared using Exiqon miRCURY LNATM miRNA microarray.The sera from 20 patients with chronic hepatitis B (CHB),20 patients hepatitis B related cirrhosis,50 patients with HBV-related liver failure and 40 healthy persons in Ruijin Hospital were collected.The relative expression of miRNA-595 was measured using quantitative real-time polymerase chain reaction (PCR).The relative expressions of miRNAs among groups were analyzed using student t test,the correlations were analyzed by Pearson and Spearman correlation.Results Microarray informed that 92 miRNAs changed significantly in patients with HBV-related liver failure,and miRNA-595 increased most significantly.The results of real-time PCR showed that the relative expressions of miRNA-595 ,miRNA-300 and miRNA-122 were 6.03 (t=3.134, P =0.003),3.12 (t=7.221 ,P <0.01)and 2.77 (t=2.671 ,P =0.021),which were higher compared to those in healthy control group.In the analysis of the relationship between miRNA-595 expression and disease prognosis in patients with HBV-related liver failure,the relative expressions of miRNA-595 in patients with CHB,hepatitis B related cirrhosis and HBV-related liver failure were 2.26 (t =3.780,P =0.001),3.32 (t = 6.111 ,P < 0.01)and 6.03 (t = 3.134,P = 0.003),respectively,which were all increased compared to that of the healthy control.The relative expression of miRNA-595 of patients with HBV-related liver failure was 2.66 times (t=2.450,P =0.043)higher than that of patients with CHB. When dividing patients according to prothrombin activity,miRNA-595 increased significantly in patients with early stage liver failure.When dividing patients according to model of end-stage liver disease (MELD) score,MELD score was positive correlated with the expression of miRNA-595 when MELD score was under 30 (r=0.673,P =0.004).The expression of serum miRNA-595 in survival group (11 .08,n=23) was higher than that in non-survival group (3.67,n = 27,t =4.309,P =0.041).Conclusions The expressions of miRNA595 ,miRNA-300 and miRNA-122 are all increased in patients with HBV-related liver failure,especially the expression of circulating miRNA-595 at early stage of the disease.The miRNA-595 may be used as a new serum biomarker for monitoring the severity of disease.
4.Digital gene expression profiling of dendritic cells treated with Seabuckthorn favones
Bingchun LIU ; Ying QIU ; Ruidong ZHAO ; Xia HAN ; Feiyu YUN ; Erxia ZHOU ; Sheng YUN
Chinese Journal of Microbiology and Immunology 2017;37(11):840-848
Objective To study the changes in morphology , phenotypes and gene expression pro-files of dendritic cells (DCs) following treatment with Seabuckthorn flavones (SF).Methods DCs were treated with 200μg/ml of SF and then cultured for 7 days.Changes in the morphology of DCs were observed under light microscope .Flow cytometry was used to detect DC surface molecules .Total RNA was extracted to construct the library for digital gene expression profiling ( DGE ) .Differentially expressed genes were screened out and further analyzed by gene ontology ( GO) enrichment analysis and Kyoto encyclopedia of genes and genomes ( KEGG ) pathway enrichment analysis .Results Compared with control group , SF treatment significantly enhanced the expression of HLA-DR, CD80, CD83 and CD86 on DCs.A total of 355 differentially expressed genes were screened out by DGE , including 176 up-regulated genes and 179 down-regulated genes .GO enrichment was mainly involved in the regulation and development of the immune sys -tem and other biological processes .KEGG pathway analysis showed that the significantly enriched pathways were closely related to inflammation , the immune system, cancer and other diseases .Conclusion SF can promote the expression of DC co-stimulatory molecules and pro-mature molecules, and regulate the expres-sion of immunity-related genes such as CD11a, SLAMF6, LMCD1, TSC22D3 and IKZF3.
5.Animal models of white matter lesions
Nana ZHAO ; Qiushi LYU ; Hongquan GUO ; Yi XIE ; Ting HU ; Ling WANG ; Ruidong YE ; Xinfeng LIU
International Journal of Cerebrovascular Diseases 2018;26(10):789-795
White matter lesion is a major subtype of cerebral small vessel disease. Its pathophysiology and mechanism remain unclear. Because the risk factors often coexist in clinical research, it is difficult to judge the relationship between certain risk factors and white matter injury. Moreover, due to the differences in animal and human brain tissue structure, there is currently a lack of reproducible animal models of white matter lesions. Therefore, establishing a practical animal model and further exploring the pathogenesis and risk factors of white matter lesions from the basic research level is crucial for the preclinical study of the treatment of white matter damage. This article reviews the characteristics, optimization measures, and application prospects of the white matter lesion models.
6.Changes of white matter in LDLR knockout hypercholesterolemic rats
Nana ZHAO ; Hongquan GUO ; Yi XIE ; Meng LIANG ; Ming WANG ; Yuhui WANG ; Guoqing LIU ; Huajuan HOU ; Ruidong YE ; Xinfeng LIU
International Journal of Cerebrovascular Diseases 2018;26(1):36-41
Objective To investigate the difference between histopathological changes of brain white matter in low-density lipoprotein receptor (LDLR) homozygous mutation rats with hypercholesterolemia and wild-type rats.Methods Thirty LDLR-/-rats and 28 wild-type rats were selected.Plasma cholesterol levels were measured by enzyme-linked immunosorbent assay at 15,18 and 26 weeks old respectively.The axonal structure of the corpus callosum area was observed by transmission electron microscopy.The myelin basic protein (MBP) of the corpus callosum area was quantitatively analyzed by Western blotting.In addition,at 26 weeks old,the myelin sheaths were stained by fast blue staining.The expression level of MBP in white matter was further detected by immunofluorescence staining,and the morphological changes of glial cells were observed.Results Compared with the wild-type rats,the plasma cholesterol concentration in LDLR-/-rats increased significantly,and it could be as high as 3.3 times at 26 weeks.The results of electron microscopy showed that the LDLR-/-rats had axonal injury at 15 weeks and aggravated gradually over time.At 26 weeks,Western blot analysis of the LDLR-/-rats showed that the MBP expression level of the corpus callosum area decreased significantly.Fast blue staining showed loosening of nerve fibers,diffuse vacuole formation,and myelinated nerve fiber loss in the corpus callosum area.In addition,it was also found that the number of oligodendrocytes in LDLR-/-rats was significantly reduced,and large numbers of astrocytes and microglia were activated.Conclusions LDLR-/-rats will have spontaneous hypercholesterolemia.Axonal injury,demyelination,decreased oligodendrocytes,as well as the abnormal activation of astrocytes and microglia are present in the early adult brain white matter area.
7.Targeted induction of human umbilical cord mesenchymal stem cells cultured with human peripheral blood serum into neural stem cells
Xia HAN ; Ruidong ZHAO ; Junli YANG
Chinese Journal of Tissue Engineering Research 2024;28(25):4000-4004
BACKGROUND:There are many kinds of cell media with different components,which have a great influence on cell growth.Several studies in and outside China have used serum-free media containing fetal bovine serum for in vitro amplification culture,but the use of media containing human peripheral blood serum to directionally induce human umbilical cord mesenchymal stem cells to neural stem cells and human peripheral blood serum to promote neural stem cells to differentiate into other nerve cells,so far there are few relevant studies. OBJECTIVE:To observe the feasibility of inducing human umbilical cord mesenchymal stem cells cultured with human peripheral blood serum into neural stem cells. METHODS:(1)Human umbilical cord mesenchymal stem cells were cultured in DMEM/F-12 culture medium containing 10%human peripheral blood serum by volume fraction.Flow cytometry analysis was performed at the third passage to identify surface markers and alizarin red staining was used to detect osteogenic differentiation function.(2)The third-generation human umbilical cord mesenchymal stem cells were induced into neural stem cells using DMEM/F-12 medium containing 0.5%N2,1.5%B27,20 ng/mL basic fibroblast growth factor,and 20 ng/mL epidermal growth factor,and their surface markers were identified.(3)Well-growing human umbilical cord mesenchymal stem cell-derived neural stem cells were taken to prepare a single cell suspension.They were evenly inoculated into 96-well plates and incubated with DMEM/F-12 culture medium containing 10%human peripheral blood serum for 8 days.Then,hematoxylin-eosin staining,microtubule-associated protein 2,and glial fibrillary acidic protein immunofluorescence staining were performed to detect the differentiation of neural stem cells into other neural cells. RESULTS AND CONCLUSION:(1)Human umbilical cord mesenchymal stem cells cultured with human peripheral blood serum grew in a spiral-like pattern and were distributed in multiple layers,with directional arrangement.The surface of human umbilical cord mesenchymal stem cells highly expressed CD44,CD105,CD29,and CD73.Cells stained with alizarin red showed a color reaction.(2)Human umbilical cord mesenchymal stem cells cultured with human peripheral blood serum could be induced to differentiate into neural stem cells,and the surface of neural stem cells was highly expressed with CD44,CD105,CD29,CD73,Nestin,NF-L,and GALC.(3)On day 8 of induced differentiation of neural stem cells,after staining with hematoxylin and eosin,it was found that the protruding protrusions were longer,with more branches and adjacent cells connected,presenting typical neural cell morphology.Immunofluorescence staining for microtubule-associated protein 2 and glial fibrillary acidic protein was positive.It is concluded that human umbilical cord mesenchymal stem cells cultured by human peripheral blood serum can be directly induced to differentiate into neural stem cells.Under the action of human peripheral blood serum,neural stem cells can differentiate into other neural cells as the culture time prolongs.
8.Effect of immune cells induced and differentiated by umbilical cord blood mononuclearcells on immune function of patients with small cell lung cancer
ZHAO Ruidong ; QIU Ying ; YUN Feiyu ; ZHOU Erxia ; HAN Xia ; LIU Bingchun ; YUN Sheng
Chinese Journal of Cancer Biotherapy 2018;25(8):810-816
Objective: To evaluate the effect of immune cells induced and differentiated by umbilical cord blood mononuclear cells (UCMCs) on the immune function of patients with small cell lung cancer (SCLC). Methods: Ninety patients with SCLC, who were admitted to the Affiliated Hospitalof InnerMongolia Medical University from January 2012 to December 2015, were randomly divided into control group (45 patients, EP regimen), study group (45 patients, EP regimen+UCMC-induced and differentiated immune cells). The study group of patients received immune cell treatment 3-5 d after chemotherapy ([1-3]×1010cells/treatment), 30 d for a cycle. The changes in T cell subsets, IFN-γ, IL-2, IL-10 and TGF-β1 in peripheral blood of patients were observed by flow cytometry at pre-treatment and 12 weeks post-treatment. Life quality and adverse events of patients were evaluated. Results: The study group, 15 cases achieved CR, 25 cases of PR and 5 cases of SD. The percent of T cell subsets in the study group was significantly higher than that in the control group (P<0.01), and the time of return to normal level was obviously shorter (P<0.05). The serum level of inflammatory cytokine IFN-γ increased or exceeded the normal range in 80.9% patients, and IL-10 and TGF-β1 levels were significantly decreased as compared with pretreatment (P<0.05). The quality of life was obviously better than that of the control group (P<0.05). Conclusion: Immune cells induced and differentiated by UCMCs can promote the recovery of immune function of patients with SCLC.