1.Intravenous transplantation of allogeneic bone marrow derived mesenchymal stromal cells at early stage of cortex ischemia significantly increases number of Iba-1+ microglia cells expressed brain-derived neurotrophic factor in the infarct area of rats
Xiaobo LI ; Haiqiang ZOU ; Chunsong ZHAO ; Renchao ZHAO ; Min HUANG ; Yao LIU ; Yunqian GUAN
Chinese Journal of Neuromedicine 2019;18(5):433-441
Objective To investigate the main cell types expressed brain-derived neurotrophic factor (BDNF) in the posterior cortical infarction area in cerebral infarction rats after early vein allograft of bone marrow mesenchymal stem cells (BM-MSCs) and the effect of BM-MSCs transplantation on their ce11 numbers and percentages.Methods (1) Fifteen SD rats were randomly divided into sham-operated group 1,ischemia control group 1,and BM-MSCs transplantation group 1 (n=5);distal middle cerebral artery occlusion (dMCA) models were used in the later two groups;1 × 106 CM-DiI labeled BM-MSCs were intravascularly transplanted into the tail vein of rats from the transplantation group 1 at one h after ischemia;all rats were sacrificed 48 h after ischemia;BM-MSCs with co-existence of CM-Dil and BDNF in the ischemia cortex areas were detected by immunofluorescence staining.(2)Fifteen SD rats were randomly divided into sham-operated group 2,ischemia control group 2,and BM-MSCs transplantation group 2 (n=5);dMCAO models were used in the later two groups;1 ×106 non-labeled BM-MSCs were intravascularly transplanted into the tail vein of rats from the transplantation group 2 at one h after ischemia;48 h after ischemia onset,all rats were sacrificed;the number of BDNF+ and CD68+ microglia cells,BDNF+ and Iba-1+ microglia cells,and BDNF+ and neuron-specific nucleoprotein (NeuN)+ neurons were measured by immunofluorescence staining.Results (1) CM-Dil red fluorescence labeled allogeneic BM-MSCs were only found in BM-MSCs transplantation group 1;the labeled cells scattered in the infarct and peri-infarct cortices;9.70%±3.47% CM-Dil labeled BM-MSCs expressed BDNF,accounting for 13.32%±4.48% of all BDNF+ cells in the infarct brain cortex.(2) In the brain tissues of cortex infarct area of BM-MSCs transplantation group 2,38.40%±9.04% BDNF+ cells were Iba-1+ microglia cells,11.65%±2.76% BDNF+ cells were CD68+ microglia cells,and 28.96%±6.99% BDNF+ cells were NeuN+ neurons;the Iba-1+ cell numbers and Iba-1+/BDNF+ double positive cell percentages in the BM-MSCs transplantation group 2 ([92.06±36.52]/mm2 and 79.21%±12.27%) were significantly increased as compared with those in the ischemia control group 2 ([31.13±10.23] mm2 and 60.15%±28.20%,P<0.05).Conclusion Allogeneic BM-MSCs is capable of migrating into the infarct cortex when intravenous transplantation of BM-MSCs is performed at the early stage after ischemia;the main sources of BDNF in these areas are microglias cells and neurons;these BM-MSCs increase both number and percentage of Iba-1+/BDNF+ double positive cells,which may be one of the underlying mechanisms of therapeutic effects.
2.Mechanism of Changyuning on PI3K/AKT Signaling Pathway in Ulcerative Colitis Rats
Haiqiang WANG ; Meng ZHANG ; Li XIONG ; Chenxi MA ; Yao WANG ; Lihong ZHENG
World Science and Technology-Modernization of Traditional Chinese Medicine 2023;25(11):3736-3743
Objective To investigate the effect of Changyuning prescription on PTEN,TLR2,Caspase-3,Caspase-9 in phosphatidylinositol-3 kinase/protein kinase B(PI3K/AKT)signaling pathway in colon tissue of 2,4,6-trinitrobenzenesulfonic acid(TNBS)-induced ulcerative colitis(UC)rats.Methods After adaptive feeding for 7 days,70 SPF rats were divided into blank group of 10 and modeling group of 60.The rats in the modeling group were given TNBS/ethanol solution enema to induce UC model.The rats that successfully bulit the model were randomly divided into model group,mesalazine group,Changyuning high-dose,medium-dose and low-dose groups,and Changyuning high-dose,medium-dose and low-dose groups,respectively,and the equivalent amount of crude drug was 4 g·mL-1,2 g·mL-1,1 g·mL-1 of Changyuning prescription solution 10 mL·kg-1,the mesalamine group was given 10 mL·kg-1 of mesalamine suspension equivalent to 0.2 g·kg-1 crude drug,the blank control group and model group were given an equal volume of 10 mL·kg-1 of normal saline was given by gavage.The rats in each group were given intragastric administration once a day for 14 consecutive days.Rats were sacrificed after the last administration.Disease activity index was evaluated;colon changes were observed by HE staining;colon tissue damage index was evaluated;The expression of PI3K,AKT,PTEN,TLR2,Caspase-3,and Caspase-9 proteins was detected by Western Blotting in colon tissue.Results Compared with the blank group,the protein expression levels of PI3K,AKT,Caspase-3,Caspase-9 and TLR2 in the model group were increased(P<0.05),and the protein expression level of PTEN was decreased(P<0.05).Compared with the model group,the expression levels of PI3K,AKT,Caspase-3,Caspase-9 and TLR2 proteins in the Changyuning high-dose group were significantly decreased(P<0.05),and the protein expression levels of PTEN were significantly increased(P<0.05);The levels of PI3K,AKT and Caspase-3 in the salazine group and Changyuning medium dose were decreased(P<0.05),the expression level of PTEN protein was increased(P<0.05),and there was no statistical significance in the expression of Caspase-9;There was no statistical significance in the expression of each protein between the Changyuning low-dose group and the model group.Conclusion Changyuning recipe has a significant effect on the treatment of UC,and its mechanism may be related to the regulation of PI3K/AKT signaling pathway and its related proteins PTEN,TLR2,Caspase-3,and Caspase-9,which can effectively relieve the symptoms of UC rats and reduce colon pathological damage.
3.Analysis of oligoclonal bands detection results of 3 217 patients with neurological disorders
Zhenyu NIU ; Haiqiang JIN ; Hongjun HAO ; Yiming ZHENG ; Jing GUO ; Yan YAO ; Feng GAO ; Zhaoxia WANG
Chinese Journal of Laboratory Medicine 2024;47(1):57-64
Objective:To study positive rates and typing of oligoclonal bands (OCB) in patients with neurological disorders, and to reveal the clinical significance and applicational value of OCB test.Methods:A retrospective analysis was performed on the detection results of 3 217 patients with neurological disorders who undertook both serum and cerebrospinal fluid OCBs in the First Hospital of Peking University from January 2012 to August 2022. According to the final diagnosis, the patients were divided into 13 groups including multiple sclerosis (479 cases), neuromyelitis optica spectrum disorders (935 cases), autoimmune encephalitis (192 cases), viral encephalitis (94 cases), nervous system complication after HSCT (232 cases), Guillain-Barré syndrome (644 cases), chronic inflammatory demyelinating polyneuropathy (157 cases), etc. Cerebrospinal fluid and serum OCBs were detected using isoelectric focusing electrophoresis combining immunofixation, then classified into Ⅰ-Ⅴ types according to the morphology. Consequently, positive rates and types were analyzed for each group. χ2 test was used for comparison between groups. Results:The positive rates of cerebrospinal fluid OCB in multiple sclerosis, nervous system complication after hematopoietic stem cell transplantation (HSCT), autoimmune encephalitis, viral encephalitis, neuromyelitis optica spectrum disorders, Guillain-Barré syndrome and chronic inflammatory demyelinating polyneuropathy were respectively 66.8% (320/479), 48.7% (113/232), 46.4%(89/192), 19.1% (18/94), 17.6% (165/935), 9.9% (64/644), 5.1% (8/157). For patients with multiple sclerosis, neuromyelitis optica spectrum disorders, viral encephalitis, and autoimmune encephalitis, Type Ⅱ bands took the majority of cerebrospinal fluid OCB-positive cases with the rates of 94.1% (301/320), 78.7% (70/89), 77.8% (14/18), and 77.6% (128/165) respectively, indicating intrathecal IgG synthesis; for patients with nervous system complication after HSCT, Guillain-Barré syndrome and chronic inflammatory demyelinating polyneuropathy, type Ⅳ bands took the majority of cerebrospinal fluid OCB-positive cases with the rates of 94.7% (107/113), 82.8% (53/64) and 100% (8/8), indicating no obvious intrathecal IgG synthesis. The positive rates of cerebrospinal fluid oligoclonal bands were significantly different among all groups (χ 2=1 268.31, P<0.001). Conclusion:The positive rates of cerebrospinal fluid oligoclonal bands are different among different neurological disorders, in which the positive rate of cerebrospinal fluid OCB is higher with type Ⅱ bands as the majority type in multiple sclerosis, which indicates that the detection and typing of cerebrospinal fluid OCB are helpful for the diagnosis of various neurological diseases, especially for multiple sclerosis.
4.The Characteristics of Constitution-Disease-Syndrome of Different Clinical Phenotypes and the Application of Three-Dimensional Diagnosis and Treatment Pattern of Bronchial Asthma
Hongsheng CUI ; Weibo BI ; Dongmei SHAO ; Haiqiang YAO ; Jia ZHU
Journal of Traditional Chinese Medicine 2024;65(10):1082-1085
This paper summarized the characteristics of constitution, disease, and syndrome of five different phenotypes of bronchial asthma including allergic asthma, cough variant asthma, chest tightness variant asthma, psychological stress asthma, and hormone-dependent asthma, and systematically expounds the application of three-dimensional diagnosis and treatment pattern, that is differentiation of constitution, disease and syndromes in the five phenotypes of bronchial asthma. It is considered that the allergic constitution is the main one of bronchial asthma, which is usually combined with the constitutions of qi-deficiency, yin-deficiency, yang-deficiency, qi-stagnation and others. In the treatment of bronchial asthma, constitution, disease and syndromes should all be taken into account but applied flexibly according to the order of the urgency and priority. The three-dimensional diagnosis and treatment pattern can reflect the essential characteristics and clinical panorama of bronchial asthma more comprehensively and objectively and provide effective intervention measures and management programs for the prevention and treatment by traditional Chinese medicine.