Could Intrathymic Injection of Myelin Basic Protein Suppress Inflammatory Response After Co-culture of T Lymphocytes and BV-2 Microglia Cells? / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>The interaction between activated microglia and T lymphocytes can yield abundant pro-inflammatory cytokines. Our previous study proved that thymus immune tolerance could alleviate the inflammatory response. This study aimed to investigate whether intrathymic injection of myelin basic protein (MBP) in mice could suppress the inflammatory response after co-culture of T lymphocytes and BV-2 microglia cells.</p><p><b>METHODS</b>Totally, 72 male C57BL/6 mice were randomly assigned to three groups (n = 24 in each): Group A: intrathymic injection of 100 μl MBP (1 mg/ml); Group B: intrathymic injection of 100 μl phosphate-buffered saline (PBS); and Group C: sham operation group. Every eight mice in each group were sacrificed to obtain the spleen at postoperative days 3, 7, and 14, respectively. T lymphocytes those were extracted and purified from the spleens were then co-cultured with activated BV-2 microglia cells at a proportion of 1:2 in the medium containing MBP for 3 days. After identified the T lymphocytes by CD3, surface antigens of T lymphocytes (CD4, CD8, CD152, and CD154) and BV-2 microglia cells (CD45 and CD54) were detected by flow cytometry. The expressions of pro-inflammatory factors of BV-2 microglia cells (interleukin [IL]-1β, tumor necrosis factor-α [TNF-α], and inducible nitric oxide synthase [iNOS]) were detected by quantitative real-time polymerase chain reaction (PCR). One-way analysis of variance (ANOVA) and the least significant difference test were used for data analysis.</p><p><b>RESULTS</b>The levels of CD152 in Group A showed an upward trend from the 3rd to 7th day, with a downward trend from the 7th to 14th day (20.12 ± 0.71%, 30.71 ± 1.14%, 13.50 ± 0.71% at postoperative days 3, 7, and 14, respectively, P < 0.05). The levels of CD154 in Group A showed a downward trend from the 3rd to 7th day, with an upward trend from the 7th to 14th day (10.00 ± 0.23%, 5.28 ± 0.69%, 14.67 ± 2.71% at postoperative days 3, 7, and 14, respectively, P < 0.05). The ratio of CD4+/CD8 + T in Group A showed a downward trend from the 3rd to 7th day, with the minimum at postoperative day 7, then an upward trend from the 7th to 14th day (P < 0.05). Meanwhile, the levels of CD45 and CD54 in Group A were found as the same trend as the ratio of CD4+/CD8 + T (CD45: 83.39 ± 2.56%, 82.74 ± 2.09%, 87.56 ± 2.11%; CD54: 3.80 ± 0.24%, 0.94 ± 0.40%, 3.41 ± 0.33% at postoperative days 3, 7, and 14, respectively, P < 0.05). The expressions of TNF-α, IL-1β, and iNOS in Group A were significantly lower than those in Groups B and C, and the values at postoperative day 7 were the lowest compared with those at postoperative days 3 and 14 (P < 0.05). No significant difference was found between Groups B and C.</p><p><b>CONCLUSIONS</b>Intrathymic injection of MBP could suppress the immune reaction that might reduce the secondary immune injury of brain tissue induced by an inflammatory response.</p>

Proteomic analysis of human fetal and adult brain cortex / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To study the differences of protein expression levels in the brain cortex of human fetus and adult with proteomics technique, and provide preliminary data on the change of proteins during brain development.</p><p><b>METHODS</b>Proteins extracted from human temporal lobes in fetal (3 month and 5 month respectively) and adult (30 years old) brain were separated by two-dimensional gel electrophoresis (2DE). The proteins were then stained with colloidal Coomassie blue to produce a high-resolution map of the proteiome. The differential protein spots were analyzed by PDQuest 7.0 software and 8 spots, which were gradually reduced or gradually increased in brain development process and the protein spots of difference over two-fold in the brain, were identified by matrix-assisted laser desorption/ionization-time of flight mass spectrometry (MALDI-TOF/TOF).</p><p><b>RESULTS</b>(1) On average, 642, 511 and 527 protein spots could be obtained in the temporal lobes of adult, 3 month and 5 month fetus. The matching rate of images was 87%. The basic proteins in adult brain were obviously much more than that in the fetus; (2) There were 172, 171 and 152 singular protein spots in temporal lobes of adult, 3 month and 5 month fetus respectively.(3) Compared with adult, there were 131 and 115 different protein spots in the 3 month and 5 month fetus respectively. There were 60 and 40 protein spots with more than 2 fold difference, among which 24 and 17 were down-regulated, and 36 and 23 were up-regulated respectively. (4) There was different expression in proteins such as serum albumin, triosephosphate isomerase, etc. in the 3 groups. Fatty acid binding protein 7 and unnamed proteins were only highly expressed in the 3 month brain; ribulose-1, 5-bisphosphate carboxylase/oxygenase large subunit and transducin beta-1 subunit were up-regulated in adult brain. Serum albumin decreases gradually with brain development. However, ATP synthase, mitochondrial F0 complex, and triosephosphate isomerase increase gradually with brain development.</p><p><b>CONCLUSION</b>The proteins of human brain cortex were obviously changed from embryonic stage to adult. The differentially displayed proteins may provide further insight into the understanding of development of human brain.</p>

Influence of temperature on extracellular glucose and lactate after traumatic brain injury / 中国医学科学院学报

<p><b>OBJECTIVE</b>To study the changes of extracellular glucose (Glu), lactate (Lac), and the ratio of lactate/pyruvate (L/P) in patients with traumatic brain injury under different body temperatures.</p><p><b>METHODS</b>Catheters for microdialysis were punctured into the penumbra zone of injured brain tissue (INJ), relatively normal brain tissue (NOR), and abdominal subcutaneous tissue (ABD) respectively in 51 patients to collect the fluid. The perfusion rate was 0.3 microl/min and one tube of fluid was collected for each hour. The average collection time was (67.10 +/- 18.27) hours. Concentrations of Glu, Lac, and pyruvate (Pyru) in the fluid were analyzed using CMA microdialysis analyzer. Patients were divided into 7 groups according to their rectal temperature (RT) values, which were RT < 33.0 degrees C, 33.0-33.9 degrees C, 34.0-34.9 degrees C, 35.0-35.9 degrees C, 36.0-36.9 degrees C, 37.0-37.9 degrees C, and > or = 38.0 degrees C.</p><p><b>RESULTS</b>The concentration of Glu in ABD was significantly higher than that in the brain tissue (P < 0.05). The Glu in NOR were significantly higher and the highest in 33.0 degrees C compared with that in the INJ when RT < 36.0 degrees C (P < 0.05). The concentration of Lac in ABD was significantly lower than that in brain (P < 0.05). The Lac in NOR were much higher than that in INJ when RT < 35.0 degrees C or > or = 37.0 degrees C (P < 0.05). The ratio of L/P decreased along with the increase of body temperature (P < 0.001). The ratio of L/P significantly decreased in an order of INJ > ABD > NOR when RT was lower than 33.0 degrees C, which was changed to the order of NOR > INJ > ABD when RT was higher than 34.0 degrees C.</p><p><b>CONCLUSION</b>Treatment of hypothermia may play more protective role when RT were between 33-34 degrees C or 36-37 degrees C.</p>