1.Effects of AMP-activated protein kinase on HMGB1 release from PC12 cells after oxygen-glucose deprivation and reoxygenation and its mediated inflammatory response in BV2 cells
Hui DANG ; Mingjia LU ; Hongyan LI ; Yi ZHU
International Journal of Cerebrovascular Diseases 2016;24(6):529-534
Objective To investigate the effects of adenosine monophosphate-activated protein kinase (AMPK) on high-mobility group box 1 (HMGB1) release from PC12 cells after oxygen-glucose deprivation and reoxygenation (OGD/R) and its mediated inflammatory response in BV2 cells.Methods PC12 and BV2 cells were cultured,respectively.The PC12 cells were used to induce a model of oxygen glucose deprivation for 12 h and reoxygenation for 24 h.After giving 5-aminoimidazole-4-carboxamide (AICAR) 5,50 and 100 μmol/L as well as Compound C 0.1,1 and 10 μmol/L activation or inhibition of AMPK phosphorylation,respectively,methyl thiazolyl tetrazolium (MTT) was used to detect the PC12 cell activity.Enzyme-linked immunosorbent assay was used to detect the HMGB1 release level in the PC12 cell culture media.After OGD/R in each group,the PC12 culture media were acted on normal cultured BV2 cells for 24 h respectively.Westem blotting and Enzyme-linked immunosorbent assay were used to detect the NFκB inhibitory protein (inhibitor of NFκB,IκB) phosphorylation level and TNF-α release level in BV2 cells,respectively.Results After OGD/R,the PC12 cell activity was decreased significantly (68.84%±6.60% vs.100.04% ± 8.82%;P < 0.01);the AMPK phosphorylation level was increased significantly (1.95 ±0.39 vs.1.00 ±0.20;P<0.05),and the extracellular HMGB1 release was increased significantly (287.66 ± 26.42 pg/μl vs.53.05 ± 9.11 pg/μl;P < 0.01).Compared with the OGD/R group,AICAR 100 μmol/L significantly increased the survival rate of PC12 cell after OGD/R (78.6% ± 3.75% vs.68.84% ± 6.60%;P < 0.05),promoted AMPK phosphorylation (3.32 ± 0.66 vs.1.95 ± 0.39;P < 0.01),and reduce the release of extracellular HMGB1 (164.06 ± 12.77 pg/μl vs.287.66 ± 26.42 pg/μl;P <0.01).In contrast,Compound C 10 μmol/L significantly reduced the cell survival rate of PC12 (40.44% ±3.79% vs.68.84% ±6.60%;P <0.01),inhibited AMPK phosphorylation (1.07 ± 0.21 vs.1.95 ± 0.39;P<0.05),and increased the release of HMGB1 (337.97 ± 18.9 pg/μlvs.287.66 ± 26.42 pg/μl;P<0.01).The conditioned medium from the AICAR 100 μmol/L group significantly inhibited IκB phosphorylation (1.68 ±0.51 vs.3.09 ± 0.10;P < 0.05) and reduced the release of TNF-α (669.53 ±38.58 pg/μlvs.841.76 ± 45.82 pg/μl;P< 0.05) in BV2 cells.The conditioned medium from the compound C 10 μmol/L group significantly promoted IκB phosphorylation (4.98 ± 1.24 vs.3.09 ± 0.10;P <0.01) and increased the release of TNF-α (1 035.32 ± 128.06 pg/μl vs.841.76 ± 45.82 pg/μl;P <0.05) in BV2 cells.Conclusions Promoting AMPK phosphorylation activation may reduce the release of HMGB1 from PC12 cells after OGD/R,and inhibit its mediated NF-κB inflammatory pathway and reduce the release of TNF-αin BV2 cells,and thus reducing neuroinflammatory injury.On the contrary,inhibiting AMPK phosphorylation may promote the release of HMGB1 from PC12 cells after OGD/R and aggravate its mediated inflammatory reaction in BV2 cells.
2.Anti-inflammation roles of Xuelian injection on mice models with Parkinson's disease
Mingjia LU ; Hui DANG ; Aishanjiang YISUPUJIANG ; Yi ZHU
Chinese Journal of Neuromedicine 2014;13(6):547-551
Objective To investigate the effect of Xuelian injection on mice models with Parkinson' s disease (PD).Methods C57BL/6 mice were given paraquat (PQ) by peritoneal injection (10 mg/kg) to induce PD models.Then,they were randomly divided into six groups:PD group,negative control group,positive controlgroup,low-dose (0.26mL/kg·d),middle-dose (0.52 mL/kg· d) and high-dose (1.04 mL/kg· d) Xuelian injection groups.Pole test and open field activity test were used for assessment of behavior 7 weeks after the injection; the dopamine (DA) content in the substantia nigra pars compacta (SNc) was observed by high-performance liquid chromatography (HPLC).The expressions of tyrosine hydroxylase (TH),integrin o (mac-l) and tumor necrosis factor α (TNF-α) in the SNc were measured by immunohistochemical staining under optical microscope.Results Seven weeks after the injection,the movement coordination disorder was significant in the PD group,and that had partial remission in the high-dose Xuelian injection group.HPLC showed that the DA content in the PD group was significantly lower than that in the negative control group (P<0.05); that in the high-dose Xuelian injection group ([10.90±1.04] μg/g) was significantly higher than that in the PD group (P<0.05).Immunohistochemical staining indicated that PD group had significantly decreased number of TH positive cells and statistically increased numbers of mac-1 and TNF-α positive cells as compared with the negative control group (P<0.05); high-dose Xuelian injection group had significantly increased number of TH positive cells (65.18±6.00) and statistically decreased numbers of mac-1 and TNF-α positive cells (93.18±5.33 and 92.73±23.21) as compared with the PD group (P<0.05).Conclusions High-dose Xuelian injection can obviously improve the movement coordination disorder and survival of DA neurons in the SNc,whose protective effect might be related to the inhabitation of microglia mediated inflammation injury and oxidative stress.
3.Outbreak of epidemic poliomyelitis in Xinjiang patients: an electromyogram analysis
Ya TUO ; Jing LI ; Mingjia LU ; Yi ZHU
Chinese Journal of Neuromedicine 2014;13(8):820-821
Objective To analyze the electromyogram (EMG) changes of patients with acute flaccid paralysis (AFP) in Hetian Xinjiang.Methods Twenty-six patients with acute poliogmyelitis,admitted to our hospital from July 2011 to September 2012,were chosen in our study; their clinical data were analyzed,and they all underwent EMG examination.Results Most patients had contact with respiratory virus or digestive virus prior to illness onset.Most of patients had flaccid paralysis,muscle strength between 0 and 2 grades,tendon reflex being abated and no sensory disturbance.Fourteen patients had positive blood stool.Motor nerve conduction velocity (MCV) was abnormal,H-reflex was decreased or not educed and sensory nerve conduction velocity was normal in lower limbs of the 26 patients; EMG showed myogenic abnormality.Nerve conduction velocity was normal or low and Fwave was normal or decreased in upper limbs; EMG showed neurogenic abnormality in five of the 26 patients.Poor prognosis was concluded in all the patients.Conclusion EMG has important diagnosis value in AFP induced by polio wild-type I.
4.Comparing one-step common bile duct exploration plus cholecystectomy with two-step endoscopic sphincterotomy plus cholecystectomy in patients over 80 years with concomitant gallbladder and common bile duct stones
Jianwen DUAN ; Lei YUAN ; Yunpeng SUN ; Da SUN ; Dapeng XU ; Mingjia XIAO ; Hongbo SHEN ; Jiajun LU
Chinese Journal of Hepatobiliary Surgery 2023;29(3):185-189
Objective:To compare safety and efficacy of one-stage laparoscopic common bile duct exploration plus laparoscopic cholecystectomy (LCBDE+ LC) with endoscopic retrodrade cholangiopancreatography plus laparoscopic cholecystectomy (ERCP+ LC) in elderly patients with concomitant gallbladder and common bile duct (CBD) stones.Methods:This is a two-center retrospective study with clinical data on 492 patients aged over 80 years diagnosed with concomitant gallbladder and CBD stones treated between January, 2014 and December, 2020 at The First Affiliated Hospital of Wenzhou Medical University and Quzhou Hospital Affiliated to Wenzhou Medical University. There were 254 males and 238 females, aged (83.9±3.0) years. These patients were divided into two groups based on their operative methods: the one-stage group (LCBDE+ LC, n=186) and the two-stage group (ERCP+ LC, n=306). Differences in surgery, stones and hospitalization costs were compared between the two groups. Results:When compared with the ERCP+ LC group, the LCBDE+ LC group had significantly higher incidences of previous gastrectomy [21.5%(40/186) vs 4.2%(13/306)], multiple stones [77.4%(144/186) vs 49.3%(151/306)], larger stone diameter [13.7(6.4, 18.6)mm vs 10.9(5.7, 16.1) mm], and increased hospitalization expenditure [(2.37±0.31) Wanyuan vs (3.26±0.44) Wanyuan] (all P<0.05). However, the rates of residual stone [2.7%(5/186) vs 1.3%(4/306)], stone recurrence [2.2%(4/186) vs 5.2%(16/306)], postoperatively overall complications [3.2%(6/186) vs 1.3%(4/306)], and total hospital stay [(10.7±6.2) d vs (11.3±5.4) d] were not significantly different between the two groups (all P>0.05). Conclusions:Allowing for the similar safety and effectiveness, and lower hospitalization expenditure, LCBDE+ LC was a preferred choice for patients aged over 80 year, especially in patients who had previous gastrectomy, multiple large CBD stones, or who could not accept endoscopic procedures for treatment of CBD stones.
5.Construction of A Macro-evaluation Tool for Dampness Syndrome Animal Model in Traditional Chinese Medicine
Chuang LI ; Peng XU ; Ruimin TIAN ; Zhaorui CAO ; Mingjia LIU ; Lei ZHANG ; Zhaoyu LU ; Taohua LAN ; Xiaowan WANG ; Wei MAO
Journal of Traditional Chinese Medicine 2024;65(14):1449-1457
ObjectiveTo construct a macro-evaluation tool for dampness syndrome (DS) animal model, which will provide a basis for experimental research on dampness syndrome in traditional Chinese medicine (TCM). MethodsConceptual framework of this study was clarified through discussions within the core working group, and dimensions of the evaluation of the animal model of DS were identified according to TCM principles. We searched CNKI, Wanfang, VIP and SinoMed databases from the inception to June 30th, 2023, on experiments involving dampness syndrome animals to create a pool of items about DS animal models. The core items were selected and extracted for factor analysis and cluster analysis. An expert importance rating questionnaire was developed based on the results of the literature review, analyzing the distribution of item scores, importance averages, and coefficient of variation. Through a comprehensive analysis of literature, expert importance scoring, and specific expert opinions, items that did not meet anyone of the criteria of average importance rating ≥2.04, coefficient of variation ≤30%, or literature eva-luation frequency ≥2% were removed, thereafter, the macro-evaluation tool for DS animal model was preliminarily constructed. ResultsSpirit and body state, autonomic activity state, body surface characteristics, diet, urination and defecation, tongue manifestation, and motor behavior assessment were constructed as the seven dimensions in the evaluation of DS animal model. A total of 348 papers about animal experiments were included and analyzed, resulting in a saturated pool of 72 items, which was refined to 38 core items of DS animal models. Factor analysis obtained 16 common factors, which were further clustered into two categories, named dampness transforming from heat syndrome and dampness transforming from cold syndrome. The expert importance scoring showed that the Kendall harmony coefficient was 0.359 (P<0.05) indicating a high level of agreement, coordination and reliability among the experts. Notably, 50% or more of the experts considered the items thick and greasy tongue coating, unclean perianal area, loose stools, lethargy, unformed stools, and listless expression as very important. The median scores for all items were 2.04(1.73, 2.37), with a coefficient of variation ranging from 19.73% to 53.38%. After expert evaluation, the macro-evaluation tool for DS animal model in TCM with 33 items and corresponding criteria for assessing the formation of DS models was finally contructed. ConclusionThe Macro-evaluation tool for DS animal model in TCM is highly scientific, credible, and operable, and can be utilized in DS animal experiments after its characteristics are actually evaluated.