1.Risk factors for death in elderly patients admitted to intensive care unit after elective abdominal surgery: a consecutive 5-year retrospective study
Shuwen LI ; Tianhui HE ; Feng SHEN ; Difen WANG ; Xu LIU ; Jingcheng QIN ; Chuan XIAO ; Wei LI ; Qing LI ; Daixiu GAO
Chinese Critical Care Medicine 2021;33(12):1453-1458
		                        		
		                        			
		                        			Objective:To investigate the risk factors that were associated with the death of elderly patients who were admitted to the intensive care unit (ICU) after elective abdominal surgery, and to find reliable and sensitive predictive indicators for early interventions and reducing the mortality.Methods:A retrospective case-control study was conducted. The clinical data of elderly (age≥65 years old) patients after elective abdominal surgery admitted to the ICU of the Affiliated Hospital of Guizhou Medical University from January 1st 2016 to December 31st 2020 were collected, including the patient's gender, age, body mass index (BMI), medical history, American Society of Anesthesiologists (ASA) grades, surgical classification, intraoperative blood loss, duration of operation, interval time between end of operation and admission to the ICU, acute physiology and chronic health evaluationⅡ(APACHEⅡ) score and the worst laboratory examination results within 24 hours of ICU admission, the first blood gas analysis in ICU, the duration of invasive mechanical ventilation, and the length of ICU stay. Postoperative abdominal infection was evaluated by the pathogenic culture of peritoneal drainage fluid and clinical symptoms and signs. The patients were divided into death group and survival group based on clinical outcomes, and clinical data were compared between the two groups. Binary multivariate Logistic regression analysis was used to screen the risk factors of death, and the receiver operator characteristic curve (ROC curve) was plotted to analyze the predictive values of these risk factors.Results:A total of 226 elderly patients with elective abdominal surgery were admitted to the ICU of our hospital during the past 5 years, of whom, two patients who did not undergo laboratory examinations within 24 hours of admission to the ICU were excluded. Finally, 224 patients met the criteria, with 158 survivors and 66 deaths. Univariate analysis showed that: compared with survival group, APACHEⅡscore, blood lactate acid (Lac) and the proportion of postoperative abdominal infection were higher in death group [APACHEⅡ score: 27.5 (25.0, 31.3) vs. 23.0 (18.0, 27.0), Lac (mmol/L): 2.9 (1.8, 6.6) vs. 1.8 (1.1, 2.8), the proportion of postoperative abdominal infection: 65.2% (43/66) vs. 35.4% (56/158), all P < 0.01], prothrombin time (PT), activated partial thromboplastin time (APTT) and interval time between end of surgery and admission to ICU were longer [PT (s): 17.20 (14.50, 18.63) vs. 14.65 (13.90, 16.23), APTT (s): 45.15 (38.68, 55.15) vs. 39.45 (36.40, 45.70), interval time between end of surgery and admission to ICU (hours): 39.2 (0.7, 128.9) vs. 0.7 (0.3, 2.0), all P <0.01], postoperative hemoglobin (Hb), platelet count (PLT), prealbumin (PA), mean arterial pressure (MAP) and oxygenation index (PaO 2/FiO 2) were lower in death group [Hb (g/L): 95.79±23.64 vs. 105.58±19.82, PLT (×10 9/L): 138.5 (101.0, 177.5) vs. 160.5 (118.5, 232.3), PA (g/L): 80.88±43.63 vs. 116.54±50.80, MAP (mmHg, 1 mmHg = 0.133 kPa): 76.8±19.1 vs. 91.6±19.8, PaO 2/FiO 2 (mmHg): 180.0 (123.5, 242.5) vs. 223.5 (174.8, 310.0), all P < 0.05]. Binary multivariate Logistic regression analysis showed that APACHEⅡscore [odds ratio ( OR) = 1.187, 95% confidence interval (95% CI) =1.008-1.294, P < 0.001], interval time between end of operation and admission to ICU ( OR = 1.005, 95% CI = 1.001-1.009, P = 0.016) and postoperative abdominal infection ( OR = 2.630, 95% CI = 1.148-6.024, P = 0.022) were independent risk factors for prognosis in these patients. MAP ( OR = 0.978, 95% CI = 0.957-0.999, P = 0.041) and PaO 2/FiO 2 ( OR = 0.994, 95% CI = 0.990-0.998, P = 0.003) were protective factors for the patients' prognosis. Lac, Hb, PLT, PA, PT and APTT had no predictive value for the prognosis of elderly patients admitted to ICU after elective abdominal surgery [ OR value and 95% CI were 1.075 (0.945-1.223), 1.011 (0.99-1.032), 1.000 (0.995-1.005), 0.998 (0.989-1.007), 1.051 (0.927-1.192) and 1.003 (0.991-1.016), respectively, all P > 0.05. ROC curve analysis showed that APACHEⅡscore, interval time between end of operation and admission to the ICU and the postoperative abdominal infection had certain predictive values for the prognosis of elderly patients, the area under ROC curve (AUC) were 0.755, 0.732 and 0.649 respectively, all P < 0.001; When the cut-off of APACHEⅡscore and interval time between end of operation and admission to the ICU were 24.5 scores and 2.15 hours, the sensitivity were 78.8% and 66.7%, respectively, and the specificity were 62.0% and 76.6%, respectively. The combined predictive value of the three variables was the highest, which AUC was 0.846, the joint prediction probability was 0.27, the sensitivity was 83.3%, and the specificity was 75.3%. Conclusion:APACHEⅡscore, interval time between end of surgery and admission to ICU, and postoperative abdominal infection may be independent risk factors for the death of elderly patients who were admitted to the ICU after elective abdominal surgery, there would be far greater predictive values when the three variables were combined.
		                        		
		                        		
		                        		
		                        	
2.Analysis of the curative effect of discectomy under Quadrant expandable channel associated with annulus repair for the treatment of adolescent lumbar disc herniation
Liang ZHANG ; Jingcheng WANG ; Zhongjun LIU ; Xinmin FENG
Journal of Chinese Physician 2021;23(4):544-547,552
		                        		
		                        			
		                        			Objective:To analyze the curative effect of discectomy under Quadrant expandable channel associated with annulus repair for the treatment of adolescent lumbar disc herniation (ALDH).Methods:10 ALDH patients received discectomy under Quadrant expandable channel associated with annulus repair (annulus repair group) and 12 patients received discectomy (control group) in Clinical Medical College of Yangzhou University from January 2013 to December 2017 were retrospectively collected. The length of skin incision, amount of intraoperative bleeding, operation time and duration of hospitalization were compared. The degree of pain was assessed by visual analogue scale (VAS) before operation, 24 h, 1 week and 1 year after operation, and the lumbar function was assessed by Oswestry disability index (ODI) before operation, 1 week and 1 year after operation. At the last follow-up, the curative effect was evaluated by MacNab Scale, and the recurrence of lumbar disc herniation during the follow-up was recorded.Results:There were no significant differences in the amount of intraoperative bleeding, operation time and duration of hospitalization between the two groups ( P>0.05). The skin incision length of the annulus repair group was less than that of control group ( P<0.05). The postoperative VAS score and ODI score at each follow up time point in both groups were significantly improved when compared with the preoperative ones ( P<0.05). There were no statistically significant difference between the two groups in the VAS score and ODI score 1 week postoperative and 1 year postoperative ( P>0.05). According to the MacNab criteria, there was no statistically significant difference between the two groups in the excellent and good rate (annulus repair group: 9/10, control group: 10/12; P>0.05). There was no recurrence case in the annulus repair group, but two cases of recurrence (one recovered by conservative treatment, the other needed second operation) in the control group ( P<0.05). Conclusions:Discectomy under Quadrant expandable channel associated with annulus repair can achieve early satisfied outcome, reduce surgery related trauma, pain in the early period postoperative and recurrence rate after operation in the treatment of ALDH.
		                        		
		                        		
		                        		
		                        	
3.Metformin inhibits apoptosis by regulating TET2-Foxo3a pathway after spinal cord injury
Jiwei ZHAO ; Zhigang MIAO ; Huihui SUN ; Le HU ; Hao SUN ; Xiaoli ZHONG ; Xinmin FENG ; Jiandong YANG ; Yuping TAO ; Jun CAI ; Liang ZHANG ; Jingcheng WANG ; Yongxiang WANG
Chinese Journal of Orthopaedics 2021;41(9):584-594
		                        		
		                        			
		                        			Objective:Through TTC staining, immunohistochemical analysis, RT-PCR and hind limb motor function evaluation and other experimental methods, to explore the regulatory mechanism of metformin on anti-apoptosis in rats with spinal cord injury (SCI).Methods:Establish a rat spinal cord injury model. Through Basso-Beattie -Bresnahan locomotor rating scale (BBB) and cant test to evaluate the recovery of hindlimb motor function in rats. The changes of necrotic area of spinal cord tissue were compared by TTC staining. Extraction of rat spinal cord tissue, by Dot blot analysis and immunohistochemical detection of the hydroxyl of DNA methylation level. By qPCR, Western Blot detection TET2mRNA and protein expression level, and the changes in the scope of spinal cord injury were detected by inhibiting the expression of TET2. The interaction between TET2 and Foxo3a was detected by immunoblotting and immunoprecipitation. Through RT-PCR assay Foxo3a downstream related changes in the level of gene expression.Results:Compared with the SCI+NS group, the necrotic area of the spinal cord tissue was reduced after metformin treatment, and the BBB score and the incline test score were higher ( P<0.05). At the same time, we found that the levels of TET2mRNA and protein increased significantly after SCI at 24 h, and the 5-hmC level of DNA increased. The levels of TET2mRNA and protein and 5-hmC increased further after the use of metformin. After using SC-1, compared with the SCI+MET group, the level of 5-hmC decreased and the area of infarction increased. After SCI, the mRNA levels of downstream genes Bim, P27kip, Bax increased significantly. After metformin treatment, the mRNA levels of Bim and Bax were lower than those in the SCI+NS group ( P<0.05). After SCI, the 5-hmC levels of downstream genes Bim, P27kip, Bax increased significantly. After metformin treatment, the 5-hmC levels of Bim and Bax were lower than those in the SCI+NS group ( P<0.05). Conclusion:Metformin can promote the interaction between TET2 and Foxo3a, increase the 5-hmC level of the overall DNA, and inhibit the activation of related apoptosis genes, thereby improving tissue damage and nerve function recovery after spinal cord injury.
		                        		
		                        		
		                        		
		                        	
4.Effects of allicin on the biological behavior of vascular endothelial cells and epidural scar
Yujie ZHANG ; Jiandong YANG ; Jingcheng WANG ; Jun CAI ; Xinmin FENG ; Shoulei ZHU ; Yuan TIAN
Chinese Journal of Orthopaedics 2021;41(10):633-643
		                        		
		                        			
		                        			Objective:To investigate the effects of allicin on the proliferation, migration and angiogenesis of rat vascular endothelial cells (RVES), and to explore the influencing mechanism of allicin on epidural fibrosis.Methods:According to the results of preliminary experiments, RVECs were divided into control group (0 mg/L), low concentration group (25 mg/L), medium concentration group (50 mg/L) and high concentration group (100 mg/L). The morphology, viability, migration rate, cell cycle, apoptosis rate and cell lumen formation ability were measured using fluorescence microscope, AnnexinV-FITC double staining, PI/RN-asestaining, scratch assay and Transwell experiments test. Western Blot was used to measure the protein expression level of JAK2, STAT3, p-STAT3, PCNA, Bax and Bcl-2 protein. Using random number method, 36 adult male SD rats were divided into sham operation group, saline group and allicin group, with 12 rats in each group. Hematoxylin-eosin (HE) staining, Masson staining and immunohistochemical staining were used to analysis the epidural fibrosis in each group.Results:With the increase of concentration of allicin, cell viability, cell migration and lumen formation ability significantly lower than that of control group ( P<0.05). With the increase of allicin concentration, the percentage of cells in the G1 and S phases gradually decreased ( P<0.05), the percentage of cells in the G2 phase and the apoptosis rate gradually increased ( P<0.05), and the cells were blocked in the G2/M phase. With the increase of allicin concentration, the protein expression levels of JAK2, STAT3, p-STAT3, PCNA and Bcl-2 were gradually down-regulated ( P<0.05), while the protein expression level of Bax was gradually up-regulated ( P<0.05), the ratio of p-STAT3/STAT3 was decreased ( P<0.05), and the ratio of Bax/Bcl-2 was increased ( P<0.05). There was no death, infection or abnormal gait in all the experimental animals. Dense scar tissue could be observed in the extradural area of the sham operation group and the epidural area of the control group, but there was obvious space between the epidural scar and the dura mater in the allicin group, and the density of collagen, the number of blood vessels, and the protein density of p-STAT3 were significantly lower than those in the control group ( P<0.05). Conclusion:Allicin inhibits angiogenesis and the severity of epidural scar after laminectomy, and the mechanism may be through inhibiting the JAK2/STAT3 signaling pathway of vascular endothelial cells.
		                        		
		                        		
		                        		
		                        	
5.Application of satisfaction index of basic medical insurance for rural and urban residents to pupils familial decision making in Kunming and Changsha City.
Xiaofang LIU ; Wenwei CHENG ; Xiaofang YAN ; Liai PENG ; Xiaoxiao SONG ; Feng JIAO ; Jingcheng SHI ; Xia XIAO
Journal of Central South University(Medical Sciences) 2020;45(7):840-848
		                        		
		                        			OBJECTIVES:
		                        			To verify the applicability and extensibility of the satisfaction index of basic medical insurance for rural and urban residents, and to explore the mechanism responsible for the satisfaction index in Kunming and Changsha City, and provide references for effective management and policy making.
		                        		
		                        			METHODS:
		                        			A stratified cluster sampling method was conducted. A total of 560 familial decision makers were randomly selected in 24 classes of 14 schools of Kunming and Changsha City. Model reliability was tested by SPSS18.0. In addition, Smart PLS 3.0 was applied to conduct model validity test, calculate the satisfaction index, and to compare the model path coefficients of Kunming and Changsha by multi-group analysis.
		                        		
		                        			RESULTS:
		                        			In the application of the satisfaction index of basic medical insurance for rural and urban residents in Kunming, Cronbach's α of the model was 0.93, split-half reliability coefficient was 0.90, and the latent variable composite reliability coefficient values were more than 0.86; the latent variable average variance extraction (AVE) values were greater than 0.66, and the square root of the AVE of each latent variable (all greater than 0.66) was larger than the correlation coefficient with other latent variables. The factor loading values were greater than 0.70, with statistical significance. The basic health insurance satisfaction index of Kunming and Changsha was 60.40 and 52.05, respectively. The difference between the path coefficient of Kunming and Changsha was not statistically significant except the path from public satisfaction to public loyalty. Perceived value had the largest direct and total effect on public satisfaction latent variable in Kunming City. While the perceived value had the largest direct effect on public satisfaction, and the perceived quality had the largest total effect on public satisfaction in Changsha City.
		                        		
		                        			CONCLUSIONS
		                        			The satisfaction index model reflects the satisfaction of pupils' basic medical insurance for urban and rural residents, and it also shows good reliability and validity in Changsha and Kunming. What's more the model can be extended to the national level to evaluate the satisfaction of basic medical insurance for urban and rural residents for primary school students. The basic health insurance satisfaction index of familial decision makers in Kunming is higher than that of Changsha. There are differences between Kunming and Changsha in the influential mechanism of the satisfaction index of for Chinese pupils with basic medical insurance for rural and urban residents, and the measures taken by the government and relevant departments to improve the satisfaction of basic medical insurance should be based on local conditions.
		                        		
		                        		
		                        		
		                        			China
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		                        			Decision Making
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		                        			Humans
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		                        			Personal Satisfaction
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		                        			Reproducibility of Results
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		                        			Rural Population
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		                        			Urban Population
		                        			
		                        		
		                        	
6.Establishment of a rat caudal disc degeneration model by percutaneous fine-needle aspiration of the fibrous ring
Tao CHEN ; Jingcheng WANG ; Zhongjun LIU ; Jiandong YANG ; Xinmin FENG ; Zenan HUANG ; Songchao BI ; Liang ZHANG
Journal of Chinese Physician 2018;20(3):347-351
		                        		
		                        			
		                        			Objective To compare the effect of different size needle gauges to the degenerative response in rat caudal discs.Methods A total of 40 Sprague Dawley (SD) rats,level 5/6,7/8 and 9/10 interverbral discs of rat caudal spine were punctured with 18 or 21 or 25-gauge needles respectively.Radiographs and magnetic resonance imaging (MRI) were obtained at 1,2,4 and 6 weeks postsurgery.At each time point,ten rats from each group were sacrificed for histological analysis.Real-time fluorescent quantitative polymerase chain reaction (qPCR) was used to examine mRNA expression level.Results Significant differences were identified in the disc height index (DHI %) and MRI grade between 18 G and normal group,MRI grade,histological score between 21 G and normal group at 2,4,and 6 weeks postsurgery.Significant differences were also identified in the histological score and mRNA expression levels between 18 G and normal group,alcian blue stain and hypoxia inducible factor-1α (HIF-1 α) mRNA expression level between 21G and normal group at all time point postsurgery.Significant differences existed in DHI%,type Ⅱ collagen and aggrecan mRNA expression levels between 21 G and normal group,all type mRNA expression levels between 25 G and normal group at 4,6 weeks.There were significant differences in MRI grade and histological score between 25 G and normal group at 6 weeks.Significant differences existed in almost all parameters compared between 18 G and 25 G at all time point.There were significant differences in DHI%,MRI grade,histological score and HIF-1α mRNA expression levels between 18 G and 21 G at 4,6 weeks.There were significant differences in type Ⅱ collagen and aggrecan mRNA expression levels between 18 G and 21 G at all time point.Significant differences exist in DHI% and HIF-1α mRNA expression level between 21 G and 25 G at 6 weeks.Compared with the 25 G group,the DHI% and Pfirrmann scores and the pathological score of each time at 2,4 and 6 weeks after operation in 18 G group have significant difference (P < 0.05).Conclusions The middle size needle (21G) is better to induce disc degeneration.The 2-week time point may be the better time frame to further experimental treatments.
		                        		
		                        		
		                        		
		                        	
7.Effect of hydrogen on blood brain barrier of mice with sepsis-associated encephalopathy
Jingcheng FENG ; Yang YU ; Xing MAO ; Mengying YAN ; Guolin WANG ; Keliang XIE ; Yonghao YU
Chinese Journal of Anesthesiology 2018;38(6):719-722
		                        		
		                        			
		                        			Objective To evaluate the effect of hydrogen on blood brain barrier of mice with sepsisassociated encephalopathy (SAE).Methods A total of 100 adult male ICR mice,aged 6-8 weeks,weighing 20-25 g,were divided into 4 groups (n =25 each) using a random number table method:sham operation group (group Sham),sham operation plus hydrogen group (group Sham+H),group SAE and SAE plus hydrogen group (group SAE+ H).Sepsis was induced by cecal ligation and puncture (CLP).Sham+H and SAE+H groups inhaled 2% hydrogen for 1 h starting from 1 and 6 h after CLP,respectively.At 24 h after CLP,Evans blue (EB) was injected via the caudal vein,and then the mice were sacrificed and brain tissues were removed for measuring the EB and water contents,for examining the pathological changes of hippocampi (with a light microscope) and for detecting the expression of occludin and VE-cadherin (by Western blot).Morris water maze test was performed at days 10-16 after CLP.Results Compared with group Sham,the contents of EB and water in brain tissues were significantly increased,the expression of occludin and VE-cadherin was down-regulated,the escape latency was prolonged,and the number of crossing the original platform was reduced in SAE and SAE+H groups (P<0.05).Compared with group SAE,the contents of EB and water in brain tissues were significantly decreased,the expression of occludin and VE-cadherin was up-regulated,the escape latency was shortened,the number of crossing the original platform was increased (P<0.05),and the pathological changes of hippocampi were significantly attenuated in group SAE+H.Conclusion The mechanism by which hydrogen mitigates SAE may be related to reducing the damage to blood brain barrier of mice.
		                        		
		                        		
		                        		
		                        	
8.Changes in FUNDC1/LC3 Ⅱ signaling pathway during sepsis-induced liver injury in mice
Mengying YAN ; Yanyan WANG ; Xing MAO ; Jingcheng FENG ; Guolin WANG ; Keliang XIE ; Yonghao YU
Chinese Journal of Anesthesiology 2018;38(6):736-738
		                        		
		                        			
		                        			Objective To investigate the changes in FUNDC1/microtubule-associated protein 1 light chain 3 Ⅱ (LC3 Ⅱ) signaling pathway during sepsis-induced liver injury in mice.Methods Tbirtytwo clean-grade healthy male C57BL/6 mice,aged 6 weeks,weighing 20-25 g,were divided into sham operation group (n =8) and sepsis group (n =24) using a random number table method.Sepsis was induced by cecal ligation and puncture.Blood samples were obtained at 24 h after operation in sham operation group and at 6,12 and 24 h after establishing the model in sepsis group for determination of concentrations of alanine aminotransferase and aspartate aminotransferase in serum.Mice were then sacrificed,and the right lobe of livers was removed for examination of the pathological changes and for determination of the expression of FUNDC1 and LC3 Ⅱ by Western blot.The mitochondria in the right lobe of livers were isolated to measure the respiratory function,and respiratory control rate was calculated.Results Compared with sham operation group,the concentrations of alanine aminotransferase and aspartate aminotransferase in serum and pathological scores were significantly increased,the respiratory control rate of mitochondria was decreased,the expression of FUNDC1 was down-regulated,and the expression of LC3 Ⅱ was up-regulated at each time point after establishing the model in sepsis group (P<0.05).Conclusion The mechanism by which sepsis induces liver injury may be related to inhibiting activation of FUNDC1/LC3 Ⅱ signaling pathway in mice.
		                        		
		                        		
		                        		
		                        	
9.Role of autophagy in dexmedetomidine-induced reduction of lipopolysaccharide-caused inflammatory responses in macrophages of mice
Xing MAO ; Mengying YAN ; Hongguang CHEN ; Jingcheng FENG ; Guolin WANG ; Keliang XIE ; Yonghao YU
Chinese Journal of Anesthesiology 2018;38(8):992-995
		                        		
		                        			
		                        			Objective To evaluate the role of autophagy in dexmedetomidine-induced reduction of lipopolysaccharide ( LPS)-caused inflammatory responses in macrophages of mice. Methods Mouse mac-rophage cell line RAW264. 7 cultured in vitro were seeded in 6-well or 96-well plates and divided into 4 groups ( n=20 each ) when cell confluence reached 60% using a random number table method: control group (group Con), LPS group, LPS plus dexmedetomidine group (group LPS+DEX), and LPS plus dexmedetomidine plus autophagy inhibitor 3-MA group (group LPS+DEX+3-MA). PBS was added and cells were cultured for 12 h in group Con. LPS at the final concentration of 1000 ng∕ml was added and cells were incubated for 12 h in group LPS. LPS at the final concentration of 1000 ng∕ml was added, and then dexmedetomidine at the final concentration of 1 μmol∕L was immediately added, and cells were incubated for 12 h in group LPS+Dex. In group LPS+Dex+3-MA, 3-MA at the final concentration of 2 mmol∕L was added and cells were incubated for 1 h, LPS at the final concentration of 1000 ng∕ml was added, and then dexmedetomidine at the final concentration of 1 μmol∕L was immediately added, and cells were incubated for 12 h. Cell viability was detected by CCK-8 assay, and the concentrations of nitrous oxide ( NO) , tumor necrosis factor-alpha ( TNF-α) and interleukin-1beta ( IL-1β) in the supernatant were determined by en-zyme-linked immunosorbent assay, and the expression of microtubule-associated protein 1 light chain 3 Ⅰ( LC3 Ⅰ) , LC3Ⅱ, P62 and Bcelin-1 was detected by Western blot. LC3Ⅱ∕LC3Ⅰ ratio was calculated. Results Compared with group Con, the cell viability was significantly decreased, the concentrations of TNF-α, IL-1β and NO and LC3Ⅱ∕LC3Ⅰratio were increased, and the expression of P62 and Beclin1 was up-regulated in group LPS (P<0. 05). Compared with group LPS, the cell viability was significantly in-creased, the concentrations of TNF-α, IL-1βand NO were decreased, LC3Ⅱ∕LC3Ⅰratio was increased, the expression of P62 was down-regulated, and the expression of Beclin1 was up-regulated in group LPS+DEX ( P<0. 05) . Compared with group LPS+Dex, the cell viability was significantly decreased, the con-centrations of TNF-α, IL-1β and NO were increased, LC3Ⅱ∕LC3Ⅰ ratio was decreased, the expression of P62 was up-regulated, and the expression of Beclin1 was down-regulated in group LPS+Dex+3-MA ( P<0. 05) . Conclusion Enhanced autophagy is involved in dexmedetomidine-induced reduction of LPS-caused inflammatory responses in macrophages of mice.
		                        		
		                        		
		                        		
		                        	
10.Role of NLRP3 inflammasome in hydrogen-rich saline-induced reduction of LPS-caused damage to mitochondria in macrophages of mice
Hongguang CHEN ; Yang ZHANG ; Huanzhi HAN ; Xing MAO ; Jing TIAN ; Jingcheng FENG ; Keliang XIE ; Yonghao YU
Chinese Journal of Anesthesiology 2018;38(10):1241-1244
		                        		
		                        			
		                        			Objective To investigate the role of nucleotide-binding oligomerization domain-like receptor family pyrin domain containing 3 (NLRP3) inflammasome in hydrogen-rich saline-induced reduction of lipopolysaccharide (LPS)-caused damage to mitochondria in macrophages of mice.Methods Macrophage line RAW264.7 of mice were routinely cultured and divided into 4 groups (n=6 each) using a random number table method:control group (group C),group LPS,hydrogen-rich saline plus LPS group (group LPS+H2) and hydrogen-rich saline plus LPS plus ATP group (group LPS+ATP+H2).LPS was given at the concentration of 1 μg/ml,and the cells were then incubated for 30 min in group LPS.LPS at the concentration of 1 μg/ml and hydrogen-rich saline at the concentration of 0.6 mmol/L were simultaneously given,and the cells were then incubated for 30 min in LPS+H2 and LPS+ATP+H2 groups.ATP at the concentration of 1 nmol/L was then given,and the cells were incubated for 6 h in group LPS+ATP+H2.Mitochondrial membrane potential (MMP) was determined by JC-1 staining,and respiratory control ratio (RCR) was measured using a Clark-type electrode.The expression of NLRP3,caspase-1 and apoptosisassociated speck-like protein containing C-terminal caspase recruitment domain (ASC) was determined by Western blot.The concentrations of INTERLEUKIN-1 BETA (IL-1β),IL-18 and IL-6 in the supernatant were determined by enzyme-linked immunosorbent assay.Results Compared with group C,MMP and RCR were significantly decreased,the concentrations of IL-1β,IL-18 and IL-6 in the supernatant were increased,and the expression of NLRP3,caspase-1 and ASC was up-regulated in group LPS (P<0.05).Compared with group LPS,MMP and RCR were significantly increased,the concentrations of IL-1β,IL-l8 and IL-6 in the supernatant were decreased,and the expression of NLRP3,caspase-1 and ASC was down-regulated in group LPS+H2 (P<0.05).Compared with group LPS+H2,MMP and RCR were significantly increased,the concentrations of IL-1β,IL-18 and IL-6 in the supernatant were decreased,and the expression of NLRP3,caspase-1 and ASC was down-regulated in group LPS+ATP+H2 (P<0.05).Conclusion Hydrogen-rich saline can reduce LPS-caused damage to mitochondria in macrophages of mice through inhibiting the activation of NLRP3 inflammasome.
		                        		
		                        		
		                        		
		                        	
            
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