Effect of miR-141-3p targeting HMGB1 on LPS-induced A549 cell injury / 安徽医科大学学报

Objective @#To investigate the effect of miR-141-3p on LPS induced A549 cell injury by targeting high mobility group protein 1 (HMGB1) ．@*Methods @#A549 cells derived from type Ⅱ alveolar epithelial cells were taken as the study object，miR-141-3p mimics，mimics NC，HMGB1 gene overexpression plasmid (pcDNA3. 1-HMGB1) and empty Vector were transfected into A549 cells respectively or co-transfected，then 10 μg / ml LPS was used for 24 h．Cell proliferation activity was detected by cell counting kit-8 ( CCK-8) ．The activity of lactate dehydrogenase ( LDH) in the supernatant of cell culture was detected by colorimetry．The apoptosis level of each group was detec- ted by flow cytometry．The levels of interleukin (IL) -1 β , IL-6 and tumor necrosis factor α (TNF-α) were detected by enzyme-linked immunosorbent assay (Elisa) ．Dual luciferase reporter gene assay verified the targeted regulatory relationship between miR-141-3p and HMGB1 . @*Results @#After treatment with LPS ，the proliferative activity of A549 cells and the expression level of miR-141-3p decreased ( P ＜0. 05 ) ，the apoptosis rate increased ( P < 0. 05) ，the levels of IL-1 β , IL-6，TNF-α and the activity of LDH in supernatant increased (P＜0. 05) ．Overex- pression of miR-141-3p increased the proliferation activity of A549 cells treated with LPS (P ＜0. 05 ) ，and de- creased the apoptosis rate and the levels of IL-1 β , IL-6，TNF-α in cells and LDH activity in supernatant (P < 0. 05) ．However，overexpression of HMGB1 gene could reverse the ameliorative effect of miR-141-3p on LPS-in- duced A549 cell injury．Dual luciferase reporter gene experiment confirmed that HMGB1 was the downstream target gene of miR-141-3p．@*Conclusion @# miR-141-3p can inhibit LPS-induced apoptosis，reduce the expression level of inflammatory factors，and improve the damage of A549 cells，which may be related to the targeted regulation of HMGB1 expression.

Impacts of miR-141-3p on pulmonary fibrosis in rats with acute respiratory distress syndrome by regulating Keap1-NRF2/ARE signaling pathway / 天津医药

Objective To investigate the effect and mechanism of miR-141-3p on pulmonary fibrosis in rats with acute respiratory distress syndrome(ARDS).Methods Rats were divided into the control group,the model group,the agomir negative control group and the miR-141-3p agomir group according to random number table,with 10 rats in each group.In addition to the control group,the ARDS rat model was established by lipopolysaccharide(LPS)infusion.Rat alveolar typeⅡepithelial cells RLE-6TN cells were divided into the NC group,the LPS group,the miR-NC group,the miR-141-3p mimics group,the miR-141-3p mimics+pcDNA group and the miR-141-3p mimics+NRF2 and Kelch-like ring associated protein 1(Keap1)group.LPS cell model was established in all groups except the NC group.The mRNA expression levels of miR-141-3p and Keap1 in lung tissue and cells were detected by qPCR.Western blot assay was used to analyze lung tissue and cell epithelial cadherin(E-cadherin),neural cadherin(N-cadherin),microtubule associated protein light chain 3B(LC3B),autophagy associated gene Beclin-1,α-smooth muscle actin(α-SMA),type I collagen(Col-Ⅰ),Keap1 and nuclear factors E2 related factor 2(NRF2)and heme oxygenase 1(HO-1).HE staining and Masson staining were used to observe pathological changes of lung tissue and to estimate the area of lung tissue injury and pulmonary fibrosis.Hydroxyproline(Hyp)in lung tissue was detected by the kit.Levels of inflammatory factor interleukin-1β,tumor necrosis factor(TNF-α)and oxidative stress index malondialdehyde(MDA)and superoxide dismutase(SOD)were detected by ELISA.Dual luciferase reporting experiment was used to verify the targeting relationship between miR-141-3p and Keap1.Results The expression of miR-141-3p was down-regulated and the expression of Keap1 was up-regulated in lung tissue and cells(P＜0.05).Overexpression of miR-141-3p can reduce the degree of pathological damage and fibrosis of lung tissue in rats,Hyp content,and up-regulate expression levels of SOD,E-cadherin,LC3B,Beclin-1,NRF2 and HO-1 in lung tissue and cells,and down-regulate the expression levels of IL-1β,TNF-α,MDA,N-cadherin,α-SMA,Col-I and Keap1(P＜0.05).Overexpression of Keap1 was able to reverse the improvement effect of overexpression of miR-141-3p on alveolar epithelial cell damage in ARDS rats(P＜0.05).Double Luciferase reporter gene experiment confirmed that miR-141-3p and Keap1 may have a targeted regulatory relationship.Conclusion Overexpression of miR-141-3p may activate the Keap1-NRF2/ARE signaling pathway,activate autophagy,inhibit inflammatory response,oxidative stress,and EMT progression,and improve pulmonary fibrosis in ARDS rats.

Investigation of infectious disease-specific health literacy among rural residents in Dongxiang Autonomous County / 预防医学

Objective@#To investigate the influencing factors of infectious disease-specific health literacy (IDSHL) among rural residents in Dongxiang Autonomous County, and to construct a nomogram-based model for prediction of IDSHL.@*Methods@#Totally 1 250 rural residents at ages of 15 years and older were sampled from Dongxiang Autonomous County using a stratified random sampling method. Participants' IDSHL was evaluated using the IDSHL Assessment Scale among Chinese Residents, and factors affecting the participants' IDSHL were identified using a multivariable logistic regression model. A nomogram-based model was created, and the predictive effectiveness of this model was evaluated using the receiver operating characteristic (ROC) curve, Hosmer-Lemeshow test and C-index.@*Results@#A total of 1 223 valid respondents were enrolled, including 687 men (56.17%) and 536 women (43.83%), and the proportion of IDSHL was 48.48%. Multivariable logistic regression analysis identified age (reference: 60 years and older; 30 to <40 years: OR=4.273, 95%CI: 2.397-7.617; 40 to <50 years: OR=3.938, 95%CI: 2.238-6.928), education level (reference: illiteracy/semi-illiteracy; primary school: OR=2.140, 95%CI: 1.456-3.144; high school/vocational high school/technical secondary school: OR=2.914, 95%CI: 1.652-5.138; junior college and above: OR=4.514, 95%CI: 2.261-9.011), healthcare seeking/medications in the past 2 weeks (reference: yes; no: OR=2.025, 95%CI: 1.346-3.046), self-rated health (reference: good; generally: OR=0.603, 95%CI: 0.376-0.966; poor: OR=0.462, 95%CI: 0.284-0.751) and daily average duration spent online (reference: no internet access; <1 h: OR=1.859, 95%CI: 1.306-3.437; 1 to <2 h, OR=1.996, 95%CI: 1.344-3.380; 2 to <3 h: OR=2.132, 95%CI: 1.109-3.116; 3 h and longer: OR=2.119, 95%CI: 1.175-3.390) as factors affecting IDSHL among rural residents in Dongxiang Autonomous County. The area under the ROC curve of the model was 0.774 (95%CI: 0.741-0.807) and the model had high calibration and differentiation levels [Hosmer-Lemeshow test: χ2=13.276, P=0.103; internal model validation (bootstrapping): mean absolute error=0.019; C-index=0.764].@*Conclusions@#Age, education level, healthcare seeking/medications in the past 2 weeks, self-rated health status and daily average duration spent online are factors affecting IDSHL among rural residents in Dongxiang Autonomous County. The nomogram model created based on these factors has a high efficiency and applicability for prediction of IDSHL among rural residents in Dongxiang Autonomous County.

Clinical Observation of Low-dose Ropivacaine Combined with Sufentanil in Upper Limb Anesthesia / 中国药房

OBJECTIVE:To observe the efficacy and safety of low-dose ropivacaine combined with sufentanil in the upper limb anesthesia. METHODS:100 patients with upper limb anesthesia were randomly divided into control group and test group. Control group was intravenously injected 20 ml 0.5% ropivacaine;test group was intravenously injected 20 ml 0.375% ropivacaine +10 μg sufentanil. The anesthesia effect,disappearance time of pain,duration of analgesia,and pain visual analogue (VAS) score before anesthesia(T0),5 min(T1),15 min(T2)and 30 min(T3)after anesthetic block,and 4 h(T4)and 8 h(T5)after surgery,and in-cidence of adverse reactions in 2 groups were observed. RESULTS:VAS scores at T1-T5 in test group and T2-T4 in control group were significantly lower than T0,test group was lower than control group at T1,T2 and T5,the differences were statistically signifi-cant(P<0.05). The total effective rate of anesthesia in test group was significantly higher than control group,disappearance time of pain was significantly shorter than control group,duration of analgesia was significantly longer than control group,incidence of adverse reactions was significantly lower than control group,the differences were statistically significant(P<0.05). CONCLU-SIONS:Both the efficacy and safety of low-dose ropivacaine combined with sufentanil in the upper limb anesthesia are good.

[Protective effect of erythropoietin on brain tissue in rats with cardiopulmonary resuscitation after asphyxia].

OBJECTIVE: To study the protective effect of erythropoietin (EPO) on brain tissue with cardiac arrest-cardiopulmonary resuscitation (CA-CPR) and its mechanism. METHODS: 120 male Sprague-Dawley (SD) rats were randomly divided into three groups (each n = 40), namely: sham group, routine chest compression group, and conventional chest compression + EPO group (EPO group). The rats in each group were subdivided into CA and 6, 12, 24, 48 hours after restoration of spontaneous circulation (ROSC) five subgroups (each n = 8). The model of CA was reproduced according to the Hendrickx classical asphyxia method followed by routine chest compression, and the rats in sham group only underwent anesthesia, tracheostomy intubation and venous-puncture without asphyxia and CPR. The rats in EPO group were given the routine chest compression + EPO 5 kU/kg (2 mL/kg) after CA. Blood sample was collected at different time points of intervention for the determination the content of serum S100 ß protein by enzyme linked immunosorbent assay (ELISA). All the rats were sacrificed at the corresponding time points, and the hippocampus was harvested for the calculation of the number of S100 ß protein positive cells, and to examine the pathological changes and their scores at 24 hours after ROSC by light microscopy. RESULTS: With prolongation of ROSC time, the serum levels of S100 ß protein (µg/L) in the routine chose compression group and the EPO group were significantly elevated, peaking at 24 hours (compared with CA: 305.7 ± 29.2 vs. 44.4 ± 6.2 in routine chest compression group, and 276.7 ± 28.9 vs. 44.7 ± 5.6 in the EPO group, both P < 0.05), followed by a fall. The levels of S100 ß protein at each time point after ROSC in EPO group were significanthy lower than those of the routine chest compression group (83.2 ± 7.5 vs. 114.3 ± 15.3 at 6 hours, 123.9 ± 20.2 vs. 184.9 ± 22.2 at 12 hours, 276.7 ± 28.9 vs. 305.7 ± 29.2 at 24 hours, 256.3 ± 26.6 vs. 283.2 ± 23.6 at 48 hours, all P < 0.05). With the prolongation of ROSC time, the S100 ß protein positive cell number in brain (cells/HP) in the routine chest compression group and the EPO group was significantly increased, peaking at 24 hours (compared with CA: 14.3 ± 2.2 vs. 6.7 ± 0.7 in the routine chest compression group, 11.3 ± 1.3 vs. 6.8 ± 0.9 in the EPO group, both P < 0.05), then it began to fall. The S100 ß protein positive cell number in brain at each time point after ROSC in the EPO group was significantly lower than that of the routine chest compression group (7.0 ± 0.9 vs. 7.9 ± 1.9 at 6 hours, 8.4 ± 1.1 vs. 10.2 ± 2.2 at 12 hours, 11.3 ± 1.3 vs. 14.3 ± 2.2 at 24 hours, 8.3 ± 0.8 vs. 10.8 ± 2.0 at 48 hours, all P < 0.05). Under the light microscope, a serious brain cortex injury was found after reproduction of the model, and the degree of injury was reduced after EPO intervention. The pathological score at 24 hours after ROSC in EPO group was lower than that of routine chest compression group (3.83 ± 0.73 vs. 4.17 ± 0.75, P < 0.05). CONCLUSIONS: The S100 ß protein level in serum and brain tissue was increased early in asphyxia CA-CPR rats. EPO intervention can reduce the expression of S100 protein and reduce the degree of brain injury.

Comparison of Therapeutic Efficacy of Desflurane and Sevoflurane for Anesthesia of Elderly Patients / 中国药房

OBJECTIVE:To compare therapeutic efficacy and safety of desflurane and sevoflurane for anesthesia of elderly pa-tients. METHODS:A randomized,parallel-controlled method was used,120 elderly patients undergoing anesthesia were random-ized into trial group and control group,with 60 cases in each group. Based on routine medication and disposal,control group was additionally given sevoflurane for anesthesia,and trial group given desflurane for anesthesia. The blood pressure and heart rate were recorded before anesthesia,2 min and 10 min after intubation,2 min and 30 min after the start of the operation;the extubation time,eye opening time,consciousness recovery time and postoperative complications were also recorded. RESULTS:10 min after intubation,the heart rate of trial group was significantly faster then that of control group,with statistical significance(P0.05). The extubation time and consciousness recovery time of trial group were significantly shorter than those of control group, with statistical significance(P<0.05). The postoperative complications mainly were hypopiesia,hypertension,bronchospasm,nau-sea and vomiting,and dysphoria;the incidence of postoperative complications in trial group and control group were 25.0% and 40.0%,with statistical significance (P<0.05). CONCLUSIONS:Desflurane is effective with less postoperative complications for the anesthesia of elderly patients.

Protective effect of erythropoietin on brain tissue in rats with cardiopulmonary resuscitation after asphyxia / 中华危重病急救医学

Objective To study the protective effect of erythropoietin (EPO) on brain tissue with cardiac arrest-cardiopulmonary resuscitation (CA-CPR) and its mechanism.Methods 120 male Sprague-Dawley (SD) rats were randomly divided into three groups (each n =40),namely:sham group,routine chest compression group,and conventional chest compression + EPO group (EPO group).The rats in each group were subdivided into CA and 6,12,24,48 hours after restoration of spontaneous circulation (ROSC) five subgroups (each n =8).The model of CA was reproduced according to the Hendrickx classical asphyxia method followed by routine chest compression,and the rats in sham group only underwent anesthesia,tracheostomy intubation and venous-puncture without asphyxia and CPR.The rats in EPO group were given the routine chest compression + EPO 5 kU/kg (2 mL/kg) after CA.Blood sample was collected at different time points of intervention for the determination the content of serum S100 β protein by enzyme linked immunosorbent assay (ELISA).All the rats were sacrificed at the corresponding time points,and the hippocampus was harvested for the calculation of the number of S100 β protein positive cells,and to examine the pathological changes and their scores at 24 hours after ROSC by light microscopy.Results With prolongation of ROSC time,the serum levels of S100 β protein (μg/L) in the routine chose compression group and the EPO group were significantly elevated,peaking at 24 hours (compared with CA:305.7 ± 29.2 vs.44.4 ± 6.2 in routine chest compression group,and 276.7±28.9 vs.44.7±5.6 in the EPO group,both P ＜ 0.05),followed by a fall.The levels of S100β protein at each time point after ROSC in EPO group were significanthy lower than those of the routine chest compression group (83.2 ± 7.5 vs.114.3 ± 15.3 at 6 hours,123.9 ± 20.2 vs.184.9 ± 22.2 at 12 hours,276.7 ± 28.9 vs.305.7 ± 29.2 at 24 hours,256.3 ± 26.6 vs.283.2 ± 23.6 at 48 hours,all P ＜ 0.05).With the prolongation of ROSC time,the S100 β protein positive cell number in brain (cells/HP) in the routine chest compression group and the EPO group was significantly increased,peaking at 24 hours (compared with CA:14.3±2.2 vs.6.7±0.7 in the routine chest compression group,11.3± 1.3 vs.6.8±0.9 in the EPO group,both P ＜ 0.05),then it began to fall.The S100 β protein positive cell number in brain at each time point after ROSC in the EPO group was significantly lower than that of the routine chest compression group (7.0±0.9 vs.7.9± 1.9 at6 hours,8.4± 1.1 vs.10.2±2.2 at 12 hours,11.3± 1.3 vs.14.3±2.2 at24 hours,8.3±0.8 vs.10.8±2.0 at48 hours,all P ＜ 0.05).Under the light microscope,a serious brain cortex injury was found after reproduction of the model,and the degree of injury was reduced after EPO intervention.The pathological score at 24 hours after ROSC in EPO group was lower than that of routine chest compression group (3.83±0.73 vs.4.17±0.75,P ＜ 0.05).Conclusions The S100β protein level in serum and brain tissue was increased early in asphyxia CA-CPR rats.EPO intervention can reduce the expression of S100 protein and reduce the degree of brain injury.

Influence of high-fat diet in intestinal flora and fecal weight in SD rats and its significance / 吉林大学学报(医学版)

Objective To investigate the influence of high-fat diet in the intestinal flora,fecal weight and its water content in rats, and to clarify the effect and significance of high-fat diet in the occurrence of obesity forming. Methods Twenty Sprague-Dawley (SD)rats were randomly divided into normal diet (ND)group and high-fat diet (HF)group (n=10).The rats in ND group were fed with normal diet,and the rats in HF group were fed with diet rich in oil and fat.The fresh feces were collected separately on days 1,15,30,and 49 for analysis of weight, water content,and intestinal flora.Results On the 49th day,the wet weight and water content of feces of the rats in ND group were (6.61 ± 0.17)g and (37.07 ± 3.04)%,respectively,while those in HF group were (4.46±0.30)g and (18.04±2.23)% (P<0.05).Compared with ND group ,the fecal pellets in HF group were increased obviously from the 7th day (P<0.05).There were obvious changes in intestinal microbial populations of HF group. The counts of enterococci, bifidobacteria, Lactobacillus, Bacteroides bacteria were significantly decreased on the day 49, but the count of Escherichia coli was increased significantly (P < 0.05 ). Conclusion High-fat diet can result in decrease for weight,water content,and pellets of feces;it can change the structure of intestinal flora. As result, there is a possibility that all of changes above can promote obesity in the future.

Dexmedetomidine intervention effects on aquaporin -1 and inflammatory cytokines in myocardial tissue of septic rat / 中国中西医结合急救杂志

Objective To study the regulation of aquaporin-1(AQP-1)changes in the heart of septic rats, compare the correlations of the AQP-1 with myocardial cytokines tumor necrosis factor-α(TNF-α),interleukin-6 (IL-6),and myocardial tissue water content,and to investigate the dexmedetomidine protective effect on myocardia in septic rats and its possible mechanism. Methods According to the random number table methods,90 male Sprague-Dawley(SD)rats were divided into sham operation group,sepsis model group and dexmedetomidine group, 30 rats in each group. The rat sepsis model was established by cecal ligation and puncture(CLP). In the sham operation group,the animal abdomen was only opened and closed without CLP. Half hour before operation in dexmedetomidine group,dexmedetomidine 1μg/kg(2μg/mL)was injected into the vein,while in the model and sham groups,saline 5 mL/kg was subcutaneously injected into the rat after the operation. At 2,12,24,48,72 hours after operation,6 rats were sacrificed and their hearts removed at one time point in a group. Enzyme linked immunosorbent assay(ELISA)was used to detect the content of AQP-1 and the levels of the TNF-α,IL-6 in the myocardial tissue homogenate at all time points,the myocardial tissue water content was detected by dry wet weight,and the correlations between AQP-1 and TNF-α,IL-6 and between AQP-1 and myocardial tissue water content were compared. Results From 2 hours after operation,the levels of the AQP-1,TNF-αand IL-6 in model group were significantly higher than those in the sham operation group;with prolongation of time,the level of AQP-1 and myocardial tissue water content were decreased, but the levels of TNF-α and IL-6 were persistently increased. From 2 hours after operation in dexmedetomidine group,all the above indexes except myocardial tissue water content at 72 hours after operation were significantly lower than those in the model group〔AQP-1(ng/g):9.29±0.15 vs. 9.73±0.26,TNF-α(pg/g):109.47±8.41 vs. 128.13±7.36,IL-6(pg/g):232.95±20.56 vs. 279.71±22.24,myocardial tissue water content:(74.82±6.37)%vs.(75.62±6.39)%,all P<0.05〕,but still higher than those of the sham operation group. The correlation analyses for the septic group showed that the change of AQP-1 was positively correlated to the myocardial water content in early stage(r=0.418,P=0.001)and later stage(r=0.235,P=0.022),and the changes of the AQP-1 in early stage (at post-operative 2 hours)were positively correlated to the concentration changes of the cytokines TNF-α(r=0.235,P=0.021)and IL-6(r=0.345,P=0.003),but in the later stage(at post-operative 72 hours)were negatively correlated with the changes of TNF-α(r=-0.408,P=0.037)and IL-6(r=-0.276,P=0.002). Conclusions In the early stage of septic rats,there is obvious myocardial injury,resulting in the over expression of AQP-1 and the occurrence of myocardial edema,dexmedetomidine can play a role in myocardial protection in such rats and its mechanism is possibly related to the reduction of the expression of AQP-1 and the levels of inflammatory cytokines, and in turn the alleviation of myocardial cell edema.

The effects of ulinastatin on the cells factor and cardiac function after cardiopulmonary resuscitation in rats / 重庆医学

Objective To investigate the protective effects of ulinastatin on the hearts of rats with anoxia-induced cardiac arrest-cardiopulmonary resuscitation(CA-CPR) and the mechanism of improving cardiac dysfunction .Methods Twenty male Sprague Dawley(SD) rats were randomly divided into three experimental groups :sham operation group (group A ,n= 8 ,only anesthesia , tracheotomy tube and vascular puncture) ,control group(group B ,n= 6 ,normal saline 4 mL · kg -1 injected via vein) ,Ulinastatin treatment group(group C ,n=6 ulinastatin 50 000 U/kg+normal saline 3 mL · kg -1 injected via vein);Factors including mean arte-rial pressure(MAP) ,left ventricular end diastolic pressure(LVEDP) ,the maximum rising and falling rates of left ventricular deep pressure(± LVdp .dt-1max) ,brain natriuretic peptide(BNP) ,cardiac troponin T(cTNT) ,IL-12 and TNF-αwere observed at setting time before and after cardiopulmonary resuscitation in rats .Results Compared with those of the group A and before CA-CPR ,the concentrations of IL-12、cTNT、TNF-α、BNP、and LVEDP increased(P<0 .01)while ± LVdp .dt-1max decreased(P<0 .01) at 6 h after CA-CPR in group B ,C .Compared with those of group B ,the concentrations of IL-12、CTNT、TNF-α、BNP and LVEDP of 6 h after CA-CPR in group C were lower and ± LVdp .dt-1max was higher(P<0 .01) ,The concentrations of MAP of 6 h after CA-CPR in group B was lower Compared with that of group A ,C and before CA-CPR(P<0 .01) .Conclusion Ulinastatin can improve cardiac dysfunction by depressing mediators of inflammation and reducing myocardial injury .

Effects of Xuebijing Injection on the Cardiac Function and Calcium Ions in Cardiac Myocytes of Rats after Anoxia-induced Cardiac Arrest-Cardiopulmonary Resuscitation(CA-CPR) / 中国药房

OBJECTIVE: To investigate the effect of Xuebijing injection on the cardiac function and calcium ions of cardiocyte in rats after anoxia-induced cardiac arrest-cardiopulmonary resuscitation(CA-CPR).METHODS: CA-CPR model was induced in rats and then the model rats were randomized to 4 groups,i.e.sham group,model group,Xuebijing injection high dose group(4 mL?kg-1),and Xuebijing injection lowdose group(Xuebijing injection 2 mL?kg-1).Mean arterial pressure(MAP),?LVdp/dt max,average fluorescence intensity of calcium and pathological changes of cardiocytes were observed at 0 and 6 hours after resuscitation,respectively.RESULTS: Compared with model group,Xuebijing ingection high dose group at 6 h showed significantly increased MAP(P