ObjectiveTo investigate the effects of HIF-1α,on mitochondrial function of cardiomyocytes stimulated by lipopolysaccharide (LPS).MethodsCultured neonatal cardiomyocytes were randomly (random number) distributed to four groups (n =5).Control group:the neonatal cardiomyocytes were cultured in normal condition without any treatmeat; LPS group:cardiomyocytes were exposed to 1 μg/ml LPS for 2 h; YC-1 + LPS group:cardiomyocytes were exposed to 4 μmol/L YC-1 for 8 h before LPS treatment; YC-1 group:cardiomyocytes were exposed to 4μmol/L YC-1 for 8 h.As indexes of mitochondrial function,MMP,the content of ATP and the activity of cytochrome oxidase were measured,COX1/2 protein expression was determined by Western blotting.Results Compared with control group,LPS decreased MMP,the content of ATP and the activity of COX obviously ( P ＜0.05 ),depressed the expression of COX- 1 protein ( P ＜ 0.05 ),induced the expression of COX-2 protein ( P ＜ 0.05 ).Compared with the LPS group,MMP,the content of ATP and the activity of COX were markedly attenuated ( P ＜ 0.01 ),the expression of COX-1 was significantly increased and the expression of COX-2 was decreased in theYC-1 +LPS group (P ＜ 0.01 ).ConclusionsYC-1 attributes to the dysfunction of mitochondrial of myocyte by alterating the expression of COX-1 and 2, HIF-1α may attenuate the mitochondrial dysfunction of cadiomyocytes stimulated by LPS.

Objective To examine the role of the reactive oxygen species (ROS) and GADD 153 protein in angiotensin Ⅱ (Ang Ⅱ)-induced cardiomyocyta apeptosis in vitro.Methods Cardiomyocytes were isolated from ventricles of healthy 1-2 day old Wistar mrs of both sexes weighing 5-10 g and cultured in high glucose culture medium in an incubator filled with 5% CO2 at 37℃.The cardiomyocytes were randamiy allocated to one of 9 groups (n=5 each): group Ⅰ control (C); group Ⅱ ,Ⅲ,Ⅳ: cardiomyocytes were exposed to Ang Ⅱ 100 nmol/L for 6 h,12 h or 24 h respectively ( Ang Ⅱ 6,Ang Ⅱ12,Ang Ⅱ24) ; group Ⅴ:cardiomyocytes were preincubated with N-acetyl-L-cysteine (NAC) 5 mmol/L for 2 h before being exposed to Aag Ⅱ 100 nmol/L for 24 h (NAC + Ang Ⅱ); group Ⅵ:cardiomyocytes were exposed to NAC 5 mmol/L for 24 h (NAG); group Ⅶ:cardiomyocytes were tranfected with GADD153 antisense oligo-denxynueleotida (onti-ODN) 10 μmol/L for 24 h before being exposed to Ang Ⅱ 100 nmol/L for 24 h; group Ⅷ: cardiomyocytes were transfected with missense oligo-deoxynucleotide (mis-ODN) 10 μmol/L for 24 h before being exposed to Ang Ⅱ 100 nmol/L for 24 h and group Ⅸ: cardiomyocytes were exposed to transfection agent of same concentration for 24 h.Viability of myocytes was measured by MTT assay; ROS production was determined by spectropbotometry; apoptosis in cardiomyocytes was detected by staining with Hoechst33342; the percentage of Annexin V positive and PI negative myocytes was measured by flow cytometry as apoptosis rote and GADD153 protein expression was determined by Western blotting.Remits The viability of myocytes was significantly lower,while the ROS production,apoptosis rate and GADD153 protein expression in myocytes were significantly higher in group Ⅱ,Ⅲ,and Ⅳ(Ang Ⅱ6,Ang Ⅱ12,AngⅡ24) than in control group (P＜0.05).The cell viability was significantly higher and the ROS production,apoptosis rate and GADD153 protein expression in myocytes were significantly lower in groupⅤ(NAC+AngⅡ)and Ⅶ(anti-ODN + Ang Ⅱ) than in group Ang Ⅱ24(Ⅳ).Conclusion Angiotensin Ⅱ induces cardiomyocyte apoptosis by increasing ROS production and up-regulating GADD153 protein expression.

AIM: To investigate the relationship between alteration of CHOP/GADD153 protein expression and cardiomyocyte apoptosis induced by angiotensin Ⅱ (AngⅡ), and inhibitory effects of CHOP/GADD153 antisense oligodeoxynucleotide (anti-ODN) on apoptosis in vitro.METHODS: Cultured neonatal cardiomyocytes were exposed to AngⅡ with or without preincubation of CHOP/GADD153 anti-ODN. Variability of myocytes was measured by MTT assay, the lactate dehydrogenase (LDH) release was also detected, the percentage of annexin V positive myocytes was monitored by flow cytometry as apoptosis rate, CHOP/GADD153, Bcl-2 and Bax expressions were determined by Western blotting. RESULTS: Compared with control group, the expression of CHOP/GADD153 was obviously increased from (0.20?0.02 to 0.75?0.06) in AngⅡ group (P

Iatrogenic vertebral artery injury is a rare complication in cervical spine surgeries,the consequences of which may be catastrophic.With the extensive development of cervical spine surgeries and application of diverse new internal fixations of cervical spine,there is a potential increase of risk of iatrogenic vertebral artery injury,which should be paid attention to.This article reviews the rate,cause and management etc.of iarogenic vertebral artery injury.

OBJECTIVETo observe the effect of sodium tanshinone II (A) sulfonate (STS) on Ang II -induced atrial fibroblast collagen synthesis and TGF-beta1 activation.

METHODAtrial fibroblasts of neonatal rats were cultured to determine the content of collagen protein. The original synthesis rate determined by the [3H]-proline incorporation method was taken as the index for myocardial fibrosis. The content of active TGF-beta1 and total TGF-beta1 in cell culture supernatants were tested and cultured by ELISA. The expression of thrombospondin-1 (TSP-1) was assessed by using Western blot.

RESULTAng II could significantly increase the content of atrial fibroblast collagen and the collagen synthesis rate, the TSP-1 expression and the concentration of active TGF-beta1, without any obvious change in total TGF-beta1. After the STS treatment, all of the indexes, apart from total TGF-beta1, were obviously down-regulated.

CONCLUSIONSTS could decrease the secretion of Ang II -induced atrial fibroblast collagen and the synthesis rate. Its mechanism is related to the inhibition of TSP-1/TGF-beta1 pathway.

Angiotensin II ; pharmacology ; Animals ; Collagen ; biosynthesis ; Fibroblasts ; cytology ; drug effects ; metabolism ; Gene Expression Regulation ; drug effects ; Heart Atria ; cytology ; Phenanthrenes ; pharmacology ; Rats ; Rats, Wistar ; Signal Transduction ; drug effects ; Thrombospondin 1 ; metabolism ; Transforming Growth Factor beta1 ; metabolism

Objective The effect of lead exposure on children is consistently associated with intellectual and other neurologic deficits.However the exact mechanism by which Pb~(2+) exerts toxic effects on developmental central nervous system remains unknown.Our group has found by gene-chip test that the expression of metabotropic glutamate receptor subtype 5 (mGluR5) mRNA was changed by lead exposure.The present study aimed to examine the effects of different level of lead exposure on the expression of mCluR5 in gestation and lactation.Methods Sprague-Dawley rats were exposed to lead acetate during gestation and lactation.Three concentrations of 0.05%,0.2%,and 0.5% lead acetate were applied and considered as low,middle and high exposure group respectively.The Pb levels of blood and hippocampus of pups were analyzed at weaning to evaluate the actual lead content at the end of the exposure.The impact of lead exposure on the expression of mGluR5 mRNA and protein in hippocampal tissue of pups was investigated by quantitative real-time polymerase chain reaction (PCR) and Western blot.The potential role of the expression of mGluR5 mRNA and protein in lead neurotoxicity were discussed.Results The levels of lead in blood and hippocampus from lead-exposed rats were significantly higher than those in the controls and positively related to the degree of lead exposure.The results of real-time PCR and Western blot showed that exposure to lead acetate decreased the expression of mCluR5 mRNA and protein with a dose-dependent manner.Conclusions Hippocampal mGluR5 might be involved in lead-induced neurotoxicity.

Objective:3D hydrogel cell model was established,and cyclic compressive loading on MC3T3-E1 cell with different intensities,frequencies and durations was applied,in order to research the suitable solution about promoting the osteoblast differentiation with cyclic compression.Methods:Cyclic compressive loading on MC3T3-E1 cell was applied with different intensity,frequency and time.After compressive loading finished,the total RNA extraction from cell-gel constructs were performed and quantified ATF4,ALP,Runx2,Osteocalcin,RANKL and RANK mRNA.Results:RANKL and RANK mRNA expression significantly with different frequencies cyclic compressive loading (P ＜ 0.05),and ALP mRNA (P ＜ 0.05) and Runx2 mRNA (P ＜ 0.01) expression significantly with different intensities and frequencies cyclic compressive loading (P ＜ 0.05).Meanwhile,Runx2 mRNA expression with 4h significant higher than 12h (P ＜ 0.05),and RANKL mRNA expression with 4h significant lower than 12h (P ＜ 0.05).Conclusion:Determine the stress intensity and frequency,1％ intensity,frequency of 0.5 Hz,4 h of cyclic compression intervention could promote the growth of osteoblasts-like cells in the 3D hydrogel model.

Objective:To investigate the risk factors of central lymph node metastasis (CLNM) in cN0 paillary thyroicl microcarcinoma (PTMC) and to establish a nomogram model for predicting the probability of cN0 PTMC CLNM.Methods:The clinicopathological data of 192 patients with cN0 PTMC admitted to the Department of General Surgery of the Second Hospital of Lanzhou University from Aug. 2016 to Aug. 2020 were retrospectively analyzed. There were 41 males and 151 females, 50 with CLNM and 142 without CLNM. The patients were divided into 2 groups according to the presence or absence of pathologically confirmed CLNM. Patient’s age, gender, tumor diameter, multiple, with Hashimoto’s disease, with nodular goiter, with or without near the posterior dorsal membrane, aspect ratio >1, with or without extratumoral infiltration, with or without lymphadenopathy, TSH levels, and TG levels were statistically analyzed. Pearson chi-square test was used to analyze the count data of hypothesis test, and the R language software package was used for Logistic multivariate analysis. The entry conditions were screened by stepwise regression to establish a nomogram prediction model, and the Bootstrap method was used for model verification. P<0.05 was considered statistically significant. Results:Multivariate logistic analysis showed that extratumoral invasion ( P=0.032) , presence of lymphadenopathy ( P=0.010) , and TG>68 μg/L ( P=0.007) were risk factors for central lymph node metastasis. The optimal model was established by stepwise regression. The factors included tumor diameter ≥0.5 cm, nodular goiter, extratumoral invasion, lymphadenopathy and TG>68 μg/L (AIC: 212.27) . The nomogram model was established according to the above risk factors. The consistency index (c-index) was 0.711. The results of calibration graph drawing and internal and external validation demonstrated its good consistency and applicability. Conclusion:Extratumoral invasion, lymphadenopathy, and TG>68 μg/L are risk factors for cN0 PTMC CLNM, and the nomogram established in the study can effectively predict the CLNM rate in patients with cN0PTMC and contribute to clinicians’ diagnosis and treatment decisions.

Objective To evaluate the role of autophagy in HL-1 cardiomyocyte injury induced by lipopolysaccharide( LPS).Methods Primary cultured HL-1 cardiomyocytes were randomly divided into 4 groups ( n =15each): normal control group( group C),LPS group,rapamycin( a autophagy inducer) group( group R) and 3-MA(a autophagy inhibitor) group.In group C cardiomyocytes were cultured continuously for 24 h.In group LPS cardiomyocytes were incubated with LPS (final concentration 1 μg/ml) for 24 h.In groups R and 3-MA,rapamycin and 3-MA was given 48 h before LPS (final concentration 1 μg/ml) incubation with final concentration of 0.2 μg/ml and 10 mmol/L respectively.The lipidated microtubule-associated protein 1 light chain 3 Ⅱ (LC3 Ⅱ ) expression,mitochondrial membrane potential,autophagosome number and optical density of mitochondria were determined,and ultrastructure of mitochondria was observed at 4 h of LPS incubation.Apoptosis rate and Caspase-3 activity were determined at 24 h of LPS incubation.Results LPS significantly increased LC3 Ⅱ expression,autophagosome number,optical density of mitochondria,apoptosis rate and Caspase-3 activity,decreased mitochondrial membrane potential in group LPS as compared with group C ( P ＜ 0.05).The LC3 Ⅱ expression,autophagosome number and mitochondrial membrane potential were higher,optical density of mitochondria,apoptosis rate and Caspase-3 activity lower in group R than in group LPS( P ＜ 0.05).The LC3 Ⅱ expression,autophagosome number and mitochondrial membrane potential were lower,optical density of mitochondria,apoptosis rate and Caspase-3 activity higher in group 3-MA than in group LPS (P ＜ 0.05).Mitochondrial histopathologic injury was reduced in group R and aggravated in group 3-MA as compared with group LPS.Conclusion Autophagy can reduce LPS-induced HL-1 cardiomyocyte injury by improving mitochondrial function and inhibiting apoptosis.

Objective To evaluate the effect of simulated training on in-service physicians electrocardiogram(ecg) theory and operational learning effect.Methods Sixty resident doctors were randomly divided into experiment group (n=30) and control group(n=30).The experimental group adopted teaching + simulation training mode to implement the theory and operation of ECG in continuing while the control group were taught only by lectures.Training effect of both groups was assessed by electrocardiogram theory and operation examination before and after training respectively,and by questionnaires at the end of the training as well.Data of the examination and questionnaires were analyzed by t test or x2 test.P＜0.05 was considered statistically significant.Results After training,the score of theory examination was (90.87 ± 6.67) points in experiment group and was (85.83 ± 5.5) points in control group,with statistical difference between these two groups(t=3.201,P=0.003).Score of operation examination was(93.40 ± 4.31) points in experiment group and was(77.03 ± 7.96)points in control group,with significant differences(t=10.204,P=0.000).The satisfaction degree of enhancing learning interest,theoretical knowledge and operation skill,and strengthening the concept of humauistic care were significantly better in the experiment group than in the control group (x2=34.737,6.405,42.088,41.713,P=0.000,0.011,0.000,0.000).Conclusions Application of simulated training in the study of electrocardiogram is obviously superior in effect to the traditional teaching model.It can improve the theoretical knowledge and operation skill,which is helpful in elevating the in-service physicians' comprehensive capability.