The aim of this study was to investigate the impact of overexpressing 70-kD heat shock pro-teins(Hsp70)on glycolysis in C2C12 cells during myogenesis and adipogenesis.Using C2C12 cells as the research material,adenovirus was used to overexpress the Hsp70 gene,and changes in the expression of glycolytic genes were detected using fluorescence quantitative PCR and Western blotting techniques.The study indicated that during C2C12 cell myogenic differentiation,the expression trend of the Hsp70 gene was consistent with that of Gsk3β,Pkm,Prkag3,Pfkm,and Hk-2 genes,suggesting a relationship between Hsp70 and the glycolytic pathway during myogenic differentiation.Overexpression of Hsp70 in the later stages of myogenic differentiation significantly upregulated the expression of Gsk3β,Pkm,Prk-ag3,and Pfkm genes(P＜0.05),with no significant impact on Hk-2 gene expression(P＞0.05).Dur-ing C2C12 cell adipogenic induction,the expression trend of the Hsp70 gene was similar to that of Gsk3β,Pkm,Prkag3,Pfkm,and Hk-2 genes,indicating a relationship between Hsp70 and the glycolytic path-way during adipogenic induction.Following Hsp70 overexpression,in the later stages of adipogenic in-duction,the number of lipid droplets was significantly higher compared to the control group,with a sig-nificant upregulation of Gsk3β,Pkm,Prkag3,and Pfkm gene expression(P＜0.05),while Hk-2 gene expression was not significantly affected(P＞0.05).In conclusion,Hsp70 in C2C12 cells in myogenic and adipogenic states promoted the breakdown of glycogen into 6-phospho-glucose,thereby enhancing the glycolytic pathway,providing insights into the functional role of the Hsp70 gene in glycolysis in C2C12 cells.

ObjectiveTo preliminarily predict the active components， action targets， and signaling pathways of Arnebia euchroma in the treatment of melanoma based on network pharmacology and molecular docking， and to verify its possible mechanism of action in in vitro experiments. MethodThe active components and related targets of A. euchroma were retrieved from the Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform （TCMSP）SwissTargetPrediction and literature， and the targets related to melanoma from the GeneCards， Online Mendelian Inheritance in Man （OMIM）， and Comparative Toxicogenomics Database （CTD）. Following the construction of the protein-protein interaction （PPI） network of active components and related targets of A. euchroma and melanoma-related targets using STRING， Cytoscape 3.8.2 was used for screening and analyzing the nodes in the network of A. euchroma against melanoma. The intersections were subjected to gene ontology （GO） and Kyoto encyclopedia of genes and genomes （KEGG） enrichment analysis using DAVID 6.8. Acetyl alkannin， the active component in A. euchroma， was docked to the target by AutoDock Vina 1.1.2. The in vitro experiments were then carried out to verify the anti-melanoma effect of A. euchroma. ResultA total of 271 common targets of A. euchroma and melanoma were harvested， among which 23 were key targets， including matrix metalloproteinase-9 （MMP-9） and Janus kinase 2 （JAK2）. As revealed by KEGG enrichment analysis， A. euchroma mainly acted on Janus kinase/signal transduction and activator of transcription （JAK/STAT）， tyrosine kinase receptor （ErbB）， and vascular endothelial growth factor （VEGF） signaling pathways to resist melanoma. According to molecular docking， acetyl alkannin exhibited a good docking activity with JAK2， STAT3， VEGF， MMP-9， and E-cadherin receptors. The results of Western blot and Real-time quantitative polymerase chain reaction （Real-time PCR） showed that acetyl alkannin at different doses inhibited the protein and gene expression of JAK2， STAT3， VEGF， MMP-9， and E-cadherin in A375 cells （P<0.05）. ConclusionA. euchroma alleviates melanoma via multiple targets and multiple pathways， and it may exert the therapeutic effects by affecting the expression of such key target proteins as JAK2， STAT3， VEGF， MMP-9， and E-cadherin and inhibiting the invasion and metastasis of melanoma cells. This study has provided an experimental basis for the treatment of tumor with A. euchroma.

OBJECTIVE: To study the relationship between preoperative plasma interleukin-1 (IL-1), interleukin-6 (IL-6), tumor necrosis factor-α (TNF-α), homocysteine (Hcy), endothelin-1 (ET-1) levels and new-onset atrial fibrillation (AF) after coronary artery bypass grafting (CABG). METHODS: In the study, 148 patients who underwent isolated CABG in Peking University People's Hospital from January 1, 2017 to December 30, 2017 were enrolled, of whom 39 had new-onset AF. The fasting venous blood was collected within 24 hours before the surgery. The preoperative plasma IL-1, IL-6, TNF-α, Hcy, ET-1 levels were detected by enzyme-linked immunosorbent assay (ELISA). The patients were divided into AF group and non-AF group according to whether new-onset AF occurred after operation. After 1 ∶1 propensity score matching (PSM), 38 people were in each group. The paired sample t-tests were performed on the five factors' concentrations of the matched AF group and the non-AF group respectively. If the concentration values did not conform to the normal distribution, the Wilcoxon signed rank sum test was performed. Conditional Logistic regression analysis was performed on the concentrations of the five indicators to explore the correlation between preoperative plasma concentrations of IL-1, IL-6, TNF-α, Hcy, ET-1 and postoperative new-onset AF after CABG. RESULTS: After a 1 ∶1 propensity score matching, the AF group was comparable to the non-AF group. The concentrations of IL-1, IL-6, TNF-α, and Hcy in the AF group were higher than those in the non-AF group[(0.867±0.589) ng/L vs. (0.742±0.262) ng/L, 21.55 (6.50, 209.90) ng/L vs. 17.95 (3.60, 86.70) ng/L, 20.30 (5.70, 361.00) ng/L vs. 21.50 (7.50, 251.80) ng/L, (0.29±0.11) μmol/L vs. (0.27±0.09) μmol/L], but the differences were not statistically significant (P=0.165, P=0.891, P=0.817, P=0.285). After the conditional Logistic regression analysis, the above four variables were not predictors of new-onset AF after CABG. The concentrations of ET-1 in the matched AF group and non-AF group were (25.80±6.20) ng/L and (29.10±8.54) ng/L, respectively. The correlation between preoperative low plasma ET-1 concentration and the new-onset AF after CABG were statistically significant (P=0.003). After conditional Logistic regression analysis, preoperative plasma ET-1 concentration was correlated with postoperative new-onset AF after CABG (P=0.039, adjusted OR=0.637, 95%CI: 0.415-0.977). CONCLUSION The levels of preoperative plasma IL-1, IL-6, TNF-α and Hcy in the patients with new-onset AF after CABG were higher than those in the patients without AF, but the difference was not statistically significant. Preoperative plasma low ET-1 concentration was statistically associated with new-onset AF after CABG.
Atrial Fibrillation/etiology* ; Coronary Artery Bypass ; Humans ; Propensity Score

Background: Hypothermic machine perfusion (HMP) is being used more often in cardiac death kidney transplantation; however, the significance of assessing organ quality and predicting delayed graft function (DGF) by HMP parameters is still controversial. Therefore, we used a readily available HMP variable to design a scoring model that can identify the highest risk of DGF and provide the guidance and advice for organ allocation and DCD kidney assessment. Methods: From September 1, 2012 to August 31, 2016, 366 qualified kidneys were randomly assigned to the development and validation cohorts in a 2:1 distribution. The HMP variables of the development cohort served as candidate univariate predictors for DGF. The independent predictors of DGF were identified by multivariate logistic regression analysis with a P < 0.05. According to the odds ratios (ORs) value, each HMP variable was assigned a weighted integer, and the sum of the integers indicated the total risk score for each kidney. The validation cohort was used to verify the accuracy and reliability of the scoring model. Results: HMP duration (OR = 1.165, 95% confidence interval [CI]: 1.008-1.360, P = 0.043), resistance (OR = 2.190, 95% CI: 1.032-10.20, P < 0.001), and flow rate (OR = 0.931, 95% CI: 0.894-0.967, P = 0.011) were the independent predictors of identified DGF. The HMP predictive score ranged from 0 to 14, and there was a clear increase in the incidence of DGF, from the low predictive score group to the very high predictive score group. We formed four increasingly serious risk categories (scores 0-3, 4-7, 8-11, and 12-14) according to the frequency associated with the different risk scores of DGF. The HMP predictive score indicates good discriminative power with a c-statistic of 0.706 in the validation cohort, and it had significantly better prediction value for DGF compared to both terminal flow (P = 0.012) and resistance (P = 0.006). Conclusion The HMP predictive score is a good noninvasive tool for assessing the quality of DCD kidneys, and it is potentially useful for physicians in making optimal decisions about the organs donated.
Adult ; Delayed Graft Function ; Female ; Humans ; Immunosuppressive Agents ; therapeutic use ; Kidney Transplantation ; adverse effects ; methods ; Logistic Models ; Male ; Multivariate Analysis ; Odds Ratio ; Organ Preservation

Objective To study virulence factors and drug resistance mechanism of linezolid-intermediate Enterococcus faecalis(E.faecalis) isolated from patients with bloodstream infection.Methods Two linezolid-intermediate E.faecalis strains,namely A and B,were isolated from two patients with bloodstream infection,the treatment of two patients was analyzed.The minimum inhibitory concentration (MIC) of linezolid and vancomycin were determined.The virulence genes (esp,asa1,gelE,ace,agg,efaA,cylA,and hyl) and linezolid resistance genes (domain V region of the 23SrRNA,cfr,cfr[B],optrA) were amplified by polymerase chain reaction (PCR).PCR products of domain V region of 23SrRNA gene were sequenced and analyzed.Results Symptoms of two patients who isolated two linezolid-intermediate E.faecalis strains were controlled after accepted linezolid therapy.Strains A and B were both susceptible to vancomycin(MICs were 1μg/mL and 4μg/mL respectively),teicoplain,ampicillin,and nitrofurantoin,while intermediate to linezolid(MIC were both 4μg/mL).Two strains both contained multiple virulence factors,strain A were negative for cylA and hyl,strain B were negative for hyl and esp,but positive for other virulence genes.There was G2621T mutation in domain V region of 23SrRNA in strain A,and no variation was found in strain B.Drug resistance genes of cfr,cfr(B),and optrA were all negative in both strain A and B.Conclusion In the present study,two linezolid-intermediate E.faecalis strains isolated from patients with bloodstream infection were susceptible to vancomycin and ampicillin,although the treatment of linezolid in two patients is effective,the utilization of linezolid therapy in clinical practice still needs to be cautious.The mutation of target site is a significant resistance mechanism,it is necessary for us to pay more attention to these clinical strains which are non-susceptible to such antimicrobial agents,and the treatment strategy needs further study.

OBJECTIVETo explore the effects of long-term exposure to particulate matter 2.5 (PM2.5) from automobile exhaust on the reproductive function of Sprague Dawley (SD) rats.

METHODSForty-five male SD rats, weighing 80 - 94 g and aged 28 days, were randomly assigned to receive intra-tracheal administration of 0.9% normal saline (control group, n = 15), PM2. 5 at 2 μg per 100 g body weight per day (low-dose PM2.5 group, n = 15), and PM2.5 at 16 μg per 100 g body weight per day (high-dose PM2.5 group, n = 15), qd, for 60 successive days. After the last 24-hour exposure, 10 rats were taken from each group for copulation with normal female ones, while the others were sacrificed, their testes removed for sperm count and deformity, pathological examination, and determination of the Connexin43 expression.

RESULTSThe conception rate was significantly decreased in the low- and high-dose PM2.5 groups as compared with that of the control (70% and 50% vs 100%), and so were the sperm count and quality. The rats in the PM2.5-exposed groups showed significantly disordered histological structure of the seminiferous tubules, reduced sperm count in the testicular lumen, some exfoliated secondary spermatocytes, downregulated Connexin43 expression in the testis, and damaged blood-testis barrier.

CONCLUSIONLong-term exposure to PM2.5 from automobile exhaust damages the reproductive function of male SD rats.

Animals ; Blood-Testis Barrier ; Body Weight ; Connexin 43 ; metabolism ; Down-Regulation ; Fertilization ; Male ; Particulate Matter ; toxicity ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Reproduction ; Seminiferous Tubules ; Sperm Count ; Spermatocytes ; Testis ; metabolism ; pathology ; Vehicle Emissions ; toxicity

Objective To investigate a reasonable and effective internal fixation method for posterolateral fracture of the tibial plateau. Methods Specimens of the tibial plateau with posterolateral fracture made from 12 adult male cadavers were randomly and evenly divided into 3 groups, and fixed by anterior 6.5 mm lag screw, lateral 4.5 mm L-shape plate, posterior 3.5 mm T-shape plate, respectively. All the specimens were loaded in turn by stress of 250, 500, 750, 1 000 N, and the corresponding axial displacement and stress were measured. Results Under the same stress, the Y-axial displacement of the anterior lag screw group was the smallest, showing a significant difference with the lateral plate group and the posterior plate group, while there was no significant difference between the lateral plate group and the posterior plate group in the Y-axial displacement. The stresses on marked points in the anterior lag screw group were evenly distributed. Conclusions For fixation of isolated posterolateral fractures of the tibial plateau, the anterior 6.5 mm lag screw can effectively increase the axial stability and balance the stress distribution around the fracture block, indicating it is an effective method for mechanical fixation. The lateral plate has certain advantage in lateral stability control, while the posterior plate has certain value to reduction of the posterior tibia plateau fracture.

OBJECTIVETo investigate regulation function of anodonta glucan HBP-A on chondrocytes through Wnt pathway in vitro.

METHODSRat chondrocytes were cultured and differentiated induced with IL-1beta (10 ng/ml) in vitro. Chondrocytes were divided into five groups:IL-13 group,IL-1beta + IWP-2 (5 microM,Wnt pathway inhibitor) group, IL-1beta + HBP-A (0.3 mg/ml) group and IL-1beta + IWP-2 + HBP-A group. Wnt-3a, beta-catenin (24 h,48 h,72 h) and MMP-13(72 h) genes expression were detected by Rt-PCR, while beta-catenin, MMP-13, Sox-9 and coll-II (48 h) protein expression were measured by Western-blot.

RESULTSAfter induction of IL-1beta, gene expression of Wnt-3a, beta-catenin and MMP-13 were increased,so were the protein expression of beta-catenin and MMP-13. In contrast,protein expression of Sox-9 and Coll-II were declined. Following addition of HBP-A, Wnt-3a, beta-catenin and MMP-13 were shown as induction of IL-1beta, but protein expression of Sox-9 and Coll-II were upgraded. Combining HBP-A with IWP-2 led to the lowest level in Wnt-3a, beta-catenin gene and beta-catenin protein expression and highest expression of Sox-9 protein.

CONCLUSIONHBP-A could not only delay the differentiation of chondrocytes through downgrading the signal expression of Wnt/beta-catenin,but also adjust the expression of Wnt-3a, beta-catenin and Sox-9 when combinated with the Wnt inhibitor.

Animals ; Anodonta ; chemistry ; Cell Differentiation ; drug effects ; Cells, Cultured ; Chondrocytes ; cytology ; drug effects ; metabolism ; Glucans ; pharmacology ; Interleukin-1beta ; metabolism ; Rats ; Wnt Signaling Pathway ; drug effects ; Wnt3A Protein ; genetics ; metabolism ; beta Catenin ; metabolism

Objective:To investigate regulation function of anodonta glucan HBP-A on chondrocytes through Wnt path-way in vitro. Methods:Rat chondrocytes were cultured and differentiated induced with IL-1β (10 ng/ml) in vitro. Chondro-cytes were divided into five groups:IL-1βgroup,IL-1β+IWP-2 (5μM,Wnt pathway inhibitor) group,IL-1β+HBP-A (0.3 mg/ml) group and IL-1β+IWP-2+HBP-A group. Wnt-3a,β-catenin (24 h,48 h,72 h) and MMP-13(72 h) genes expres-sion were detected by Rt-PCR,whileβ-catenin,MMP-13,Sox-9 and coll-Ⅱ (48 h) protein expression were measured by Western-blot. Results:After induction of IL-1β,gene expression of Wnt-3a,β-catenin and MMP-13 were increased,so were the protein expression ofβ-catenin and MMP-13. In contrast,protein expression of Sox-9 and Coll-Ⅱwere declined. Follow-ing addition of HBP-A,Wnt-3a,β-catenin and MMP-13 were shown as induction of IL-1β,but protein expression of Sox-9 and Coll-Ⅱwere upgraded . Combining HBP-A with IWP-2 led to the lowest level in Wnt-3a ,β-catenin gene andβ-catenin protein expression and highest expression of Sox-9 protein. Conclusion:HBP-A could not only delay the differentiation of chondrocytes through downgrading the signal expression of Wnt/β-catenin,but also adjust the expression of Wnt-3a,β-catenin and Sox-9 when combinated with the Wnt inhibitor.

ObjectiveTo investigate the activation of MAPK signal pathway in multiple myeloma and the regulation of BLyS expression levels through MAPK signal pathway; preliminarily study the role of MAPK signal pathway in the up-regulation of BLyS expression levels induced by IFN-γ.MethodsActivated MAPK pathway were detected by Western blot,while the expression of BLyS were detected with RT-PCR and Western blot,and Western blot investigated the effect of MAPK pathway on BLyS expression levels induced by IFN-γ.ResultsIn addition to the expression of ERK,JNK,p38,p-JNK was also expressed in MM cell lines,the MAPK pathway inhibitor targeting JNK SP600125 can down-regulate the expression of BLyS,and its activator anisomycin can up-regulate the expression of BLyS.SP600125 restrained the proliferation and survival of MM cells.ConclusionJNK/SAPK pathway was activated in MM cells; The activated degree of JNK/SAPK pathway and the expression level of BLyS was positively correlated.JNK/SAPK pathway play an important role in the up-regulation of BLyS expression levels induced by IFN-γ.