Animals ; Carps ; physiology ; Galvanic Skin Response ; drug effects ; physiology ; Skin ; cytology ; Strychnine ; pharmacology ; Synaptic Transmission ; drug effects ; physiology

Objective To evaluate the changes of cardiac function and myocardial perfusion by Gated 99Tcm-MIBI myocardial perfusion imaging after autologous mesenchymal stem cell implantation in patients with acute myocardial infarction at high altitude area.Methods 33 patients with anteroseptal myocardial infarction were ran- domly divided into two groups.18 patients (control group) underwent percutaneous tranluminal coronary angioplasty (PTCA) and 14 cases (transplantation group) received additional mesenchymal stem cell transplantation.Myocardi- al perfusion imaging were performed in all patients before and at 6 and 12 months after treatment.Results Com- pared to pre-implantation,LVEF of transplantation group was improved 8%～9%after 6 months.The improving lev- els of control group were lower.However,there were not statistical differences among all data.Conclusion Mesen- chymal stem cell transplantation could improve myocardial systolic function and myocardial perfusion.

Objective To induce mouse embryonic stem(ES)cells to differentiate into insulin-secreting cells by means of a 5-step model system.Methods E14.1 mouse ES cells were cultured in the presence of leukemia inhibitory factor(LIF)for 2 days(step 1),then the cells were cultured in hanging drops to form embryonic bodies(EBs)and the resulting EBs were cultured in suspension for 6 days in the presence of basic fibroblast growth factor bFGF(step 2).Subsequently the EBs were cultured in the medium containing glucagon- like peptide 1(GLP-1),hepatocyte growth factor(HGF),nerve growth factor(NGF)and nicotinamide for 10 days(step 3).After that,the EBs were dissociated into single cells,and the cells were cultured in monolayer in the presence of GLP-1,betacellulin,activin A,bFGF and nicotinamide for 10 days(step 4).Finally,the cells were cultured in low-glucose medium containing nicotinamide for 4 days(step 5).Insulin and some other islet- related genes expressions were investigated using RT-PCR and insulin expression was also investigated by DTZ- staining and immunohistochemistry.The percentage of insulin-secreting cells was evaluated by flowcytometry and insulin concentrations were measured by RIA.Results mRNA expression of insulin became visible at step 3 and more evident at step 5.Additionally,at step 5,mRNAs of glucagon,somatostatin,pancreatic polypeptide(PP), pancreatic duodenal homeobox 1(PDX-1),beta-cell E box transactivator 2(Beta2)and neurogenin 3(Ngn3) were detected.DTZ-staining positive cells and insulin immunohistochemical staining positive cells were observed. The percentage of insulin-positive cells was(24.0?2.5)%(n=6).In the presence of 5.6 mmol/L and 25 mmol/L glucose,insulin concentrations were(0.05?0.01)?g/L and(0.13?0.02)?g/L respectively(n= 6).Conclusion E14.1 mouse ES cells can be induced to differentiate into insulin-secreting cells by the 5-step model system.Insulin-secreting cells can release insulin into culture medium when treated with glucose,and insulin concentrations increase with rising concentration of glucose.

Abstract: Objective　To detect the distribution of CYP2A6∗2, CYP2A6∗10, CYP2A6∗17, CYP2B6∗4, CYP2B6∗6, and CYP2B6∗18 loci affecting the metabolism of artemisinins in Kazak population in Xinjiang. To explore the pharmacogenetic background of the Kazak population in Xinjiang for artemisinin drugs and provide clinical decision support for the treatment and prevention of malaria based on artemisinin drugs. Methods　Six SNPs including CYP2A6∗2, CYP2A6∗10, CYP2A6∗17, CYP2B6∗4, CYP2B6∗6, and CYP2B6∗18 were selected for the sequencing experiment. 330 whole blood samples were collected from the Kazak population in Xinjiang. After extracting the whole blood DNA genome, multiplex PCR and high-throughput sequencing were used for genotyping. The allele frequencies were analyzed using the Hardy-Weinberg equilibrium. Results　In this study all SNPs follow the Hardy-Weinberg equilibrium (P>0.05), there was no significant difference in the distribution of SNPs between different genders (P>0.05). The number of successfully sequenced samples of CYP2A6∗2, CYP2A6∗10, CYP2A6∗17, CYP2B6∗4, CYP2B6∗6, and CYP2B6∗18 were 326, 319, 328, 318, 322 and 328 respectively. The frequencies of variant alleles of CYP2A6∗2, CYP2A6∗10, CYP2A6∗17, CYP2B6∗4, CYP2B6∗6, and CYP2B6∗18 in Kazak population are: 0.61%, 0%, 0%, 30.97%, 22.98%, 0%. Conclusions　Mutation alleles affecting the metabolism of artemisinins exist in the Kazak population in Xinjiang. When using artemisinins, the relationship between the drug effect and individual pharmacogenetic background should be further explored.

Cannabinoid receptors are one of the most expressed G protein-coupled receptors in the central nervous system, which are potential drug targets for inflammation, pain and drug abuse. Cannabinoid receptors are composed of type 1 receptor (CB1R), type 2 receptor (CB2R) and other receptors, of which CB1R plays a vital role in regulating central memory, cognition, and motor function. Therefore, screening CB1R agonists has potential value in treating nervous system diseases. In this study, the intracellular loop 3 (ICL3) domain of CB1R was replaced with a circular-permutated enhanced green fluorescent protein (cpEGFP). After infecting HEK 293T cells with lentivirus particles, we obtained a stable cell line that was overexpressed human CB1R-cpEGFP after puromycin selection. The interaction between receptor agonists and CB1R led to the change of receptor conformation, resulting in de-protonation of the EGFP, and enhancing the fluorescence intensity. Therefore, active CB1R compounds could be verified by measuring the fluorescence intensity. Using CB1R agonist arachidonyl-2′-chloroethylamide (ACEA) as a positive control to evaluate the reliability of this model, studies have shown that ACEA could induce receptor activation and increase fluorescence intensity, while antagonist rimonabant inhibited receptor activation with unchanged fluorescence intensity. In conclusion, this study successfully constructed a fluorescent probe screening model for CB1R agonists.

Background:Previous studies of contrast-induced acute kidney injury (CI-AKI) were mostly based on selective percutaneous coronary intervention (PCI) cases,and risk factors of CI-AKI after emergency PCI are unclear.The aim of this study was to explore the risk factors of CI-AKI in a Chinese population undergoing emergency PCI.Methods:A total of 1061 consecutive patients undergoing emergency PCI during January 2013 and June 2015 were enrolled and divided into CI-AKI and non-CI-AKI group.Univariable and multivariable analyses were used to identify the risk factors of CI-AKI in emergency PCI patients.CI-AKI was defined as an increase in serum creatinine ≥25％ or ≥0.5 mg/dl (44.2 tmol/L) above baseline within 3 days after exposure to contrast medium.Results:The incidence of CI-AKI in patients undergoing emergency PCI was 22.7％ (241/1061).Logistic multivariable analysis showed that body surface area (BSA) (odds ratio [OR] 0.213,95％ confidence interval [CI]:0.075-0.607,P =0.004),history of myocardial infarction (MI) (OR 1.642,95％ CI:1.079-2.499,P =0.021),left ventricular ejection fraction (LVEF) (OR 0.969,95％ CI:0.944-0.994,P =0.015),hemoglobin (Hb) (OR 0.988,95％ CI:0.976-1.000,P =0.045),estimated glomerular filtration rate (OR 1.027,95％ CI:1.018-1.037,P ＜ 0.001),left anterior descending (LAD) stented (OR 1.464,95％ CI:1.000-2.145,P =0.050),aspirin (OR 0.097,95％CI:0.009-0.987,P =0.049),and diuretics use (OR 1.850,95％ CI:1.233-2.777,P =0.003) were independent predictors of CI-AKI in patients undergoing emergency PCI.Conclusion:History of MI,low BSA,LVEF and Hb level,LAD stented,and diuretics use are associated with increased risk of CI-AKI in patients undergoing emergency PCI.

Objective:To explore the effect of hyperbaric oxygen (HBO) on the blood-brain barrier via the silent information regulator 1 (SIRT1)/Forkhead box O1(FoxO1) signaling pathway after cerebral ischemia and reperfusion using a rat model.Methods:Forty Wistar rats were randomly assigned into sham, cerebral ischemia-reperfusion (CIR), CIR+ HBO and CIR+ HBO+ EX527 groups, each of 10. The cerebral ischemia-reperfusion model was established in all groups except the sham group by right middle cerebral artery occlusion using the modified thread-occlusion method. The sham group was not ligated. Both the CIR+ HBO and CIR+ HBO+ EX527 groups were given HBO 1, 9, 21, 45 and 69 hours after the reperfusion. The CIR+ HBO+ EX527 group was additionally injected with 5mg/kg of EX527(a SIRT1inhibitor) peritoneally 4, 12, 24, 48 and 72 hours after the reperfusion. Then 2% Evens blue (EB) was injected into the tail vein an hour before the rats were sacrificed. The content of EB and the expression of SIRT1, FoxO1, ZO-1, Occludin, Claudin-5 mRNA and their proteins were determined using spectrophotometry, reverse transcription-polymerase chain reactions and Western blotting.Results:The average EB content of the hippocampal brain tissue from the CIR, CIR+ HBO and CIR+ HBO+ EX527 rats was significantly greater than the Sham group′s average 72h after reperfusion. The average expression of SIRT1, FoxO1, ZO-1, Occludin and Claudin-5 mRNA and their proteins was significantly lower, with the CIR + HBO + EX 527 group′s average significantly lower than that of the CIR+ HBO group.Conclusions:HBO can increase the expression of tight junction protein via the SIRT1/FoxO1 pathway. It helps to protect the blood-brain barrier in CIR injury situations.

Objective To explore the significance of urine deoxypyridinoline to diagnosis on osseous metastasis of lung neoplasms.Methods.182 cases with lung carcinoma was divided into two groups.One group was case with osseous metastasis,the other group was case without osseous metastasis,uDPD/uCr, uCa/Cr,sCa and sAKP in two groups were respectively compared.Sensitivity and specificity of these indexes to diagnosis on osseous metastasis of lung cancer were also acalculated and compared.80 cases without osseous metastasis were follow-up for 6 months.Results The ratio of uDPD/uCr with osseous metastasis group[(12.35?2.65)nmol/mmol]was significantly higher than that of without osseous metastasis group [(7.76?2.11)nmol/mmol](t=2.46,P

Objective In this study,we investigated the relationship between oxidative stress,selenoproteins level and onset of Keshan disease (KD) through detecting the expression of 8-hydroxy-2-deoxy guanosine (8-OH-dG),thioredoxin reductase 1 (TrxR1) and glutathione peroxidase 1 (GPx1) in myocardial tissue.Methods Myocardium samples of autopsy patients including 8 cases of KD (KD group included 4 acute KD and 4 chronic KD) and 9 cases of non-KD (control group) were immunohistochemically stained for 8-OH-dG,TrxR1 and GPx1.The staining intensities subsequently quantified by using Olympus Image-Pro Plus 6.0 software.Results The positive rate of 8-OH-dG expression in myocardial nuclei was higher in the case group[(68.6 ± 20.4)％] than that of the control group[(2.4 ± 1.5)％,t =8.515,P ＜ 0.05].In addition,the positive rate of 8-OH-dG expression in acute KD[(91.7 ± 3.7)％] was significantly higher than that of chronic KD[(53.2 ± 7.9)％,t =6.409,P＜0.05].The distribution of TrxR1 and GPx1 was not associated with the distribution of myocardial damage.The expression of these two selenoproteins in KD group (401340 ± 59865,497590 ± 197082) were both lower than that of control group(2790300 ± 379298,1348400 ±615840; t =-28.493,-6.016,respectively,all P＜0.01).Conclusions Oxidative damage is detected in myocardium tissue of KD,and 8-OH-dG expression is associated with the degree of myocardial damage in KD.Selenoproteins,TrxR1 and GPx1,may be closely related to the pathogenesis of KD.

PURPOSE: To evaluate the impact of five different tooth preparation designs on the marginal and internal fit discrepancies of cobalt-chromium (CoCr) crowns produced by computer-aided designing (CAD) and selective laser melting (SLM) processes. MATERIALS AND METHODS: Five preparation data were constructed, after which design crowns were obtained. Actual crowns were fabricated using an SLM process. After the data of actual crowns were obtained with structural light scanning, intaglio surfaces of the design crown and actual crown were virtually superimposed on the preparation. The fit-discrepancies were displayed with colors, while the root means square was calculated and analyzed with one-way analysis of variance (ANOVA), Tukey’s test or Kruskal-Wallis test (α =.05). RESULTS: The marginal or internal color-coded images in the five design groups were not identical. The shoulder-lip and sharp line angle groups in the CAD or SLM process had larger marginal or internal fit discrepancies compared to other groups (P < .05). In the CAD process, the mean marginal and internal fit discrepancies were 10.0 to 24.2 µm and 29.6 to 31.4 µm, respectively. After the CAD and SLM processes, the mean marginal and internal fit discrepancies were 18.4 to 40.9 µm and 39.1 to 47.1 µm, respectively. The SLM process itself resulted in a positive increase of the marginal (6.0 – 16.7 µm) and internal (9.0 – 15.7 µm) fit discrepancies. CONCLUSION The CAD and SLM processes affected the fit of CoCr crowns and varied based on the preparation designs. Typically, the shoulder-lip and sharp line angle designs had a more significant effect on crown fit. However, the differences between the design groups were relatively small, especially when compared to fit discrepancies observed clinically.