Cardiomyopathy, Dilated ; genetics ; Humans ; Molecular Biology

Insulin resistance model of 3T3-L1 adipocytes were prepared with plamotic acid.Adipocytes with generated insulin resistance were cultured with different concentrations of globular domain of adiponectin(gAd:250,500,1 000 ng/ml).The cell culture medium glucose content was detected with the glucose oxidase method,the mRNA expressions of insulin receptor substrate-1 (IRS-1),phosphatidylinositol-3 kinase(PI3K),and protein kinase B(PKB) were detected with real-time quantitative PCR method.The phosphorylation of IRS-1 was detected by Western blot.Compared with the control group,the experimental group showed significantly increased glucose consumption (P ＜ 0.01),and with the increasing gAd concentration,glucose consumption was gradually increasing.IRS-1 phosphorylation was increased gradually with the increasing concentration of gAd.These results suggest that gAd can promote glucose uptake by 3T3-L1 adipocyte model with generated insulin resistance.This may be correlated with promoting insulin signal transduction and improving insulin resistance in adipocytes.

The aim of this paper was to investigate the effect of terpene penetration enhancers on membrane fluidity and membrane potential using HaCaT keratinocytes, and study the potential mechanisms of these terpene compounds using as natural transdermal penetration enhancer. Six terpene compounds, namely menthol, limonene, 1,8-cineole, menthone, terpinen-4-ol and pulegone, were chosen in this study on account of their good penetration-enhancement activities. The cytotoxicity of these terpene compounds was measured using an MTT assay. The fluorescence recovery after photobleaching (FRAP) technique was employed to measure the change of membrane fluidity of HaCaT cells. The flow cytometer was used to study the alteration of membrane fluidity of HaCaT cells, and investigate the effect of terpene compounds on intracellular Ca2+. It was found that 6 terpene compounds possessed low cytotoxicity in comparison to the well-established and standard penetration enhancer azone. Those terpene compounds could significantly enhance HaCaT cells membrane fluidity and decrease HaCaT cells membrane potentials. Meanwhile, after treated with various terpene compounds, the Ca2(+)-ATPase activity and intracellular Ca2+ of HaCaT cells was decreased significantly. Terpene penetration enhancers perhaps changed the membrane fluidity and potentials of HaCaT cells by altering the Ca2+ balance of the cell inside and outside, resulting in the low skin permeability to increase the drug transdermal absorption.
Cell Line ; Drugs, Chinese Herbal ; pharmacokinetics ; Humans ; Keratinocytes ; drug effects ; metabolism ; Membrane Fluidity ; drug effects ; Skin Absorption ; drug effects ; Terpenes ; pharmacokinetics

ObjectiveTo investigate the feasibitity of detecting S100A6 expression at mRNA level in endoscopic ultrasonography guided fine needle aspiration (EUS-FNA) pancreatic ductal adenocarcinoma (PDA) specimens and its diagnostic value in PDA.MethodsA total of 18 PDA specimens and 22 normal pancreatic specimens were collected. RNA was extracted for reverse transcription.The expression of S100A6 gene was examined by real-time polymerase chain reaction.The cut-off value of S100A6 expression at mRNA level in PDA diagnosis was established through receiver operating characteristic (ROC) analysis. 28 patients with pancreatic head masses were selected for EUS-FNA examination,and the value of S100A6 mRNA expression level in PDA diagnosis was prospectively evaluated. The expression of S100A6 protein in PDA tissue was determined by immunohistochemistry staining.ResultsS100A6 mRNA expression in EUS-FNA and surgical PDA specimens (0.05023±0.10120,0.02083 ± 0.02848) was significantly higher than that of normal pancreatic tissues (0.00164±0.00202),both P＜0.01.The expression of S100A6 in 22 EUS-FNA PDA specimens was significantly higher than that of 6 pancreatic benign disease biopsy specimens (0.00193 ± 0.00278,P =0.0009). There was no significant difference in S100A6 expression between 6 pancreatic benign disease biopsy specimens and normal surgical pancreatic samples (P=0.6143).When S100A6 mRNA expression in EUS-FNA specimens over 0.00525 was taken as positive diagnostic value,the sensitivity,specificity and accuracy in prospective pancreatic cancer diagnosis were 90.01％,100 ％ and 92.85 ％,respectively.ConclusionThe high expression of S100A6 mRNA in EUS-FNA specimens of PDA has good preoperative diagnostic value.

Calcium hydroxide (CH) is applied to improve disinfection of root canals in most root canal retreatment. This study aimed to analyze the CH removal efficacy using 7 different root preparing files (K file, pre-curved K file, EndoActivator, Ultrasonic file, pre-curved ultrasonic file, F file and needle irrigation alone) with apical transportation. Standardized models of curved canal with such apical transportation or not were set up before applying CH to root canal for 7 days. Seven techniques described above were used for its removal. Then the roots were disassembled and digital photos were taken. The ratio of residual CH in the overall canal surface was calculated using the image analyzer image pro plus 6.0. The data were analyzed using one-way ANOVA with post hoc Tukey test. Results revealed that CH was effectively removed (P<0.05) by using all 6 mechanical methods except irrigation alone. In curved root canals with apical transportation, EndoActivator, pre-curved ultrasonic file and F file were found to be more effective in removing CH than the other four file (P<0.001), while there was no significant difference among EndoActivator, pre-curved ultrasonic file and F file groups (P>0.05). The percentage of residual CH in the canal with apical transportation was higher than that in the canal without apical transportation (P<0.05). In conclusion, CH can be hardly removed completely. Canal with apical transportation will result in insufficient CH removal. EndoActivator, pre-curved ultrasonic file and F file are more effective in the curved root canal with apical transportation.

Objective To study interleukin-1 receptor(IL-1R) and connexin(Cx36) gene expression following Mg 2+-free-induced seizures in cultured developing neuron. Methods Rat embryo cortical neurons cultured for 6 and 17 days were exposed to Mg 2+-free media to induce seizure. At different time after Mg 2+-free treatment, real-time RT-PCR was used to detect IL-1R and Cx36 mRNA expression. Results 1. IL-1R mRNA expression transiently decreased after Mg 2+-free treatment in neurons cultured for 6 and 17 days in vitro. Then the levels of IL-1R mRNA expression recovered in neurons cultured for 6 days, but IL-1R mRNA expression were increased in neurons cultured for 17 days compared with control group and the peak was at 24 hours. 2. In neurons cultured for 6 days in vitro, Cx36 mRNA expression increased after Mg 2+-free treatment compared with control group, the peak was at 24 hours. But in neurons cultured for 17 days in vitro, Cx36 mRNA expression decreased at 6 hours after Mg 2+-free treatment compared with control group, the peak was at 24 hours. Conclusions IL-1R mRNA and Cx36 mRNA expression following Mg 2+-free-induced seizures are different between the neurons cultured for 6 and 17 days in vitro. This is possibly related to the different neuron injury between 6 and 17 days in vitro following seizures.

Objective To optimize extraction process of flavonoids from Toosendan Fructus.Methods Three parameters that significantly affect the extraction rate of flavonoids, ratio of material to water, extraction time and ethanol concentration, were taken into consideration for extraction ratio of flavonoids from Toosendan Fructus. Based on the results of single-factor experiment, the extraction parameters of flavonoids from Toosendan Fructus were optimized by Box-Behnken of response surface methodology (RSM).Results The optimum extraction conditions were as follow: material to water ratio was 1:35; extraction time should be 3 h; ethanol concentration was 70%. The extraction ratio was up to 0.587 3% in the optimum extraction conditions.Conclusion The actual value is highly matching the theoretically value of RSM model. The results indicated that this method can rationally optimize extraction process of flavonoids from Toosendan Fructus.

Glomerular hypertrophy is the main pathological characteristic in the early stage of diabetic nephropathy (DN), and its regulatory mechanism is closely related to mammalian target of rapamycin (mTOR) signaling pathway activity. mTOR includes mTOR complex 1 (mTORC1) and mTOR complex 2(mTORC2), in which, the upstream pathway of mTORC1 is phosphatidylinositol-3-kinase (PI3K)/serine-threonine kinase(Akt)/adenosine monophosphate activated protein kinase(AMPK), and the representative signaling molecules in the downstream pathway of mTORC1 are 4E-binding proteins(4EBP) and phosphoprotein 70 S6Kinase(p70S6K). Some Chinese herbal extracts could improve cell proliferation via intervening the expressions of the key molecules in the upstream or downstream of PIK/Akt/mTOR signaling pathway in vivo. As for glomerular mesangial cells(MC) and podocyte, mTOR plays an important role in regulating glomerular inherent cells, including adjusting cell cycle, energy metabolism and matrix protein synthesis. Rapamycin, the inhibitor of mTOR, could suppress glomerular inherent cell hypertrophy, cell proliferation, glomerular basement membrane (GBM) thickening and mesangial matrix deposition in model rats with DN. Some Chinese herbal extracts could alleviate glomerular lesions by intervening mTOR signaling pathway activity in renal tissue of DN animal models or in renal inherent cells in vivo and in vitro.
Animals ; Diabetic Nephropathies ; drug therapy ; enzymology ; genetics ; pathology ; Drugs, Chinese Herbal ; administration & dosage ; Humans ; Hypertrophy ; drug therapy ; enzymology ; genetics ; pathology ; Kidney Glomerulus ; drug effects ; metabolism ; pathology ; Signal Transduction ; drug effects ; TOR Serine-Threonine Kinases ; genetics ; metabolism

Objective To investigate the drug compliance and related factors in patients after intracoronary stent implantation. Methods Investigated 81 patients after intracoronary stent implantation self-designed questionnaire by drug compliance scale, depress scale and family function scale and analyze the results.Results Drug compliance of patients after intracoronary stent implantation was related to depress status (r =-0. 292,P ＜0. 01 ) and main diagnosis( r = -0. 253, P ＜ 0. 05 ), number of stent was related to depress status (r =0. 368 ,P ＜0. 01 ) and family function(r = -0. 259,P ＜0. 05 ). Conclusions Except for the patients, nurse should also enhance the health education to their family members for promoting patients' drug compliance out of hospital.

Objective To investigate the effects of different doses of pituitary adenylate cyclase- activating polypeptide(PACAP)on the functional and morphological outcome in a mice model of Parkinson' s disease(PD)rendered by 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine(MPTP).Methods Male mice were treated with PACAP 0.02, 0.20 or 2.00 ?g by iv bolus for 7 days after MPTP was administered, and were compared with the saline-treated mice.The immunohistochemistry and Western blot were used to detect the alterations of PD biomarker including tyrosine hydroxylase(TH), dopamine transporter(DAT)and vesicular monoamine transporter2(VAMT2).In addition, monoamine neurotransmitters in the striatum of mice were measured by the high performance liquid chromatography (HPLC).Results TH immunohistochemistry indicated that the number of TH-positive neurons in the substantia nigra was increased in all PACAP-treated mice(PACAP(0.02 ?g/d)group was 93.33?4.87, F=85.85,P