Objective:To evaluate the role of microRNA-149-5p (miR-149-5p) in resveratrol-induced reduction of lipopolysaccharide (LPS)-induced cardiomyocyte injury in rats.Methods:Rat cardiomyocyte cell line H9C2 was cultured and then divided into 5 groups ( n=27 each) using a random number method: control group (C group), LPS group, resveratrol group (RSV group), miR149-5p inhibitor negative control group (LRN group), and miR149-5p inhibitor group (LRI group). A cardiomyocyte injury model was prepared by incubating cells with culture medium containing 10 μg/ml LPS for 24 h. RSV group was incubated with resveratrol (final concentration of 10 μmol/L) for 24 h, followed by incubation with culture medium containing 10 μg/ml LPS for another 24 h. LRN group and LRI group were transfected with miR149-5p inhibitor negative control and miR149-5p inhibitor, respectively, and then the other treatments were similar to those previously described in RSV group. The cell viability was measured by CCK-8 assay, the apoptosis rate by flow cytometry, the concentration of lactate dehydrogenase (LDH) and content of glutathione (GSH) in the supernatant by microplate method, the content of malondialdehyde (MDA) by TBA reaction method, the activity of superoxide dismutase (SOD) by WST-1 method, the level of reactive oxygen species (ROS) by DCFH-DA fluorescent probe, the concentrations of tumor necrosis factor-alpha (TNF-α) and interleukin-6 (IL-6) in the supernatant by enzyme-linked immunosorbent assay, and the expression of miR-149-5p by quantitative real-time polymerase chain reaction. Results:Compared with C group, the expression of miR-149-5p was significantly down-regulated, the cell viability was decreased, the concentrations of LDH, TNF-α and IL-6 in supernatant, apoptosis rate, ROS level and MDA content were increased, and the GSH content and SOD activity were decreased in LPS group ( P<0.05). Compared with LPS group, the expression of miR-149-5p was significantly up-regulated, the cell viability was increased, the concentrations of LDH, TNF-α and IL-6 in supernatant, apoptosis rate, ROS level and MDA content were decreased, and the GSH content and SOD activity were increased in RSV group ( P<0.05). Compared with RSV group or LRN group, the expression of miR-149-5p was significantly down-regulated, the cell viability was decreased, the concentrations of LDH, TNF-α and IL-6 in supernatant, apoptosis rate, ROS level and MDA content were increased, and the GSH content and SOD activity were decreased in LRI group ( P<0.05). Conclusions:The mechanism by which resveratrol alleviates LPS-induced cardiomyocyte injury is associated with the up-regulation of miR-149-5p expression and inhibition of cell apoptosis, oxidative stress and inflammatory responses in rats.

Objective:To observe any effect of low-frequency whole body resonant stimulation on the ba-lance and walking ability of hemiplegic stroke survivors.Methods:Sixty-six stroke survivors with hemiplegia were randomly divided into a low-frequency resonance training group, a high-frequency vibration training group and a control group, each of 22. All received routine exercise training at individualized intensities. All three groups underwent five 1-minute cycles of 7Hz, 15Hz or 1Hz stimulation twice a day, five days a week for eight weeks. Before and after the intervention, balance and walking ability were evaluated using the Berg Balance Scale, the timed up and go test and a 10m walking test. Step length, step frequency and step speed were also measured.Results:There were no significant differences among the three groups before the training. Afterward, significant improvement was observed in all of the groups in terms of all of the measurements. The average results of the low-frequency resonance training group were at that point significantly better than the other two groups′ averages, while the high-frequency vibration training group′s results were superior to those of the control group.Conclusion:Resonance training at 7Hz is the most effective in improving the balance and walking ability of stroke survivors with hemiplegia.

Objective To confirm untoward reactions in uterine benign disease caused by the application of high intensity fo‐cused ultrasound (HIFU) is lighter .Methods Following‐up 877 cases of patients with uterine benign disease who received HIFU treatment from September 2014 to May 2016 in Zigong Fourth People′s Hospital ,the intra‐and post‐operative untoward reactions were summarized and analyzed .Results We summarized the occurrence of untoward reactions ,treatment measures and prognosis :98% of untoward reactions were SIR A to B level ,and fully recovered after treatment immediately to 1 week without special inter‐vention;2% of untoward reactions were SIR C to D ,also fully recovered after hospital and nursing treatment ;no serious complica‐tions(SIR D and above) occurred .Conclusion HIFU can be used in the treatment of uterine benign diseases in our hospital ,which could provide references for the treatment of uterine benign diseases by using HIFU .

Objective:To investigate the influence of inhibitor of kappa B kinase ( IKK) inhibitor on carotid artery restenosis in rat by balloon catheter injury. Methods: A total of 32 male Wistar rats were randomly divided into control group, injured group and med?icine group. The balloon catheter injury was performed on left common carotid artery of rat by imitating the process of angioplasty. In?jured group and control group were injected saline, medicine group were injected IKK inhibitor. After 15 days, the process of neointi?mal and media hyperplasia was observed by HE staining and the expression level of NF?κB, IKK, inhibitor of nuclear factor kappa B ( IκB) was detected by Western blot, the content of IL?6, TNF?αwas detected by radioimmunoassay, and the content of MMP?1 was determined by MMP?1 kit. Results: Arterial neointima hyperplasia hapened in 3 groups. There was significantly difference between control group and medicine group and injured group in intimal area, the expression level of NF?κB, IKK, IκB, and the content of IL?6, TNF?α and MMP?1 ( P<0?05 or 0?01) . There was no significance in these indexes between control group and medicine group ( P>0?05) . Conclusion:IKK inhibitor can reduce the carotid artery restenosis after balloon injury by inhibiting inflammatory reaction induced by IKK?IκB?NF?κB pathway.

In order to enhance the antitumor efficacy of recombinant Newcastle disease virus, rNDV-IL15 was rescued in this study. Recombinant plasmid prNDV-IL15 was constructed, and BHK21 cells were transfected with the recombinant plasmid. Finally, the recombinant Newcastle disease virus rNDV-IL15 was successfully rescued. The growth curves of these two recombinant viruses were determined. Murine melanoma B16F10 cells were infected with rNDV-IL15 at MOI of 0.1, and the expression level of IL15 in the supernatant was detected by ELISA. The antitumor efficacy of rNDV-IL15 and rNDV was compared in vitro and in vivo. Results showed that prNDV-IL15 was constructed and recombinant virus rNDV-IL15 was successfully rescued. The growth curve of rNDV-IL15 showed that the growth of rNDV-IL15 had not been changed after insertion of IL15 gene. Results showed that there was high level of IL15 expression in the supernatant of rNDV-IL5-infected B16F10 cells (1 044.3 +/- 27.7 ng x mL(-1)). rNDV-IL15 and rNDV significantly inhibited the growth of B16F10 cells in vitro in a time-dependent manner. However, there was no significant difference between them. In animal experiments, rNDV-IL15 efficiently suppressed tumor growth in vivo when compared with rNDV, and the difference was statistically significant. The results suggested that rNDV-IL15 is a more effective antitumor agent.

In order to obtain the lead compound for treatment of rheumatoid arthritis (RA), in this study, therapeutic efficacy of three bispecific antibodies (BsAB-1, BsAB-2 and BsAB-3) against both hIL-1beta and hIL-17 were compared on CIA model mice. First, by ELISA method we compared the binding capacity of the three bispecific antibodies to the two antigens. The results showed that all three antibodies could simultaneously bind both antigens, among these antibodies, BsAB-1 was superior over BsAB-2 and BsAB-3. CIA model was established with chicken type II collagen (CII) and developed RA-like symptoms such as ankle swelling, skin tight, hind foot skin hyperemia. The CIA mice were treated with three antibodies once every two days for total of 29 days. Compared with the CIA model mice, the RA-like symptoms of the antibody treated-mice significantly relieved, while the BsAB-1 treated-mice were almost recovered. CII antibody level in the serum and cytokines (IL-2, IL-1beta, IL-17A and TNF-alpha) expression in the spleen were examined. Compared with the CIA model mice, all three antibodies could significantly reduce CII antibody and cytokine expression levels. BsAB-1 antibody was more potent than BsAB-2 and BsAB-3. In summary, BsAB-1 is superior over BsAB-2 and BsAB-3 in amelioration of RA symptoms and regulation of CII antibody production and pro-inflammatory cytokine expression, therefore, BsAB-1 can be chosen as a lead compound for further development of drug candidate for treatment of RA.

Objective To investigate the effects of remifentanil on sinoatrial (SA) node autorhythmicity in rabbits.Methods Twenty-four healthy rabbits of both sexes weighing 1.8-2.2 kg were sacrificed.Their hearts were removed and sinoatrial nodes were dissected and placed in Tyrode solution saturated with 95 ％ O2-5 ％ CO2 at 36 ℃.The action potentials of the sinus node pacemaker cells were recorded by intracellular glass microelectrode technique.The experiment was performed in 3 parts (n =8 each).Part Ⅰ:the sinoatrial node was exposed to remifentanil 2,4,8,16 and 32 ng/ml respectively.The action potentials were recorded after the sinoatrial nodes were exposed to each concentration of remifentanil for 15 min.Part Ⅱ and Ⅲ:the sinoatrial nodes were first exposed to Ca2+ channel agonist Bay K8644 0.5 μmol/L or K+ channel blocker TEA 20 nmol/L for 15 min.Then remifentanil was added until the concentration reached 16 ng/ml (final concentration) and 15 min later the action potentials were recorded.The action potential parameters included,amplitude of action potential (APA),rate of pacemaker firing (RPF),action potential duration at 90％ repolarization (APD90) and velocity of diastolic depolarization (VDD).Results Remifentanil significantly decreased,APA,RPF,VDD and prolonged APD90 in a concentration dependent manner as compared with the baseline values.Pretreatment with Bay K8644 could block the effects of remifentanil on SA node pacemaker cells,while TEA did not affect the electrophysiologic effects of remifentanil on SA node pacemaker cells.Conclusion Remifentanil exerts a negative chronotropic action on SA node pacemaker cells.These effects are likely produced by decrease in Ca2+ current,while opening of K + channels is not involved in these effects.

Objective To determine the median effective plasma concentration of remifentanil (EC50) inhibiting cardiovascular response to tracheal intubation and skin incision when combined with propofol in patients of Uighur nationality.Methods Thirty ASA Ⅰ or Ⅱ Uighurs aged 20-60 yr with body mass index of 18-30 kg/m2 undergoing elective laparoscopic cholecystectomy were enrolled in this study.Anesthesia was induced and maintained with propofol infusion.BIS value was maintained at 40-50.TCI of remifentanil was then started.The initial target plasma concentration (Cp) was 6.0 ng/ml.EC50 of remifentanil was determined by modified Dixon' s upand-down sequential experiment.Each time Cp increased/decreased by 0.5 ng/ml.Tracheal intubation was facili.tated with cisatracurium 0.2 mg/kg at 2 min after a new Cp was set when target effect-site concentration (Ce) was balanced with Cp.Positive cardiovascular response was defined as increase in SBP by 15％ and/or HR ＞ 90 bpm lasting＞15 s.The EC50 (95％ confidence interval) of remifentanil blunting responses to tracheal intubation and skin incision was calculated.Results EC50 (95 ％ confidence interval) of remifentanil inhibiting cardiovascular response to tracheal intubation and skin incision when combined with propofol was 3.4 (2.3-4.5) ng/ml and 3.8 (2.8-4.9) ng/ml respectively.Conclusion When depth of anesthesia was maintained at BIS value 40-50 with propofol infusion,the EC50 of remifentanil inhibiting cardiovascular response to tracheal intubation and skin incision is 3.4 and 3.8 ng/ml respectively.

FGF21 (fibroblast growth factor 21) is a recently described member of the FGF family. It has been previously demonstrated that FGF21 is a potent regulator of glucose homeostasis. To improve stability of FGF21 for better efficacy, a new form of recombinant FGF21 was generated by fusion of a full length FGF21 gene and the Fc fragment of human IgG4 with flexible linker sequence. To examine the glucose regulation activity of FGF21-L-Fc, 3T3-L1 pre-adipocytes were differentiated into adipocytes, and glucose uptake activity of FGF21-L-Fc was examined by glucose oxidase and peroxidase (GOD-POD) assay. The results showed that in comparison with wild type FGF21, FGF21-L-Fc was more potent in stimulation of glucose uptake by 3T3-L1. In vivo studies on the modified protein demonstrated that FGF-L-Fc had a better efficacy in lowering blood glucose of the STZ-induced diabetic animals and controlled glucose level for a longer time. The results provided a sound basis for further studies.

OBJECTIVETo study on the separation from essential oil-in-water emulsion of Citri Reticulatae Pericarpium Viride by ultrafiltration and acetoacetate extraction methods respectively, and the comparison of the oil yields and chemical compositions.

METHODEssential oil-in-water emulsion of Citri Reticulatae Pericarpium Viride was separated by ultrafiltration and acetoacetate extraction methods respectively, and the chemical compositions were analyzed and compared by GC-MS.

RESULTUltrafiltration method could enrich essential oil more and its chemical compositions were more similar to the essential oil prepared by steam distillation method.

CONCLUSIONUltrafiltration method is a good medium to separate essential oil from essential oil-in-water emulsion of Citri Reticulatae Pericarpium Viride.

Chemical Fractionation ; methods ; Citrus ; chemistry ; Emulsions ; Gas Chromatography-Mass Spectrometry ; Oils, Volatile ; analysis ; isolation & purification ; Plant Extracts ; analysis ; isolation & purification ; Ultrafiltration ; methods