LncRNA TUG1 promotes proliferation of vascular smooth muscle cell and atherosclerosis through regulating miRNA-21/PTEN axis.

OBJECTIVE: Atherosclerosis is a major risk factor for cardiovascular disease, but its mechanism of progression remained unclear. However, many long non-coding RNAs (lncRNAs) have recently been implicated in different processes for cardiovascular disease. In this study, we mainly focused on the role of lncRNA TUG1 in atherosclerosis. PATIENTS AND METHODS: qRT-PCR was used to detect the expression of lncRNA TUG1 in atherosclerosis patients and animal model. Moreover, the expression of TUG1 in vascular smooth muscle cell dysfunction model was also measured. Proliferation ability was tested by CCK-8 and cyclin D1 assay, through loss- and gain-of function approaches. Western-blot was used to measure the expression of PTEN, when TUG1 was in different levels. RESULTS: We found that the lncRNA TUG1 was highly expressed in serum samples from 38 patients with atherosclerosis, compared with 24 healthy volunteers.  LncRNA TUG1 was dramatically upregulated in atherosclerotic plaques of ApoE-/- mice. We also found that the expression of TUG1 was upregulated in vascular smooth muscle cell injury model. Through loss- and gain-of function approaches, we showed that TUG1 promotes cell proliferation and induces apoptosis in vitro. What's more, TUG1 expression level was reversely correlated with PTEN expression in patients with atherosclerosis. LncRNA TUG1 could compete with PTEN for miR-21 binding. CONCLUSIONS: We found that lncRNA TUG1 was closely related to the progression of atherosclerosis, which could be a potential target for treating atherosclerosis.

Comparison of anaesthetic and analgesic effects of emulsified isoflurane used alone or combined with lidocaine and fentanyl in dogs.

AIM: The objective of this study was to investigate the effects of a combination of emulsified isoflurane, lidocaine, and fentanyl (EI-L-F) compared with the efficacy of emulsified isoflurane alone (EI), a combination of emulsified isoflurane and lidocaine (EI-L) or emulsified isoflurane and fentanyl (EI-F) for anaesthetising dogs. METHODS: Eight mongrel dogs were anesthetised with EI (8 mL/kg/hour), EI-L (3 mg/kg/hour lidocaine and 6 mL/kg/hour of emulsified isoflurane), EI-F (1.5 µg/kg/hour fentanyl and 6 mL/kg/hour of emulsified isoflurane), and EI-L-F (5 mL/kg/hour of emulsified isoflurane, 1 µg/kg/hour of fentanyl and 2.4 mg/kg/hour of lidocaine). Each dog received all four treatments and there was a 15-day washout period between the treatments. The dogs' anaesthesia and analgesia scores and physiological parameters were determined before and 5, 10, 20, 30, 40, 50, 60, 70 and 80 minutes after the administration of anaesthetic agents. RESULTS: The dogs in each of the four groups became laterally recumbent within 1 minute. Respiration rate and heart rate increased (p<0.05) during the first 5 minutes of anaesthesia in all groups. Respiration rate in the EI-F-L group was higher (p=0.037) than other groups from 30 to 50 minutes. Heart rate was higher in the EI than EI-F-L group (p=0.018) from 10 to 20 minutes, then returned to near baseline. Arterial oxygen saturation decreased during the period of anaesthesia but was higher (p=0.032) from 10 to 50 minutes in EI-F-L group than in other groups. The total anaesthesia scores in the EI-L-F group were higher than the EI and EI-L groups (p<0.05). The mean time to body movement was 5 (SD 2), 5 (SD 2), 7 (SD 2) and 8 (SD 2) minutes for the EI, EI-L, EI-F and EI-F-L groups, respectively. The mean time to standing was 8 (SD 2), 9 (SD 2), 10 (SD 2) and 13 (SD 3) minutes for the EI, EI-L, EI-F and EI-F-L groups, respectively. No excitement was observed during recovery after anaesthesia. CONCLUSIONS: The EI-F-L combination that was used in this study provided an adequate anaesthesia effect in dogs, which was characterised by adequate analgesia and muscle relaxation without any complications.

Interface modification of the InGaN/GaN quantum wells: the strain pre-relief effect.

Interface modification by inserting an ultrathin low-temperature GaN layer prior to the growth of high-temperature GaN barriers followed by an annealing process was employed to improve the properties of the InGaN/GaN quantum wells. By detailed studies and comparisons of the surface morphology, photoluminescence and the surface compositions of the InGaN/GaN quantum wells at different growth stages with and without the interface modification, we find that with the interface modification the surface morphology was significantly improved with better smoothness, and smaller and shallower pits of lower density compared with that without interface modification; further, the indium aggregation and phase separation were suppressed. The physical phenomena are attributed to the 'strain pre-relief effect' by the formation of quasi-dots (approximately 20 nm in diameter) prior to temperature ramping and growth of high-temperature GaN barriers. Furthermore, the ultrathin low-temperature GaN layers have a good protection property as confirmed by PL and XPS measurements.

The use of ion-selective electrodes for evaluating residence time distributions in expanded bed adsorption systems.

The suitability of ion-selective electrodes (ISE) for the determination of residence time distribution (RTD) in turbid, cell-containing fluids was examined. The electrodes were found to give reproducible signals in biomass-containing feedstock with up to 20% wet weight of solids. The enhanced feedstock compatibility of IES, when compared to other tracer sensing devices, allows the study of expanded bed system hydrodynamics under relevant operating conditions. Within the linear range of the corresponding ISE-tracer pair, both examined ISE (Li(+)- or Br(-)-selective) showed to be insensitive against the range of flow rate and pH normally employed during expanded bed adsorption (EBA) of proteins. Analyzing the RTD obtained after a perfect ion tracer pulse in terms of the PDE model (PDE, axially dispersed plug-flow exchanging mass with stagnant zones) gave a quantitative description of the underlying hydrodynamic situation during EBA processing. These data provided a powerful tool to make predictions on the adsorptive global process performance with a defined feedstock type and composition. The link between the hydrodynamic events during feedstock application and the actual process performance was shown when applying intact yeast cell suspensions at different biomass content (up to 7.5% wet weight) and buffer conductivity (5-12 mS) onto an EBA column filled with the adsorbent Streamline Q XL as fluidized phase. On the basis of our experimental results, a guideline for the successful application of the ISE/RTD method to EBA process design is presented.

Minimising biomass/adsorbent interactions in expanded bed adsorption processes: a methodological design approach.

Expanded bed adsorption (EBA) is an integrated technology for the primary recovery of proteins from crude feedstock. Interactions between solid matter in the feed suspension and fluidised adsorbent particles influence bed stability and therefore have a significant impact on protein adsorption in expanded beds. In order to design efficient and reliable EBA processes a strategy is needed, which allows to find operating conditions, where these adverse events do not take place. In this paper a methodological approach is presented, which allows systematic characterisation and minimisation of cell/adsorbent interactions with as little experimental effort as possible. Adsorption of BSA to the anion exchanger Streamline Q XL from a suspension containing S. cerevisiae cells was chosen as a model system with a strong affinity of the biomass towards the stationary phase. Finite bath biomass adsorption experiments were developed as an initial screening method to estimate a potential interference. The adhesiveness of S. cerevisiae to the anion exchanger could be reduced significantly by increasing the conductivity of the feedstock. A biomass pulse response method was used to find optimal operation conditions showing no cell/adsorbent interactions. A good correlation was found between the finite bath test and the pulse experiment for a variety of suspensions (intact yeast cells, E. coli homogenate and hybridoma cells) and adsorbents (Streamline Q XL, DEAE and SP), which allows to predict cell/adsorbent interactions in expanded beds just from finite bath adsorption tests. Under the optimised operating conditions obtained using the prior methods, the stability of the expanded bed was investigated during fluidisation in biomass containing feedstock (up to 15% yeast on wet weight basis) employing residence time distribution analysis and evaluation by an advanced model. Based on these studies threshold values were defined for the individual experiments, which have to be achieved in order to obtain an efficient EBA process. Breakthrough experiments were conducted to characterise the efficiency of BSA adsorption from S. cerevisiae suspensions in EBA mode under varying operating conditions. This allowed to correlate the stability of the expanded bed with its sorption efficiency and therefore could be used to verify the threshold values defined. The approach presented in this work provides a fast and simple way to minimise cell/adsorbent interactions and to define a window of operation for protein purification using EBA.