Objective To investigate the effects of FKBP12.6 gene transfection on the contractility of ventricular myocytes of rats with heart failure.Methods Rat model of heart failure was established by a surgical operation of abdominal aortic stenosis.The enzymatic hydrolysis was employed to isolate the rats' ventricular myocytes.Mediated by adenovirus the FKBP12.6 gene was transfected into ventricular myocytes.Ad-GFP infected cardiac myocytes from heart failure rats and normal rats were used as control.Western blotting and reverse transcriptase-polymerase chain reaction(RT-PCR) were used to detect the specific overexpression of FKBP12.6.Transfected ventricular myocytes were loaded with the membrane-permeant Ca2+ dye X-rhod-1-AM.Under the condition of field stimulation,the laser confocal microscope in line-scan and plane-scan mode was used to detect the Ca2+ transients and myocytes contraction.Results The mRNA and protein levels of FKBP12.6 in the myocytes of rats with heart failure were significantly lower than that in normal myocytes(0.47?0.08 vs 0.96?0.12,0.25?0.04 vs 0.48?0.07,P

Hamartoma ; Humans ; Laryngeal Diseases ; Male ; Middle Aged

AIM:To study the effects of FK506 binding protein 12.6(FKBP12.6) overexpression on the performance of sarcoplasmic reticulum(SR) in ventricular myocytes of rats with heart failure.METHODS:The adenovirus(Ad)-mediated gene transfer was used to overexpress FKBP12.6 in ventricular myocytes of rats with heart failure.Western blotting and reverse transcriptase-polymerase chain reaction(RT-PCR) analysis were used to reveale specific overexpression of FKBP12.6.X-rhod-1-AM was used as the Ca2+ indicator,and cells were viewed under a confocal microscope.RESULTS:Adenovirus mediated overexpression of FKBP12.6 resulted in a 5-fold increase in relative FKBP12.6 mRNA levels and a 4-fold increase in relative FKBP12.6 protein levels at 48 h after transfection compared with control.The amplitude of the fluorescence [Ca2+]i transient was significantly increased in Ad-FKBP12.6-GFP cardiomyocytes compared with Ad-GFP myocytes(peak F/F0,3.16?0.42 vs 1.43?0.38,P

ObjectiveTo explore the effect of brain wave-biofeedback on attention deficit hyperactivity disorder.Methods29 children with attention deficit hyperactivity disorder used VBFB3000 Brain Wave-Biofeedback system to control the 4～8 Hz brain wave and activate the 12～16 Hz wave twice a week.Results84.6% children primarily with attention deficit became normal,as well as 100% with hyperactivity,91.6% with mixed appearing.ConclusionBrain Wave-Biofeedback is effective on any types of attention deficit hyperactivity disorder.

Through scale-up cultivation of Eriobotrya japonica suspension cells using WAVE bioreactor, the cell growth and ursolic acid (UA) accumulation were studied. The comparison test was carried out in the flask and the reactor with cell dry weight (DW) and UA content as evaluation indexes. The culture medium, DW and UA content were compared in 1 L and 5 L working volumes of bioreactor. The orthogonal test with main actors of inoculation amount, speed and angle of rotation was developed to find the optimal combination, in 1 L working volume of bioreactor. DW of the cell growth and the UA content in bioreactor were higher than those of the shaker by 105.5% and 27.65% respectively. In bioreactor, the dynamic changes of elements in the fluid culture, the dry weight of the cell growth and the UA content in 1 L and 5 L working volumes were similar. Inoculation of 80 g, rotational speed of 26 r · min(-1), and angle of 6 ° was the optimal combination, and the cell biomass of 19.01 g · L(-1) and the UA content of 27.750 mg · g(-1) were achieved after 100 h cultivation in 1 L working volume of bioreactor. WAVE Bioreactor is more suitable than flasks for the E. japonica cell suspension culture, and culture parameters can be achieved from 1 L to 5 L amplification.
Biomass ; Bioreactors ; Cell Culture Techniques ; instrumentation ; methods ; Culture Media ; chemistry ; metabolism ; Eriobotrya ; chemistry ; growth & development ; metabolism ; Triterpenes ; analysis ; metabolism

The positive ratio is 74 percent using the plate anaerobe culturing device made by author, higher than using common anaerobic box and anaerobic jar.

ObjectiveTo explore the effect of metalloproteinases (MMPs) inhibitor on expression of MMP-3 in degenerated lumbar intervertebral disc.MethodsThe animal model of degenerated lumbar intervertebral disc was established with rats. The 32 rats were randomly divided into the experimental group and control group with 16 animals in each group. From the fourth week after operation, the animals of the experimental group were injected with tetracycline 25 mg per day, those of the control group with saline. Two weeks late, the tissue of degenerated lumbar intervertebral disc was taken and the expression of MMP-3 was tested by immumohistochemistry and Western-bloting.ResultsThe expression of MMP-3 in the experimental group decreased and significantly different from the control group ( P<0.05).ConclusionMMPs inhibitor can decrease expression of MMP-3 in degenerated lumbar intervertebral disc.

OBJECTIVE: To investigate the effect of asymmetric dimethylarginine (ADMA), an endogenous inhibitor of nitric oxide synthase, on the activation of hepatic stellate cells (HSCs) and its mechanism. METHODS: Primary HSCs isolated from SD rats were cultured and treated with different concentrations (1, 3 or 10micromol/L) of ADMA for various periods (12 approximately 48h). Expression of alpha-smooth muscle actin (alpha-SMA) and synthesis of type-I collagens in HSC were determined. Messenger RNA levels of the transforming growth factor-beta1 (TGF-beta(1)) in the HSCs were determined using RT-PCR. Intracellular reactive oxidant species (ROS) production was measured using oxidant-sensitive fluorescent indicator. Activation of nuclear factor-kappaB (NF-kappaB) was detected by electrophoretic mobility shift assay (EMSA). RESULTS: ADMA could increase alpha-SMA-positive cells ratio and Type I collagens production of HSCs in a concentration- and time-dependent manner, concomitant with the increase of the TGF-beta(1) mRNA level. Treatment with ADMA (10micromol/L) significantly increased the intracellular ROS production and activated NF-kappaB. Such effects of ADMA on the level of TGF-beta(1) mRNA could be markedly attenuated by pretreatment with antioxidant pyrrolidine dithiocarbamate (25micromol/L). CONCLUSION ADMA can induce the HSC activation by increasing TGF-beta(1) expression through ROS-NF-kappaB-dependent pathway. Therefore, ADMA should be a novel and endogenous activator of HSC, which may be involved in the development of liver fibrosis.
Actins ; biosynthesis ; Animals ; Arginine ; analogs & derivatives ; pharmacology ; Cells, Cultured ; Collagen Type I ; metabolism ; Dose-Response Relationship, Drug ; Gene Expression ; drug effects ; Hepatocytes ; cytology ; drug effects ; metabolism ; Male ; NF-kappa B ; metabolism ; Nitric Oxide ; metabolism ; Nitric Oxide Synthase ; metabolism ; RNA, Messenger ; genetics ; metabolism ; Rats ; Rats, Sprague-Dawley ; Reactive Oxygen Species ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction ; Transforming Growth Factor beta ; genetics

Hippo signaling pathways are evolutionarily conserved signal transduction mechanisms mainly involved in organ size control, tissue regeneration, and tumor suppression. However, in mammals, the primary role of Hippo signaling seems to be regulation of immunity. As such, humans with null mutations in STK4 (mammalian homologue of Drosophila Hippo; also known as MST1) suffer from recurrent infections and autoimmune symptoms. Although dysregulated T cell homeostasis and functions have been identified in MST1-deficient human patients and mouse models, detailed cellular and molecular bases of the immune dysfunction remain to be elucidated. Although the canonical Hippo signaling pathway involves transcriptional coactivator Yes-associated protein (YAP) or transcriptional coactivator with PDZ motif (TAZ), the major Hippo downstream signaling pathways in T cells are YAP/TAZ-independent and they widely differ between T cell subsets. Here we will review Hippo signaling mechanisms in T cell immunity and describe their implications for immune defects found in MST1-deficient patients and animals. Further, we propose that mutual inhibition of Mst and Akt kinases and their opposing roles on the stability and function of forkhead box O and β-catenin may explain various immune defects discovered in mutant mice lacking Hippo signaling components. Understanding these diverse Hippo signaling pathways and their interplay with other evolutionarily-conserved signaling components in T cells may uncover molecular targets relevant to vaccination, autoimmune diseases, and cancer immunotherapies.

Objective To observe the clinical efficacy of acupuncture at cranial sutures plus rehabilitation training in treating post-stroke spastic palsy.Method Sixty patients with post-stroke spastic palsy were randomized into a treatment group and a control group, 30 cases each. The treatment group was intervened by acupuncture at cranial sutures plus rehabilitation training, while the control group was intervened by rehabilitation training alone. The Modified Ashworth Scale (MAS), Fugl-Meyer Assessment, and activities of daily living (ADL) in the Modified Barthel Index (MBI) were observed before and after the treatment for the two groups.Result After the intervention, there was a significant difference in MAS score between the two groups (P<0.05). The FMA and ADL scores were changed significantly after the treatment in both groups (P<0.05). The FMA and ADL scores in the treatment group were significantly different from those in the control group after the treatment (P<0.05).Conclusion Acupuncture at cranial sutures plus rehabilitation is an effective approach in treating post-stroke spastic palsy.