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1.
Stem Cell Rev Rep ; 18(5): 1822-1833, 2022 06.
Article in English | MEDLINE | ID: mdl-35089463

ABSTRACT

Highly efficient gene knockout (KO) editing of CRISPR-Cas9 has been achieved in iPSCs, whereas homology-directed repair (HDR)-mediated precise gene knock-in (KI) and high-level expression are still bottlenecks for the clinical applications of iPSCs. Here, we developed a novel editing strategy that targets introns. By targeting the intron before the stop codon, this approach tolerates reading frameshift mutations caused by nonhomologous end-joining (NHEJ)-mediated indels, thereby maintaining gene integrity without damaging the non-HDR-edited allele. Furthermore, to increase the flexibility and screen for the best intron-targeting sgRNA, we designed an HDR donor with an artificial intron in place of the endogenous intron. The presence of artificial introns, particularly an intron that carries an enhancer element, significantly increased the reporter expression levels in iPSCs compared to the intron-deleted control. In addition, a combination of the small molecules M3814 and trichostatin A (TSA) significantly improves HDR efficiency by inhibiting NHEJ. These results should find applications in gene therapy and basic research, such as creating reporter cell lines.


Subject(s)
CRISPR-Cas Systems , Induced Pluripotent Stem Cells , Recombinational DNA Repair , CRISPR-Cas Systems/genetics , DNA End-Joining Repair/genetics , Introns/genetics , Pyridazines , Quinazolines
2.
Article in English | MEDLINE | ID: mdl-34633931

ABSTRACT

An assessment of shoulder muscle coordination patterns is important to gain insight into muscle fatigue during wheelchair propulsion. The objective of the present study was to quantify muscle coordination changes over time during fatiguing wheelchair propulsion, as the muscles go through distinct levels of fatigue, a) non-fatigued, b) transiting to fatigue and c) fatigued to exhaustion. We recorded surface electromyography (sEMG) signals of the anterior deltoid (AD), middle deltoid (MD), posterior deltoid (PD), infraspinatus (IS), upper trapezius (UT), sternal head of the pectoralis major (PM), biceps brachii (BB), and triceps brachii (TB) during a wheelchair incremental exercise test. Nine wheelchair users with a diagnosis of spina bifida or T6-T12 spinal cord injury volunteered for the study. Oxygen uptake and SmartWheel kinetic parameters were also recorded during the test. EMG signals were processed by wavelet and principal component analysis (PCA), allowing for an assessment of how wheelchair users modify their muscle coordination patterns over time. Analyses of covariance (ANCOVA) were conducted to identify the main effect of fatigue levels on muscle coordination patterns by controlling for the effect of increased workload as covariate. A significant effect of fatigue levels on the PC1 and PC3 loading scores was found after controlling for the effect of increasing workloads (with both cases). In addition, PC3 reflects the most dominant fatigue effect on muscle coordination patterns which are not affected by increased ergometer workload. PC3 indicates muscle imbalance when muscles are fully fatigued and muscle co-contraction when muscles are beginning to fatigue. We conclude that fatigue-related changes in neuromuscular activity during wheelchair propulsion contribute to muscle imbalance and reflect a strategy of stiffening the shoulder joint.


Subject(s)
Wheelchairs , Electromyography , Humans , Muscle Fatigue , Muscle, Skeletal , Principal Component Analysis , Shoulder
3.
Nucleic Acids Res ; 46(19): 10195-10215, 2018 11 02.
Article in English | MEDLINE | ID: mdl-30239926

ABSTRACT

Genome editing of human induced pluripotent stem cells (iPSCs) is instrumental for functional genomics, disease modeling, and regenerative medicine. However, low editing efficiency has hampered the applications of CRISPR-Cas9 technology in creating knockin (KI) or knockout (KO) iPSC lines, which is largely due to massive cell death after electroporation with editing plasmids. Here, we report that the transient delivery of BCL-XL increases iPSC survival by ∼10-fold after plasmid transfection, leading to a 20- to 100-fold increase in homology-directed repair (HDR) KI efficiency and a 5-fold increase in non-homologous end joining (NHEJ) KO efficiency. Treatment with a BCL inhibitor ABT-263 further improves HDR efficiency by 70% and KO efficiency by 40%. The increased genome editing efficiency is attributed to higher expressions of Cas9 and sgRNA in surviving cells after electroporation. HDR or NHEJ efficiency reaches 95% with dual editing followed by selection of cells with HDR insertion of a selective gene. Moreover, KO efficiency of 100% can be achieved in a bulk population of cells with biallelic HDR KO followed by double selection, abrogating the necessity for single cell cloning. Taken together, these simple yet highly efficient editing strategies provide useful tools for applications ranging from manipulating human iPSC genomes to creating gene-modified animal models.


Subject(s)
CRISPR-Cas Systems/physiology , Gene Editing/methods , Induced Pluripotent Stem Cells/metabolism , bcl-X Protein/genetics , Animals , Cells, Cultured , Genome, Human/genetics , HEK293 Cells , Humans , Jurkat Cells , K562 Cells , Mice , Transfection , Up-Regulation/genetics
4.
Can J Neurol Sci ; 43(4): 518-22, 2016 Jul.
Article in English | MEDLINE | ID: mdl-26889714

ABSTRACT

OBJECTIVE: To study the relationship of Nε-(carboxymethyl)-lysine level (CML) with microstructure changes of white matter (WM), and cognitive impairment in patients with type 2 diabetes mellitus (T2DM) and to discuss the potential mechanism underlying T2DM-associated cognitive impairment. METHODS: The study was performed in T2DM patients (n=22) with disease course ≥5 years and age ranging from 65 to 75 years old. A control group consisted of 25 sex- and age-matched healthy volunteers. Fractional anisotropy (FA) of several WM regions was analyzed by diffusion tensor imaging scan. Plasma CML levels were measured by enzyme-linked immunosorbent assay, and cognitive function was assessed by Mini-Mental State Examination and Montreal cognitive assessment (MoCA). RESULTS: The total Mini-Mental State Examination score in the patient group (25.72±3.13) was significantly lower than the control group (28.16±2.45) (p<0.05). In addition, the total MoCA score in the patient group (22.15±3.56) was significantly lower than the control group 25.63±4.12) (p<0.01). In the patient group, FA values were significantly decreased in the corpus callosum, cingulate fasciculus, inferior fronto-occipital fasciculus, parietal WM, hippocampus, and temporal lobes relative to corresponding regions of healthy controls (p<0.05). Plasma CML level was negatively correlated with average FA values in the global brain (r=-0.58, p<0.01) and MoCA scores (r=-0.47, p<0.05). CONCLUSIONS: In T2DM, WM microstructure changes occur in older patients, and elevations in CML may play a role in the development of cognitive impairment.


Subject(s)
Cognition Disorders/etiology , Diabetes Mellitus, Type 2/blood , Diabetes Mellitus, Type 2/complications , Leukoencephalopathies/etiology , Lysine/analogs & derivatives , Aged , Anisotropy , Case-Control Studies , Cognition Disorders/diagnostic imaging , Diabetes Mellitus, Type 2/diagnostic imaging , Diffusion Magnetic Resonance Imaging , Female , Humans , Image Processing, Computer-Assisted , Leukoencephalopathies/diagnostic imaging , Lysine/blood , Male , Mental Status Schedule , Neuropsychological Tests
5.
J Ginseng Res ; 39(3): 221-9, 2015 Jul.
Article in English | MEDLINE | ID: mdl-26199553

ABSTRACT

BACKGROUND: Minor ginsenosides, those having low content in ginseng, have higher pharmacological activities. To obtain minor ginsenosides, the biotransformation of American ginseng protopanaxadiol (PPD)-ginsenoside was studied using special ginsenosidase type-I from Aspergillus niger g.848. METHODS: DEAE (diethylaminoethyl)-cellulose and polyacrylamide gel electrophoresis were used in enzyme purification, thin-layer chromatography and high performance liquid chromatography (HPLC) were used in enzyme hydrolysis and kinetics; crude enzyme was used in minor ginsenoside preparation from PPD-ginsenoside; the products were separated with silica-gel-column, and recognized by HPLC and NMR (Nuclear Magnetic Resonance). RESULTS: The enzyme molecular weight was 75 kDa; the enzyme firstly hydrolyzed the C-20 position 20-O-ß-D-Glc of ginsenoside Rb1, then the C-3 position 3-O-ß-D-Glc with the pathway Rb1→Rd→F2→C-K. However, the enzyme firstly hydrolyzed C-3 position 3-O-ß-D-Glc of ginsenoside Rb2 and Rc, finally hydrolyzed 20-O-L-Ara with the pathway Rb2→C-O→C-Y→C-K, and Rc→C-Mc1→C-Mc→C-K. According to enzyme kinetics, K m and V max of Michaelis-Menten equation, the enzyme reaction velocities on ginsenosides were Rb1 > Rb2 > Rc > Rd. However, the pure enzyme yield was only 3.1%, so crude enzyme was used for minor ginsenoside preparation. When the crude enzyme was reacted in 3% American ginseng PPD-ginsenoside (containing Rb1, Rb2, Rc, and Rd) at 45°C and pH 5.0 for 18 h, the main products were minor ginsenosides C-Mc, C-Y, F2, and C-K; average molar yields were 43.7% for C-Mc from Rc, 42.4% for C-Y from Rb2, and 69.5% for F2 and C-K from Rb1 and Rd. CONCLUSION: Four monomer minor ginsenosides were successfully produced (at low-cost) from the PPD-ginsenosides using crude enzyme.

6.
Guang Pu Xue Yu Guang Pu Fen Xi ; 35(1): 44-7, 2015 Jan.
Article in Chinese | MEDLINE | ID: mdl-25993817

ABSTRACT

Extreme ultraviolet lithography is one of the most promising technologies on the next generation of high-capacity integrated circuit manufacturing. However, techniques for ion debris mitigation have to be considered in the application of extreme ultraviolet source for lithography. In our paper the dynamics of ion debris from Sn plasma by using dual ns laser pulses were investigated. The results show that debris from plasma greatly depends on the energy of pre-pulse and the delay time between the two laser pulses. The energy of Sn ions debris was efficiently mitigated from 2. 47 to 0. 40 keV in the case of dual laser pulses, up to 6. 1 times lower than that by using single laser pulse. We also found that Sn ions debris can be mitigated at all angles by using the dual laser pulses method.

7.
Int J Neurosci ; 125(4): 298-306, 2015 Apr.
Article in English | MEDLINE | ID: mdl-24901953

ABSTRACT

PURPOSE: Motor impairment is an important index for assessing the extent of brain injury. The present study uses a new method, the movement capture analysis (MOCA) system, for assessing motor damage after acute ischemia. MATERIALS AND METHODS: Forty rats were divided into four groups: standard ischemia, sham-operated, Dizocilpine (MK-801), and Ginkgo biloba extract (GBE) groups. Brain ischemia was induced using the temporary right middle cerebral artery occlusion model. Longa score and MOCA were used to assess motor injury one day after ischemia. Infarct volume was delineated with 2% 2,3,5-triphenyltetrazolium chloride (TTC) staining. The correlation of infarct volume with Longa score and MOCA data was calculated. RESULTS: Compared with the sham-operated group (0.10 ± 0.31), Longa scores of MK-801 (2.33 ± 0.73), GBE (1.80 ± 0.58), and standard (2.88 ± 0.83) groups showed a statistical difference (p < 0.05); however, it was unable to discern the difference between MK-801 and standard groups. MOCA was able to clearly discern the differences in motor disparity among the four groups, standard (1.00 ± 0.19), sham-operated group (0.17 ± 0.02), MK-801 (0.79 ± 0.08), GBE (0.38 ± 0.05) (p < 0.05). Both MK-801 (18.03 ± 0.96%) and GBE (10.82 ± 1.93%) treatment reduced infarct size compared with the standard ischemia group (25.88 ± 1.16%) (p < 0.05). The MOCA data showed a more significant correlation with infarct size than Longa score (r = 0.85:0.53). CONCLUSIONS: MOCA system proved to be more sensitive than the Longa score. It may potentially be more accurate method for behavioral evaluation in clinical trials.


Subject(s)
Brain Injuries/complications , Movement Disorders/diagnosis , Movement Disorders/etiology , Animals , Brain Infarction/etiology , Brain Injuries/etiology , Disease Models, Animal , Infarction, Middle Cerebral Artery/complications , Male , Neurologic Examination , Rats , Statistics as Topic
8.
Guang Pu Xue Yu Guang Pu Fen Xi ; 33(2): 531-4, 2013 Feb.
Article in Chinese | MEDLINE | ID: mdl-23697148

ABSTRACT

Laser induced plasma spectroscopy of alloy steel was produced by Nd : YAG pulsed laser at 1 064 nm, and the spectral signal was detected by high resolution and width controlled ICCD. Several Fe atomic spectral lines such as 404.581, 414.387, 427.176 and 438.355 nm were chosen for analysis, and the effects of different experimental parameters on LIBS spectral signal intensity were investigated. It is shown that the experimental parameters such as pulse energy, laser focus location and laser delay time have great influence on the LIBS signal. LIBS signals with high spectral intensity and signal-background ratio (SBR) as well as the optimum experiment conditions were obtained by optimizing these experiment parameters so as to make composition analysis of the alloy steel.

9.
Chemistry ; 18(44): 14037-46, 2012 Oct 29.
Article in English | MEDLINE | ID: mdl-22996598

ABSTRACT

A 10-hydroxycamptothecin-encapsulated magnetic nanovehicle (HEMN) was fabricated by coencapsulating Fe(3)O(4) nanoparticles and 10-hydroxycamptothecin (HCPT) into a micelle core self-assembled from the amphiphilic copolymer methoxy-poly(ethylene glycol)-poly(d,l-lactide-co-glycolide) through a facile dialysis method. A satisfactory drug-loading content of (9.03 ± 0.67) % and a relatively high encapsulation efficiency of (53.52 ± 6.46) % were achieved. In vitro drug release was performed by membrane dialysis and a pH-dependent release behavior was observed. In comparison with free HCPT dissolved in dimethylsulfoxide, HEMNs showed a greatly improved in vitro antitumor efficacy against three different human cancer cell lines-HeLa, A549, and HepG2-and lower IC(50) values were measured. The mechanism of cell death was investigated, and it was clearly demonstrated that the apoptosis process was triggered. An in vitro wound-healing assay and a transwell assay indicated that HEMNs exerted much stronger activity in inhibiting HeLa cell migration. The cellular uptake of HEMNs in a desired area can be significantly enhanced by an external magnetic field. These results demonstrate HCPT-encapsulated magnetic nanovehicles might have important potential in clinical applications for inhibiting tumor metastasis and for targeted drug delivery.


Subject(s)
Antineoplastic Agents, Phytogenic/chemistry , Camptothecin/analogs & derivatives , Magnetite Nanoparticles/chemistry , Antineoplastic Agents, Phytogenic/toxicity , Apoptosis/drug effects , Camptothecin/chemistry , Camptothecin/toxicity , Cell Line, Tumor , Cell Movement/drug effects , Drug Carriers/chemistry , Ferrosoferric Oxide/chemistry , HeLa Cells , Hep G2 Cells , Humans , Magnetic Fields , Micelles
11.
Neurorehabil Neural Repair ; 25(4): 359-68, 2011 May.
Article in English | MEDLINE | ID: mdl-21343527

ABSTRACT

BACKGROUND: After hemiparetic stroke, coordination of the shoulder flexor and elbow extensor muscles during a reaching movement is impaired and contributes to poor performance. OBJECTIVE: The aim was to determine whether functional coupling between electromyographic signals of synergist muscles during reaching was weakened in stroke patients who had poor motor coordination. METHODS: Surface electromyography (EMG) from the anterior deltoid, triceps brachii, biceps brachii, pectoralis major, supraspinatus, and latissimus dorsi of the affected upper limb in 11 stroke patients (mean Fugl-Meyer upper extremity score 27 ± 8) and in the dominant arm of 8 healthy controls were measured. RESULTS: Coherence between the EMG of the anterior deltoid and triceps brachii, 2 synergists for reaching, was lower in patients compared with controls, in the 0- to 11-Hz range. Detailed segmented frequency-range analysis indicated significant differences in the coherence between groups in 0- to 3.9-Hz and 4- to 7.9-Hz ranges. CONCLUSIONS: This weakened functional coupling may contribute to poor reaching performance and could be a consequence of a loss of common drive at the frequency bands as a result of interruption of information flow in the corticospinal pathway.


Subject(s)
Arm/physiopathology , Muscle Contraction/physiology , Muscle Weakness/physiopathology , Muscle, Skeletal/physiopathology , Paresis/physiopathology , Stroke/physiopathology , Aged , Arm/innervation , Female , Humans , Male , Middle Aged , Muscle Weakness/diagnosis , Muscle Weakness/etiology , Muscle, Skeletal/innervation , Paresis/diagnosis , Paresis/etiology , Stroke/complications
12.
Eur J Pharmacol ; 654(3): 320-5, 2011 Mar 11.
Article in English | MEDLINE | ID: mdl-21236251

ABSTRACT

Microvascular complications are much earlier and common in diabetes. Advanced glycation end products (AGEs), together with high glucose, play a key role in the endothelial dysfunction of diabetic vascular complications. So it is of more significance to expedite the therapies to block the formation and/or the effects of AGEs. Berberine has been showed to have anti-diabetic effects, however the effects on diabetic complications were less explored, especially the effects on the microvascular complications and the formation and pathways of AGEs which have not been reported. Therefore, the present study established an in vitro model of diabetic microendothelial (microEC) injury by the combination of high glucose and AGEs to mimic the clinical situations and examine the effects and mechanisms of berberine on high glucose-AGEs-induced microEC injuries and on the formation of AGEs. We prepared AGEs, established the high glucose-AGEs injured microEC models by MTT assay, which was further supported by significantly decreased nitric oxide (NO) release, NO synthase (NOS) and thrombomodulin production with ELISA, western blot and RT-PCR analysis. Berberine treatments showed significant improvements as indicated by significantly increased NO release, NOS and thrombomodulin production. Moreover, we also observed significant inhibition effects of berberine on AGEs formation. We concluded that the in vitro model of diabetic microEC injury could be established by the combination treatments of high glucose and AGEs, while berberine could improve the diabetic microvascular injury in vitro and inhibit the formation of AGEs, suggesting the potential clinical therapies with berberine for diabetes and its vascular complications.


Subject(s)
Berberine/pharmacology , Diabetes Complications/drug therapy , Endothelium, Vascular/drug effects , Endothelium, Vascular/injuries , Glucose/pharmacology , Glycation End Products, Advanced/pharmacology , Vascular System Injuries/drug therapy , Animals , Berberine/therapeutic use , Cell Extracts , Cell Line , Diabetes Complications/chemically induced , Diabetes Complications/metabolism , Diabetes Complications/pathology , Dose-Response Relationship, Drug , Drug Interactions , Endothelium, Vascular/metabolism , Enzyme-Linked Immunosorbent Assay , Mice , Nitric Oxide/metabolism , Nitric Oxide Synthase/metabolism , Thrombomodulin/genetics , Thrombomodulin/metabolism , Vascular System Injuries/chemically induced , Vascular System Injuries/metabolism , Vascular System Injuries/pathology
13.
Phytother Res ; 25(1): 137-41, 2011 Jan.
Article in English | MEDLINE | ID: mdl-20632296

ABSTRACT

Ginkgo biloba extracts (GBE) have long been used as a traditional herbal medicine for treating central nervous system diseases and peripheral vascular diseases, but the underlying mechanisms have yet to be elucidated. Furthermore, traditional GBE is in the form of microsomes and only dissolves in organic solvents; its clinical applications have been greatly limited. Therefore, in the present study, nanometer GBE (nGBE) was prepared utilizing supercritical anti-solvent (SAS) upon CO(2) -supercritical fluid extraction (CO(2) -SPF). Using whole-cell patch clamp techniques, the effects of different preparations of GBE on N-methyl-D-aspartate (NMDA)-activated currents (I(NMDA) ) from acutely isolated rat hippocampal neurons were investigated and the difference in protective potency between nGBE and mGBE evaluated. The results showed that the inward current activated by NMDA could be depressed by mGBE and nGBE. The inhibitory rates were 40% ± 17% and 64% ± 15%, and the half-inhibition concentrations (IC(50) ) were 0.0210 ± 0.0055 and 0.0262 ± 0.0038 mg/mL, respectively. In comparison, the modulatory effect of nGBE (dissolved in extracellular solution) on NMDA-activated current was significantly greater than that of mGBE (dissolved in DMSO) (p < 0.05). This indicated that the modulatory effects of GBE on NMDA-activated current may contribute to the neuroprotective effects of GBE and the modulatory effect of nGBE on NMDA-activated current was greater than that of mGBE.


Subject(s)
Ginkgo biloba/chemistry , Hippocampus/drug effects , N-Methylaspartate/pharmacology , Neurons/drug effects , Neuroprotective Agents/pharmacology , Plant Extracts/pharmacology , Animals , Neurons/physiology , Neuroprotective Agents/isolation & purification , Particle Size , Patch-Clamp Techniques , Plant Extracts/chemistry , Plant Extracts/isolation & purification , Rats , Rats, Sprague-Dawley , Receptors, N-Methyl-D-Aspartate/drug effects , Receptors, N-Methyl-D-Aspartate/physiology , Solubility
14.
Chin Med J (Engl) ; 122(21): 2560-6, 2009 Nov 05.
Article in English | MEDLINE | ID: mdl-19951570

ABSTRACT

BACKGROUND: A five-year follow-up study of intensive multifactorial intervention was undertaken to assess the changes of circulating serum amyloid A (SAA) levels and the incidence of atherosclerosis (AS) in patients with short-duration type 2 diabetes mellitus (T2DM) without macroangiopathy, and whether intensive multifactorial intervention could prevent or at least postpone the occurrence of macroangiopathy. METHODS: Among 150 patients with short-duration T2DM, 75 were assigned to receive conventional outpatient treatment (conventional group) and the others underwent intensive multifactorial integrated therapy targeting hyperglycemia, hypertension, dyslipidemia and received aspirin simultaneously (intensive group). RESULTS: Plasma SAA levels were higher in diabetic patients than those in healthy control subjects, and decreased obviously after intensive multifactorial intervention. The levels of SAA were positively correlated with body mass index (BMI), waist hip ratio (WHR), triglyceride (TG), high sensitive C-reactive protein (hs-CRP) and common carotid intima-media thickness (CC-IMT). The standard-reaching rates of glycemia, blood pressure and lipidemia were significantly higher in intensive group than those of conventional group. The incidence of macroangiopathy decreased by 58.96% in intensive group compared with conventional group. CONCLUSIONS: Intensive multifactorial intervention may significantly reduce the SAA levels and prevent the occurrence of AS in short-duration patients with T2DM. SAA might be one of the risk factors of T2DM combined with AS.


Subject(s)
Diabetes Mellitus, Type 2/complications , Diabetes Mellitus, Type 2/metabolism , Diabetic Angiopathies/etiology , Serum Amyloid A Protein/metabolism , Adult , Aged , Antihypertensive Agents/pharmacology , Antihypertensive Agents/therapeutic use , Blood Glucose/metabolism , C-Reactive Protein/metabolism , Diabetes Mellitus, Type 2/drug therapy , Female , Humans , Hypoglycemic Agents/pharmacology , Hypoglycemic Agents/therapeutic use , Hypolipidemic Agents/pharmacology , Hypolipidemic Agents/therapeutic use , Male , Middle Aged , Multivariate Analysis , Triglycerides/blood , Tunica Media/drug effects
15.
World J Gastroenterol ; 15(15): 1829-35, 2009 Apr 21.
Article in English | MEDLINE | ID: mdl-19370779

ABSTRACT

AIM: To investigate the hepatoprotective activity of tea polyphenols (TP) and its relation with cytochrome P450 (CYP450) expression in mice. METHODS: Hepatic CYP450 and CYPb(5) levels were measured by UV-spectrophotometry in mice 2 d after intraperitoneal TP (25, 50 and 100 mg/kg per day). Then the mice were intragastricly pre-treated with TP (100, 200 and 400 mg/kg per day) for six days before paracetamol (1000 mg/kg) was given. Their acute mortality was compared with that of control mice. The mice were pre-treated with TP (100, 200, and 400 mg/kg per day) for five days before paracetamol (500 mg/kg) was given. Hepatic CYP2E1 and CYP1A2 protein and mRNA expression levels were evaluated by Western blotting, immunohistochemical staining and transcriptase-polymerase chain reaction. RESULTS: The hepatic CYP450 and CYPb(5) levels in mice of TP-treated groups (100, 200 and 400 mg/kg per day) were decreased in a dose-dependent manner compared with those in the negative control mice. TP significantly attenuated the paracetamol-induced hepatic injury and dramatically reduced the mortality of paracetamol-treated mice. Furthermore, TP reduced CYP2E1 and CYP1A2 expression at both protein and mRNA levels in a dose-dependent manner. CONCLUSION: TP possess potential hepatoprotective properties and can suppress CYP450 expression.


Subject(s)
Acetaminophen/toxicity , Analgesics, Non-Narcotic/toxicity , Cytochrome P-450 CYP1A2/metabolism , Cytochrome P-450 CYP2E1/metabolism , Flavonoids/therapeutic use , Liver/drug effects , Phenols/therapeutic use , Tea/chemistry , Animals , Cytochrome P-450 CYP1A2/genetics , Cytochrome P-450 CYP2E1/genetics , Dose-Response Relationship, Drug , Isoenzymes/genetics , Isoenzymes/metabolism , Liver/metabolism , Liver/pathology , Male , Mice , Microsomes, Liver/drug effects , Microsomes, Liver/enzymology , Polyphenols
16.
J Neurosci Methods ; 177(1): 232-40, 2009 Feb 15.
Article in English | MEDLINE | ID: mdl-18977246

ABSTRACT

This study investigates time-dependent associations between source strength estimated from high-density scalp electroencephalogram (EEG) and force of voluntary handgrip contraction at different intensity levels. We first estimate source strength from raw EEG signals collected during voluntary muscle contractions at different levels and then propose a functional random-effects model approach in which both functional fixed effects and functional random-effects are considered for the data. Two estimation procedures for the functional model are discussed. The first estimation procedure is a two-step method which involves no iterations. It can flexibly use different smoothing methods and smoothing parameters. The second estimation procedure benefits from the connection between linear mixed models and regression splines and can be fitted using existing software. Functional ANOVA is then suggested to assess the experimental effects from the functional point of view. The statistical analysis shows that the time-dependent source strength function exhibits a nonlinear feature, where a bump is detected around the force onset time. However, there is the lack of significant variations in source strength on different force levels and different cortical areas. The proposed functional random-effects model procedure can be applied to other types of functional data in neuroscience.


Subject(s)
Brain/physiology , Electroencephalography , Models, Biological , Muscle, Skeletal/physiology , Nonlinear Dynamics , Adult , Analysis of Variance , Brain Mapping , Female , Humans , Male , Muscle Contraction/physiology , Time Factors , Young Adult
17.
Brain Res ; 1250: 101-12, 2009 Jan 23.
Article in English | MEDLINE | ID: mdl-19028460

ABSTRACT

OBJECTIVE: Recent research has shown dissociation between changes in brain and muscle signals during voluntary muscle fatigue, which may suggest weakening of functional corticomuscular coupling. However, this weakening of brain-muscle coupling has never been directly evaluated. The purpose of this study was to address this issue by quantifying EEG-EMG coherence at times when muscles experienced minimal versus significant fatigue. METHODS: Nine healthy subjects sustained an isometric elbow flexion at 30% maximal level until exhaustion while their brain (EEG) and muscle (EMG) activities were recorded. The entire duration of the EEG and EMG recordings was divided into the first half (stage 1 with minimal fatigue) and second half (stage 2 with severer fatigue). The EEG-EMG coherence and power spectrum in each stage was computed. RESULTS: The power of both EEG and EMG increased significantly while their coherence decreased significantly in stage 2 compared with stage 1 at beta (15-35 Hz) band. CONCLUSIONS: Despite an elevation of the power for both the EEG and EMG activities with muscle fatigue, the fatigue weakens strength of brain-muscle signal coupling at beta frequency band. SIGNIFICANCE: Weakening of corticomuscular coupling may be a major neural mechanism contributing to muscle fatigue and associated performance impairment.


Subject(s)
Brain/physiology , Muscle Fatigue/physiology , Muscle, Skeletal/physiology , Adult , Arm/physiology , Beta Rhythm , Electroencephalography , Electromyography , Female , Humans , Isometric Contraction/physiology , Male , Middle Aged
18.
Chin J Integr Med ; 14(2): 117-22, 2008 Jun.
Article in English | MEDLINE | ID: mdl-18679602

ABSTRACT

OBJECTIVE: To investigate the relationship between the excitotoxicity and serum-inducible kinase (SNK) and spine-associated Rap GTPase-activating protein (SPAR) pathway in primary hippocampal neuron injury induced by glutamate and furthermore, to explore the molecular mechanism of neuroprotection of Zibu Piyin Recipe (ZBPYR) and the relationship between ZBPYR and the morphological regulation of dendritic spines. METHODS: The serum containing ZBPYR was prepared by seropharmacology. Reverse transcription and polymerase chain reaction (RT-PCR) was used to detect the expression of mRNA for SNK, SPAR, postsynaptic density protein 95 (PSD-95) and N-methyl-D-aspartate (NMDA) receptor subunits (NR1, NR2A and NR2B) in primary rat hippocampal neuron cultures after pretreatment with 10 micromol/L glutamate and ZBPYR serum. RESULTS: ZBPYR serum pretreatment resulted in a significant down-regulation of glutamate-induced SNK mRNA expression (P<0.05). Significant up-regulation was seen on the mRNA expression of SPAR and PSD-95 (P<0.05). All these changes were dose-dependent. The mRNA expression of NR1, NR2A and NR2B was down-regulated to different degrees (P<0.05). CONCLUSION: The mechanism of effect of ZBPYR on glutamate-induced excitotoxicity may be related to the regulation of SNK-SPAR signal pathway. ZBPYR may play a role in protecting and maintaining the normal morphology and structure of dendritic spines, which may be achieved by inhibiting the excessive activation of NMDA receptors.


Subject(s)
Drugs, Chinese Herbal/pharmacology , GTPase-Activating Proteins/metabolism , Glutamic Acid/toxicity , Neurons/enzymology , Neurons/pathology , Protein Kinases/metabolism , Animals , Cells, Cultured , Disks Large Homolog 4 Protein , GTPase-Activating Proteins/genetics , Gene Expression Regulation/drug effects , Hippocampus/drug effects , Hippocampus/enzymology , Hippocampus/pathology , Intracellular Signaling Peptides and Proteins/genetics , Intracellular Signaling Peptides and Proteins/metabolism , Membrane Proteins/genetics , Membrane Proteins/metabolism , Neurons/drug effects , Protein Kinases/genetics , Protein Serine-Threonine Kinases , RNA, Messenger/genetics , RNA, Messenger/metabolism , Rats , Rats, Sprague-Dawley , Receptors, N-Methyl-D-Aspartate/genetics , Receptors, N-Methyl-D-Aspartate/metabolism , Reverse Transcriptase Polymerase Chain Reaction , Serum
19.
Chin Med J (Engl) ; 121(13): 1165-8, 2008 Jul 05.
Article in English | MEDLINE | ID: mdl-18710632

ABSTRACT

BACKGROUND: Tanis was reported as a putative receptor for serum amyloid A (SAA) involving glucose regulated protein in insulin regulated resistance. It was found to be dysregulated in diabetic rats (Psammomys obesus, Israeli sand rat) and its homologue for humans is SelS/AD-015. The present study analyzed mRNA expression of SelS in omental adipose tissue biopsies from patients with type 2 diabetes mellitus (T2DM), and age- and weight-matched nondiabetic patients, the relationship of SelS mRNA with Homa-IR and serum SAA level. METHODS: Human omental adipose tissues from ten cases of type 2 diabetic patients and twelve cases of nondiabetic individuals were analyzed for the expression level of SelS mRNA by semiquantitative polymerase chain reaction (PCR), Homa-IR estimated by standard formula and SAA level by enzyme-linked immunosorbent assay (ELISA). RESULTS: SelS mRNA expression, Homa-IR and serum SAA were higher in T2DM sufferers than in nondiabetic control group. SelS mRNA level was positively correlated with Homa-IR and SAA level in each group. CONCLUSIONS: SelS protein may be involved in insulin resistance in Chinese with T2DM by acting as the SAA receptor, thus playing an important role in the development of T2DM and atherosclerosis.


Subject(s)
Adipose Tissue/metabolism , Diabetes Mellitus, Type 2/metabolism , Insulin Resistance , Membrane Proteins/genetics , Omentum/metabolism , RNA, Messenger/analysis , Selenoproteins/genetics , Serum Amyloid A Protein/analysis , Adult , Aged , Base Sequence , Female , Humans , Male , Molecular Sequence Data
20.
Brain Res ; 1228: 229-40, 2008 Sep 04.
Article in English | MEDLINE | ID: mdl-18590710

ABSTRACT

Little is known about the association between brain white matter (WM) structure and motor function in humans. This study investigated complexity of brain WM interior shape as determined by magnetic resonance imaging (MRI) and its relationship with upper-extremity (UE) motor function in patients post stroke. We hypothesized that (1) the WM complexity would decrease following stroke, and (2) higher WM complexity in non-affected cortical areas would be related to greater UE motor function. Thirty-eight stroke patients (16 with left-hemisphere lesions) underwent MRI anatomical brain scans. Fractal dimension (FD), a quantitative shape metric, was applied onto skeletonized brain WM images to evaluate WM internal structural complexity. Wolf Motor Function Test (WMFT) and Fugl-Meyer Motor Assessment (FM) scores were measured to assess motor function of the affected limb. The WM complexity was lower in the stroke-affected hemisphere. The FD was associated with better motor function in two subgroups: with left-subcortical lesions, FD values of the lesion-free areas of the left hemisphere were associated with better FM scores; with right-cortical lesions, FD values of lesion-free regions were robustly associated with better WMFT scores. These findings suggest that greater residual WM complexity is associated with less impaired UE motor function, which is more robust in patients with right-hemisphere lesions. No correlations were found between lesion volume and WMFT or FM scores. This study addressed WM complexity in stroke patients and its relationship with UE motor function. Measurement of brain WM reorganization may be a sensitive correlate of UE function in people recovering from stroke.


Subject(s)
Brain/pathology , Fractals , Motor Skills/physiology , Stroke/physiopathology , Upper Extremity/physiopathology , Adult , Aged , Aged, 80 and over , Analysis of Variance , Dominance, Cerebral/physiology , Female , Humans , Image Processing, Computer-Assisted , Magnetic Resonance Imaging/methods , Male , Middle Aged , Paresis/pathology , Paresis/physiopathology , Recovery of Function/physiology
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