AIMTo investigate membrane electro-physiological features in vestibular ganglion neuronal population using a voltage-sensitive dye and optical recording technique.

METHODSDissociated and cultured mouse vestibular ganglion neurons were stained with an absorption voltage-sensitive dye, RH 155, and were imaged in 16 x 16 elements Photodiode arrays (PDA) optical recording system.

RESULTSWhen the cells were depolarized during perfusion with 150 mmol/L potassium solution, optical absorption of the dye that bound to the external surface of neuron membranes increased. The relative ratio (delta I/I) of optical absorption change was 0.23% +/- 0.08% (x +/- s, n = 28). These optical responses were wavelength dependent. Under our experimental conditions, photo toxicity and pharmacological effects of the dye were either absent or insignificant.

CONCLUSIONOur results suggest that optical recording provides a new, practical and less toxic method to simultaneously monitor changes in membrane potential from several cultured vestibular ganglion cells.

Action Potentials ; physiology ; Animals ; Cells, Cultured ; Female ; Fluorescent Dyes ; Male ; Membrane Potentials ; physiology ; Mice ; Mice, Inbred ICR ; Neurons ; physiology ; Patch-Clamp Techniques ; methods ; Vestibular Nerve ; physiology

Nanoporous ZnO was used as a carrier to prepare drug solid dispersion, the mechanism of which to improve the drug dissolution was also studied. Nanoporous ZnO, obtained through chemical deposition method, was used as a carrier to prepare indomethacin and cilostazol solid dispersions by melt-quenching method, separately. The results of scanning electron microscope, surface area analyzer, fourier transform infra-red spectroscopy, differential scanning calorimeter and X-ray diffraction showed that drugs were implanted into nanopores of ZnO by physical adsorption effect and highly dispersed into nanopores of ZnO in amorphous form, moreover, these nanopores strongly inhibited amorphous recrystallization in the condition of 45 degrees C and 75% RH. In addition, the results of the dissolution tested in vitro exhibited that the accumulated dissolutions of indomethacin and cilostazol solid dispersions achieved about 90% within 5 min and approximately 80% within 30 min. It was indicated in this study that the mechanism of drug dissolution improvement was associated with the effects of nanoporous ZnO carrier on increasing drug dispersion, controlling drug in nanopores as amorphous form and inhibiting amorphous recrystallization.
Anti-Inflammatory Agents, Non-Steroidal ; administration & dosage ; chemistry ; Calorimetry, Differential Scanning ; Drug Carriers ; Indomethacin ; administration & dosage ; chemistry ; Microscopy, Electron, Scanning ; Nanostructures ; Phosphodiesterase 3 Inhibitors ; administration & dosage ; chemistry ; Solubility ; Spectroscopy, Fourier Transform Infrared ; Tetrazoles ; administration & dosage ; chemistry ; X-Ray Diffraction ; Zinc Oxide ; chemistry

Our previous results have demonstrated that insulin reduces myocardial ischemia/reperfusion (MI/R) injury and increases the postischemic myocardial functions via activating the cellular survival signaling, i.e., phosphatidylinositol 3-kinase (PI3-K)-Akt-endothelial nitric oxide synthase (eNOS)-nitric oxide (NO) cascade. However, it remains largely controversial whether c-Jun NH2-terminal kinase (JNK) is involved in the effects of insulin on MI/R injury. Therefore, the aims of the present study were to investigate the role of JNK, especially the cross-talk between JNK and previously expatiated Akt signaling, in the protective effect of insulin on I/R myocardium. Isolated hearts from adult Sprague-Dawley rats were subjected to 30 min of regional ischemia and followed by 2 or 4 h of reperfusion (n=6). The hearts were pretreated with PI3-K inhibitor LY294002, or phosphorylated-JNK inhibitor SP600125, respectively, then perfused retrogradely with insulin, and the mechanical functions of hearts, including the heart rate (HR), left ventricular developed pressure (LVDP) and instantaneous first derivation of left ventricular pressure (+/-LVdp/dt(max)) were measured. At the end of reperfusion, the infarct size (IS) and apoptotic index (AI) were examined. MI/R caused significant cardiac dysfunction and myocardial apoptosis (strong TUNEL-positive staining). Compared with the control group, insulin treatment in MI/R rats exerted protective effects as evidenced by reduced myocardial IS [(28.9 +/- 2.0)% vs (45.0 +/- 4.0) %, n=6, P<0.01], inhibited cardiomyocyte apoptosis [decreased AI: (16.0 +/- 0.7) % vs (27.6 +/- 1.3) %, n=6, P<0.01] and improved recovery of cardiac systolic/diastolic function (including LVDP and +/-LVdp/dt(max)) at the end of reperfusion. Moreover, insulin resulted in 1.7-fold and 1.5-fold increases in Akt and JNK phosphorylation in I/R myocardium, respectively (n=6, P<0.05). Inhibition of Akt activation with LY294002 abolished, and inhibition of JNK activation with SP600125 enhanced the cardioprotection by insulin, respectively. And the abolishment by LY294002 could be partly converted by SP600125 pretreatment. In addition, SP600125 also decreased the Akt phosphorylation (n=6, P<0.05). These results demonstrate that insulin simultaneously activates both Akt and JNK, and the latter further increases the phosphorylation of Akt which attenuates MI/R injury and improves heart function; this cross-talk between Akt and JNK in the insulin signaling is involved in insulin-induced cardioprotective effect.
Animals ; Apoptosis ; Heart ; Insulin ; metabolism ; JNK Mitogen-Activated Protein Kinases ; MAP Kinase Signaling System ; Myocardial Infarction ; Myocardial Ischemia ; Myocardial Reperfusion Injury ; Myocardium ; Myocytes, Cardiac ; Nitric Oxide Synthase Type III ; Phosphatidylinositol 3-Kinase ; metabolism ; Phosphatidylinositol 3-Kinases ; metabolism ; Phosphorylation ; Proto-Oncogene Proteins c-akt ; Rats ; Rats, Sprague-Dawley ; Reperfusion Injury ; Signal Transduction

This study is to investigate the protective effect of longistyline A against corticosterone-induced neurotoxicity in PC12 cells. While PC12 cells were exposed to 100 micromol x L(-1) corticosterone for 48 h, cell survival rate was reduced and lactate dehydrogenase (LDH) release increased. In parallel, corticosterone caused significant elevations of DNA fragmentation, [Ca2+]i and caspase-3 activity. However, when the PC12 cells were incubated with longistyline A (4.0, 8.0 and 16.0 micromol x L(-1)) in the presence of 100 micromol x L(-1) corticosterone for 48 h, the effects were evidently alleviated, but dose-dependent manner was not obvious. In summary, longistyline A could generate a neuroprotective effect against corticosterone-induced neurotoxicity in PC12 cells possibly by decreasing [Ca2+]i and caspase-3 activity.
Animals ; Cajanus ; chemistry ; Calcium ; metabolism ; Caspase 3 ; metabolism ; Cell Survival ; drug effects ; Corticosterone ; toxicity ; DNA Fragmentation ; drug effects ; L-Lactate Dehydrogenase ; metabolism ; Molecular Structure ; Neuroprotective Agents ; isolation & purification ; pharmacology ; PC12 Cells ; Phenols ; isolation & purification ; pharmacology ; Plant Leaves ; chemistry ; Plants, Medicinal ; chemistry ; Rats

OBJECTIVETo detect the IgH-MMSET fusion gene resulted from t (4;14) translocation in multiple myeloma and illuminate its significance.

METHODSIgH-MMSET fusion gene was detected in bone marrow specimens of 25 multiple myeloma (MM) patients and MM cell line NCI-H929 using reverse-transcription PCR (RT-PCR) assay followed by nested PCR to increase the sensitivity. The purified PCR products were cloned into pGEM-T vector and then sequenced using M13 forward primers. The fragment sequences were compared with that in GenBank to find matched sequences.

RESULTSOnly a 438 base pair long fragment was obtained after RT-PCR assay and was confirmed by sequencing to be a fusion gene product of IgH gene and MMSET gene in MM cell line NCI-H929. The breakpoints were located within the C micro region of IgH gene on chromosome 14 and intron 3 of MMSET gene on chromosome 4. IgH-MMSET hybrid transcripts were detected in 3 of 25 MM patients through nested PCR assay. The amplified fragments of the 3 patients were 237 base pairs (bp), 239 bp and 239 bp in length, respectively. The breakpoints on chromosome 4 were identical to that of NCI-H929 cell.

CONCLUSIONSThe formation of IgH-MMSET fusion gene is resulted from t (4;14) translocation in MM. The incidence rate is 12.0%. The presence of IgH-MMSET fusion gene may predict poor prognosis.

Adult ; Aged ; Base Sequence ; Chromosomes, Human, Pair 14 ; Chromosomes, Human, Pair 4 ; Female ; Humans ; Male ; Middle Aged ; Molecular Sequence Data ; Multiple Myeloma ; genetics ; Oncogene Proteins, Fusion ; genetics ; Protein-Tyrosine Kinases ; Receptor, Fibroblast Growth Factor, Type 3 ; Receptors, Fibroblast Growth Factor ; genetics ; Reverse Transcriptase Polymerase Chain Reaction ; Translocation, Genetic

OBJECTIVETo investigate the influence of the vascular endothelial growth factor( VEGF) antibody targeted vascular therapy on the expression of human collagen type I in hyperplastic scar of nude mice.

METHODSThe hyperplastic scar from one female burn patient with 1% TBSA deep-partial thickness burns were implanted into subcutaneous skin of scapular region of 48 nude mice. Three weeks later, the nude mice were divide into large dose (LA) , medium dose (MD) , small dose (SD) and control groups, with 12 mice in each group. The mice in LA,MD and SD groups were injected with 200 microl of 15,10, 5 microg/ml VEGF monoclonal antibody diluted in 0.01 mol/L PBS, respectively in the scar twice a week for 3 weeks, while those in C group were injected with equal amount of 0. 01 mol/L PBS. The area and volume of the scar in each group were calculated and histological changes were observed, and the expression of collagen type I mRNA and its protein in each group were determined 3 days after treatment.

RESULTSThe volume of scar in LA, MD, SD and C groups were (55.3 +/-4.1, 67.9 +/-5.7, 78.9 +/-5.5, 85.0 +7.3) mm(3), respectively. Compared with that in C group, the volume of the scar were significantly decreased in AD and MD groups ( P <0.05). A few number of vessels and fibroblasts were observed in LD, MD groups, with decreased number of collagen fibers arranged in order. Compared with that in C group ,The expression of procollagen type I mRNA and its protein in C group was obviously higher than those in LD and MD groups ( P < 0. 05) , but it was similar to those in SD group.

CONCLUSIONVEGF targeted vascular therapy is beneficial for the inhibition of the angiopoietins of hyperplastic scar, the expression of collagen , and the growth of scar.

Adult ; Animals ; Antibodies, Monoclonal ; therapeutic use ; Cicatrix, Hypertrophic ; metabolism ; therapy ; Collagen Type I ; metabolism ; Disease Models, Animal ; Female ; Gene Expression ; Humans ; Hyperplasia ; metabolism ; therapy ; Mice ; Mice, Inbred BALB C ; Mice, Nude ; Neovascularization, Pathologic ; metabolism ; RNA, Messenger ; metabolism ; Vascular Endothelial Growth Factor A ; immunology

Adolescent ; Adult ; Aged ; Cardiomyopathy, Hypertrophic ; surgery ; Catheter Ablation ; methods ; Child ; Follow-Up Studies ; Heart Septum ; surgery ; Humans ; Middle Aged

OBJECTIVETo compare the safety and efficacy of myocardial contrast enhancement (MCE)-guided and angio-pressure (AP)-guided transcoronary ablation of septal hypertrophy (TASH) for patients with hypertrophic obstructive cardiomyopathy (HOCM).

METHODSTASH was performed under MCE-guide (n = 47, group I) or AP-guide (n = 25, group II) for drug-refractory patients with HOCM. Myocardial perfusion imaging (MPI) data as well as other clinical data were compared.

RESULTSTASH both under MCE-guide or AP-guide resulted in similar and significant reduction of left ventricular outflow tract gradient (PG) and associated with significant symptom improvement (all P < 0.001). Dosage of ethanol use, peak-level of CK-MB and ablated myocardial area and incidence of arrhythmia were also similar between the two groups.Similar left ventricular/atrial dimension changes post TASH were observed in the 2 groups during follow-up. However, the first selected septal vessels were changed under MCE in 6 patients.

CONCLUSIONSOur data demonstrated that the MCE-guided TASH was not superior to AP-guided TASH in safety and efficacy. However, MCE-guided TASH can avoid the misplace of ethanol to avoid innocent myocardial ablation.

Adult ; Cardiac Catheterization ; methods ; Cardiomyopathy, Hypertrophic ; diagnostic imaging ; therapy ; Catheter Ablation ; methods ; Female ; Humans ; Male ; Middle Aged ; Myocardial Perfusion Imaging ; Ultrasonography

OBJECTIVETo provide reliable evidence of "J in three-needle therapy" for treatment of stroke.

METHODSMulti-central randomized controlled trials were adopted, 180 hemiplegia patients of ischemic stroke were randomly divided into a fin three-needle group (90 cases) and a routine acupuncture group (90 cases). Two groups were both treated with basic neurology therapies, and J in three-needle group was treated with J in three-needle therapy, three acupoints of tempora, hand and foot etc. were selected; the routine acupuncture group was treated with traditional acupuncture, Quchi (LI 11), Huantiao (GB 30), Futu (ST 32) etc. were selected. Both groups were treated with acupuncture for 5 weeks. The cognitive function score of functional comprehensive assessment scale (FCA), the scores of mini-mental state examination scale (MMSE) and modified Barthel index (BI) were compared before and after treatment between two groups. Results After treatment, the scores of FCA, MMSE and BI in both groups were significantly improved compared to those before treatment (P < 0.01, P < 0.05); the improvement of FCA score, MMSE score and BI score in the J in three-needle group were superior to those of the routine acupuncture group after treatment (P < 0.01, P < 0.05). The total effective rate of 85.4% in the J in three-needle group was superior to tohat of 70.0% in the routine acupuncture group (P < 0.05).

CONCLUSIONJ in three-needle acupuncture treatment can obviously improve the cognitive function and activity ability of daily life of hemiplegia patients after stroke, and the therapeutic effect of J in three-needle therapy is superior to that of traditional acupuncture treatment.

Activities of Daily Living ; Acupuncture Therapy ; Adult ; Aged ; Cognition ; Female ; Hemiplegia ; etiology ; psychology ; rehabilitation ; therapy ; Humans ; Male ; Middle Aged ; Stroke ; complications

BACKGROUNDThe temporal bone has the most complicated anatomic feature among the whole human body, which always challenges otolaryngologists. This study was to study three-dimensional (3D) morphology of the temporal bone and the ear by means of a computer image processing technique, for the purpose of providing a 3D image to help in pathological, diagnostic and surgical procedures.

METHODSForty sets of temporal bone celloidin serial sections with reference points were prepared and the contours of selected structures and reference points were entered into a graphics programme. The technique of computer-aided 3D reconstruction was applied to obtain 3D images and parameters of the temporal bones and the ears. Stereo views of the ossicles (n = 5), the facial nerves (n = 11), the posterior tympanic sinuses (n = 11), the posterior ampullary nerves (n = 4), the endolymphatic ducts and sacs (n = 5), and the bony and membranous labyrinth (n = 1) were reconstructed.

RESULTSThree-dimensional images, including the cochlea, the ossicles, the nerves, the tendons and the endolymphatic fluid system in the temporal bone, were obtained. Stereo picture pairs and 3D parameters of spatial dimensions, angle and volume for these reconstructed structures were calculated. The arrangement of the ossicles, spatial relationship of the bony and membranous labyrinth, the whole course of the facial nerves, the endolymphatic sac and posterior tympanic cavity were clearly observable. Stereo picture pairs made the spatial relationships among the above-mentioned structures much clearer. The operation of the posterior ampullary nerve transection was designed and simulated on the graphic computer based on 3D anatomic investigations.

CONCLUSIONThe technique of computer-aided 3D reconstruction provides a new tool to observe the morphology of the temporal bone and thus may allow design and study of new surgical approaches.

Ear ; anatomy & histology ; Facial Nerve ; anatomy & histology ; Humans ; Image Processing, Computer-Assisted ; Imaging, Three-Dimensional ; Temporal Bone ; anatomy & histology