Amino Acids ; metabolism ; Animals ; Brain ; metabolism ; Electromagnetic Fields ; Electrophysiological Phenomena ; Glutamates ; metabolism ; Male ; Neurotransmitter Agents ; metabolism ; Pain ; physiopathology ; Pain Threshold ; Rats ; Rats, Sprague-Dawley ; gamma-Aminobutyric Acid ; metabolism

Animals ; Electromagnetic Fields ; Embryo, Mammalian ; metabolism ; Female ; Mice ; Pregnancy ; Proto-Oncogene Proteins c-fos ; biosynthesis ; genetics ; RNA, Messenger ; biosynthesis ; genetics ; Reverse Transcriptase Polymerase Chain Reaction

Objective To analyze the cognitive changes and influencing factors in patients with lacunar cerebral infarction after carotid artery intervention therapy. Methods Sixty lacunar cerebral infarction combined with carotid stenosis patients treated with artery intervention therapy (intervention therapy group) and 68 lacunar cerebral infarction without carotid stenosis patients treated with drug therapy (drug therapy group) were selected. The neuropsychological test was completed at entry and 1, 6, 12 months after entry, and the results were compared with 60 healthy controls (control group). The cognitive changes were observed. The neuropsychological test included mini mental state examination (MMSE), Montreal cognitive assessment scale (MoCA) and cognitive field test. Results There were statistical differences in other scores except the Stroop test C section and Wechsler adult intelligence scale (WAIS-RC) picture arrangement subtest at entry in intervention therapy group and drug therapy group compared with control group (P<0.05). There were no statistical differences in the all scores at entry between drug therapy group and intervention therapy group (P>0.05). In intervention therapy group, the MMSE scores, MoCA total score, Rey-Osterrieth complex figure test (ROCFT), auditory verb learning test (AVLT), and the WAIS-RC picture arrangement subtest, verbal fluency test, WAIS-RC digit span backwards subtest of performing function 12 months after entry were significantly better than those at entry, and there were statistical differences (P<0.05). MMSE score, MoCA total score, long-time delayed recall of ROCFT, the immediate recall, long-time delayed recall and short delayed recall of AVLT, semantic category fluency test of performing function and digit span backwards subtest of WAIS-RC 6 months after entry were significantly better than those at entry:(27.8 ± 2.2) scores vs. (26.4 ± 1.9) scores, (20.7 ± 2.3) scores vs. (19.3 ± 2.0) scores, (12.4 ± 3.2) scores vs. (10.8 ± 2.6) scores, (54.3 ± 10.6) scores vs. (49.9 ± 10.9) scores, (12.4 ± 2.0) scores vs. (11.2 ± 2.8) scores, (12.9 ± 2.0) scores vs. (10.6 ± 2.6) scores, (17.5 ± 4.0) scores vs. (15.4 ± 3.4) scores and (4.0 ± 0.9) scores vs. (3.5 ± 0.9) scores, and there were statistical differences (P<0.05). In drug therapy group, there were no statistical differences in the all scores 1 and 6 months after entry, compared with that at entry (P>0.05);the MMSE score, MoCA total score, ROCFT, the immediate recall, long-time delayed recall and short delayed recall of AVLT, WAIS-RC picture arrangement subtest, verbal fluency test, WAIS-RC digit span backwards subtest of performing function and digit span backwards subtest of WAIS-RC 12 months after entry were significantly better than those at entry, and there were statistical differences (P<0.05). There were no statistical differences in all scores 12 months after entry between intervention therapy group and drug therapy group (P>0.05). In patients intervention therapy group, Logistic regression analysis showed that the MoCA score was related with age, hypertension and low education level (P<0.01 or<0.05), but was not related with smoking, diabetes and interventional treatment (P>0.05). Conclusions Cognitive impairment in patients with lacunar cerebral infarction and carotid stenosis is severe and extensive, but most cognition disorders can improve to normal level 12 months after artery intervention therapy.

OBJECTIVE:To establish an RP-HPLC method for content determination of s-omeprazole sodium and its related substances.METHODS:The separation of s-omeprazole sodium and the related substances was carried out on a Phenomenex Luna C18 column,the mobile phase consisted of methanol-0.033 mol?L-1 ammonium dihydrogen phosphate-triethylamine (58∶41.8∶0.2,adjusted to pH 7.0 by phosphate acid).The detection wavelength was 302 nm,the flow rate was 1.0 mL?min-1,the column temperature was 25 ℃,and the sample size was 20 ?L.RESULTS:The linear range of omeprazole sodium was 10～500 mg?L-1 (r=0.999 7).The average recovery rate was 100.27% (RSD=0.74%).The average content of the related substances in samples was 0.42%.CONCLUSION:This method is simple,accurate,specific and applicable for content determination of s-omeprazole sodium and its related substances.

OBJECTIVE:To establishment the method for the determination of content of vitamin A palmitate(VAP) and its related substances by HPLC. METHODS:The HPLC conditions were consisted of Phenomenex Luna C18 column with a mobile phase of a mixture of acetonitrile-isopropanol (90∶10) ,the detection wavelength of 328 nm,the column temperature of 30 ℃ and the flow rate of 1.0 mL?min-1. RESULTS:VAP was completely separated from impurities,the linearity range was 90～400 mg?L-1(r=0.999 2). The average recovery rate was 99.60% (RSD=1.32%). The average content of the related substances were lower than 2.53% . CONCLUSION: This accurate and reliable HPLC method is applicable for the quality control of VAP.

OBJECTIVETo investigate the effect of the extremely low frequency pulsed electromagnetic field (PEMF) on the proliferation and differentiation of osteoblast-like cells.

METHODSThe MC3T3-E1 cell and the primary osteoblast cell derived from 2-day-old Sprague Dawley (SD) rat calvaria were exposed to PEMF with a magnetic flux density of 1.55 mT at 48 Hz for 24 or 48 h. MTS was applied to analyze cell proliferation and flow cytometry to detect cell cycle. The intracellular alkaline phosphatase (ALP) activity was measured by colorimetry.

RESULTSPEMF of 1.55 mT at 48 Hz decreased significantly the cell percentage of S or G(2)M phase (P < 0.05), but did not affect cell number of MC3T3-E1 cells. Although the number of the primary osteoblast cells did not alter by MTS assay after exposure to PEMF for 24 h continuously, the cell percentage of G(2)M phase increased significantly (P < 0.01). When the culture time extended to 48 h, the cell number increased greatly (P < 0.01) and the cell percentage of G(2)M phase decreased significantly despite of the exposure type (P < 0.01). After the primary osteoblast cells were exposed to PEMF for 24 h continuously, the ALP activity decreased significantly (P < 0.05), whereas it increased significantly after exposure to PEMF for 48 h continuously (P < 0.05).

CONCLUSIONPEMF of 1.55 mT at 48 Hz does not affect proliferation and differentiation of MC3T3-E1 cell, but it promotes proliferation of primary osteoblast cell, inhibits differentiation at proliferation stage and promotes differentiation at differentiation stage of primary osteoblast cell.

Animals ; Cell Differentiation ; radiation effects ; Cell Proliferation ; radiation effects ; Cells, Cultured ; Dose-Response Relationship, Radiation ; Electromagnetic Fields ; adverse effects ; Mice ; Osteoblasts ; cytology ; metabolism ; radiation effects ; Rats ; Rats, Sprague-Dawley

OBJECTIVETo study the effect of extremely low frequency magnetic fields on intracellular calcium concentration ([Ca(2+)]i).

METHODSFura-2 loaded HepG2 cells were exposed to 1.55 mT (average value), 16 Hz pulsed magnetic fields for 60 min and to 300 mT, 2 Hz rotating magnetic fields for 5 min, and then [Ca(2+)]i was measured by fluorescence spectrophotometer. [Ca(2+)]i of HepG2 cells was also measured when they were exposed to 0.9 mT [root mean square (rms)], 16 Hz sinusoidal magnetic fields in real time.

RESULTSThe R values (F(340) nm/F(380) nm) of the control and the exposed group were 2.4519 +/- 0.2378 and 2.5266 +/- 0.2915 respectively after HepG2 cells were exposed to 1.55 mT, 16 Hz magnetic fields, 1.365 0 +/- 0.0626 and 1.3602 +/- 0.0771 respectively to 300 mT, 2 Hz rotating magnetic fields. The ratios of the trendline slope [r((501 - 1,000)) / r((0 - 500))] from the data of R values were 1.1213 +/- 0.4559 and 1.0727 +/- 0.1971 respectively (P > 0.05), and the ratios of the intercept [b((501 - 1,000)) / b((0 - 500))] from the trendline were 0.9912 +/- 0.0098 and 0.9979 +/- 0.0060 (P > 0.05) when HepG2 cells were exposed to the 0.9 mT, 16 Hz sinusoidal magnetic fields.

CONCLUSIONThe effect of extremely low frequency magnetic fields on [Ca(2+)]i of HepG2 cells under the experimental condition has not been found.

Calcium ; metabolism ; Cell Line, Tumor ; drug effects ; metabolism ; radiation effects ; Chelating Agents ; pharmacology ; Egtazic Acid ; pharmacology ; Electromagnetic Fields ; Humans ; Ion Transport ; drug effects ; radiation effects ; Octoxynol ; pharmacology ; Spectrometry, Fluorescence ; Time Factors

OBJECTIVETo study the effects of low frequency pulsed magnetic field on the proliferation and differentiation of HepG2 cells.

METHODS3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium (MTS) colorimetry method and ELISA assay of alpha-fetoprotein (AFP) were used to determine the cell proliferation and differentiation after the cells were exposed to pulsed magnetic fields with different frequency but the same field intensity.

RESULTSThere were no significant differences in cell proliferation between sham and treated groups exposed to the field of 80 Hz, 1.55 mT for 1, 4, 8, 12, 24 h (P > 0.05). There were also no significant differences in cell proliferation and AFP secretion between sham and treated groups exposed to 16 Hz, 1.55 mT pulsed magnetic fields for 1, 4, 8, 24 h (P > 0.05).

CONCLUSIONThere were no "window effects" found in HepG2 cells proliferation or AFP secretion at 16 Hz and 80 Hz pulsed magnetic fields.

Cell Differentiation ; radiation effects ; Cell Division ; radiation effects ; Cell Line, Tumor ; cytology ; metabolism ; radiation effects ; Electromagnetic Fields ; Humans ; alpha-Fetoproteins ; analysis

Abstract:By using PVX derived vector pGR107, the effect of BYDV-MP nuclear localization signal on the movement of PVX was studied. BYDV-MP was cloned into pGR107 using GFP as an indicator. BYDV-MP was then shown to induce the systemic infection and exacerbate the symptom of PVX through infecting Nicotiana benthamiana. When the PVX gene encoding 25kD protein, which functioned as a systematic movemnet protein,was deleted and the above experiment was repeated, the result showed that BYDV-MP could compensate the systemic movement of PVX. A serial mutants with substitutions on the fifth, sixth and seventh amino acids of BYDV-MP nuclear localization signal was further constructed. It was found that the mutants at the fifth, sixth amino acids in BYDV-MP nuclear localization signal could only delay or weaken systemic movement of PVX whereas the mutant at seventh amino acid could entirely inhibit systemic movement of PVX.
Amino Acid Sequence ; Green Fluorescent Proteins ; genetics ; Luteovirus ; physiology ; Molecular Sequence Data ; Nuclear Localization Signals ; chemistry ; physiology ; Plant Viral Movement Proteins ; physiology ; Potexvirus ; genetics ; physiology

Objective To study the neurotoxicity of homocysteine (HCY) on routine hippocampal neuronal HT22 cells and investigate the mechanisms. Methods Differentiated HT22 cells were treated with different concentrations of HCY (0, 0.5, 1.0, 1.5, 2.0, and 2.5 mmol/L) in the presence or absence of NMDA antagonists MK801 and memantine. The cell viability was tested using MTS cytotoxicity assay. Hoechst 33342 and PI staining were used to observe the morphological changes of the cells. Results HCY induced concentration-dependent toxicity in HT22 cells with the 50% effective concentration (EC50>) of about 1.25 mmol/L. Administration of MKS01 and memantine significantly increased the cell viability, which reached the highest level after treatment with MKS01 and memantine at the concentration of 10 μmol/L (P<0.05). Hoechst 33342 and PI staining revealed obvious cell apoptosis at low HCY concentrations (1.0 mmol/L), while cell necrosis was obvious at a higher concentration (2.0 mmol/L). Conclusion HCY induced obvious concentration-dependent neurotoxicity in HT22 cells largely through the activation of NMDA receptors, and the interaction between HCY and the NMDA receptors may provides an important pathway for research of neuronal cell loss.