OBJECTIVETo investigate the influence of fluorosis on nicotinic acetylcholine receptors (nAChRs) in protein and gene levels in SH-SY5Y cells and the mechanism of the receptor modification.

METHODSSH-SY5Y cells, a human neuroblastoma cell line, were incubated with different concentrations of fluoride or with antioxidant for 48 hours. The functions of cells were measured by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazoliumbromide (MTT) method, and protein oxidation detected by carbonyl content; the alpha3 and alpha7 nAChR subunits in protein level were measured by Western blotting and in mRNA level by RT-polymerase chain reaction (RT-PCR).

RESULTSIn high-dose group as compared to the control, the decreased MTT (49%), increased protein oxidation (72%), and lower expression of alpha3 (51%) and alpha7 (47%) nAChR subunit proteins were obviously observed in SH-SY5Y cells. There were no changes in expression of nAChR subunit mRNAs between the cells treated with fluoride and those un-treated in controls. Prior treatment with antioxidant resulted in preventing the decrease of nAChR protein in cells exposed to the high doses of fluoride.

CONCLUSIONFluorosis should result in damage of cells and the declined expression of nAChRs in protein levels, but no influences on gene expression of the receptors in human neuroblastoma neurons. The decreased nAChR proteins might be involved in the mechanism of oxidative stress induced by fluorosis.

Cell Line, Tumor ; Fluoride Poisoning ; metabolism ; Fluorides ; toxicity ; Humans ; Neuroblastoma ; metabolism ; pathology ; Protein Processing, Post-Translational ; drug effects ; Proteins ; metabolism ; RNA, Messenger ; biosynthesis ; genetics ; Receptors, Nicotinic ; biosynthesis ; genetics

OBJECTIVETo compare the painless effect of four anesthetic methods during opening pulp cavity and undergoing pulpectomy for acute or chronic pulpitis.

METHODS80 teeth of 80 patients were randomly allocated into four groups. Each group had 20 teeth. Anesthetic methods applied four different groups included block anesthesia of nerve, supraperiosteal infiltration, periodontal membrane injection and intrapulpal injection. Anesthesia doses were recorded and the pierced points, the zones of pain, the time of anesthesia action, the time of anesthesia persistence and the degrees of anesthesia were evaluated with four levels synthetic evaluation standard of anesthesia.

RESULTSCompared with periodontal membrane injection and intrapulpal injection, block anesthesia of nerve and supraperiosteal infiltration had the later time of anesthesia action and the longer time of anesthesia persistence (P<0.05). In four anesthetic methods, block anesthesia of nerve had the best painless effect (P<0.05).

CONCLUSIONFour anesthetic methods have their own superiorities, and we should select proper anesthetic methods in clinical work.

Adult ; Anesthesia, Dental ; Anesthetics ; Anesthetics, Local ; Bicuspid ; Female ; Humans ; Injections ; Lidocaine ; Male ; Mandibular Nerve ; Nerve Block ; Periodontal Ligament ; Pulpectomy ; Pulpitis

OBJECTIVEIn this report are reviewed two unrelated patients with typical normokalemic periodic paralysis (normoKPP) features and the results of screening the SCN4A gene for the disease-related mutation.

METHODSTwo sporadic cases with normoKPP were screened for previously known mutations in SCN4A gene (T704M, A1156T, M1360V, I1495F, M1592V) that lead to hyperKPP; denaturing high performance liquid chromatography (DHPLC) was used. Then the rest exons of SCN4A gene were screened by DHPLC, and sequence analysis was performed on those with DHPLC chromatogram variation when compared with unaffected control.

RESULTSTwo cases and one patient's father were detected with V781I, which was proved to be a singular missense mutation in SCN4A gene.

CONCLUSIONThe mutation V781I exists in Chinese patients with normoKPP and may be responsible for normoKPP.

Adult ; Amino Acid Sequence ; Base Sequence ; Child ; Chromatography, High Pressure Liquid ; methods ; DNA ; analysis ; genetics ; Exons ; Female ; Humans ; Male ; Molecular Sequence Data ; Mutation, Missense ; NAV1.4 Voltage-Gated Sodium Channel ; Paralyses, Familial Periodic ; genetics ; Point Mutation ; Sequence Analysis, DNA ; Sodium Channels ; genetics

OBJECTIVETo establish a new method for the determination of fangchinoline and tetrandrine in Stephania tetrandra and Fengtongan capsule by noanqueous capillary electrophoresis.

METHODSeparation was carried out in an uncoated fused capillary (50 cm x 75 microm i.d.) with a running buffer containing 50 mmol x L(-1) ammonium acetate, 1.0% acetic acid and 20% acetonitrile in methanol. A separation voltage of 20 kV and a UV detector wavelength at 214 nm were adopted. Sample was introduced from the anode.

RESULTThe calibration ranges were 1.00, 500 mg x L(-1) for both analytes. Under the optimum conditions, the relative standard deviation (RSD, n = 6) for the migration time of each analyte were 0.09%, 1.9% (intra-day) and 0.63%, 1.9% (inter-day); The RSD for the peak area of each analyte were 0.45%, 5.9% (intra-day) and 2.3%, 5.6% (inter-day), respectively. The contents of the analytes were determined easily with average recoveries 102% for fangchinoline and 105% for tetrandrine in S. tetrandra and 94.6% for fangchinoline and 98.7% for tetrandrine in Fengtongan capsules, respectively.

CONCLUSIONThe proposed method is simple, rapid, accurate and higher repeatable, and can be used to control of the quality of S. tetrandra and Fengtongan capsules.

Benzylisoquinolines ; analysis ; Calibration ; Capillary Electrochromatography ; methods ; Capsules ; Drugs, Chinese Herbal ; chemistry ; isolation & purification ; standards ; Plant Roots ; chemistry ; Plants, Medicinal ; chemistry ; Quality Control ; Reproducibility of Results ; Stephania tetrandra ; chemistry

OBJECTIVETo compare the difference between digital subtraction angiography (DSA) and type B ultrasonography in the evaluation of vascular injury in patients inflicted with high voltage electrical injury.

METHODSNineteen patients with high voltage electrical injury of upper limbs were enrolled in the study as burn group, and another 12 healthy volunteers as controls. The endovascular membrane, vascular wall thickness, intra-vascular blood flow and endovascular thrombosis formation of ulnar and radial arteries at wound site and in regions 5, 10 and 15 cm proximal to the wounds were examined by DSA and type B ultrasonography and compared with imagings of healthy volunteers as control. The injury degree of the ulnar and radial arteries was examined during operation for evaluation to corroborate with DSA and ultrasonography findings. Necrotic and/or thrombotic vessels were excised and sent for pathomorphological examination.

RESULTSBy DSA images abnormal signs as thrombosis, vascular lumen stenosis and blood flow deceleration were found in 14 ulnar and 11 radial arteries, and the signs were more pronounced in ulnar arteries. By type B ultrasonography, abnormal signs as roughing of tunica intima, swelling or exfoliation, thickening of vascular wall, lumen stenosis, decreased blood flow, even necrosis of vascular wall and thrombosis were identified in 19 ulnar and 16 radial arteries in burn group (P < 0.05 approximately 0.01). The blood flow in ulnar artery 5 cm to the approximal part of the wound edge was obvious lower than that of the control (31.60 +/- 13.90 ml/min vs 47.70 +/- 9.60 ml/min, P < 0.05).

CONCLUSIONType B ultrasonography and DSA could be helpful in the evaluation of vascular injury in patients inflicted with high voltage electrical injury.

Adult ; Angiography, Digital Subtraction ; methods ; Burns, Electric ; diagnostic imaging ; Female ; Humans ; Male ; Middle Aged ; Radial Artery ; diagnostic imaging ; injuries ; Ulnar Artery ; diagnostic imaging ; injuries ; Ultrasonography, Doppler, Color ; methods

OBJECTIVETo explore the application of ultrasonography in the diagnosis of deep electric injury.

METHODSHP-IPHX high resolution color and pulse doppler ultrasonography was employed in the study. The hemodynamic indices were determined in the burn wound area and tissues 5 - 15 cm proximal to the wound in 12 patients with deep electric injury. At the same time, injuries to subcutaneous and muscular tissue and blood vessels (fifty-six blood vessels detected) were detected.

RESULTS1. It was found by two-dimentional ultrasonography that the injury degree in different tissue after deep electric injury was different, i.e. blood vessels were most liable to injury followed by muscles and subcutaneous tissue. In the burn wound area, endothelium was not visualized in 7 blood vessels and endothelial swelling was identified in 12 blood vessels. Furthermore, vascular occlusion was found in 4 blood vessels and thrombosis found in 5 vessels. 2. It was also demonstrated by color ultrasonography that change in course of blood vessel and tortuesity were observed in 12 blood vessels, stenosis of lumen in 21 vessels and widened intravascular space in 11 vessels, All these findings were confirmed in the subsequent operations. 3. It was revealed by pulse Doppler that the top blood flow speed increased during vascular contraction period in narrowed blood vessels with decreased blood flow per minute.

CONCLUSIONBeing an non-invasive examination, ultrasonography could directly demonstrate the morphological changes in subcutaneous tissue, muscle and blood vessels after a deep electric injury, which might help determine the injury degree and the hemodynamic changes in the injured site.

Adolescent ; Adult ; Blood Vessels ; diagnostic imaging ; injuries ; Burns, Electric ; diagnosis ; diagnostic imaging ; physiopathology ; Female ; Hemodynamics ; Humans ; Male ; Middle Aged ; Muscles ; diagnostic imaging ; injuries ; Skin ; diagnostic imaging ; injuries ; Ultrasonography, Doppler ; methods

OBJECTIVETo study the protective impact of tea polyphenols (TP) on the injury of fibrinolytic functions induced by high-methionine dietary in rats.

METHODS50 male Wistar rats were divided by stratified based on body weight into 5 groups with 10 in each group: namely control group, model group, low-dose TP group, medium-dose TP group and high-dose TP group. The rats in model group and TP groups were fed with 3% methionine dietary, control group rats with routine diet. In addition, rats in low-dose, medium-dose and high-dose TP groups were treated with TP at 50, 100 and 200 mg/kg dosage respectively by gavages every day, control group and model group rats were given with same amount distilled water. The animals were sacrificed after 8 weeks. The levels of tissue-type plasminogen activator (t-PA) and type-1 plasminogen activator inhibitor (PAI-1) in plasma were determined by ELISA assays, mRNA levels of t-PA and PAI-1 in aortic arch were detected by RT-PCR, t-PA and PAI-1 expression in aortic arch were detected by immunohistochemistry strept-avidin-biotin complex (SABC).

RESULTSAfter experiment, the t-PA expression of aortic arch in control group, model group, low-dose TP group, medium-dose TP group and high-dose TP group were 133.03 ± 10.14, 95.46 ± 11.08, 111.97 ± 11.91, 130.23 ± 10.80, 139.39 ± 9.41 (F = 14.15, P < 0.01), respectively, and the PAI-1 expression were 90.91 ± 8.67, 166.76 ± 12.18, 139.63 ± 12.71, 134.66 ± 13.19, 109.49 ± 10.82 (F = 31.44, P < 0.01). The t-PA concentration of plasma were (10.69 ± 1.26), (6.13 ± 0.92), (8.56 ± 1.19), (9.69 ± 0.92), (11.97 ± 1.08) ng/ml, respectively (F = 41.98, P < 0.01), and the PAI-1 concentration of plasma were (6.31 ± 0.81), (16.98 ± 1.27), (11.39 ± 0.82), (8.46 ± 0.67), (8.08 ± 0.91) ng/ml, respectively (F = 207.74, P < 0.01). The mRNA levels of t-PA in aortic arch were 1.12 ± 0.02, 0.75 ± 0.14, 1.01 ± 0.09, 0.95 ± 0.08, 1.05 ± 0.13 (F = 5.77, P < 0.05), and the mRNA levels of PAI-1 in aortic arch were 1.25 ± 0.11, 1.74 ± 0.06, 1.23 ± 0.05, 1.09 ± 0.14, 1.23 ± 0.04 (F = 23.56, P < 0.01).

CONCLUSIONThe results indicate that TP seems to have regulatory function on transcription and protein levels of t-PA and PAI-1, in addition to maintaining the balance between PAI-1 and t-PA and healing the injury of fibrinolytic functions in rats induced by high-methionine dietary.

Animals ; Diet ; Fibrinolysis ; drug effects ; Male ; Methionine ; adverse effects ; Plasminogen Activator Inhibitor 1 ; blood ; Polyphenols ; pharmacology ; Rats ; Rats, Wistar ; Tea ; chemistry ; Tissue Plasminogen Activator ; blood

OBJECTIVEMutation screening was performed to a Chinese family with hypokalaemic periodic paraiysis(HOKPP) for locating the corresponding mutations of gene and for specifying the clinical features associated with mutations.

METHODSThe cilnical features of patients from HOKPP family were summurized. Techniques of target exon PCR and direct sequencing were used to screen the mutation in CACNA1S and SCN4A genes in all numbers of the family.

RESULTSTwo patients of the family showed the typical features of HOKPP: the age of disease onset is during the childhood, acetazolamide is effective to patients treated. A heterozygous point mutation 3716 (G>A) causing R1239H was found in exon 30 of CACNA1S gene of the patients, but not found in normal members of the family.

CONCLUSIONThe mutant R1239H in CACNA1S gene exists in Chinese patients with familial hypokalaemic periodic paralysis.

Adolescent ; Adult ; Base Sequence ; Calcium Channels ; genetics ; China ; DNA Mutational Analysis ; Family Health ; Female ; Humans ; Hypokalemic Periodic Paralysis ; genetics ; Male ; Mutation ; Pedigree ; Polymerase Chain Reaction

Acrylamide ; toxicity ; Animals ; Behavior, Animal ; drug effects ; Blotting, Western ; Cytoskeletal Proteins ; blood ; Dose-Response Relationship, Drug ; Electrophoresis, Polyacrylamide Gel ; Gait Ataxia ; blood ; chemically induced ; Male ; Motor Activity ; drug effects ; Neurotoxicity Syndromes ; blood ; etiology ; Rats ; Rats, Wistar

OBJECTIVETo clone the entire coding sequence and analyze the function of P2X7 receptor of J6-1 human leukemia cells.

METHODSThe entire coding sequence of P2X7 receptor was amplified by RT-PCR and then inserted into pTARGET plasmid to construct an eukaryotic expressing plasmid followed by DNA sequencing. HEK293 cells stably expressing P2X7 receptor were obtained after transfection and screening, and confirmed by RT-PCR and Western blotting. The bleb formation upon agonist stimulation was observed under phase contrast microscope.

RESULTSThe entire coding sequence of P2X7 receptor of J6-1 cells was successfully cloned. DNA sequencing analysis revealed a substitution of G559, for A559, causing a substitution of Glu187 for Gln187. The P2X7 receptor derived from J6-1 cells could be functionally expressed in HEK293 cells, in which bleb formation could be detected upon stimulation.

CONCLUSIONSThe entire coding sequence of P2X7 receptors was successfully cloned from J6-1 leukemia cells. Other unknown mechanism may contribute to the dysfunction of P2X7 receptor in these cells.

Cell Line, Tumor ; Cloning, Molecular ; DNA, Complementary ; genetics ; Gene Expression ; Humans ; Leukemia ; genetics ; metabolism ; Receptors, Purinergic P2 ; genetics ; physiology ; Receptors, Purinergic P2X7 ; Reverse Transcriptase Polymerase Chain Reaction ; Transfection