Objective To detect the expression of growth factor independence 1 (GFI-1) in peripheral blood of patients with Sézary syndrome and normal persons, so as to provide a theoretical basis for developing GFI-1 gene target therapy. Methods CD4+ CD7- Sézary cells (SS cells) were separated and purified from peripheral blood of 7 patients with Sézary syndrome by flow cytometry, CD4+ T cells from peripheral blood of 10 normal persons, Sézary syndrome-derived cell line Hut78 and human acute T cell leukemia cell line Jurkat as controls. The mRNA and protein expressions of GF-1were detected by qPCR and Western blotting, respectively. Then after interferon-a-2b (IFN-a2b) was used to induce Hut78 cell apoptosis, the cell proliferation wasmeasured by MTS, themRNA expression of GFI-1, cell cycle-dependent protein kinase inhibitor P21, tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) and Caspasi-3 was detected by qPCR, and the cell apoptosis was detected by flow cytometry. Results The expression of GFI-1 mRNA in the SS cells was significantly higher than that in the Jurkat and CE4+ T cells (all P<0.05). The expression of GF-1 protein in the SS cells and Hut78 cells was significantly higher than that in the Jurkat and CD4+ T cells (all P<0.05). IFN-a2b significantly mhibited the proliferation of Hut78 cells, and the effect was concentration-dependent and time-dependent The mRNA expression of GFI-1 ln Hut78 cells was significantly decreased in a time-dependent manner at 12 h and 24 h treated with IFN-a2b, while the mRNA expressions of P21, TRAIL and Caspase-3 were significantly increased (P〈0. 05). The apoptosis of Hut78 cells was significantly increased at 12 h and 24 h treated with IFN-α2b (P〈0. 05). Conclusion The expression of GFI-1 gene in peripheral blood SS cells of patients with Sézary syndrome is increased and can be inhibited by IFN-a2b, indicating that GFI-1 gene may play an important regulatory role in tumor proliferation of SS cells in patients with Sézary syndrome.

Objective To investigate the effect of calcium antagonist verapamil on the function of rat?- cells and tolbutamide (D860)-induced insulin release.Methods Insulin released from isolated islets were measured in control,verapamil,D860,and verapamil+D860 groups.Furthermore,intravenous glucose tolerance test (IVGTT) was conducted in acute experiments treated with verapamil and D860 respectively to assess?-cell function in rats with the same allocation as in vitro.Another IVGTT was performed in the end of 4 weeks' treatment.The insulin contents in pancreas were assayed and pancreas islets morphology were observed with immunohistochemistry.Results Verapamil could inhibit insulin release from isolated islets.Verapamil group was [(1.244?0.082)ng?ml~(-1)?islet~(-1)]and control group (2.623?0.226) ng?ml~(-1)?islet~(-1)(P0.05).Also,similar results were obtained in normal rats during acute experiments and verapamil reduce the hypoglycemic effect promoted by D860. However,above results were not observed in the end of 4 weeks experiments,and no difference for insulin content and morphological change in islets was found among four groups.Conclusion Treatment of verapamil chronically does not impair islet function and interfere with the hypoglycemic effect of D860 in rats .

Arthroplasty, Replacement, Knee ; methods ; Bone Transplantation ; Bone Wires ; Female ; Humans ; Middle Aged ; Tibia ; surgery ; Transplantation, Homologous

AIMTo observe the effects of Ca2+ -activated K+ channel of primary cultured fetal SD rat cortex neurons in the veratridine triggered neuronal damage.

METHODSThe patch clamp technique of cell-attach and inside-out mode for these two kinds of single channel recordings were used.

RESULTSExtracellular veratridine activated the Kca. In Ca2+ bath solution of cell-attach mode, Vp + 30 mV, when the concentration (micromol/L) of veratridine were 15,25,50 and 75, the open probabilities of the channel were 0.014 +/- 0.003, 0.085 +/- 0.010, 0.132 +/- 0.016 and 0.059 +/- 0.006 (P < 0.01) respectively. It appeared concentration-dependent within 50 micromol/L veratridine. In Ca2+ free bath solution of cell-attach mode, Vp = +50 mV, when the concentration (micromol/L) of veratridine were 15, 40,60 and 100, the open probabilities of the channel were 0.014 +/- 0.010, 0.113 +/- 0.006, 0.141 +/- 0.004 and 0.295 +/- 0.009 (P < 0.05) respectively. In the 6 cases of inside-out mode patch clamp, Vp = +40 mV, when the concentration of veratridine were 0, 25 micromol/L and 50 micromol/L, the open probabilities of the channel were 0.011 +/- 0.008, 0.010 +/- 0.010 and 0.012 +/- 0.007 (P > 0.05) respectively. There were no significant difference on open probabilities, average open/close times and amplitudes at different intracellular veratridine concentration.

CONCLUSIONVeratridine can affect the activation of the Kca channel through regulating the concentration of cytoplasmic free Ca2+. The opening of Kca activated by increase of intracellular Ca2+ during the early stage of anoxia may be a protection reaction of ischemic neurons.

Animals ; Animals, Newborn ; Calcium ; metabolism ; Cells, Cultured ; Neurons ; cytology ; drug effects ; physiology ; Patch-Clamp Techniques ; Potassium Channels, Calcium-Activated ; metabolism ; Rats ; Rats, Sprague-Dawley ; Veratridine ; pharmacology

A rapid, sensitive and simple liquid chromatography-tandem mass spectrometry (LC-MS/MS) method was developed and validated for the simultaneous determination of clevidipine butyrate and its primary metabolite clevidipine acid in dog blood. After one-step protein precipitation with methanol, the chromatographic separation was carried out on an Ecosil C18 column (150 mm x 4.6 mm, 5 µm) with a gradient mobile phase consisting of methanol and 5 mmol · L(-1) ammonium formate. A chromatographic total run time of 13.0 min was achieved. The quantitation analysis was performed using multiple reaction monitoring (MRM) at the specific ion transitions of m/z 454.1 [M-H]- --> m/z 234.1 for clevidipine butyrate, m/z 354.0 [M-H]- --> m/z 208.0 for clevidipine acid and m/z 256.1 [M-H]- --> m/z 227.1 for elofesalamide (internal standard, IS) in the negative ion mode with electrospray ionization (ESI) source. The linear calibration curves for clevidipine butyrate and clevidipine acid were obtained in the concentration ranges of 0.5-100 ng · mL and 1-200 ng · mL(-1), separately. The lower limit of quantification of clevidipine butyrate and clevidipine acid were 0.5 ng · mL(-1) and 1 ng · mL(-1). The intra and inter-assay precisions were all below 12.9%, the accuracies were all in standard ranges. Stability testing indicated that clevidipine butyrate and clevidipine acid in dog blood with the addition of denaturant methanol was stable under various processing and/or handling conditions. The validated method has been successfully applied to a pharmacokinetic study of clevidipine butyrate injection to 8 healthy Beagle dogs following intravenous infusion at a flow rate of 5 mg · h(-1) for 0.5 h.
Animals ; Butyrates ; blood ; pharmacokinetics ; Calibration ; Chromatography, Liquid ; Dogs ; Infusions, Intravenous ; Pyridines ; blood ; pharmacokinetics ; Tandem Mass Spectrometry

OBJECTIVETo explore the mechanism of Chinese medicinal therapy for cooling blood and removing stasis (CBRS) in treating psoriasis of blood-heat syndrome type by observing its clinical efficacy and impact on serum level of cytokines.

METHODSChanges of psoriasis area and severity index (PASI) scores, scores of syndrome and serum levels of tumor necrosis factor alpha (TNF-alpha, Th1 cytokine) and interleukin-10 (IL-10, Th2 cytokine) in 32 psoriatic patients before and after treatment were observed.

RESULTSThe total effective rate after 4 weeks of treatment and 8 weeks of treatment was 65.63% (21/32 cases) and 75.00% (24/32 cases) respectively. Scores of PASI and syndrome all improved after treatment (P < 0.05). And comparison of TNF-alpha level before and after treatment also showed significant difference (P < 0.05) , whereas that of IL-10 was insignificant (P > 0.05). Moreover, a positive correlation was found between PASI scores and secreting level of TNF-alpha in patients with psoriasis (r = 0.3709, P < 0.05).

CONCLUSIONChinese therapy for CBRS could play its effect in treating psoriasis by way of reducing TNF-alpha level.

Adolescent ; Adult ; Aged ; Case-Control Studies ; Cytokines ; blood ; Drugs, Chinese Herbal ; therapeutic use ; Female ; Humans ; Male ; Middle Aged ; Psoriasis ; blood ; drug therapy ; immunology ; pathology ; Th1 Cells ; immunology ; Th2 Cells ; immunology ; Young Adult

Objective To investigate the clinical and genetic characteristics of a case with non-classical 21-hydroxylase deficiency(210HD).Methods Clinical features and laboratory data were obtained from a patient with non-c]assical 21OHD,and the promoter and coding areas of CYP21 gene were sequenced.Results The old female patient presented with hypertension.The laboratory examinations showed that plasma androstenedione, testosterone,progesterone and 17-hydroxy progesterone(17OHP)were increased.CT scan revealed bilateral adrenal nodular enlargement.Furthermore,rapid ACTH stimulation test showed that the plasma 17OHP concentration was further increased up to 68.3?g/L.Sequencing analysis showed a C1187T(R356W) substitution at exon 8 and the C-125T,G-112A,T-109C variations in the promoter of CYP21 gene,which was not previous reported.Conclusion The combined heterozygous mutations,Cl187T at exon 8 and C-125T,G -112A,T-109C in promoter,seem to be associated with non-classical 21OHD phenotype.

The study aims to explore the effects and mechanisms of water extract of Potentilla anserina (PA) on myelosuppression mice induced by cyclophosphamide based on metabonomics. The myelosuppressive mouse model was established by injected with cyclophosphamide and treated with water extract of PA. Thymus and spleen indexes, peripheral hemogram and bone marrow nucleated cells of each group was detected. Bone marrow pathology analysis was performed by hematoxylin-eosin staining. The levels of interleukin 3 (IL-3), interleukin 6 (IL-6), erythropoietin (EPO), granulocyte colony stimulating factor (GM-CSF), malondialdehyde (MDA), superoxide dismutase (SOD) and catalase (CAT) in serum were measured. The changes of biomarkers and related metabolic pathways were analyzed by UPLC-Q-TOF/MS-based metabonomics. Animal experiments were approved by the Animal Ethics Committee of Southwest Minzu University. The high doses of PA could significantly improve the decrease of white blood cell (WBC), red blood cell (RBC) counts and hemoglobin (HGB) levels of mice induced by cyclophosphamide (P < 0.05), and significantly increase the number of nucleated cells and the area of hematopoietic tissue in femoral bone marrow. The medium and high doses of PA could significantly improve the serum levels of SOD, CAT, MDA, IL-6 and GM-CSF (P < 0.05), and have no significant effect on the expression of IL-3 and EPO (P > 0.05). Serum metabolomics analysis showed that the aqueous extracts of PA could alleviate myrosuppression by regulating the aminoacyl-tRNA, valine, leucine and isoleucine biosynthesis mediated by 13 different metabolites such as valine, leucine, asparagine and hydroxyisohexic acid. PA improve the inhibition of hematopoietic function in myelosuppression mouse, and its mechanisms may be related to anti-oxidation and promoting the expression of hematopoietic-related cytokines and regulating the related metabolic pathways.

OBJECTIVETo investigate stress distribution of different material restored post-cores in dentine and provide a theoretical guidance for clinical use.

METHODSA three-dimensional finite element model of maxillary central incisor restored with post-core and PFM crown was constructed by SCT scan technology. Based on this model, stress distribution in dentine was analyzed before and after post-core restorations with 6 different materials, including cast Ni-Cr alloy, cast titanium alloy, cast gold alloy, glass fiber reinforced composite, polythene fiber reinforced composite, and common composite resin.

RESULTSWhen the tooth was restored with cast Ni-Cr alloy post and PFM crown, the maximum tensile stress and Von Mises stress in dentin at post apex increased 152% and 162% respectively, compared with a tooth restored only with PFM crown. If polythene fiber reinforced composite was used as post material, the stress distribution did not significantly change. When the other materials were used for the post, the stress distribution changed greatly. The elastic modulus of post-core materials affected the stress distribution pattern in dentine.

CONCLUSIONThe materials with elastic modulus similar to that of dentin, such as polythene fiber reinforced composite, may be suitable for post restoration.

Chromium Alloys ; Dental Materials ; Dental Restoration, Permanent ; Dental Stress Analysis ; methods ; Dentin ; Finite Element Analysis ; Humans ; Incisor ; physiology ; Materials Testing ; Post and Core Technique ; Tensile Strength