A total of 126 patients with type 2 diabetes mellitus were randomized into two groups:one received glimepiride 1 mg twice daily and the other 2 mg once daily.Fasing blood glucose(BG),BG 2 h after meals(breakfast,lunch and dinner)and HbA_(IC)were tested,△and standard deviation of the 4 point BG were calculated.It was found that two kinds of administration of glimepiride were equally effective in decreasing BG and once daily aministration could ease better the fluctuation of BG.

OBJECTIVETo investigate CD4+ CD25+ regulatory T cell frequencies in the peripheral blood of poor or non-responsiveness to Hepatitis B vaccine, and try to understand the relationship between CD4+ CD25+ regulatory T cell and poor or non-responsiveness to Hepatitis B vaccine.

METHODSFlow cytometric analysis was employed for CD4+ CD25+ regulatory T cell frequencies in the peripheral blood of 25 cases of non-responsiveness, 30 cases of poor-responsiveness, and collected 20 cases of responsiveness as control.

RESULTSCD4+ CD25+ regulatory T cell frequencies of responsiveness was (4.32 +/- 1.21)%, poor-responsiveness was (7.01 +/- 1.06)% and non-responsiveness was (12.75 +/- 2.01)%. It was found that non and poor-responsiveness showed a high percentage of CD4+ CD25+ regulatory T cell as compared with responsiveness (t = 8.426, t = 3.289, P<0.01).

CONCLUSIONThe poor and non-responsiveness should be related with the increase of CD4+ CD25+ regulatory T cell and this might be considered as an important cause of poor and non-responsiveness.

Adolescent ; Adult ; Blood ; immunology ; CD4 Antigens ; immunology ; Female ; Flow Cytometry ; Hepatitis B Vaccines ; immunology ; Humans ; Interleukin-2 Receptor alpha Subunit ; immunology ; Male ; Middle Aged ; T-Lymphocyte Subsets ; immunology ; T-Lymphocytes, Regulatory ; immunology ; metabolism ; Young Adult

Metabonomics was employed to investigate the effect of Angelica sinensis volatile oil (ASVO) to the endogenous metabolites of normal rats, and to reveal the possible ways of metabolism in rats caused by ASVO. The fifty male Waster rats were randomly divided into five groups (each consists of 10 rats), such as control group, high dose group of ASVO, middle dose group of ASVO, low dose group of ASVO, and Aspirin group. They were given 0.9% saline, 0.352 mL x kg(-1) ASVO, 0.176 mL x kg(-1) ASVO, 0.088 mL x kg(-1) ASVO and ASP respectively with the equal volume of 0.2 mL. Drugs and vehicle were given for 3 successive days. The urine was collected at 12, 24, 36, 48 h after modeling with metabolic cages. Rat urine metabolic fingerprint in different stages was analyzed using GC-MS, based on which the principal component analysis (PCA)and orthogonal partial least-squares discriminant analysis (OPLS-DA) models were established for metabonomic analysis. Potential biomarkers were screened by using variable importance in the projection (VIP) and T test. It was revealed that the middle dose of ASVO at 36 h induces a substantial change in rat urine. Compared with control group, seven kinds of endogenous metabolites in ASP group and ASVO group change significantly (P < 0.05), among which aconitic acid, succinic acid, citric acid, alpha-ketone glutaric acid, glycine and malic acid content had an upward trend (P < 0.05) and prostaglandin content had a downward trend (P < 0.01). The mechanism of ASVO and ASP have the similarity. It is likely that ASVO intervenes the metabolic process by affecting the energy, amino acid and lipid metabolism. Our work also indicates that rats administrated with ASVO can increase the energy metabolism of the body, induce the production of inflammatory substances and strengthen the body's immune ability. The result has also provide a proof for futher interpret ASVO pharmacological effects.
Angelica sinensis ; chemistry ; metabolism ; Animals ; Dose-Response Relationship, Drug ; Drugs, Chinese Herbal ; analysis ; metabolism ; pharmacology ; Energy Metabolism ; drug effects ; Lipid Metabolism ; drug effects ; Male ; Metabolomics ; Oils, Volatile ; analysis ; metabolism ; pharmacology ; Plant Oils ; analysis ; metabolism ; pharmacology ; Rats ; Rats, Wistar ; Urine ; chemistry

OBJECTIVETo investigate the developmental profiles on surface expression and co-localization of NMDA receptor clusters and AMPA receptor clusters on dendrite in cultured hippocampal neurons of rats.

METHODSGreen fluorescent protein tagged GluR2 subunit (GFP-GluR2) and FLAG tagged NR2B subunit (FLAG-NR2B) were transfected into cultured hippocampal neurons at 5 days in vitro (DIV5). FLAG-NR2B containing NMDA receptor clusters and GFP-GluR2 containing AMPA receptor clusters expressed on membrane surface were then labeled in living neurons using anti FLAG mAb/Cy3-conjugated anti-mouse antibody and anti-GFP pAb/Alex488-conjugated anti-rabbit antibody.

RESULTThe numbers of receptor cluster per 100 microm dendrite in the neurons at DIV7 and DIV19 were 39.7+/-5.0 and 64.7+/-6.1 (P<0.01) for NR2B-NMDAR, 59.1+/-3.3 and 99.7+/-6.4 (P<0.01) for GluR2-AMPAR, and 29.9+/-4.5 and 37.5+/-2.5(P<0.05) for the co-localized, respectively. At DIV7 and DIV19, 75.4% and 57.9% NR2B-NMDAR clusters were co-localized with GluR2-AMPAR; and 50.6% and 37.6% GluR2-AMPAR clusters were co-localized with NR2B-NMDAR, respectively.

CONCLUSIONThe density of NR2B-NMDAR containing and GluR2-AMPAR containing receptor clusters increases during development of hippocampal neurons in culture. Although the co-localized clusters are increased as well in an unit length of dendrite, the extent to which the two receptor clusters are co-localized decreases. These data imply a possible change in the partnership of AMPA receptor subtype and NMDA receptor subtype at newly formed synapses during development.

Animals ; Cells, Cultured ; Dendrites ; chemistry ; Hippocampus ; chemistry ; Rats ; Rats, Sprague-Dawley ; Receptors, AMPA ; analysis ; Receptors, N-Methyl-D-Aspartate ; analysis

Objective To observe the changes of serum S100B and MBP in rats with liquid petroleum gas poisoning. Methods Fifty-four healthy adult male SD rats were randomized into normal control group (n=6), 20% LPG poisoning group (n=24) and 50% LPG poisoning group (n=24). Rat models of liquid petroleum gas poisoning were established in the later 2 groups, and controls were given the same volume of air. The rats of each group(n=6) were scored with neurological severity scale (NSS) and the blood serum was collected on the 1st, 2nd, 3nd and 7th d of poisoning, respectively. The levels of S100B and MBP were detected by euzymelinked immunosorbent assay (ELISA).Results As compared with the scores of NSS in the normal control group, those on the 1st d of poisoning in the 20% LPG poisoning group and those on the 1st and 2nd d of poisoning in the 50% LPG poisoning group were significantly higher (P＜0.05). The levels of Sl00B and MBP in 20% and 50% LPG poisoning groups were higher than those in the control group on the 1st and 2nd d of poisoning (P＜0.05); the levels of S100B and MBP in 50% LPG poisoning group were higher than those in 20% LPG poisoning group on the 1st and 2nd d of poisoning (P＜0.05). The levels of S 100B and MBP in 50% LPG poisoning group were higher than those in 20% LPG poisoning group and normal control group on the 3rd of poisoning (P＜0.05). The NSS scores and the levels of S100B and MBP in rats with LPG poisoning enjoyed the highest scores or levels on the 1st d of poisoning and those decreased after that. Conclusion The levels of S100B and MBP of rats with LPG poisoning increase, indicating that gliocytes participate in the mechanism of nervous system injury in rats with liquid petroleum gas poisoning.

OBJECTIVETo study the molecular mechanism of antithrombin (AT) gene C2759T (Leu99Phe) mutation causing AT deficiency.

METHODSA mutated AT cDNA expression plasmid ATM2759 was constructed by mega-primer method. ATM2759 and wild type AT cDNA expression plasmid ATN were transfected into COS7 cells or CHO cells by using Superfect reagent respectively for in vitro expression study and immunofluorescence assay.

RESULTSThe antigen levels of AT (AT:Ag) in the cell lysate of ATM2759 transfected COS7 cells and the cell culture supernatant were 174.97% and 35.63% of that of ATN transfected COS7 cells respectively, whereas the AT activity in the cell culture supernatant was 47.73% of the control's. Immunofluorescence analysis showed that the fluorescence intensity was significantly higher in ATM2759 transfected CHO cells than in those transfected with ATN.

CONCLUSIONSLeu99Phe substitution may not affect the binding capacity of AT with heparin. Secretion defect and intracellular accumulation of the mutated AT protein might be the mechanisms of this mutation causing AT deficiency.

Animals ; Antithrombin III ; genetics ; metabolism ; Antithrombin III Deficiency ; genetics ; CHO Cells ; COS Cells ; Cercopithecus aethiops ; Cricetinae ; Cricetulus ; Fluorescent Antibody Technique ; Mutation ; Plasmids ; genetics ; Transfection

OBJECTIVETo investigate the distribution of -63A/C polymorphism of human glutathione S-transferase M3(GSTM3) gene in Chinese Han population and the association of -63A/C polymorphism with essential hypertension (EH).

METHODS-63A/C polymorphism of GSTM3 gene was examined by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) in 234 patients with EH and 328 healthy controls. The genotypes were confirmed partially by DNA sequencing.

RESULTSThe distribution of GSTM3 -63A/C was in Hardy-Weinberg equilibrium. The CC genotype frequency in EH group (6%) was significantly higher than that in control group(1.8%) (P < 0.05). After adjustment for age, body mass index, and other risk factors, the CC genotype was independently associated with hypertension (OR=3.447, 95%CI: 1.19-7.63; P=0.04).

CONCLUSIONThe GSTM3 -63A/C polymorphism was associated with EH in Chinese Han population, the C allele might be a risk factor for EH in Chinese Han nationality.

Adult ; Aged ; Asian Continental Ancestry Group ; genetics ; Base Sequence ; Case-Control Studies ; Ethnic Groups ; genetics ; Female ; Gene Frequency ; Genotype ; Glutathione Transferase ; genetics ; Humans ; Hypertension ; genetics ; pathology ; Isoenzymes ; genetics ; Male ; Middle Aged ; Polymorphism, Restriction Fragment Length ; Polymorphism, Single Nucleotide ; genetics ; Regression Analysis

Recent studies have shown that astrocytes play important roles in ATP degradation and adenosine (a well known analgesic molecule) generation, which are closely related to pain signaling pathway. The aim of this study was to investigate whether morphine, a well known analgesic drug, could affect the speeds of ATP enzymolysis and adenosine generation in rat astrocytes. Intracellular calcium concentration ([Ca(2+)](i)) of astrocyte was measured by flow cytometry, and the time points that morphine exerted notable effects were determined for subsequent experiments. Cultured astrocytes were pre-incubated with morphine (1 μmol/L) and then were incubated with substrates, ATP and AMP, for 30 min. The speeds of ATP enzymolysis and adenosine generation were measured by high performance liquid chromatography (HPLC). The results showed that both 1.5 and 48 h of morphine pre-incubation induced maximal ATP enzymolysis speed in astrocytes among all the time points, and there was no statistical difference of ATP enzymolysis speed between morphine treatments for 1.5 and 48 h. As to adenosine, morphine pre-incubation for 1.5 h statistically increased adenosine generation, which was degraded from AMP, in cultured astrocytes compared with control group. However, no difference of adenosine generation was observed after 48 h of morphine pre-incubation. These results indicate that treatment of morphine in vitro dynamically changes the concentrations of ATP and adenosine in extracellular milieu of astrocytic cells. In addition, astrocyte can be regarded as at least one of the target cells of morphine to induce changes of ATP and adenosine levels in central nervous system.
Adenosine ; biosynthesis ; Adenosine Triphosphate ; metabolism ; Analgesics, Opioid ; pharmacology ; Animals ; Animals, Newborn ; Astrocytes ; cytology ; drug effects ; metabolism ; Calcium ; analysis ; metabolism ; Cells, Cultured ; Cerebral Cortex ; cytology ; Morphine ; pharmacology ; Rats ; Rats, Sprague-Dawley

OBJECTIVETo assess the clinical value of ultrasound-guided vacuum-assisted Mammotome (MMT) system for surgical resection of benign breast disease.

METHODSThis retrospective study was conducted among 1267 patients who underwent minimally invasive surgery with ultrasound-guided MMT system for benign breast disease at our center between January, 2009 and January, 2014. The resection rate, incidence of complication, recurrence rate, patients' satisfaction, clinical follow-up results and risk factors were analyzed. The patients were followed up at 1 month, 6 months and every 6 months thereafter for up to 2 years with a median follow-up of 22 months.

RESULTSOf the total of 1267 patients, 1259 (99.36%) had complete resection of the breast lesions, and residual lesions were found in 8 cases 1 month after the operation. The resection rate was significantly associated with lesion size (P=0.003) but not with the patients'age, pathology, BI-RADS classification, or the number or location of the lesions (P>0.05). Eighty-nine (7.02%) patients showed postoperative complications, and hematoma occurred in 70 (5.52%) patients after the operation. The complication rate was significantly associated with the number and location of lesions (P=0.000) but not with age, pathology, BI-RADS classification or the lesion size (P>0.05). A total of 193 (15.23%) patients had recurrence after the operation, including 65 (5.13%) with in situ recurrence and 128 (10.1%) with new lesions. The recurrence rate was significantly associated with the number and size of lesions (P=0.000) but not with age, pathology, BI-RADS classification or location of lesions(P>0.05). Six patients were not satisfied with the appearance of the incision, and the overall satisfaction rate of the patients was 99.52%.

CONCLUSIONs Ultrasound-guided vacuum-assisted MMT excision is a safe and effective procedure for benign breast disease with a low surgical complication rate, a high resection rate and a low recurrence rate. This technique results in good postoperative appearance for treatment of benign and high-risk breast lesions, especially multiple benign breast lesions.

OBJECTIVETo identify the factor XI gene mutation in a Chinese pedigree of congenital factor XI deficiency.

METHODSThe peripheral blood samples were collected from the proband and her family members and the plasma FXI:C and FXI:Ag were assayed. All the exons and their adjacent intron sequences of factor XI were amplified with PCR and sequenced thereafter.

RESULTSTwo novel nonsense mutations TGG-->TGA (Trp228stop) and TGG-->TAG (Trp383stop) were identified in the family.

CONCLUSIONThe compound heterozygous Trp228stop and Trp383stop may attribute to the pathogenesis of the congenital factor deficiency.

Adult ; Asian Continental Ancestry Group ; Codon, Nonsense ; Factor XI ; genetics ; Factor XI Deficiency ; congenital ; genetics ; Female ; Humans ; Male ; Middle Aged ; Pedigree ; Polymerase Chain Reaction ; Sequence Analysis, DNA