OBJECTIVE To establish a convenient method for detecting plasmid-mediated AmpC beta-lactamases in Enterobacteriaceae lacking chromosomal AmpC beta-lactamases and to investigqte their genotype.METHODS Fifteen isolates of Klebsiella pneumoniae were collected and the diameters of inhibitory zone of cefoxitin in combination with or without cloxacillin were measured separately by standard disc diffusion test.The M3D assay was used as control method.And the activity of AmpC beta-lactamases of all isolates was simultaneously assayed.Multiplex PCR was performed to determine the genotype of AmpC beta-lactamases.RESULTS Among 15 isolates,8 isolates were identified to have plasmid-mediated AmpC beta-lactamases.The sensitivity and specificity of AmpC beta-lactamases phenotypic confirmatory test were 100%.Eight of 15 isolates were identified to be DHA-1 beta-lactamases by multiplex PCR.CONCLUSIONS The new AmpC beta-lactamases phenotypic confirmatory test is a reliable method for detecting plasmidmediated AmpC beta-lactamases in Enterobacteriaceae lacking chromosomal AmpC beta-lactamases.DHA-1 beta-lactamases are the main genotypes of plasmid-mediated AmpC beta-lactamases in Hubei Province of China.

BACKGROUND: Parameters of platelet can reflect functions of coagulation. Granule membrane protein-140 (GMP-140), which is also called P-Selectin, in α-granules of platelets can reflect activities of platelet. Dysfunction of blood sugar and blood lipid in diabetic patients can result in complications such as thrombotic diseases.OBJECTIVE: To analyze the variation regulation of the platelet parameters and GMP-140 in patients with type 2 diabetes mellitus.DESIGN: A case-control study.SETTING:Department of Clinical Laboratory and Department of Endocrinology, Union Hospital Affiliated to Tongji Medical College, Huazhong University of Science and Technology.PARTICIPANTS: From March to April 2005, forty-five patients with type 2 diabetes mellitus, who were treated at the Department of Endocrinology,Union Hospital Affiliated to Tongji Medical College, Huazhong University of Science and Technology, were enrolled as diabetic group, and 45 healthy people, who were checked with health examination at the same period,were selected as control group. They all joined the experiment voluntarily. METHODS: Changes of quantity of blood platelet, mean platelet volume (MPV), platelet distribution width (PDW) and platelett-large cell ratio (PLCR) were analyzed with blood cell counter. GMP-140 was measured by enzyme-linked immunosorbent assay (ELISA), meanwhile, compared with those of the healthy people.MAIN OUTCOME MEASURES: Contents of the parameters of platelets and GMP-140 of subjects in the two group s.RESULTS: A total of 90 cases without drop out, 45 cases in the diabetic group and control group, respectively, were involved in the result analysis.①Comparison of the content of GMP-140 in the two groups: It was significantly higher in the diabetic group than that in the control group [(28.8±10.3) ,(9.1±5.6)μg/L(t=2.37,P ＜ 0.01 )]. ②Comparison of parameters of blood platelet of subjects of the two groups: The MPV, PDW and P-LCR in patients of the diabetic group were markedly higher than those in the control group (t=2.02, 2.02, 2.02, P ＜ 0.01 ), while the blood platelets (PLT)was dramatically lower than that in the control group (t=2.15, P ＜ 0.01 ).CONCLUSION: The MPV, PDW, P-LCR and the content of GMP-140 of patients with type 2 diabetes mellitus significantly increase, while the PLT decreases, which indicate that platelet activity enhances and the consumption of platelet increases in patients with type 2 diabetes mellitus.

With the great success of imatinib mesylate(IM) and other tyrosine kinase inhibitor (TKI),chronic myelogenous leukemia (CML) is taken as a successful model of targeted therapies for human cancer.Although the emergence of TKI has greatly improved the prognosis of CML,the long-term TKI treatment may bring the problems of drug resistance,serious side effects,poor compliance,and heavy economic burden.Recently,people focus on whether patients would be cured after discontinuation of TKI treatment.Here,the progresses on discontinuation of TKI therapy in CML presented in the 54th American Society of Hematology (ASH) annual meeting were reviewed.

Objective To summarize the key point of collaboration during the surgery of endoscopic guided vitrectomy for complicated ocular trauma.Methods From October 2010 to March 2013,a total of 25 cases(25 eyes) of complicated ocular trauma were treated by endoscopic guided vitrectomy.The the key point of collaboration was summarized.Results Surgeries of 25 cases (25 eyes) went smoothly,visual acuity improved,without enucleation cases.Postoperative complications were mainly secondary glaucoma and vitreous hemorrhage,and all were improved after treatment.Conclusions Endoscopic guided vitrectomy for the treatment of complicated ocular trauma is one of the effective means,and the advantage is that the surgery could be done when the refractive medium is opacity.

The changes in shape, structure and function of red blood cells (RBC) after cold storage were observed by using slow cold storage technique. Before and after cold storage, RBC mean corpuscular volume (MCV), RBC fragility (RF), whole blood viscosity (WBV), RBC deformity (RBCD), LFU, SatO2, hematocrit (HCT), RBC recovery rate (RBCRR) and RBC immune function were measured. The result showed that after resuscitation, RBC had vital resistance to hypotonic solution. There was no significant difference in MCV, RBCD and LFU (P>0.05). HCT and WBV were decreased and RBCRR reached 84.1 %(all P<0.01). SatO2 was increased from 30.5 % to 98.2 %. RBC immune function showed no significant difference (P>0.05). It was suggested that quality of RBC didn't change after cold storage and RBC could be cryopreserved for a long time.

Objective:To establish a method for the determination of total chloride in oral rehydration salts powder (Ⅲ) by ion chromatography (IC). Methods:The analysis was performed on an IonPac AS 14A (250 mm × 4 mm) column with an IonPac AG 14A (50 mm × 4 mm) guard column and 9mmol·L-1 Na2 CO3 solution as the mobile phase. The flow rate was 1. 0 ml·min-1 with isocratic elution and the sample size was 25 μl. The suppressor was AERS 500(4mm) with 45mA suppression current, and an ECD was used for the detection. The quantity of chloride ion ( Cl-1 ) was calculated by the peak area with an external standard method. Re-sults:The linear range of Cl-1 was 0. 05-20. 00 mg·L-1(r=0. 9999);the recovery was 98. 89％(RSD=0. 56％, n=6). Conclu-sion:The results demonstrated that the method has the advantages of high sensitivity, simple sample pretreatment, promising accuracy and good reliability, which is suitable for the determination of total chloride in oral rehydration salt powder (Ⅲ) .

In this study, we examined the effects of aspirin on the growth rates, subcellar distribution of beta-catenin protein, the expression of beta-catenin/TCF signaling pathway target gene cyclinD1 mRNA, and cell cycle of Jurkat cell line (Human T-acute lymphoblastic leukemia). Our results showed that the treatment with aspirin inhibited the growth of Jurkat cell line. Jurkat cells treated with 3 mmol/L of aspirin could significantly decrease nuclear localization of beta-catenin, and at 5 mmol/L of aspirin, the nuclear localization of beta-catenin was undetectable. QRT-PCR showed that the target gene cyclinD1 mRNA expression was gradually decreased with the dosage of aspirin. Aspirin induced G0/G1 cell cycle arrest in Jurkat cells. We are led to conclude that aspirin acts through beta-catenin-independent mechanisms. The effects of aspirin include down-regulation of beta-catenin nuclear localization and G0/G1 cell cycle arrest, which might serve as a means of growth inhibition in aspirin-treated human Jurkat cell line.

To investigate the effect of autoimmune regulator (AIRE) on phagocytic clearance of apoptotic cells, a recombinant expression vector containing full-length human AIRE cDNA was transfected into 16HBE cells. After incubation with transfected 16HBE cells, engulfment of apoptotic HL-60 cells induced by camptothecin was detected by myeloperoxidase (MPO) staining. The change in the expression of Rac 1 in transfected 16HBE cells was determined by RT-PCR and Western blotting. The results showed that the phagocytosis percentage of the experimental group, the mock transfection group and the negative control group (non-apoptotic cells) was (25.50+/-3.67)%, (6.25+/-1.58)% and (1.0+/-0.67)%, respectively. Moreover, the expressions of Rac 1 mRNA and protein were up-regulated in AIRE-transfected 16HBE cells, suggesting that AIRE may function as a regulator in the phagocytic clearance of apoptotic cells by promoting the expression of Rac 1.

Objective:To set up a real-time fluorescent RT-PCR and to quantitate the expression levels of hTERT mRNA in peripheral blood mononuclear cells from patients with AML and to observe the correlation between the expression of hTERT mRNA and occurrence and relapse of AML and to probe into the correlation between the expression of hTERT mRNA and telomerase activity.Methods:Real time fluorescent RT-PCR and Lightcycler PCR system were used to quantitate expression levels of hTERT mRNA.PCR-ELISA was used to quantitate telomerase activity.Results:①N_(hTERT) from AML at initial presention,AML at relapse,AML at complete remission and health examinee was 299.2?292.8,550.1?441.3,14.0?9.2 and 12.3?6.7 respectively and the expression levels of hTERT mRNA from AML at initial presention and AML at relapse elevated signficantly comparing to that from AML at complete remission and health examinee,moreover the expression levels from AML at relapse was higher than that from AML at initial presention significantly.②The telomerase activity from AML at initial presention,AML at relapse,AML at complete remission and health examinee was 32.8%?24.3%,48.6%?31.4%,7.4%?5.1% and 7.6%?3.6% respectively and the telomerase activity from AML at initial presention and AML at relapse elevated signficantly comparing to that from AML at complete remission and health examinee,moreover telomerase activity from AML at relapse was higher than that from AML at initial presention significantly.③There was a strong correlation between telomerase activity and the expression of hTERT mRNA and the correlation coeffecients was 0.78.Conclusion:The up-regulation of the expression levels of hTERT mRNA and telomerase activity are one of important factors during occurrence and relapse of AML,moreover there is a strong correlation between telomerase activity and the expression of hTERT mRNA.

0.05) but quite different from European and African's(P