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.

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.

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.

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.

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.

T cell immunoglobulin mucin (TIM) family plays a key role in regulating immune responses. In this study, the interactions of human TIM family with apoptotic cells were evaluated in order to provide a foundation for further study on the roles of human TIM genes in apoptosis. Nine kinds of pEGFP-N1 eukaryotic expression vectors containing different lengths of the three members of human TIM genes for the expression of TIM-EGFP and the vectors for the expression of TIM-Fc fusion proteins were constructed. It was found that human TIM proteins could recognize and bind to apoptotic cells directly, but not to viable cells. The interactions of sTIM-1-EGFP, sTIM-3-EGFP and sTIM-4-EGFP with apoptotic cells were blocked by TIM-1-Ig, TIM-3-Ig and TIM-4-Ig fusion proteins respectively. In addition, human TIM proteins mediated the recognition of apoptotic cells and bound to apoptotic cells directly via the IgV domains. In conclusion, the TIM family may play a key role in the regulation of apoptosis. Our data also suggest that human TIM proteins probably serve as novel proteins for the detection of the early cellular apoptosis.

Objective To study the significance of anti-cyclic citrullinated peptide (anti-CCP),anti keratin antibody (AKA) and anti-RA33 autoantibodies (RA33)in the diagnosis of elderly-onset rheumatoid arthritis (EORA) and to explore the role of the united detection of anti-CCP, AKA and RA33 in the diagnosis of EORA. Methods The above three kinds of autoantibodies were detected in 69 patients with EORA, 73 patients with polymyalgia rheumatica (PMR) and 65 patients with osteoarthritis (OA). The anti-CCP and RA33 were measured by enzyme-linked immunosorbent assay (ELISA),and AKA was measured by indirect immunofluorescence (IIF). Results In the group of patients with EORA, the sensitivity and specificity of anti-CCP were 55.1% and 94.3%, the sensitivity and specificity of AKA were 31.3% and 91.5%, and the sensitivity and specificity of RA33 were 36.2% and 95.4% respectively, which were significantly higher than those in the groups of patients with PMR and OA. United-detection of anti-CCP, AKA and RA33 reduced the sensitivity,hut increased the specificity (100%) with higher positive predictive value. Conclusions Anti-CCP,AKA and anti-RA33 can emerge in the patients with EORA and the anti-CCP displays higher sensitivity and specificity. United detection of anti-CCP, AKA and RA33 can increase the specificity and positive predictive value (100%) and has high value in the diagnosis of EORA.

ObjectiveTo test the antifungal activity of berberine (BBR) against Candida albicans in vitro. MethodsAccording to the National Committee for Clinical Laboratory Standards (NCCLS) M27-A protocol, broth microdilution test was performed to assess the susceptibility of 30 clinical isolates of Candida albicans to BBR and fluconazole alone or in combination. ResultsThe geometric mean minimum inhibitory concentration (MIC) was 248.18 mg/L and 2.64 mg/L for BBR and fluconazole alone, respectively. When combined with BBR of 320, 160 and 80 mg/L, the geometric mean MIC decreased to 0.03, 0.35 and 1.35 mg/L respectively for fluconazole. Fluconazole and BBR showed synergistic effects in 26.7％ of these isolates including 3 fluconazoleresistant strains, additive effects in 56.7％, indifferent effects in 16.7％, antagonistic effects in none. Conclu sionIn vitro, BBR has antifungal activity against Candida albicans and can enhance the activity of fluconazole against Candida albicans.

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) .