Insulin autoimmune syndrome (IAS) is rare.A patient with IAS was herewith reported and 50domestic cases reported in the last two decades were reviewed to discuss the approach to this syndrome.In hypoglycemic patients with high insulin level and with Graves' disease and taking tapazole,insulin autoantibody detection is mandatory to avoid misdiagnosis and unnecessary surgery.

Objective To investigate the changes of nuclear factor?B activity in peripheral blood mononuclear cell(PMNC)in newly diagnosed type 2 diabetic patients.Methods Thirty normal individuals and 30 newly diagnosed type 2 diabetics were selected.Phosphorylation status of NF-?B P65 of PMNC was determined by immunoblotting with phosphor-specific antibodies to NF-?B P65(Ser 536 ).Serum hs-CRP was assayed by enzyme linked immunosorbent assay(ELISA).Results The level of NF-?B P65 phosphorylation was increased in newly diagnosed type 2 diabetic patients compared with control subjects[0.85(0.49,1.02)vs 0.47(0.26,0.67),P

Objective: To explore the psychological status and therapeutic efficacy of peri-operative psychological intervention among patients with oral carcinoma.Method: psychological statuses of 60 patients with oral carcinoma were investigated,and specific peri-operative psychological interventions were performed according to features and initiating factors of their psychological crisis,in order to provide references for the therapy of oral carcinoma.Result: Common psychological crises among patients with oral carcinoma such as anxiety and fear have been dramatically decreased by proper psychological investigation and peri-operative psychological intervention.Conclusion: Psychological investigation contributes to a great improvement of psychological status of patients with oral carcinoma.Therefore,in addition to routine peri-operative medical treatment,an effective psychological investigation is also crucial for patients with oral carcinoma to cure the disease and regain health.

Objective:To explore the function of risk management in outpatient blood collection work under the conditions of informationization. Method:This paper retrospectively reviewed the nursing risk management in out-patient blood collection work from January 2014 to January 2015 in our hospital, analyzed the causes of risk in out-patient blood collection work, evaluated the possible adverse outcomes, and put forward the measures to prevent and control risks. Results:Through the nursing risk management, the nurses′ risk prevention consciousness was enhanced, as well, both nurse and patient satisfaction was improved. Conclusion:Application of risk management in outpatient blood collection work could improve the quality of nursing, conform to the ethical requirements of guaranteeing patient safety, and effectively reduce the incidence of medical risks and accident.

Objective To design and apply ventilator-associated pneumonia prevention checklist in ICU.Methods The ventilator-associated pneumonia prevention checklist was designed by referring to guidelines and related literature.The checklist was applied to ventilator-dependent patients.The head of nursing group evaluated prevention status of ventilator-associated pneumonia performed by nurses every day.Results After application of the checklist,the average indwelling time of tracheal catheter and the incidence of ventilator-associated pneumonia decreased among ICU patients.Conclusion Using the ventilator-associated pneumonia prevention checklist in quality management in the prevention of ventilator-associated pneumonia can shorten the average indwelling time of tracheal catheter,and reduce the incidence of ventilator-associated pneumonia,and the checklist can be promoted and applied.

Objective To investigate the effect of rh-resistin on lipid metabolism in HepG2 cells and to elucidate its relation to AMPK pathway. Methods We treated the HepG2 cells with 50 ng/ml rh-resistin and 0.5 mmol/L palmitic acid,used siRNA technique to inhibite α2 subunite expression of AMPK in HepG2 cells and quantitative RT-PCR to detect ACC1,ACC2,and HL mRNA expression levels of related lipid metabolism genes. The P-AMPK-Thr172 of AMPK and P-ACC-Ser79 of ACC were determined by Western blotting. The Lipid accumu-lation in cells was determined by images of Laser Scanning Confocal Microscope after Nile red staining. Results Rh-resistin decreased the AMPKα2,HL mRNA expressions and the phosphorylation level of AMPK and ACC in both basal and insulin-stimulated conditions (P < 0.05),had no influence on ACC2 mRNA expressions (P >0.05),while it increased ACC2 mRNA expressions and cytoplasmic lipid droplets in the same conditions (P <0.05). Conclusion Rh-resistin may affect lipid metabolism via AMPK pathway with increase of fatty acid synthe-sis and inhibition of triglyceride catabolism,which leading to lipid accumulation in HepG2 cells.

Objective To investigate the effect of rh-resistin on glucose metabolism in HepG2 cells and to elucidate whether the underlying mechanisms are related to AMPK pathway. Methods Cells transfected with control siRNA or AMPKα2 siRNA were cultured in 6-well plates and then treated with 50 ng/mL rh-resistin for 24 hours , while untransfected cells were treated with or without 50 ng/mL rh-resistin on the same conditions , followed by serum-starving in glucose-free DMEM for 3 ~ 5 hours in the continued absence or presence of rh-re-sistin. Then the cells were treated with or without insulin for 2 hours. AMPKα2, G6Pase, PEPCK and Glut2 mRNA expression levels were determined by quantitative RT-PCR. The phosphorylation state of AMPK was deter-mined by Western blotting. Glycogen synthesis was measured by the incorporation of D-[U-14C] glucose to glycogen. Results Rh-resistin suppressed the AMPKα2, Glut2 mRNA expressions, and reduced the phosphory-lation level of AMPK and glycogen synthesis on both basal and insulin-stimulated conditions (P < 0.05), while it accelerated G6Pase and PEPCK mRNA expressions on the same conditions (P < 0.05). The mRNA expression levels of G6Pase , PEPCK , Glut2 and the phosphorylation level of AMPK and glycogen synthesis were signifi-cantly different between the rh-resistin group and the rh-resistin in conjunction with AMPKα2 siRNA-treated group. Conclusion Rh-resistin may affect glucose metabolism in HepG2 cell via AMPK pathway.

BACKGROUND: The experimental results showed that insulin sensitivity and glucose uptake in skeletal muscle could be improved by activating adenosine monophosphate-activated protein kinase a2 (AMPKα2). AMPKa2 is expected to become a new physiological and pharmacological target for the prevention and treatment of type 2 diabetes mellitus. OBJECTIVE: To clone human AMPKa2 subunit gene and to construct its wild-type and mutant eukaryotic expression vectors. DESIGN: A single sample observation.TIME AND SETTING: The experiment was performed in the Clinical Molecular Biology Laboratory, the Second Affiliated Hospital of Sun Yet-sen University from April 2007 to January 2008.MATERIALS: QuikChange II Site-Directed Mutagenesis Kit was produced by Stratagene. Eukaryotic expression vector pcDNA3.1(+) and E. coll DH5a were provided by the laboratory. Human skeletal muscle tissue was from patients who received amputation surgery in the Second Affiliated Hospital of Sun Yat-sen University. Informed consent was obtained from the patients, and fresh samples were collected and frozen in liquid nitrogen.METHODS: The human AMPKo2 subunit gone was amplified from human skeletal muscle by RT-PCR, cloned into T vector, and the recombinant plasmid was confirmed by sequencing. In vitro site-directed mutagenesis was carded out with Quickchange site-directed mutagenesis kit. The wild-type and mutant coding genes were subcloned into eukaryotic expression vector pcDNA3.1, and the recombinant plasmids were validated by enzyme digestion and sequencing.MAIN OUTCOME MEASURES: ①The cloning of aim gone; ②site-directed mutagenesis; ③ eukaryotic expression plasmid. RESULTS: The human AMPKα2 subunit gene (about 1700 bp) was successfully cloned, with 99％ homology to the reported AMPK α2 gene. A GenBank accession number was EF056019, The achieved mutation of the 45<'th> Lysine (AAA) was found to Arginine(AGA). The wild-type and mutant pcDNA-AMPKα2 recombinant plasmids were constructed successfully. CONCLUSIONS: The human AMPKα2 subunit gene was cloned successfully from human skeletal muscle and its wild-type and mutant eukeryotic expression vectors were constructed successfully in the experiment.

BACKGROUND: Hundreds of mouse embryonic stem cell lines are established presently. Most of these cell lines are collected from 129, C57BL/6J and BALB/C mice. Success rate of creating embryonic stem cell lines with Kunming mice is very low. OBJECTIVE: To explore the method of isolation and culture of murine embryonic stem cells of kunming species to increase the efficiency of establishment of embryonic stem cell lines from Kunming species mouse. DESIGN, TIME AND SETTING: The cell experiment was performed at the Chongqing Key Laboratory of Neurology from April 2006 to June 2007. MATERIALS: Blastocysts of 3.5 days and 4 days from Kunming species mouse were collected. METHODS: The blastocysts of 3.5 days and 4 days from Kunming species mouse were cultured on the fibroblast cell feeder layers for 4-5 days, and the cells from inner cell mass were isolated and subsequently cultured in vitro in 2.5 g/L trypase-0.4 g/L ethylenediamine tetraacetic acid (EDTA) and 1 g/L trypase-0.2 g/L EDTA at room temperature for 2-4 minutes. Embryonic stem cell colony was mechanically straggled. MAIN OUTCOME MEASURES: Colony growth was observed and determined by alkaline phosphatase staining, OCT-4 staining and karyotype analysis. RESULTS: Adherence rate, inner cell mass frequency and cloning efficiency of 4 day post coitum (dpc) embryo were higher than those of 3.5 dpc embryo (P

Objective To explore the effect of embryonic stem cells(ESCs) differentiating into cardiomyocytes(CM) by cardiomyocytes.Methods The blastocytes of 3.5 d from Kunming mouse,cultured on mouse embryonic fibroblaste cell feeder layers for 4～5 d,the mouse ESCs were isolated and subsequently cultured.Day 2～3 embryoid bodies(EBs) were derived from ESCs of passage 3 to 5 and then incubated with neonatal cardiomyocyte(CM) to induce into cardiomyocytes and the expression of cardiac specific cardiac troponin-T(TnT) and ?-actin were detected by immunofluoresence.Results The rhythmic beating embryoid bodies were observed on day 3 and reached 93% of rhythmic beating embryoid bodies on 12 day in group 2.All the beating cardiomyocytes derived from ESCs expressed cardiac-specific proteins for TnT and ?-actin,and the group induced by direct cell contact acquired the highest differentiating ratio as 56.5%,and was the highest differentiating ratio compared with other groups(P