Objective:To establish a quantitative method for determination of CK19 mRNA with TaqMan real time quantitative RT-PCR.Methods: A 230 bp fragment of CK19 mRNA was amplified from the total RNA of gastric cancer cells using RT-PCR methods and was introduced into pMD 18-T Simple vector.The plasmid was purified and the fluorescent standard PCR product was prepared.The expression levels of CK19 mRNA in standard PCR product,5 tumor tissue specimens and 30 healthy subjects were observed.Results: A 230 bp fragment of CK19 mRNA was successfully cloned into the pMD 18-T Simple vector and was verified by sequence analysis.A stable standard for detection of CK19 mRNA was established,that is,when C_(T) was set within 35 cycles,negative specimen was defined when the result was lower than 100 copies.Conclusion: TaqMan real time quantitative RT-PCR is stable and reliable in quantitative detection of CK19 mRNA in peripheral blood.

Animals ; Humans ; Phytotherapy ; Rheumatic Diseases ; drug therapy ; Stilbenes ; therapeutic use

Amino Acid Metabolism, Inborn Errors ; etiology ; therapy ; Glutarates ; blood ; Humans ; Infant, Newborn ; Male

Age-related macular degeneration(AMD) is a macular disease and its incidence is gradually increasing with age.It is thought that its pathogenesis is related to the age,heredity,smoking,diet,oxidative stress,immune inflammation reaction and cardiovascular disease and so on.Oxidative stress is closely related to AMD.Antioxidant therapy provides a new strategy for the prevention and treatment of AMD.Also,it provides a new way to alleviate and prevent AMD.

Objective To analyze the transformation of the function of research management department in medical institutions based on the knowledge management.Methods The value chain is introduced to portray the functions of research management department, and a survey is employed to collect existing problems in research management.Results Lack of knowledge management is main reason that limits the research development in medical institutions.Conclusions It is very important to re define the strategy for conducting the research in the medical institutions and to improve the knowledge management in those medical institutions.

Objective: To explore expression of EBV-LMPI and ZEBRA in B cell in SLE patients. Methods: Labeled by immunofluoresence indirectly, determined by flow cytometry. Results: Expression of EBV-LMP1 and ZEBRA in SLE patients was higher than normal control P 0.05) . Conclusion: EBV may be induce SLE occurrence, reproduce of EBV can promote the development of SLE. Detection expression of EBVLMP1 and ZEBRA can use as a predicator of SLE activity.

Objective To investigate the effects of ?-aescin on nuclear factor-?B (NF-?B) activities and tumor necrosis factor-? (TNF-?) protein expression in the rat brain tissue following acute traumatic brain injury (TBI). Methods A total of 62 SD rats were subjected to a lateral cortical impact injury caused by a free-falling object and divided randomly into four groups, ie, sham operation group (Group A), injury group (Group B), ?-aescin treatment group (Group C) and pyrrolidine dithocarbamate (PDTC) treatment group (Group D). Group C was administered with ?-aescin and Group D treated with PDTC immediately after injury. A series of brain samples were obtained directly 6, 24 hours and three days respectively after operation in four groups. The NF-?B activation of rat brain was determined by electrophoretic mobility shift assay (EMSA) and the levels of TNF-? protein in rat brain measured by radio-immunoassay (RIA). In the meantime, the water content of rat brain was measured and pathomorphological observation carried out. Results Compared with Group A, NF-?B activities, the levels of TNF-? protein and the water content of the rat brain were significantly increased (P

Objective To investigate the span and frequency of physical symptoms of panic attack in patients with panic disorder.Methods Panic symptoms presented at the time of panic attacks were assessed and categorized in forty-eight consecutive outpatients with panic disorder during August 2000 to November 2002.Results Palpitations (44/48,91.7%), shortness of breath (36/48,75.0%), sweating (34/48,70.8%), feel suffocated (33/48,68.8%), faint (31/48,64.6%), dizziness (27/48,56.3%) and fear of dying (27/48,56.3%) were the most frequent symptoms reported by the patients with panic disorder. If categorized according to system, the most frequent symptoms were presented in cardiovascular, neurological, respiratory and autonomic nervous systems of patients with panic disorder.Conclusions Physical symptoms varies when panic attack occurs. Panic disorder was often misdiagnosed and underdiagnosed because of wide and non-specific spectrum of physical symptoms.

Objective The changing patterns of pathogenic isolates and antibiotic susceptibility in Chongqing's neonates between 2010 and 2015 were investigated for the purpose to provide evidence for rational use of antibiotics and control of nosocomial infections.Methods The distribution of pathogenic bacteria and antibiotic susceptibility were analyzed.Identification and antibiotic susceptibility testing were carried out using BD Phoenix 100 automated system and the conventional Kirby-Bauer method.The results were interpreted in accordance with the breakpoints of the Clinical and Laboratory Standards Institute.Results A total of 10 569 pathogenic bacterial strains were isolated during the period,most of which were gram-negative bacteria (80.8 ％,8 540/10 569),primarily Klebsiella pneumoniae (29.3 ％),followed by Escherichia coli (16.7 ％),Acinetobacter baumanmii (9.9 ％),Enterobacter cloacae (8.6 ％) and Pseudornonas aeruginosa (3.3 ％).Gram-positive strains accounted for 14.1％ (1 490/10 569),mainly Staphylococcus aureus (7.8％),Staphylococcus epidermidis (2.2 ％),and Staphylococcus haemolyticus (1.8 ％).Imipenem and meropenem showed high activity against Enterobacteriaceae (＜ 10％ resistant),followed by P.aeruginosa (＞ 10 ％ resistant),and A.baumannii (＞20％ resistant).The prevalence of carbapenem-resistant strains was 8,4 ％ in K.pneumoniae and 2.9 ％ in E.coli isolates,No gram-positive isolates were resistant to vancomycin,teicoplanin or linezolid.Conclusions K.pneumoniae was the most frequently isolated pathogen in the neonates treated in Children's Hospital of Chongqing Medical University.The prevalence of A.baumannii isolates is increasing.Carbapenem-resistant Enterobacteriaceae strains are emerging.

Objective: To study the expression levels of forkhead transcription factor(FoxO1) mRNA and protein in the insulin resistance (IR) HepG2 cells model (HepG2/IR) and IR reversal HepG2 cells model (HepG2/IR-PH), and to explore its mechanism in IR.Methods:The HepG2/IR was induced with different doses of insulin (1×10-10, 1×10-9, 1×10-8, 1×10-7, 1×10-6 and 1×10-5 mol·L-1) for different time(24, 36 and 48 h)in the HepG2 cells.The cells in control group were not treated with insulin.The glucose levels in supernant were determined by glucose oxidase method, and the glucose consumption in HepG2 in various groups were calculated to confirm the optimum induction conditions of HepG2/IR.The HepG2/IR-PH was induced with different doses of pioglitazone hydrochloride (PH) (0.156, 0.313, 0.625, 1.250, 2.500, 5.000, 10.000 and 20.000 mmol·L-1) in the HepG2 cells, and control group was set up at the same time. The proliferation activities of cells were observed by MTT assay to confirm the optimum reversal concentration of PH.The FoxO1 mRNA and protein expression levels were detected by Real-time PCR and Western blotting methods.Results: The glucose consumption decreased by 45.84% in HepG2/IR after treated with 1×10-7 mol·L-1 insulin for 36 h, and there was significant difference compared with control group(P<0.05), the proliferation activity of cells had no change in the HepG2/IR-PH after treated with 1.25 mmol·L-1 PH for 24 h(P>0.05).Compared with control group, the expression levels of FoxO1 mRNA and protein in HepG2/IR were significantly increased(P<0.01);compared with control group, the expression levels of FoxO1 mRNA and protein in HepG2/IR-PH had no significant differences(P>0.05).Conclusion:The IR of HepG2/IR is associated with the FoxO1 mRNA expression.The detection of FoxO1 mRNA seems to be an indicator to evaluate the efficacy of insulin sensitizer, and inhibiting the expression of FoxO1 mRNA may be developed as a potential therapy for type 2 diabetes.