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

To explore the cognitions and behavior of cough etiquette of 556 residents in Longhua Community and self-compare the changes of cognitions and behavior before and after health education and training.The results showed that awareness rate of cough etiquette was low before training (senior group:25.4％,children:11.9％,young white-collar:36.1％,immigrants:14.3％,P ＜ 0.05) and increased significantly (senior group:97.2％,children:99.2％,young white-collar:100.0％,immigrants:98.6％,P ＜ 0.05).Except for no difference in compliance aspects of immigrants (P ＞ 0.05),the other three groups improved after training.And the difference was statistically significant before and after training (P＜0.05).

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

Objective To study the curative effect and medchanism of action about prostaglandin E1 and magnesium sulfate in the treatment of chronic pulmonary heart disease in the aggravation period. Methods Fifty-three patients with chronic pulmonary heart disease in the aggravation period were accepted emergencyward in the Guangzhou Red Cross Hospital during June 2007 to May 2008,they were randomly divided into the treatment group (28 case) and control group (25 case) . Both groups were given low flux breathe in oxygen and dissipate phlegm and relieve a cough and resist infection and spasmo]ysis and calm down asthma and strive heart and diuresis colligate therapy. Mean while,patients in the treatment group,beside the colligate therapy,were treated with prostaglandin E1 10 ml and magnesium sulfate 10 ml ,the course of treatment lasted 2 weeks. Then we investigate the amelioration of clinical symptom and alteration of blood gas analysis in the two groups before and after the treatment. Results The rate of clinical efficacy were 89.3% (25/28) and 76.0% (18/25),with superiority in the treatment group (χ2=1.87,P<0.05) . Whole blood viscosity,fibrinogen,PaO2 and PaCO2 were improved in both groups,amelioration of blood gas analysis observation superiority in the treatment group compared with control group (P<0.05). Conclu-sions Prostaglandin E1 and magnesium sulfate can depress pulmonary artery pressure and abate the afterload of right ventricle,which have better treatment effect in chronic pulmonary heart disease in the aggravation period.

Parenteral nutrition associated cholestasis (PNAC) mainly occurs in children. Currently identified risk factors include premature infants, lack of enteral feeding, repeated infections, and toxicity or nutrient deficiency of parenteral nutrition solution. Recent studies have shown that nuclear receptor-mediated bile acid transporter may be the key site of PNAC pathogenesis, in which some cytokines play important roles. New anti-cholestatic therapy based on the regulation of expression of these molecules may prevent end-stage liver disease caused by PNAC.

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.

The involvement of oxidization of peroxid in the antigen-antibody reaction makes the cell membrane permeability enhanced, so the fluorescence-labeled antibody stain can infiltrate into the cell easily and contact the antigen rapidly and diffusely, and then the antigen-antibody crosslinking can be formed efficiently. With the enhanced staining efficiency, shortened test duration, simplified operation, increased positive detection rate and accuracy, the new technology lays a foundation of clinical definite, curative effect view and prognosis.

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.

Objective To investigate the suppressive effects of Maytansine on the proliferation,migration and invasion of nasopharyngeal carcinoma (NPC) cells.Methods MTT assay was employed to study the effect of Maytansine on the viability of 5-8F and SUNE1 cell lines.Wound healing assays and transwell were used to assess the effects of Maytansine on the migration and invasion capacities of NPC cells.Furthermore,an integrated bioinformatics analysis was conducted to clarify the potential mechanisms of Maytansine.Results Time course analysis of Maytansine treated NPC cells indicated significant decrease in cell viability,assessed by MTT.Furthermore,we identified that Maytansine reduced the migration and invasion capacities of NPC cells.Moreover,bioinformatics analysis showed that Maytansine suppressed the proliferation,migration and invasion via the upregulation of CDKN1B,BMP2,HBEGF,SMAD6,ADM,VEGFA,CTGF and IL6 in NPC cells.Conclusion Maytansine plays a significant suppressive role in the proliferation,migration and invasion of NPC cells and may be explored further for its potential therapeutic effects on NPC in humans.