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

Objective To investigate the therapeutic effect of bundle treatments for critically ill patients with pulmonary tuberculosis and respiratory failure (RF).Methods A prospective study was conducted, including 56 patients with pulmonary tuberculosis and RF necessary for invasive mechanical ventilation admitted into Department of Critical Care Medicine of the Fourth People's Hospital of Nanning City from January 2013 to December 2014 as the observation group to be treated by bundle treatments. A series of treatments and cares were given to the critically ill patients, such as invasive mechanical ventilation, application of antibiotics by experience within the first hour, supportive treatment targeted to hemodynamics in early stage, correction of brain dysfunction, effective therapy for tuberculosis, establishment of enteral nutrition in early stage and prevention of ventilator-associated pneumonia (VAP), etc. All the above treatments were completed one by one in 6 hours to 12 hours. Meanwhile, 42 patients who hospitalized from January 2011 to December 2012 and treated with conventional targeted therapy were designed as the control group. The changes of vital signs, blood routine test, respiration, liver, kidney, etc organ functions, the improvement of acute physiology and chronic health evaluation Ⅱ (APACHE Ⅱ) score, expected mortality and sequential organ failure assessment (SOFA) score, the incidence of VAP, cure and improvement rate, mortality, duration of invasive mechanical ventilation, the length of stay in intensive care unit (ICU) were compared between the two groups after treatments.Results There were no statistically significant differences in the comparisons between the two groups before and after treatments in temperature, white blood cell count (WBC), blood glucose, total bilirubin (TBil), alanine transaminase (ALT), albumin (Alb) and cholesterol level (allP > 0.05), except the platelet (PLT) count in observation group was higher than that in control group before treatments. After treatments for 72 hours, in the two groups, the heart rate (HR), respiration rate (RR), APACHE Ⅱ score and expected mortality were lower than those before treatments, while the arterial partial pressure of oxygen (PaO2) and oxygenation index were higher than those before treatments. There were no statistically significant differences in pH value, PLT, arterial partial pressure of carbon dioxide (PaCO2) and SOFA score before and after treatments in the control group (allP > 0.05). In the observation group, after treatments, the pH value was increased compared with that before treatments, while PLT, PaCO2 and SOFA score were decreased compared with those before treatments (allP < 0.05). The degrees of improvement of PaCO2 and oxygenation index in the observation group were superior to those in the control group [PaCO2 (mmHg, 1 mmHg = 0.133 kPa): 43.32±9.10 vs. 56.10±9.39, oxygenation index (mmHg): 330.60±100.98 vs. 245.65±83.20, bothP < 0.05]. After bundle treatments, compared with control group, the incidence of VAP was decreased [16.07% (9/56) vs. 33.33% (14/42),P < 0.05], improvement and cure rate was increased [78.57% (44/56) vs. 59.52% (25/42),P < 0.05] and mortality was decreased significantly in observation group [10.71% (6/56) vs. 28.57% (12/42),P < 0.05]. The duration of invasive mechanical ventilation (days: 9.15±3.59 vs. 16.96±13.44) and the length of stay in ICU (days: 13.30±4.24 vs. 23.00±15.03) in the observation group were shorter than those in the control group, but no statistically significant differences were found (bothP > 0.05).Conclusion The bundle treatments can effectively reduce the incidence of VAP and elevate the improvement and cure rate in patients with pulmonary tuberculosis complicated with RF.

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 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.

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 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 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.