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

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.

Objective To select antiplatelet regimen according to the results of CYP2C19 gene polymorphism,and then compare the major adverse cardiac events,bleeding events and the incidence of adverse reactions between two antiplatelet regimens. Methods Two hundred and seven patients who were diagnosed with acute coronary syndrome(ACS)and underwent elective PCI were tested for CYP2C19 genetic polymorphism,and 94 cases with CYP2C19 intermediate metabolism were randomly divided into high-dose clopidogrel group and ticagrelor group(47 cases each).High-dose clopidogrel group was given clopidogrel 150 mg once daily,and ticagrelor group ticagrelor 180 mg twice daily.Major adverse cardiac events,bleeding events and the incidence of adverse reactions were observed between two groups one month later. Results The average declined platelet aggregation rate was significantly different between the two groups[(6.27±5.65)% and(12.30±10.23)%,P<0.01];Adverse drug reactions ,the incidence of bleeding events and major adverse cardiac events of two groups were not significantly different. Conclusion Ticagrelor has stronger antiplatelet aggregation effects than high-dose clopidogrel.There is no difference in short-term clinical outcomes between the two groups.

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 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.

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

The fetal period is the critical time in the development of various organs.The changes in the intrauterine growth environment can cause fetal and neonatal multiple organ system underdevelopment and damage,which is also closely related to the adult diseases,including nervous system damage (such as brain development disorders,functional abnormalities and cerebral palsy,etc.),cardiovascular system damage(such as coronary heart disease,high blood pressure,etc.),growth disorders and metabolic disorders(such as insulin resistance,type Ⅱ diabetes,obesity,growth and development disorders),urinary system damage (such as interstitial renal disease,renal hypertension,etc.),respiratory system damage(such as respiratory distress syndrome,bronchial dysplasia,etc.),etc.In this paper,we introduced this to help clinicians to understand and strengthen the intervention measures to improve the quality of life.

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 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.