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 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 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

AIM: To study the efficacy and safety of Ultra Q- YAG vitreolysis for vitreous floaters.
METHODS: Retrospective case series study. From September 2014 to May 2015 in Beijing Aier - Intech Eye Hospital, 263 cases (340 eyes) with vitreous floaters were involved. All patients underwent visual acuity, intraocular pressure, slit lamp, mydriatic fundus, B ultrasonic examination, and recorded the form of a vitreous opacity excluded pathological fundus lesions. All the patients were divided into two groups: Group A (<30 years old) 78 cases (82 eyes); the morphology of vitreous opacity were dot, filiform and mesh. Group B ( > 45 years old), 185 cases (258 eyes), the morphology of vitreous opacity was Weiss ring, translucent flocculent clouds or dense fibrous membrane. Patients in 30 - 45 years old were eliminated because of the untypical opacity factor. The treatment was done by the same physician. Vision changes was analyzed before and after the treatment.
RESULTS: Questionnaire survey was done. According to the scores of the questionnaires, patients were divided into 3 groups: no improvement ( 1 - 2 points), partial improvement (3-5 points), significant improvement (6-10 points ). At 1mo after treatment, Group A: no improvement in 9 eyes (11. 0%), partial improvement in 57 eyes (69. 5%) and significant improvement in 16 eyes (19. 5%); Group B: no improvement in 0 eyes, partial improvement in 23 eyes ( 8. 9%) and significant improvement in 235 eyes (91. 1%); all the patients had no complications.
CONCLUSION: The treatment with YAG vitreolysis for vitreous floaters is safe and effective, especially for the patients more than 45 years old whose vitreous floaters caused by posterior vitreous detachment.

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.

Objective To investigate the effect of combination therapy with cognitive behavior on the quality of life of patients with gastroduodenal ulcer.Methods The patients with gastroduodenal ulcer treated in our hospital from April 2015 to April 2017 were randomly divided into intervention group and control group according to the order of treatment.Both groups were treated with bismuth-containing quadruple therapy and routine nursing was performed.The intervention group received cognitive behavior on this basis.SF-36 scale was used to evaluate the quality of life of the two groups of patients, and the prognosis of the patients was evaluated.The patient satisfaction was investigated by the self-designed questionnaire of patient satisfaction questionnaire.The clinical efficacy, treatment compliance, quality of life improvement, prognosis and satisfaction were compared between the two groups.Results The total effective rate was 96.67%(58/60) in the intervention group and 81.67%(49/60) in the control group.The total effective rate of the intervention group was significantly higher than that of the control group(P<0.05).The scores of SF-36 scale in the intervention group were higher than those in the control group(P<0.05).The incidence of complications in the intervention group was lower than that in the control group, and the satisfaction rate was higher than that of the control group(P<0.05).Conclusion Cognitive behavioral intervention combined with bismuth quadruple therapy for gastroduodenal ulcer can improve the patient's compliance and efficacy, significantly improve the quality of life of patients, improve patient satisfaction, with clinical application value.