Objective To study the effects of knock down midkine(MK)by siRNA on chemosinsitivity in bladder cancer cells. Methods Three MK siRNAs were designed and constructed. After transfected with MK siRNA or scrambled siRNA for different time, cultured cells were harvested to carry on the next experiments. Expression of MK was determined by real-time RT-PCR and western blot, and apoptosis were evaluated by caspase-3 activity and TUNEL assay. MTT was performed to examine the inhibition effect of Paclitaxel (PTX) on cells. Results MK siRNA could down-regulate the MK expression in a dose- and time-dependent manner. The MTT results showed that the inhibit rates were (18.21±0.36)%, (18.19±0.29)%, (17.89±0.33)%, (1.86±0.52)%, (32.56±0.53) %, (53. 83±0.38) % and (78. 95±0.55) % in different groups PTX alone(0.2μmol/L), ConA +PTX(0.2 μmol/L), Con-B +PTX(0.2 μmol/L), siRNA alone(12. 50 nmol/L), siRNA(3. 125nmol/L) + PTX (0. 2 μmol/L), siRNA (6. 25 nmol/L) + PTX (0. 2 μmol/L) and siRNA (12. 50nmol/L)+PTX(0.2 μmol/L), respectively. The TUNEL results showed that apoptosis index was (1.81 ±0. 36)%, (1. 89±0. 38)%, (5. 56±0. 58)%, (9. 68±0.55)% and (15. 25±0.56)% in different groups (Con-A, Con-B, siRNA (3. 125 nmol/L), siRNA (6. 25 nmol/L) and siRNA ( 12. 5nmol/L), respectively. The activity of caspase-3 increased significantly in transefected cells with a dose-and time-dependent manner. Conclusion MK siRNA could sensitize human bladder cancer cells to chemotherapy which might be through the apotosis.

ObjectiveTo determine optimal positive end expiratory pressure (PEEP) in mechanical ventilation in children with severe asthmatoid disease based on the quasistatic pressure-volume (P-V) curve.MethodsA serf-control study was done on 23 children with severe asthmatoid disease in the pediatric intensive care unit( PICU ).Quasistatic lung P-V curve of these patients was analyzed and the lower inflection point (LIP) from P-V curve was determined.Three different PEEP (0 cm H2O,LIP,LIP+2 cm H2O,1 mm H2O =0.098 kPa) were given to the patients.The effects of PEEP at different levels on gas exchange,hemodynamic and airway pressure were observed.ResultsThe quasistatic LIP were (2.70 ±2.00)cm H2O.When PEEP was increased to the level of LIP + 2 cm H2O,PaO2 / FiO2 and lung compliance improved significantly (P ＜ 0.01 ) and dynamic lung compliance was the highest,peak inspiratory pressure was (22.30 ± 3.00) cm H2O and mean airway pressure was( 14.11 ± 1.01 ) cm H2O,without obvious adverse effects on mean arterial blood pressure and heart rate.There was no difference in PaCO2,when compared PEEP =0 cmH2O to PEEP =LIP + 2 cmH2O.ConclusionThe application of PEEP is safe.LIP + 2 cm H2O from quasistatic P-V curve could be set as the optimal PEEP under which mechanical ventilation has the best efficacy and do not aggravate CO2 retention and abnormality of hemodynamics in children with severe asthmatoid disease.

Objective To study the relationship between MRI and clinical profiles of periventricular leukomalacia (PVL) in children Methods The clinical and MRI findings in 34 cases with PVL were retrospectively analyzed Results (1) Periventricular hyperintensity on T 2WI was more prominent in the preterm group than that in the term group, and P value was 0 000; (2) Cortical lesion and subcortical leukomalacia was seen in 9 of 19 cases in the children with PVL born at term, but detected in only 1/15 in the preterm group P value was 0 020; (3)Seizure was common in term children P value was 0 036; (4) The degree of reduction of periventricular white matter correlated with motor impairment and mental retardation in all children, and P values were 0 002 and 0 000, respectively The thinning of the corpus callosum also correlated with mental retardation and P value was 0 012 The degree of reduction of periventricular white matter correlated with visual impairment in preterm group Conclusion The end stage PVL can been clearly displayed by MRI, and gestational age and clinical manifestation were closely related to the findings of MRI

Amyloid beta-Peptides ; analysis ; Animals ; Frontal Lobe ; drug effects ; physiology ; Hippocampus ; drug effects ; physiology ; Learning ; drug effects ; Memory ; drug effects ; Rats ; Rats, Wistar ; Sodium Azide ; pharmacology ; Space Perception

Animals ; Electric Stimulation ; Neurons ; pathology ; physiology ; Pain Threshold ; Rats ; Rats, Sprague-Dawley ; Red Nucleus ; pathology ; physiology

Objective To establish an HPLC method for the determination of metoclopramide (MCP) and its related substances in MCP nasal spray .Methods Chromatographic separation was performed on an Agilent TC-C18 column (250 mm ×4.6 mm,5 μm) using acetonitrile and phosphate buffer solution (0.05 mol/L potassium dihydrogen phosphate solution, added with 5 ml of triethylamine and adjusted to pH 4.0 with phosphoric acid)(19∶81) as the mobile phase at 1.0 ml/min.The detection wavelength was 275 nm and the column temperature was set at 30℃.Results and Conclusion Related substances were completely separated from MCP .For MCP,the linearity of determination was over the range of 10-200 μg/ml and the recovery of the method ranged from 100.3%to 101.6%.The relative standard deviation was 0.68%(n=9).The method is accurate, reliable, repeatable, and could be readily utilized as a quality control method for MSP nasal spray .

Objective To perform the MR findings and clinic characteristics of a series cases with malformations of cortical organization and to have a better understanding of malformations of cortical organization.Methods The clinical records and MRI studies of 5 cases with malformations of cortical organization were retrospectively reviewed.Results (1) 3 of 5 cases were polymicrogyria and 2 cases were schizencephaly. (2) Most patients with malformations of cortical organization suffered from epilepsy.Conclusion Malformations of cortical organization were recognized as the important causes of developmental delay and epilepsy and should be paid more attentions to them.

Objective To examine the role of noggin on the expression of nestin and glial fibrillary acidic protein (GFAP) in the hippocampus of adult rats. Methods Antisense oligodeoxynucleotide (ASODN) technique was employed to inhibit endogenous noggin expression and immunohistochemistry was used to detect the expressions of Nestin and GFAP in the hippocampus of adult rats. Results It was observed that the number of nestin and GFAP immunoreactive cells in the dentate gyrus (DG) of hippocampus was increased in adult rats treated with antisense oligodeoxynucleotide to noggin. Moreover, the number of GFAP immunoreactive cells was increased in the subventricular zone of the rats treated with antisense oligodeoxynucleotide to noggin. Conclusion The results in the present study indicates that noggin may play a role in the differentiation of neural stem cells in the adult hippocampus, and it promotes the differentiation of neural stem cells in the DG to neuronal fate.

The high-copy-number plasmid pcDNA3.1+ is unstable within S almonella typhimurium. A novel plasmid pmcDNA3.1+ was constructed by removin g the promoter sequence of ampicillin resistance gene (bla gene) in plasmid pcDNA3.1+. In contrast to pcDNA3.1+, pmcDNA3.1+ was stable within Salmonel la typhimurium SL7207 in LB medium with or without ampicillin. Further experi ments showed the ?-lactamase activity of Salmonella typhimurium SL7207(pmc DNA3.1+) was apparently lowered than that of Salmonella typhimurium SL7207( pcDNA3.1+) and the high ampicillin concentration was maintained longer in LB me dium culturing Salmonella typhimurium SL7207(pmcDNA3.1+). When mice were a dministered with Salmonella typhimurium SL7207(pmcDNA3.1+) intraperitoneall y, more than 95% of Salmonella cells separated from the spleen still harbore d the plasmid pmcDNA3.1+ 7 days later; but 99% of Salmonella cells lost the plasmid pcDNA3.1+ at day 3 in mice innoculated with Salmonella typhimurium SL7207(pcDNA3.1+). By lowering the expression of bla gene, the rapid deco mposition of ampicillin in LB medium was avoided and the metabolic pressure was relieved for the host cells. This method offers a solution for the problem of t he instability of high-copy-number plasmid within Salmonella typhimurium.

Objective To investigate the characteristic and its clinical value of tumor necrosis factor (TNF)-α and its receptor, interleukin (IL)-1β and its receptor in serum and bronchoalveolar lavage fluid(BALF) in patients with pulmonary tuberculosis and to determine the role of them in the immunopathogenesis of tuberculosis. Methods The concentrations of TNF-α,soluble TNF receptor (sTNF-R) Ⅰ, IL-1β and IL-1 receptor were measured using sandwish ABC-enzyme-linked immunosorbent assay (ELISA) method in serum and BALF of 46 patients with active tuberculosis and 21 patients with inactive tuberculosis, and in the serum of 20 cases of healthy control. Meanwhile the above-mentioned cytokine levels in serum and BALF of 19 patients with active tuberculosis were followed up. Differences between groups were assessed for significance by t test. Results The TNF-α,sTNF-R Ⅰ, IL-1β and IL-1 receptor levels and TNF-α/sTNF-R Ⅰ ratios in BALF of active tuberculosis group were (286.2±96.3) pg/L,(2 431.5±1 124.6) pg/L,(58.6±3.2) pg/L,(162.4±17.1) pg/L and 0.06±0.01, respectively, which were all significantly higher than those with inactive tuberculosis group (t=3.36,3.25,2.95,2.27 and 3.12 respectively; P<0.05). The TNF-α,sTNF-R Ⅰ,IL-1β and IL-1 receptor levels and TNF-α/sTNF-R Ⅰ ratios in BALF of cavernous tuberculosis group were (381.4±106.4) pg/L,(2 824.7±1 318.5) pg/L,(66.4±4.6) pg/L,(176.4±18.7) pg/L and 0.07±0.01, respectively,which were all significantly higher than those of non-cavernous tuberculosis group (t= 3.46,2.37, 3.19, 2.99 and 3.22, respectively; P<0.05). After 2-month' antituberculosis treatments, among 19 cases, the TNF-α,sTNF-R Ⅰ,IL-1β and IL-1 receptor levels and TNF-α/sTNF-R Ⅰ ratios in BALF of 16 cases were significantly lower than those at the beginning of treatments (t= 3.26,3.17, 3.28, 2.92 and 3.12 respectively; P<0.01). Meanwhile, their clinical symptoms improved, sputum smear negative, lesions on chest X-ray resolved and the cavity shrinked or closed. Conclusions TNF-α, sTNF-R Ⅰ, IL-1β and IL-1 receptor are likely to be involved in the immunopathogenesis of tuberculosis. Detection of TNF-α, sTNF-R Ⅰ, IL-1β and IL-1 receptor levels in the serum and BALF is helpful to understand the activity of disease, determine the clinical pattern of disease,assess the prognosis of disease and monitor the therapeutic effect in patients with pulmonary tuberculosis.