Objective To investigate p16 promoter methylation in premature rats with chronic lung disease induced by hyperoxia. Methods Eighty premature Wistar rats were randomly divided into two groups: hyperoxia group (fraction of inspiratory oxygen) 0. 90 and control group (fraction of inspiratory oxygen 0. 21), 40 rats for each group. Semi-nested methylation specific polymerase chain reaction and methylation specific polymerase chain reaction were applied respectively to detect p16 promoter methylation in lung tissues. Additionally, p16 mRNA and protein expressions in lung tissue were detected by reverse transcription- polymerase chain reaction, Western blot and immunohistochemistry method. Results The methylation was not found in control group by seminested methylation specific polymerase chain reaction and methylation specific polymerase chain reaction, while was found in different aged rats of the hyperoxia group. The methylation detection rate was higher by using the semi-nested methylation-specific polymerase chain reaction (52.5%, 21/40) than that by methylation specific polymerase chain reaction (42.5%, 17/40) in the hyperoxia group,but there was no statistically significant difference between the two methods. The p16 mRNA in the hyperoxia group were significantly lower than in the control group at day 7, 14 and 21(1.73 ± 0.40 vs 2.11±0. 37,1.29±0. 19 vs 1.60±0. 27,0. 95±0.25 vs 1.72±0. 34, t=2.19, 2.95 and 10. 43,P＜0. 05). The p16 protein expressions by western blot in the hyperoxia group were significantly lower than in the control group at day 7, 14 and 21 also (88. 1±8. 7 vs 95.0±4.1,65.7±4.5 vs 83. 5±13.6 and 50.4±4.9 vs 86.7±11.9, t=2.27,3.95 and 13.40,P＜0.05). The expression of p16 mRNA (1.06±0.61) and protein (62.32±25.65) in lung tissues of rats with methylation was lower than that without methylation (1.63±0.62 and 94.93±22.21, respectively) (t=2.95, OR=0. 86;t=4.28, OR=0. 85,P＜0.01, respectively). Conclusions Exposure to hyperoxia might induce p16 promoter methylation in lung tissues in premature rats. Methylation risk increases as exposure time extends. p16 promoter methylation induced by hyperoxia might participate in the mechanism of lowering p16 mRNA and protein expression, but might not result in p16 gene silence.

Tuberculosis is an infectious disease caused by Mycobacterium tuberculosis .The rapid development of laboratory diagnostic technology provides a variety of options for diagnosis and treatment of tuberculosis .Nucleic acid detection is to use the theory and technology of molecular biology ,through direct detection of status or defect of particular segments ,then make diagnosis of the state of human body and disease .In recent years ,the development of nucleic acid diagnostic technique has greatly promoted the rapid diagnostic of laboratories .This article reviews the new methods of bacterial nucleic acid and host mi‐croRNAs detection technologies .

Objective To evaluate the left ventricular function in patients with different extents of coronary stenosis by the left ventricular pressure volume loop Methods In 65 patients who were undergoing coronary angiography and left ventriculography examinations, materials of clinical and cardiac catheter examinations were collected In right anterior oblique 30 degrees the left ventriculography was progressed The pressure curves of the left ventricle and the aorta were recorded continuously and volumes of the left ventricles were measured with the dot tracing method Pressure volume loop was set up and ventricular energy indexes embodied by the pressure volume loop were measured and calculated Results In normal group, pressure volume loop was located in the left and lower portion of the coordinate system Along with the levels of the coronary stenosis becoming more severe, pressure volume loops moved to the right and upper portion of the coordinate system Little changes occurred in ejection fraction except that there was a decreasing in patients with lesions of three branches; Stroke work showed no obvious changes; Filling energy became larger; Total energy increased obviously in both groups with lesions of double branches and three branches; End systolic energy increased gradually while energy efficiency decreased gradually Conclusion The ventricular pressure volume loop can be obtained in routine ventriculography which can reflect many indexes of ventricular function quantitatively The ventricular energy indexes change correspondingly with coronary artery lesions and may be useful to assess ventricular function in patients with different levels of coronary stenosis

OBJECTIVEThis research aims to study the changes in pain threshold and glial cell line-derived neurotrophic factor (GDNF) in a Sprague Dawley (SD) rat model oftrigeminal neuralgia.

METHODSA total of 36 male SD rats were randomly divided into three groups: operative, sham-operative, and control. In the operative group, a chronic constriction injury (CCI) was caused by placing loose chromic gut ligatures around the right infraorbital nerve (ION). In the sham-operative group, the right ION was subjected to the same procedure, but without ligation. In the control group, the right ION was not subjected to any treatment. The pain thresholds of the three groups were recorded at different times after the operation. The GDNF expression in each group was analyzed via immunohistochemical staining.

RESULTSAn allodynia to mechanical stimulation in the region of the ligated ION was observed starting on the 2nd week after operation. Pain thresholds started to increase gradually from the 6th week and returned to the original level at the 10th to 12th week after operation. Cells that expressed the GDNF markedly increased in number in the operative group with changes observed at different times.

CONCLUSIONWe use chronic constriction injury to the infraorbital nerve (CCI-ION) to establish a trigeminal neuralgia-like animal model in SD rats. GDNF may play a role in regulating pain by promoting the restoration and regeneration of nerve fibers.

Animals ; Constriction ; Disease Models, Animal ; Glial Cell Line-Derived Neurotrophic Factor ; Glial Cell Line-Derived Neurotrophic Factors ; Hyperalgesia ; Male ; Pain Threshold ; Rats ; Rats, Sprague-Dawley ; Trigeminal Neuralgia

The standardized training for pediatric residents is a necessary route to promote their clinical ability and make them grow up.With the aim to improve the residents' basic theory,clinical skills and medical quality,we set up training plans according to the requirement of standardized training in shanghai,earnestly implement the related training in the process of training content,method,time node and stage appraisal system,formulating personalized training plan and implementing one to one teaching.Besides,we attach importance to the cultivation of scientific research and teaching ability of residents and set up a perfect evaluation system.Residents are required to obtain a certificate of training through the unified comprehensive examination before graduation.Here,the problems existing in the process of pediatric resident standardization training and preliminary countermeasures are also discussed.

OBJECTIVE:To establish a method for the contents determination of tanshinol,protocatechuic aldehyde,ferulic ac-id and salvianolic acid B in Yixinshu capsule. METHODS:Dual-wavelength HPLC was performed on the column of Eclipse XDB-C18 with mobile phase of 0.5%phosphoric acid-methanol-acetonitrile(gradient elution)at the flow rate of 1.0 ml/min,the de-tection wavelength was 280 nm(tanshinol,protocatechuic aldehyde,salvianolic acid B)and 320 nm(ferulic acid),column tempera-ture was 30℃and volume was 10 μl. RESULTS:The linear range of tanshinol,protocatechuic aldehyde,ferulic acid and salvianolic acid B were respectively 9-144μg/ml(r=0.999 9),0.5-8μg/ml(r=0.999 9),0.65-10.4μg/ml(r=0.999 9)and 221.25-3 540μg/ml (r=0.999 9);RSDs of precision,stability and reproducibility tests were no more than 1.90%;the average recovery was respective-ly 100.8%(RSD=1.65%,n=9),100.1%(RSD=2.87%,n=9),100.1%(RSD=3.01%,n=9) and 99.4%(RSD=2.05%,n=9). CONCLUSIONS:The method is simple and reproducible,and can be used for the quality control of Yixinshu capsule.

OBJECTIVE To analyze the isolation rate and four consecutive years′(2004-2007) antibiotic-resistance trend of extended-spectrum ?-lactamases(ESBLs) producing Escherichia coli in urinary tract infection and to provide evidence for anti-infection therapy.METHODS The anti-microbial susceptibility test results of E.coli cultured from the midstream urine of outpatients and inpatients of our hospital from 2004 to 2007 were collected.Whonet 5.4 statistical analysis software was used for analysis.RESULTS During four years,no apparent change has been shown about antibiotic resistant rate of ESBLs producing E.coli.The isolation rate of this kind of E.coli increased year by year from 31% to 44% and showed high resistance to third generation cephalosporins and quinolones antibiotics,but was 100% sensitive to imipenem.CONCLUSIONS With the ESBLs-producing E.coli increasing year by year,the antibiotic-resistance rate of E.coli isolated from urinary tract infection is apparently increased and thus screening of antibiotics sensitive to enzyme-producing bacterial is the key point when the empirical therapy especially when the third and fourth generation cephalosporins are ineffective.

OBJECTIVE:To establish a method for simultaneous determination of tanshinol,protocatechuic aldehyde,paeoni-florin,ferulic acid and salvianolic acid B in Shenshao oral liquid. METHODS:RP-HPLC was performed on the column of Eclipse XDB C18 with mobile phase of 0.5%phosphoric acid-methanol-acetonitrile(gradient elution)at the flow rate of 1.0 ml/min,the de-tection wavelength was 280,230 and 320 nm,column temperature was 30 ℃ and volume was 10 μl. RESULTS:Under the chro-matographic conditions,5 kinds of components could be completely separated,the linear range of tanshinol,protocatechuic alde-hyde,paeoniflorin,ferulic acid and salvianolic acid B were respectively 24-384 μg/ml(r=0.999 9),1.25-20 μg/ml(r=0.999 9), 40.5-648 μg/ml(r=0.999 8),1.5-24 μg/ml(r=0.999 9),145-2 320 μg/ml(r=0.999 9);RSDs of precision,stability and reproduc-ibility tests were no more than 2.2%;the average recovery was respectively 100.7%(RSD=1.23%,n=9),100.0%(RSD=2.19%,n=9),99.6%(RSD=0.87%,n=9),100.3%(RSD=1.11%,n=9) and 99.3%(RSD=2.46%,n=9). CONCLUSIONS:The method is specific with good precision and reproducibility,and can be used for the content determination of 5 components in Shenshao oral liquid.

OBJECTIVE:To establish a method for simultaneous determination of danshensu,salvianolic acid B,protocatechu-ic aldehyde,paeoniflorin and ferulic acid in Jingzhi guanxin granule. METHODS:HPLC was performed on the column of Zorbax Eclipse XDB-C18 with mobile phase of acetonitrile-methanol-0.5% H3PO4(gradient elution)at a flow rate of 1.0 ml/min,the detec-tion wavelength was 280 nm(for danshensu,protocatechuic aldehyde,salvianolic acid B),230 nm(for ferulic acid)and 320 nm (for paeoniflorin),column temperature was 30 ℃,injection volume was 10 μl. RESULTS:The linear range was 1.19-478.34 μg/ml for danshensu(r=0.999 9),0.11-44.93 μg/ml for protocatechuic aldehyde(r=0.999 9),7.49-995.20 μg/ml for salvianolic acid B (r=0.999 7),0.95-379.39 μg/ml for paeoniflorin (r=0.999 9) and 0.01-3.12 μg/ml for ferulic acid (r=0.999 5);the limits of quantitation were 1.91 ng,0.36 ng,150.00 ng,2.74 ng and 0.10 ng,limit of detection were 0.96 ng,0.10 ng,45.00 ng,1.52 ng and 0.03 ng;RSDs of precision,stability and reproducibility tests were lower than 2%;recoveries were 98.06%-99.47%(RSD=0.52%,n=6),98.01%-99.49%(RSD=0.70%,n=6),98.44%-99.45%(RSD=0.37%,n=6),96.94%-100.71%(RSD=1.27%,n=6)and 95.44%-100.44%(RSD=1.90%,n=6). CONCLUSIONS:The method is simple and accurate,and suitable for the simultaneous determination of danshensu,salvianolic acid B,protocatechuic aldehyde,paeoniflorin and ferulic acid in Jingzhi guanxin granule.

Objective To estimate the significance of E-selectin and P-selectin in nodular vasculitis.Methods The EnVision two-step method was used to measure the expression of E-selectin and P-selectin in skin samples from the lesions of 70 patients with nodular vasculitis and normal skin of 24 human controls. The differences between the patients and controls in the expression of E-selectin and P-selectin and relationship between their expression levels in nodular vasculitis lesions were assessed. Results The expression of E-selectin was detected in all the specimens of nodular vasculitis, and most of the expression level was moderately intensive (++); while E-selectin was absent in all of the control specimens. All the specimens of nodular vasculitis stained postivive for P-selectin, which was strongly (+++) expressed in most of the specimens; while only 2 control specimens stained weakly positive for P-selectin. A significant difference was observed in the expression of E-selectin and P-selectin between the specimens from the patients and controls (both P< 0.01), but not among specimens from patients at different ages and between specimens from female and male patients (all P > 0.05). In addition, the expression of E-selectin and P-selectin was well correlated with each other in lesions of nodular vasculitis (P < 0.01). Conclusions The expression of E-selectin and P-selectin is correlated with each other in lesions of acute nodular vasculitis, and is associated with the development of nodular vasculitis.