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

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.

AIM: To observe whether metallothionein plays a role in cardiac protective effect of basic fibroblast growth factor on anoxic/reperfusion (A/R) injury in cultured cardiomyocytes and study the possible mechanism of cardiac protection by bFGF.METHODS: The present study made the model of myocyte A/R injury after having a 24 h incubation by bFGF( 10-10、10-9、10-s mol/L) and bFGF( 10-9 mol/L) + PD098059 respectively. We measured the levels of MT and MDA in myocytes, and the changes of LDH and protein in cultured medium. We also counted the number of viable cell in groups. RESULTS: The contents of myocardial MT were significantly increased after treatment by bFGF. The levels of MT in I0-l0 mol/L、10-9 mol/L and 10-8 mol/L bFGF treated groups increased 54 %、 62%、 76% respectively, compared with the A/R group, and the number of viable cell were also greatly increased, LDH and protein leakage in cultured medium and MDA contents in myocyte were dramatically decreased in bFGF treated groups. All the protection were completely disappeared with the inhibition of MT production with PD098059, theinhibitor of mitogen- activated protein kinase(MAPK). CONCLUSION: MT involves in the protection of bFGF in cultured cardiomyocytes. It might be related with activation of MAPKase.

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 investigate the influence of a combination of Chinese herbs,Astragalus membranaceus var.mongholicus and A ngelica sinensis (A&A)on the production and elimination of reactive oxygen species (ROS)and the underlying mechanism during the process of renal interstitial fibrosis in the obstructive kidneys of rats with unilateral ureteral obstruction(uuo).Method Male Wistar rats were randomly divided into control,sham,UUO and UAA (UUO+A&A)groups.The rats in UAA group were administered with A&A(14 g/kg)by oral gavage once daily:the ones in sham and UUO group were given with equal volumes of water.Three days after setting up models,pathological injury of renal tissue was evaluated.Level of total antioxidant capacity(T-AOC)and activity of CuZn superoxide dismutase(CuZn-SOD)in renal homogenates were measured bv spectrophotometry.Expression of NADPH oxidase subunits p47-phox,p22-phox and nintrotyrosine was analyzed by Western blot. Results Severe interstitial inflammatory cells infiltration, mild tubular atrophy and interstitial fibrosis were found in UUO group, which were alleviated by A&A administration. Compared to sham group, T-AOC of UUO group was not significantly changed, but the expression of the nitrotyrosine, NADPH oxidase subunits p47-phox and p22-phox was increased significantly (P<0.05). After A&A administration, T-AOC level was increased (2.5±1.1 vs 1.5±0.5, P<0.05) and the expression of nitrotyrosine was decreased (P< 0.05) in UAA group compared with UUO group. Additionally, the expression of p47-phox was reduced at day 3 (P<0.05), accompanied with a reduced expression of p22-phox (P< 0.05). CuZn-SOD activity was not significantly changed among the groups. Conclusion The inhibition of A&A in NADPH oxidase subunits p47-phox and p22-phox, which is responsible for reduction of oxidative stress, is associated with the alleviation of renal fibrosis in obstructive rat kidney.

Objective To study the cardioprotection effects of heme oxygenase-1(HO-1)on cardiopulmonary resuscitation(CPR).Method Male Sprague-Dawley rats were asphyxiated for 9 minutes and resuscitated.Rats wefe randomly divided into 4 groups,namely,sham asphysiation group,CPR group,Hemin group and Heroin +ZnPP group(zinc protoporyphyrin IX).Resuscitated groups were further divided into two subgroups according to various intervals:6 hours and 24 hours after resuscitation.Hemodynamic was observed.Serum creatine phosphokinase-MB(CPK-MB)and lactate dehydrogenase were determined.HO-1 in heart tissue homogenates was assayed.Ultrastructure of rats hearts was examined.Statical evaluation was performed with analysis of variance.Results The mean blood pressure(MBP)in resuscitated groups was significantly reduced after resuscitation,hadn't any difference between supgroups.The scores of dp/dt 40 and-dp/dt were significantly decreased in CPR group and Hemin+ZnPP group after resuscitation(all P＜0.05),but dP/dt40 in Heroin group did nol differ significantly after resuscitation.and-dp/dt decreased only 0.5 hours and one hour after resuscitation and returned to baseline values two hours after resuscitation.The scores of dp/dt 40and-dp/dt in Heroin group at different intervals after resuscitation were significantly higher than those in CPR group and Hemin+ZnPP group(all P＜0.05).Serum CPK-MB and LDH in CPR group and Hemin+ZnPP group at 6 hours and 24 hours after resuscitation were significantly higher than those in Hemin group(all P＜0.05).The cardiac tissue ultrastructure of rats in Hemin group was more intact than that of CPB group and Hemin+ZnPP goup.HO-1 levels in heart tissue homogenates of Hemin group at 6 hours and 24 hours after resuscitation were significantly higher than those in CPR group and Heroin+ZnPP group(all P＜0.05).Conclusions HO-1 expression induced by Heroin can effectively improve post-resuscitation myocardial dysfunction,alleviate cardiac injury,keep the ultrassructure integrity of cardiac myocytes.It may be a new approach to treat myocardial dysfunction after resuscitation.

Objective To investigate imaging and clinical findings of severe adenoviral pneumonia in children.Methods The imaging and clinical findings of severe adenoviral pneumonia in three children were retrospectively analyzed. All cases underwent radiography and CT examinations of chest. Results Two patiens showed fairly homogeneous opacity with air bronchograms in segmental or lobar distribution in left lung,in company with volume decreased in one case. There was interstitial change in 1 case,pleural effusion in one case and heart shadow enlarged in 3 cases. All of patients had cough and fever (range,38.4℃～40.3℃). Physical examinations showed decreased breathing sound and crackles in all cases. Laboratory findings included abnormally elevated aspartate aminotransferase,lactic dehydrogenase, hydroxybutyric dehydrogenase and creatine kinase and positive adenovirus examination in all cases. Conclusion Lobar pneumonia in company with liver and heart dysfunction should be suspected as the severe adenoviral pneumonia in children.

AIM: To investigate the effects of Shuguan Capsule on myocardial blood flow (MBF) and myocardial oxygen consumption (MOC) in anesthetized dogs. METHODS: Coronary blood flow (CBF) of twenty-five thoracotomized dogs was detected by electromagnetic flow meter and the MBF was calculated. While the oxygen content in the artery (AO2 ) and in the coronary venous sinus (VO2) was determined with blood oxygen analysis.Moreover, the other cardiac hemodynamic parameters, such as heart rate (HR), femoral arterial blood pressure (BP), were observed by physiological polygraph. RESULTS: It was found that Shuguan Capsule (48.5 mg/kg and 194 mg/kg) could significantly increase the MBF, and then decease the coronary artery resistance. Furthermore, Shuguan Capsule could also lower the AO2, but increase the VO2, which led to the decreased MOC. CONCLUSION: Shuguan Capsule exhibits the effects to keep the balance between blood supply and oxygen consumption in the heart by modulating the coronary resistance and by reducing MOC in dogs.