OBJECTIVE　To develop a method for the determination of theophylline in serum with concomitant use of ofloxacin.METHODS　Theophylline was serum was extracted with chloroform-hexane(7∶3,v/v) after some ammonium sulfate was added.It was re-extracted with NaOH solution (0.1 mol*L-1)and detected with a UV spectrophotometer at 275 nm and 299.5 nm respeitively.RESULTS　The method was linear over the range of 5.0～40.0 mg*L-1(r=0.9997,n=7).The average recovery of methodology was 100.3%(RSD=1.2%),and the extraction recovery from serum was 87.7%(RSD=4.6%).CONCLUSION　This practical method for the determination of serum theophylline can eliminate the interference of ofloxacin and other antibiotics.

Objective To evaluate the changes of left ventricular (LV) bulk rotation and untwisting in transplanted hearts using 2-dimensional speckle tracking imaging(STI).Methods Basal and apical LV short-axis images were acquired in 15 heart transplant recipients 3 months post surgery(HT group) and 56 healthy control subjects.Basal and apical rotation versus time profiles were drawn using 2-dimensional STI software.Appropriate values were chosen from the dataset obtained and compared between two groups.Results ①Compared with the control group,the heart rate,anterior-posterior diameter of left atrium,enddiastolic interventricular septum thickness,left ventricular posterior wall thickness,isovolumic relaxation time and E/e ratio were significantly increased,e and a values were decreased significantly in HT group (P ＜ 0.05).② No significant difference was noticed in the peak degrees of LV bulk rotation,the degrees of LV bulk rotation at the time of aortic valve closure and mitral valve opening (P =0.700,0.984,0.495,respectively) between 2 groups.In both groups,systolic rotation reached its maximum at end-systole [(96.1 ± 8.4) ％ in HT group vs (100.5 ± 6.3) ％ in control group,P =0.065].③Significant decreases in untwisting rate and trend untwisting variables were observed in the HT group(P ＜0.001).Conclusions 3 months after transplanted,left ventricular bulk rotation of cardiac allografts remained normal,and significant decreases in both untwisting rate and trend untwisting variables showed that the diastolic function of cardiac allografts was impaired.

Objective To explore the value of two-dimensional strain echocardiography for quantitative assessing the change of regional left ventricular myocardial function in rats following acute myocardial infarction. Methods Sixty Wistar rats were randomly divided into two groups. The study group consisted of 50 rats with occlusion of LAD for 30-45 minutes and the sham-operated group consisted of 10 rats without occlusion of LAD. Echocardiography were performed before operation, which was defined as baseline, and 1 week, 4 weeks and 8 weeks after operation. Left ventricular internal diameter at diastole ( LVIDd) and systole < LVIDs), fractional shortening( FS), ejection fraction (EF) and left ventricular mass(LVM) were measured by anatomical M-model echocardiography. High frame rate two-dimensional images were recorded in the left ventricular short-axis views at the papillary muscle level. Peak systolic radial strain(PRS) and circumferential strain(PCS) of each segment were measured using 2-dimensional strain software. The rats were sacrificed and the infarcted size of each segment was measured using triphenyl tetrazolium chloride (TTC) after echocardiography was performed. Fibrosis of left ventricular myocardium was displayed using Van Gieson stain in 1 weeks after infarction. Results Based on the TTC findings,the left ventricle of the study group was divided into three regions:infarcted,peri-infarct and remote myocardial regions. Van Gieson stain showed fibrosis existed in all the three regions. Compared with baseline and sham-operated group, PRS and PCS of infarcted, peri-infarct and remote myocardial regions of the study group significantly decreased within 1 week after operation ( P <0. 01) and persisted for 8 weeks. PCS and PRS of infarcted, peri-infarct and remote myocardial regions of the study group in 4 weeks and 8 weeks after operation showed no significant difference when compared with those in 1 week after operation ( P >0. 01). Compared with baseline and sham-operated group,LVIDd,LVIDs and LVM of study group all increased significantly ( P <0. 05) in 4 weeks and 8 weeks after operation,and FS and EF reduced significantly ( P <0. 05). Two-dimensional strain obtained in interobserver and intraobserver both showed high agreement. Conclusions Two-dimensional strain echocardiography can assess regional function of myocardium with different perfusion in rats following acute myocardial infarction, and provides a sensitive and reliable method to follow up the process of left ventricular remodeling after myocardial infarction.

Objective To investigate the location and expression of hypoxia inducible factor (HIF) subunits in the remnant kidney of 5/6 nephrectomy rats. Methods Remnant kidneys were produced in adult male SD rats by 5/6 nephrectomy. The renal function and histopathological changes were evaluated at week 1, 2, 4, 6, 8 and 12 after operation. Tissues of remnant kidneys were collected to detect the location and expression of HIF-1α and HIF-2α by immunohistochemistry staining and Western blotting. The mRNA levels of HIF targeted genes vascular endothelial growth factor (VEGF) and heme oxygenase-1 (HO-1) were determined by RTPCR. Results (1) 5/6 nephrectomy rats underwent one week of acute renal failure at first[Scr (122.8±22.1) μmol/L] and then developed compensative chronic renal failure [(66.0±3.7)-(66.4±8.4) μmol/L], but the level of Scr increased quickly after week 6 [(66.4±8.4)-(127.8±22.7) μmol/L],concomitantly with progressive tubulointerstitial fibrosis in remnant kidney cortex. (2) In cortex, HIF-1α was expressed only in the atrophic and dilated tubular cells while HIF-2α was located in endothelial, interstitial fibroblasts, and vascular smooth muscle cells. The semiquantitative results of imunohistochemistry and Western blotting revealed that HIF-1α and HIF-2α were both gradually up-regulated during the early stage of remnant kidney, peaked at week 4 and 6, and then gradually down-regulated. (3) The mRNA levels of HIF targeted genes VEGF and HO-1 transiently peeked at week 4 and 6, and then decreased gradually. Conclusions The increased stabilization of HIF-αprotein and transcription of HIF targeted genes at the early stage of this model is a compensation reaction towards hypoxia. The mechanism of decreased expression of HIF-α at the end stage of chronic kidney disease deserves further investigation.

ObjectiveTo find specific biomarkers related to HAART treatment in plasma samples of AIDS patients for clinical therapeautic efficacy evaluation and guidance for the prognosis of HIV treatment.MethodPlasma samples of AIDS patients were collected from Infectious Disease Department 1 of Shanghai Public Health Clinical Center in June of 2008 to February of 2009,including 11 successfully HAART treated cases (HIV load ＞ 50 copies/ml) and 11 unsuccessfully HAART treated cases (HIV load ＜50 copies/ml).Patients' age ranged from 22 to 63.Plasma samples were treated by Bio-rad AurumTM Serum Protein Mini Kit to remove high abundant proteins:albumin and immunoglobulin were removed.The treatedplasmaproteinswereseparatedbytwo-dimensionalelectrophoresisandanalyzedby electrophoretogram using Imagemaster software to find differentially-expressed proteins related to therapeutic efficacy.After digestion by trypsin,the differentially-expressed proteins were identified by online reversed-phasenano-flow liquid chromatography coupled with electrospray ionization ion trap mass spectrometry.ResultsLow abundant proteins were efficiently enriched after the AurumTM Serum Protein Mini Kit treatment.Six differentially-expressed proteins were detected while comparing successfully and unsuccessfully HAART treated group.These proteins were accurately identified by tandem Mass spectrometry (MS), including serum transferrin, serum β-fibrinogen, etc.ConclusionsOur proteomic research revealed that the differentially-expressed proteins such as transferrin,which is related to plasma virus loading in AIDS patients in the process of treatment,might be potential biomarkers evaluating HAART therapeutic efficacy.

Objective To explore the differential liver plasma membrane (PM) proteins that may be related to the occurrence,development and reversal process of hepatitis and to understand the pathogenesis of hepatitis and the new drug targets by performing a comparative proteomics research of liver PM between hepatitis B surface antigen (HBsAg) transgenic mice and wild-type C57 mice.Methods A 6-month-old HBsAg transgenic mouse model was established.The pathological examination was performed to observe the pathological changes of transgenic mice and wild-type C57 mice.The PM from liver tissue of 6-month-old transgenic mouse and the control mouse were purified through twice sucrose density grade centrifugation combined with second antibody magnetic bead enrichment.The purity of extracted PM was verified by Western blot.Differential proteome expression analysis was performed by using two-dimensional electrophoresis (2-DE) and ImageMaster software analysis.The differentially expressed proteins were lysed by trypsin and identified by liquid chromatography combined with tandem mass spectrometry (LC-MS/MS) analysis.Results The pathological examination results showed that hepatitis was observed in the transgenic mouse group,while no abnormity was found in the controls.The PM was successfully enriched and the mitochondria contamination was reduced by sucrose density grade centrifugation combined with second antibody magnetic bead purification treatment.Thirty differential mice liver PM protein spots were visualized,in which 11 non-redundant proteins were successfully identified by LC-MS/MS in transgenic mouse group,including 9 up regulated protein spots and 2 down-regulated protein spots.These differentially expressed proteins included keratin,cardiac Ca2+ release channel,cytochrome B5,ATP synthase subunit alpha,etc.Conclusions A batch of HBsAg gene expression related differential proteins are identified in mouse liver plasma.These proteins might be new drug targets for anti-HBV treatment.This study will guide further investigation on the mechanism of HBV infection induced hepatitis.

Objective To investigate human plasma glycoproteomie changes related to human immunodeficiency virus (HIV) infection,and to identify glycoproteins with potential anti-HIV activity or anti-HIV drug targets. Methods Plasma proteins with lower abundance were enriched through affinity purification to remove albumin and IgG in clinical samples (HIV-positive patient, n= 10, and healthy controls, n= 20). Proteins were separated by two-dimensional electrophoresis (2-DE) and stained by Pro-Q emerald glycoprotein stain kits. The 2-DE image was analyzed by ImageMaster software to find differential glycoproteins. Furthermore, the depleted HIV-positive and healthy control plasma proteins were digested by PNGase F. Glycoproteins were deglycoliszed, separated by 2-DE and analyzed by ImageMaster software. Differential glycoproteins were identified by liquid chromatography combined with high capacity ion trap mass spectrometry (HCT). Results The pretreatment of HIV-positive plasma prior to 2-DE could efficiently remove the high aboundant albumin and IgG in plasma and improve the detection of proteins with low-abundance. High revolution 2-DE gel images of glycoproteins from HIV positive and healthy control plasma samples were obtained. Glycoproteins were successfully deglycolized through PNGase F treatment. Thirteen differential glycoproteins were identified by liquid chromatography combined with mass spectrometry. These proteins included alphalantitrypsin precursor and serine/threonine-protein kinase N1. Conclusions Potential HIV infection related proteins,such as alphal-antitrypsin precursor are successfully identified. Our study may offer some help to understand the molecular mechanism of HIV infection and select new drug targets for preventing HIV infection.

Objective To investigate the value of two-dimensional speckle tracking imaging (2D-STI) in assessing left ventricular global and regional myocardial function in normal rats. Methods Echocardiography was performed during low-dose dobutamine stress in 20 normal adult male Wistar rats. High frame rate of 2D images were recorded from the left ventricular short-axis views at the papillary muscle level. Peak systolic radial strain (PRS) and circumferential strain (PCS) of each segment, left ventricular global peak systolic radial strain (G_(PRS)) and circumferential strain (G_(PCS)) were measured at EchoPAC work station. Left ventricular internal diameter at diastole (LVIDd), systole (LVIDs), fractional shortening (FS) and ejection fraction (EF) were measured with anatomical M-model echocardiography. Dynamic changes of each index during stress experiment were observed. The correlations between EF and left ventricular global two-dimensional strain (G_(PRS), G_(PCS)) were analyzed respectively. Results PRS was similar in all segments of mid-ventricular in short-axis view (P>0.05) at baseline, while PCS of each segment showed heterogeneity, with the anteroseptal and anterior wall showing the largest value and the inferior wall showing the lowest value (P<0.05). G_(PRS), G_(PCS), PRS and PCS of each segment increased correspondingly when the dobutamine dose increasing, as well as FS and EF, whereas LVIDd and LVIDs decreased. When dobutamine dose was 10.0 μg/(kg·min), these parameters were not significantly different compared with those of 5.0 μg/(kg·min) (P>0.05). Conclusion With good reproducibility, 2D-STI can noninvasively and easily assess global and regional left ventricular myocardial deformation of normal rats at rest and during low-dose dobutamine stress.

Objective To investigate the prevalence and the related factors of allergic rhinitis in Xinjiang College students.Methods Five universities'students in Urumqi were chosen as the research subjects,the question-naire survey was conducted by the method of cluster sampling,and the results were analyzed.Results 1 609 ques-tionnaires were valid from 2 000 questionnaires.The prevalence rate of allergic rhinitis was 36.8% in the five univer-sities in Urumqi.Prevalence rate increased with grade.The prevalence rates of drinking,regular cleaning nasal cavity, scrubbing their nose in allergic rhinitis were 43.2%,40.7%,53.8% respectively,which were higher than those in not drinking,not regular cleaning nasal cavity,not scrub nose,the differences were statistically significant (χ2 =6.945,9.679,100.845,all P <0.05 ).There was no significant differences in the prevalence of allergic rhinitis among college students and gender,nationality and BMI(all P >0.05).Conclusion Allergic rhinitis not only closely related to the grades and the environmental factors,but also has some connections with living habits(drink,clean nasal cavity and scrub noses).

Objective To study correlation between systemic sclerosis(SSc)and plasma D-dimer and to reveal the probably rules of the fibrinolytic systems in SSc.Methods In 2013 January to 2014 January,cellected 32 patients with SSc and 35 with healthy controls to detected level of plasma D-dimer.Logistic and t student test were used for statistical analysis for correlation between SSc and pulmonary artery hypertension.Results When compared to healthy controls (0.28±0.04μg/ml),the level of plasma D-dimer were significantly increased in SSc patient (0.31±0.05μg/ml,t=1.997,P=0.008).After stratifying SSc patients according to disease subset,whereas patients with diffuse subset displayed substantially increased values (0.41±0.06μg/ml,t=2.051,P<0.001).The level of plasma D-dimer was associated with pulmonary artery hyper-tension (OR=4.38,95%CI=2.59~8.91,P=0.008).Conclusion Demonstrated that SSc patients with diffuse subset are characterized by increased plasma D-dimer values,reflecting a potential activation of fibrinolytic cascaded,which might finally predispose these patients to thrombotic complications and pulmonary artery hypertension.