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 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.

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.

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 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 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 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 investigate the effects of changes of miR-126 and spouty related EVH,domain containing proteinl(SPRED1) after transient ischemic attack(TIA)on prognostic value for pathogenesis of secondary cerebral infarction.Methods Retrospective analysis of the clinical data of 106 patients with TIA was performed.The expression levels of miR-126,SPRED1 and vascular endothelial growth factor(VEGF)in peripheral blood were detected at 3 h,6 h and 12 h after TIA onset respectively.The specificity and sensitivity of miR 126 and SPRED1 in the diagnosis of TIA were analyzed.The miR-126 and SPRED1 levels versus ABCD2 score were compared for evaluating their predictive value in the diagnosis of secondary cerebral infarction within 30 days after TIA onset.Results The miR-126 level was declined after TIA onset at 3 h(9.41±1.04),especially at 12 h(6.59 ±2.78),versus in healthy control (9.35±1.76)(t =-7.764,P=0.000).The SPRED1 level after TIA onset was increased at 3 h(58.05 ± 17.53)pg/L,12 h(82.64 ± 18.60)pg/L versus in healthy control(52.38 ± 13.24)pg/L(t=12.374,P =0.000).A closely negative correlation was found between levels of miR 126 and SPRED1 at 12 h point but not at 3 h and 6 h(r=-0.278,P=0.004).Both miR-126 and SPRED1 levels at 12 h after TIA were implied to sensitivity and specificity evaluation.Additionally,VEGF was significantly increased at 3 h (345.61 ± 76.76) pg/L,6 h (461.65 ±103.87)pg/L and 12 h (519.22 ± 103.55)pg/L after TIA onset as compared with healthy control (107.77± 26.04) pg/L(t =26.569,29.756,34.699,all P =0.000).The decrease of miR-126 and increase of SPRED1 at 12h after TIA indicated high incidences of cerebral infarction but their significance was less than ABCD2 score.Combination of miR 126,SPRED1 and ABCD2 score significantly improved the prediction for cerebral infarction(Z=2.105,P =0.035).Conclusions After the onset of TIA,levels of miR-126 and SPRED1 expression in combination of ABCD2 score can improve predictive value for cerebral infarction development.

This study evaluated the change in regional left ventricular myocardial function in rats following acute occlusion of the left anterior descending coronary artery (LAD) by using two-dimensional speckle tracking imaging (2D-STI). Sixty Wistar rats were randomly divided into two groups, a myocardial infarction (MI) group, in which 50 rats were subjected to LAD occlusion for 30-45 min, and a sham-operated (SHAM) group that contained 10 rats serving as control. Echocardiography was performed at baseline and 1, 4 and 8 week(s) after the operation. High frequency two-dimensional images of left ventricular short axis at papillary muscle level were recorded. Peak systolic radial strain (PRS) and circumferential strain (PCS) were measured in the mid-ventricle in short-axis view by using EchoPAC workstation. 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. Infarct size was measured using triphenyl tetrazolium chloride (TTC) staining 1 week and 8 weeks after the operation. Fibrosis of left ventricular myocardium was displayed using Van Gieson staining 1 week after the infarction. In terms of the TTC staining results, the left ventricle fell into three categories: infarcted, peri-infarcted and remote myocardial regions. Compared with those at baseline and in the SHAM group, (1) PRS and PCS in the infarcted, peri-infarcted and remote myocardial regions were significantly decreased in the MI group within 1 week after the operation (P<0.05) and the low levels lasted 8 weeks; (2) Compared with those at baseline, LVIDd, LVIDs, FS, EF and LVM in the MI group showed no significant difference 1 week after the operation (P>0.05). However, LVIDd, LVIDs and LVM were increased significantly 4 and 8 weeks after the operation (P<0.05), and FS and EF were decreased substantially (P<0.05). Van Gieson staining showed that fibrosis developed in all the three myocardial regions to varying degrees. It is concluded that 2D-STI is non-invasive and can be used to assess regional function of myocardium with different blood supply in rats following acute occlusion of the LAD, and can be used as a sensitive and reliable means to follow up the process of left ventricular remodeling.

The effects of epigenetic modification on the differentiation of islet cells and the expression of associated genes (Pdx-1, Pax4, MafA, and Nkx6.1, etc) were investigated. The promoter methylation status of islet differentiation-associated genes (Pdx-1, Pax4, MafA and Nkx6.1), Oct4 and MLH1 genes of mouse embryonic stem cells, NIH3T3 cells and NIT-1 cells were profiled by methylated DNA immunoprecipitation, real-time quantitative PCR (MeDIP-qPCR) techniques. The histone modification status of these genes promoter region in different cell types was also measured by using chromatin immunoprecipitation real-time quantitative PCR methods. The expression of these genes in these cells was detected by using real-time quantitative PCR. The relationship between the epigenetic modification (DNA methylation, H3 acetylation, H3K4m3 and H3K9m3) of these genes and their expression was analyzed. The results showed that: (1) the transcription-initiation-sites of Pdx-1, MafA and Nkx6.1 were highly methylated in NIH3T3 cells; (2) NIH3T3 cells showed a significantly higher level of DNA methylation modification in the transcription-initiation-site of Pdx-1, Pax4, MafA and Nkx6.1 genes than that in mES cells and NIT-1 cells (P<0.05); (3) NIT-1 cells had a significantly higher level of H3K4m3 modification in the transcription-initiation-site of Pdx-1, Pax4, MafA and Nkx6.1 genes than that in mES cells and NIH3T3 cells (P<0.05), with significantly increased level of gene expression; (4) NIH3T3 cell had a significantly higher level of H3K9m3 modification in the transcription-initiation-site of Pdx-1, Pax4, MafA and Nkx6.1 genes than that in mES cells and with NIT-1 cell (P<0.05), with no detectable mRNA expression of these genes. It was concluded that histone modification (H3K4m3 and H3K9m3) and DNA methylation might have an intimate communication between each other in the differentiation process from embryonic stem cells into islet cells.