BACKGROUND: Breviscapine, extracted from a Chinese herb Erigeron breviscapus (Vant.) Hand-Mazz (Bre), has remarkable activities against platelets and thrombus formation. It can protect against ischemic brain damage through eliminating free radicals and apoptosis.OBJECTIVE: To study the protective effects of Breviscapine on brain damage in rats after ischemia-reperfusion injury, taking MgSO4 as control.DESIGN: Randomized controlled study and analysis of variance.SETTING: College of Life Science and Technology, Xiaogan University.MATERIALS: This experiment was conducted in the College of Life Science and Technology of Xiaogan University between May 2004 and November 2004. Forty male Wistar rats were selected and randomized into five groups: sham-operation group, cerebral ischemia-reperfusion (IR)group, MgSO4 group, 50 mg/kg Breviscapine group and 75 mg/kg Breviscapine group with 8 in each group.METHODS: Rats were subjected to cerebral ischemia-reperfusion induced by inserting a nylon thread into the internal carotid artery to block the origin of middle cerebral artery and removing the thread later. Normal saline of 20 mL/kg was administrated in sham-operation group and IR group, while 50 mg/kg, 75 mg/kg Breviscapine and 30 mg/kg MgSO4 were administrated in other groups respectively 10 minutes after the onset of ischemia. Neurological scoring was performed 1 hour, 2 hours, 5 hours and 23 hours after cerebral ischemia-reperfusion (Five points in total: 0 as unobvious neurological symptom; 1 as inability to completely extend left anterior claw; 2 as rotation toward the left; 3 as inclining to the left side atwalking; 4 as inability to walk by itself. The higher scale, the more severe the behavioral disturbance was). Brain infarcted area was assayed 1 hour and 23 hours after cerebral ischemia-reperfusion, and was expressed as the percentage of stained area to non-stained area. Neuron apoptosis was detected by the terminal deoxynucleotidyl transferase (TdT)-mediated dUTPfluorescence nick end labeling (TUNEL) method to assess DNA fragments of apoptotic cells 1 hour after cerebral ischemia, and 2 hours, 5 hours and 23 hours after reperfusion. The apoptotic cells were counted under the fluorescence microscope. The percentage of apoptosis cells (%) = (apoptotic neurons ÷ hippocampal neurons) ×100%. The protein expression of caspase-3 was detected by immunohistochemical staining. The positive cells were identified and counted under the light microscope. The percentage of positive cells of caspase-3 (%) = (positive neurons of caspase-3 ÷ hippocampal neurons) ×100%.cells of caspase-3 1 hour after cerebral ischemia, and 2, 5 and 23 hours after reperfusion in each group.scoring: Neurological score of rats in 50 mg/kg, 75 mg/kg Breviscapine groups and MgSO4 group was 1.2±0.4, 0.5±0.4, and 1.3±0.4, respectively,which were all lower than that in IR group [(2.2±0.6)] 23 hours after cerebral ischemia-recirculation (F=6.09, P=0.001). However, it was lower in farcted area in rats of 50 mg/kg and 75 mg/kg Breviscapine groups and MgSO4 group was (0.18±0.03)%, (0.10±0.02)%, and (0.28±0.02)%, respectively, which were all lower than that of IR group [(0.43±0.05) %] 23 hours after cerebral ischemia-reperfusion (F=2.3, P=0.001). It was smaller in hippocampal apoptotic cells: It was (27.2±4.3) %, (20.6±3.6)%, and (35.4±5.5)% in 50 mg/kg and 75 mg/kg Breviscapine groups and MgSO4 group,which were all lower than that of IR group [(60.4±6.2)%] 23 hours after cerebral ischemia-reperfusion (F=6.17, P=0.000 7). It was lower in Brevispositive cells of caspase-3: It was (34.2±5.3)%, (21.6±3.5)%, and (47.4±4.5)% in 50 mg/kgand 75 mg/kg Breviscapine groups and MgSO4 group,which were all lower than that of IR group [(76.3±6.2)%] 23 hours after cerebral ischenia-reperfusion (F=6.88, P=0.000 1). It was lower in Breviscapine group than in MgSO4 group (P ＜ 0.01).CONCLUSION: Breviscapine can protect brain damage induced by cerebral ischemia-reperfusion by markedly decreasing the neurological score,area of cerebral infarction, number of hippocampal apoptotic cells and number of positive cells of caspase-3, and it has better effect than MgSO4.

Objective To observe the changing levels of serum sFas before and after intravenous i mmunoglobulin (IVIG) treatment of incomplete Kawasaki disease (IKD),to explore the roles of sFas in the pathogenesis of IKD and IVIG treatment mechanism.Methods Thirty eight cases of IKD children were selected as experimental group and 20 examples of the same age of children as the control group.The IKD children were treated by IVIG in combination with aspirin (ASP) ; and blood test was performed before treatment,3 days after treatment,and 14 days after treatment,respectively.Dual-resistant sandwich enzyme-linked immunosorbent assay (ELISA) was used to detect the levels of serum sFas,plasma Fibrinogen (PT-D),d-dimer (D-D),and c-reactive protein (CRP).Results The levels of serum sFas,PT-D,D-D,and CRP were significantly higher than the control group for IKD children before treatment[(0.55 ± 0.14)ng/L vs (0.24 ±0.04) ng/L,(552.3 ± 147.2) mg/dl vs (277.3 ±82.5)mg/dl,(649.0 ±201.6) μg/L vs (315.4 ±91.8)μg/L,and(72.2 ±28.7)mg/L vs (7.2 ±2.9)mg/L; t' =12.41,9.11,8.64,13.82;All P ＜ 0.05] ;3 days after treatment,compared with those before treatment and control group,the sFas level of IKD children at the third day after treatment was significantly decreased compared to that before treatment and control groups,respectively [(0.43 ± 0.09) ng/L vs (0.55 ± 0.14) ng/L,(0.24 ± 0.04) ng/L,F =47.624,All P ＜0.05] ;For the level of sFas at the 14th day after treatment,no statistical significance was found between IKD children and the control group[(0.24 ±0.05) ng/L vs (0.24 ±0.04) ng/L,t =0.596,P ＞ 0.05].Conclusions The abnormally increased serum sFas level before IVIG treatment suggests that dysfunction of apoptosis be involved in the pathogenesis of the IKD.Intravenous immunoglobulin treatment may be involved in the apoptosis process.

Objectives To explore the change of interleukin-17 (IL-17) in Kawasaki disease (KD). Methods Fourty KD pediatric patients, among them 12 patients with echocardiographic abnormalities in acute phase, 25 age-matched non-KD patients were enrolled. The level of serum IL-17 was measured by enzyme linked immunosorbent assay in acute and convalescent phase of KD patients and non-KD patients. At the same time, C-reactive protein (CRP), globulin, albumin were detected. Results In acute phase of KD patients, the level of serum IL-17 were signiifcantly higher than that in convalescent phase of KD patients and non-KD patients (P<0.05). The level of serum IL-17 was no signiifcant differences in convalescent phase of KD patients and non-KD patients (P>0.05). In acute phase of KD patients with echocardiography abnormalities, the level of serum IL-17 was signiifcantly higher than that with non-echocardiography abnormalities (P<0.05). The level of serum IL-17 in acute phase of KD patients were positively correlated with CRP and globulin (r=0.750, 0.750, P<0.05), and negatively correlated with albumin (r=-0.779, P<0.05). Conclusions IL-17 may be involved in KD immune pathogenesis. Serum IL-17 is one of the activity index of KD, which associ-ated with cardiovascular damages.

The nitrate-N content in KNO3 solution and soil was rapidly predicted using techniques of mid-infrared spectroscopy, in which 15 NO-3 and 14 NO-3 were distinguished and predicted. The results showed that the characteristic band of nitrate in solution and soil was 1200-1500 cm-1 , and compared with 14 NO-3 , the red shift of characteristic band of 15 NO-3 was about 35 cm-1 . In the characteristic band of nitrate, absorption band increased with the nitrate nitrogen concentration with less interference absorption. The linear regression was made between the first principal component of characteristic band and nitrate-N content, and correlation coefficient was more than 0 . 9840 , indicating that the technique of mid-infrared attenuated total reflectance spectroscopy could be applied for rapid monitoring of nitrate in solution and soil. Meanwhile, based on the red shift characteristic of 15 NO-3 absorption band, the method of partial least squares were involved to predict the nitrate-N of different N-isotope labeled in solution and soil, resulting that all the prediction models reached excellent levels. For 14 NO3-N and 15 NO3-N in solution, the correlation coefficients ( R2 ) were 0. 9980 and 0. 9982 respectively, and ration performance to standard deviations ( RPD ) were 6. 44 and 4. 76, respectively. While for 14 NO3-N and 15 NO3-N in soil, the correlation coefficients ( R2 ) were 0. 9794 and 0. 9679, and RPD were 5. 75 and 4. 78, respectively. Therefore, the technique of mid-infrared attenuated total reflectance spectroscopy can be applied for rapid monitoring different N-isotope labeled nitrate in solution and soil, to provide a new in situ and fast time method to study nitrification process in soil.

Objective To evaluate the potential value of IgG antibodies against recombinant PPE65 protein (rPPE65) of Mycobacterium tuberculosis in serodiagnosis of tuberculosis.Methods The gene encoding PPE65 protein of M.tuberculosis was cloned into the PET-28a vector and then expressed in Escherichia coli.The rPPE65 was purified with Ni-NTA affinity and ion exchange chromatography.After dialysis renaturation, the concentration of rPPE65 was determined using Lowry assay.ELISA was used to detect the levels of specific IgG against rPPE65 and recombinant PstS1 protein (rPstS1) in sera from 144 patients with pulmonary tuberculosis (PTB patients), 144 health controls, and 56 patients with non-tuberculosis pulmonary diseases.ROC curves were used to determine cut-off values with the results of IgG antibodies against rPPE65 and rPstS1 for 144 PTB patients and 97 controls with negative PPD skin test.The sensitivity, specificity, positive predictive value (PPV), negative predictive value (NPV), and accuracy of rPPE65 and the combination of rPPE65 and rPstS1 were counted.Results The PPE65 protein of M.tuberculosis was successfully expressed in E.coli. The purity and concentration of rPPE65 were 95% and 0.5 mg/ml, respectively.ROC analysis showed that the cut-off of ELISA using rPPE65 was 0.64.The sensitivity, specificity, PPV, NPV, and accuracy of rPPE65 were 34.7%(50/144), 93.5%(187/200), 79.4%(50/63), 66.5%(187/287), and 68.9%(237/344), respectively.The sensitivity, specificity, PPV, NPV, and accuracy of the combination of rPPE65 and rPstS1 were 59.0%, 91.0%, 82.5%, 75.5%, 77.6%, respectively.Conclusions The rPPE65 of M.tuberculosis appears to be a candidate antigen for serodiagnosis of tuberculosis.Detection of IgG antibodies against the combination of rPPE65 and rPstS1 can increase the sensitivity of serological test for tuberculosis.

Objective To investigate the state of perceived discrimination,mental health knowledge and self-discrimination of the public,and analyze the relationship among these variables,and explore the mediating effect of mental health knowledge between the perceived discrimination and self-discrimination.Methods A total of 1 088 residents in Guangzhou City were assessed with the Perceived Devaluation-Discrimination Scale (PDDS),Attitudes about Mental Illness Associated Stigma Scale-Chinese Version (AMIASS-C) and Mental Heahh Knowledge Questionnaire (MHKQ).Results The average score of PDDS was 2.63±0.35.The rate of mental health knowledge was 72.4％(788/1 088) and the score of AMIASS-C was 2.58±0.45.The perceived discrimination,mental health knowledge and self-discrimination were significantly correlated with each other,r=0.320,-0.240,P＜0.01.The mental health knowledge played a negative mediating role between perceived discrimination and self-discrimination.Conclusions The mental health knowledge was a mediator between perceived discrimination and self-discrimination.

Objective To research the effects of progressive time phase selective light (PTPSL)therapy on shift work disorder patients.Methods 24 shift work disorder patients were randomlv divided into PTPSL group and control group.Both groups received treatment for 12 days including 6 days during the night shift time and 6 days during the day shift time continuously.The clinical effects were evaluated by Karolinska Sleepiness Scale (KSS),Leeds sleep evaluation questionnaire (LSEQ) and core body temperature rhythm.Results (1) During the night shift time,PTPSL group patients were better in score LSEQ and KSS(discerned for 39.80 ± 2.89,6.59 ±0.48) than control group' s(discerned for 36.86 ± 3.88,6.98 ± 2.03) ; temperature curve peak phase in PTPSL group moved backward greater than that of control group((8.35 ± 0.82) h vs (7.45 ± 1.05) h) ; and the difference was statistical significance(P ＜ 0.05).During the day shift time,PTPSL group patients were better in score LSEQ and KSS(discerned for 45.57 ± 2.77,6.33 ± 0.48) than control group' s (discerned for 40.35 ± 4.19,6.82 ± 1.03),temperature curve peak phase in PTPSL group moved forward greater than that of control group((7.37 ± 0.94)h vs (6.31 ±1.37)h),and the difference was statistical significance(P＜0.05).Conclusion PTPSL can adjust shift sleep disorder patients' mussily sleep rhythm system in order,and make the inner rhythm svstem and external schedules tend to be consistent,and provides a new treatment choice for shift sleep disorders.

Objective To study the expression levels of N-terminal pro-brain natriuretic peptide (NT-proBNP),serum albumin of Kawasaki' s disease (KD),incomplete Kawasaki' s disease (IKD),and children whose fever were unexplained and to explore the clinical significance of the levels of NT-proBNP and serum albumin in the early diagnosis of IKD.Methods The levels of NT-proBNP of 246 cases of KD (KD group),61 cases of IKD (IKD group) and 301 cases of children with unexplained fever (fever group)were measured by the enzyme-linked fluorescence analysis (ELFA) at the day of admission,meanwhile,the levels of albumin were tested in KD,and IKD children were underwent ECG and echocardiography.Based on the test results,patients were further divided into the group with cardiovascular damage and the group without cardiovascular damage.SPSS 19.0 was used for statistical analysis.The t test was used to compare the parameters between each group,the variance analysis and association analysis were carried out with Pearson's correlation analysis.The ROC curve analysis was done to identify the cardiovascular damage threshold.Results ① The level of plasma NT-proBNP of the KD group,the IKD group was significantly h igher than the fever group [(789.1±4.7) ng/L,(824.8±4.4) ng/L vs (92.5±2.3) ng/L,F=230.736,all P＜0.05];② The level of albumin of the KD group and the IKD group was significantly lower than that of the fever group [(33.9±2.8) g/L,(33.8±3.1) g/L vs (40.8±3.6) g/L,F=355.648,all P＜0.05]; ③ The levels of NT-proBNPs between the cardiovascular damage group and the groups without cardiovascular damage among the KD group,and those of the IKD groups were compared.In the KD group,the NT-proBNPs level of the two subgroups was (2948±3) g/L (n=103) vs (305±3) g/L,n=143; while in the IKD group,the NT-proBNPs of the two subgroups was (1454±4) g/L (n=38) vs (323±4) g/L (n=23).The dif-ferences were statistically significant (t=16.464,4.356,all P＜0.05).④ The plasma NT-proBNP level higher than 933.5 ng/L was identify as the indicator for cardiovascular damage in both KD and IKD children.Its sensi-tivity was 88.1％,and its specificity was 89％.⑤ When the level of NT-proBNP was higher than 250 ng/L,the sensitivity for diagnosis in the KD,the IKD was 80.9％,85.2％ respec-tively,and the specificity was 85.7％.When the level of NT-proBNP was higher than 250 ng/L and that of albumin was lower than 35 g/L,the sensitivity for diagnosis of KD,IKD was 67.5％,70.5％ respectively,the specificity was 99.7％.Conclusion The level of plasma NT-proBNP (＞250 ng/L) accompanied by decreased albumin (＜35 g/L) may be specific markers for early diagnosis of IKD.In addition,the level of NT-proBNP ≥933.5 ng/L can be used as a diagnostic threshold,which has good sensitivity and specificity for identifica-tion of cardiovascular damage in the KD and IKD in children.

The aim of the study was to investigate the effect of oleic acid(OA)and isosorbide mononitrate(ISMN)on the phase diagrams of glyceryl monooleate(GMO)liquid crystalline systems and release of ISMN from them. Liquid crystalline systems of GMO/H2O, GMO/H2O/ISMN and GMO/H2O/OA were prepared and their phase diagrams were plotted. Investigation of water absorption of GMO and ISMN release profiles shown that the release mechanism of ISMN was diffusion-controlled. Increased drug loading accelerated the release rate. However, the increased loading of OA decreased ISMN release. The result could offer practical basis for drug loading and the selection of additives in GMO liquid crystalline system.

OBJECTIVETo revalidate the conversion factor（CF）for the conversion of BCR-ABL （P210）transcript levels to the international scale（BCR- ABLIS）in chronic myeloid leukemia（CML） which validated before.

METHODSPeking University People's Hospital（PKUPH）prepared the exchange samples for revalidation of CFs of 15 laboratories which validated nine or eighteen months ago. The fresh BCR-ABL（P210）（+）bone morrow or peripheral blood nucleated cells were diluted with BCR-ABL （P210）（-）cells to achieve different BCR- ABL levels, totally 16 sets and 24 samples per set were prepared. TRIzol reagent was added in each tube. Each laboratory tested BCR-ABL transcript levels of one set of samples. Agreement between BCR-ABLIS of each laboratory and PKUPH was assessed by the Bland- Altman method. For laboratories which did not meet the criteria of revalidation, linear regression equation was derived after the samples with maximum BCR-ABL deviation were removed until R²>0.98, then new CF was calculated.

RESULTS10 laboratories met the revalidation criteria with both bias within ±1.4 fold and 95% limits of agreement within ±6 folds, and their CFs still could be used for accurately conversion of BCR-ABLIS. New CFs were recalculated as of 1.8-6.3 folds of their previous CFs in 5 laboratories not met the criteria.

CONCLUSIONRevalidation of CF by sample exchange among laboratories was necessary for accurate and continuous application of BCR-ABLIS, which not only tested the validity of CF acquired before but also calculated new available CFs for those with invalid CFs.