Objective:To establish a detection method for residual toluene in diclofenac sodium raw material by GC. Methods:An HP-5 column (0. 53 mm × 30 m, 2. 65 μm) with nitrogen as the carrier gas was employed for the separation. The temperature of the column, injection port and FID detector was set at 50℃, 200℃ and 250℃, respectively. An internal standard method with isooctane as the internal standard was used to assay the residual amount after the sample was dissolved in DMSO. Results:The linear calibration curve was obtained over the range of 9. 27-111. 19μg·ml-1(r=0. 999 4). The mean recovery was 97. 27%(RSD=2. 80%,n=9). The LOD and LOQ was 0. 59μg·ml-1 and 1. 46μg·ml-1 , respectively. The residual contents of toluene in 3 batches of the products all met the relevant regulations. Conclusion:The established method is reliable and accurate, and can be used in the quality control of diclofenac sodium.

BACKGROUND: It is very important to evaluate vascularization of postoperative hydroxyapatite (HA) orbital implants in human eyes. Recently, methods for evaluating vascularization of HA orbital implants are limited; however, ultrasonic contrast is a novel technique to high-precisely measure blood flow.OBJECTIVE: To investigate the feasibility of ultrasonic contrast to evaluate the vascularization of HA orbital implants, and to compare with enhanced MRI.DESIGN, TIME AND SETTING: The comparative observation was performed at Departments of Ophthalmology and Ultrasound,Jiangyin People's Hospital between January 2008 and January 2009.PARTICIPANTS: Ten patients including 8 males and 2 females were treated with eye ball extirpation and HA orbital implants. The patients were aged 24-61 years, with the mean age of (45.0±10.2) years. HA orbital implants were crossly coated using anterior part of pedicle scleral flap.METHODS: PHILIPS IU22 color Doppler ultrasound was used in this study, and SonoVue (59 mg) was the major contrast medium. Sulphur hexafluoride was dissolved in saline to make suspension. A 2.4-mL suspension was injected through peripheral vein of elbow. Enhanced MRI was additionally used in this study: Siemens Magnetom Avanto 1.5T, standard head coil, horizontal axis, 2.0-3.0 mm thickness, no interval, FOV 160 mmxi80 mm, 256x256 matrix, and SE sequence T1WI (TR 500 ms, TE 12 ms).Dimeglumine gadopentetate injection was used as a contrast medium.MAIN OUTCOME MEASURES: Vascularization characteristics and degrees between ultrasonic contrast and enhanced MRI at different time points after implantation.RESULTS: Ultrasonic contrast showed a well dynamic contrast-enhanced image of the anterior part of HA in early stage of post-operation, and showed distributing instance and density of blood vessel. Later stage of post-operation (about 3-6 months),contrast-enhanced image was unconspicuous. The enhanced MRI showed a clear vascularization of HA orbital implants at the stage of post-operation.CONCLUSION: Ultrasonic contrast is a effective investigative way to evaluate vascularization of HA orbital implants in early stage of implantation, and it is limited at the later stage of post-operation than enhanced MRI.

Objective To analyze the detection rates,species distribution and drug-resistance of urinary fungal infection in elderly patients at Beijing Hospital from 2011 to 2013,in order to provide the basis for the reasonable clinical use of anti-epiphyte medicines.Methods Totally 263 patients with an average of 79.6 years old were collected from Beijing Hospital.The urine from freshly voided midstream or bladder puncture was collected under aseptic condition for fungal culture,then the strains of epiphytes were identified by using API 20C AUX.The drug sensitivity was tested with ATB fungus3.Results 263 strains of epiphytes were isolated from the 2 983 urine samples,of which 92 were C.tropicalis,85 were C.glabrata,77 were Candida albican,and 9 were other fungus candida.The rates of drug resistance to fluconazole were 14.1 ％ (13 strains),37.6 ％ (32 strains) and 15.6％ (12 strains),and to itraconazole were 16.3％(15 strains),35.3％(30 strains) and 9.1％(7 strains),respectively.All of the 263 strains were not found to have drug resistance to amphotericin.Conclusions The isolation rate of urinary fungal infections is 8.8％ in Beijing Hospital.The majority of the tested fungal are C.glabrata,C.tropicalis and Candida albican,the former has higher resistance rate to azoles,and the two latter have better sensitivity to azole,and all of them have the sensitivity to amphotericin.

Objective To study the effect of Na-2SeO-3 on experimental carcinogenesis of stomach and on p53 and p16 expression. Methods Weaning male Wistar rats were divided into four groups randomly:high selenium group(4mg/L),low selenium(2mg/L)group,experiment control group and normal control group.Wistar rat gastric cancer was induced by N-methyl-N′-nitro-N-nitrosoguanidine(MNNG,20mg/kg)given daily for 10days.Na 2SeO 3 was given by piped drinking before one week of MNNG administration.Rats were killed at the 43th week.The surface characters of gastric mucosa were observed with noked eyes.Histopathologic changes were observed by methods of HE staining and Alcian Blue Periodic acid Schiff reaction(AB-PAS).Changes of p53 and p16 were detected by SP immunohistochemical method.The immunohistochemical results were quantitatively analysed by image analysis.Statistical analysis was taken by SPSS. Results Dietary Na-2SeO-3(2mg/L,4mg/L)aggravated gastric erosion and hemorrhage and promoted intestinal metaphasia of gastric cancer (P0.05).Conclusion These results suggest that dietary Na-2SeO-3 by piped drinking might not decrease incidence of Wistar rat gastric cancer induced by MNNG.The mechanism may be associated with the mutations of p53 and abnormal expression of p16 gene.

Objective In this study,we elucidated the role of high mobility group box chromosomal protein 1 (HMGB1) in collagen-induced arthritis (CIA) rat and the antagonist role of ethyl pyruvate by using a rat model of CIA as the research object by comparing the expression of HMGB1 in normal control group,CIA model group and ethyl pyruvate group.Methods Thirty-six rats were randomly divided into 3 groups (n=12):normal control group,CIA group and ethyl pyruvate group.Then the 6 rats were dissected at the 6th,9th week respectively.Thc expression of HMGB1 was analyzed by immunohistochemistry and Pathology-image analysis software in the cytoplasma.The expression of HMGB1 mRNA with real time-polymerse chain reaction (PCR) was evaluate,and the HMGB1 expression of each group were compared with t-test.Results The immunohistochemical results of HMGB1 showed that the expression intensity in the normal control group,CIA model group and ethyl pyruvate group was 2.1±0.6,7.3±1.2,6.0±1.2 respectively at the 6th week; and 2.2±0.7,12.4±4.5,5.5±1.0 at the 9th week respectively.The HMGB1 mRNA real time-PCR results had shown that the relative quantification of the normal control group and CIA model group were 1,2.865,2.602respectively at the 6th week and 1.005,4.694,1.729 at the 9th week.At those two points, the HMGB1 expressions of HMGB1 antagonist group were significantly higher than those of the normal controls (P＜0.05).In addition,there was statistical significant difference(P＜0.05) in the HMGB1 expression when compared with the placebo group.Furthermore, when the degree of HMGB1 expression among the three groups was compared,the HMGB1 antagonist group was decreased significantly (P＜0.05).Conclusion The results has demonstrated that HMGB1 could induce inflammation in the synovial tissue of CIA rats,and has provided the rationale that HMBG 1 could be the target of treating rheumatoid arthritis (RA).The results of this study have shown that ethyl pyruvate could antagonize the effect of HMGB1.This finding may provide a new therapeutic target for the treatment of RA.

Objective To explore the comparative advantage of Team based learning (TBL) in common diseases teaching among pre-selected trainees for medical non-commissioned officer. Methods 200 trainees were randomly divided into two groups of equal size (each having 100 trainees), one TBL group and another lecture based learning (LBL) group as the control group. The teaching content was both Chapters of High Altitude Illness and Hypertension. In the LBL control group, the traditional lec-ture based teaching and learning manner was followed. In contrast, the trainees in the TBL group were assigned pre-lecture self-learning outline and questions to discuss in advance, team-based group dis-cussion was conducted, and scene simulation with typical disease case analysis were adopted in class. In both groups, the teaching effectiveness were evaluated by both exam test and questionnaire manner. Results It turned out the team based learning model resulted in significantly improved overall trainee assessment scores (88.90 ± 5.28 vs. 76.10 ± 5.12, P<0.05) and higher overall teacher satisfaction scores (8.63 ± 0.85 vs 7.18 ± 0.72, P<0.05), compared to the LBL group. Conclusion TBL model is more effective in improving trainees' capabilities of cultivating team work spirit, self-confidence, learn-ing enthusiasm, problem analysis and knowledge application. TBL model significantly brings more advantage in teaching pre-selected trainees for medical non-commissioned officer.

Objective To test the predicative roles of neutrophil gelatinase-associated lipocalin (NGAL) and kidney injury molecule-1( KIM-1 ) for acute kidney injury (AKI) in critically ill patients. Methods Children from pediatric intensive care unit, were randomly divided into four groups:critically ill patients with AKI (group 1 ), critically ill patients with non-AKI (group 2) , chronic kidney disease group (group 3), healthy control group (group 4). 1.5 ml venous blood and urine specimens were collected and kept under-70°C. Serum creatinine , urine NGAL and urine KIM-1 were analyzed. Results Compared with group 2, group 3 and group 4, the urine NGAL and urine KIM-1 increases obviously in group 1 (P<0.05). There is no signiifcance of urine NGAL and urine KIM-1 between group2, group3 and group 4 (P>0.05). The concentration of urine NGAL increased more than 10 times of base-line level 2 days before the diagnosis of AKI under the Acute Kidney Injury Network standard with area under curve (AUC) 0.955 (P<0.05) , and the concentration of urine KIM-1 increased more than 5 times of base-line level 1 day before AKI with AUC 0.878 (P<0.05). The AUC was 0.984 (P<0.01) when they were combined. There is negative correlation between the increased times of urinary KIM-1, urinary NGAL and vally value of creatin clearance rate. Conclusions The concentrations of urine NGAL and urine KIM-1 are useful early biomarkers for predicting AKI, especially when they were combined.

Aim To observe whether DHEA has enhancing effect on DbcAMP -induced differentiation of NG108-15 cells, including neurite outgrowth, and study its possible mechanisms. Methods NG108-15 cells (a h ybrid cell line of mouse neuroblastoma and rat glioma) were used as a substitute for primary culture neuron in vitro. The morphology of NG108-15 cells was o bserved and neurite outgrowth was determined in an inversed microscope after treatme nt with various drugs. Gelatin-substrate gel electrophoresis was used to detect gelatinases (MMP-9 and MMP-2). Results ① DHEA and DbcAMP inhibited NG108-15 proliferation.②DHEA had enhancing effect on the promoting activity of neuronal differentiation and neurite outgrowth by DbcAMP. DbcAMP could increase neurite elongation of NG108-15 cells. Compared with this, the combined treatment with DHEA and DbcAMP significantly enhanced the neurite outgrowth of NG108-15 cells, including neurite length and numbers of cells with neurite, in a DHEA dose-dependent manner. ③ MMPs were involved in neuronal differentiation. DbcAMP induced the increase in MMP-9 and MMP-2 activities and such elevation was enhanced by DHEA in a dose-dependent manner. Conclusion DHEA enhances the effect of DbcAMP in promoting the neurite outgrowth of NG108-15 cells, which might be related to the increase in MMP-9 and MMP-2 activities.

Objective:To explore the expression of peptidyl prolyl cis-trans isomerase (Pin1) activity in peripheral blood of patients with rheumatoid arthritis (RA) and the value and correlation of T helper cell 17/regulatory cells (Th17/Treg) cells, and to analyze the effect and influence of all-transretinoic acid (ATRA) on it.Methods:① Comparing the difference of Pin1 expression and absolute counts of Th17 and Treg between RA patients before and after treatment and healthy control group, Kruskal-Wallis rank sum test was used for analysis. ② To analyze the correlation between the expression of Pin1 and its general data, activity indicators [such as erythrocyte sedimentation rate (ESR), C-reactive protein (CRP) and disease activity score 28 joints (DAS28) scores], Th17, Treg and some cytokines in RA patients, and to use Pearson and Spearman correlation tests. ③ To analyze the difference of Pin1 expression and Th17/Treg in peripheral blood of RA patients treated with low-dose all-trans retinoic acid (10 mg twice a week) and traditional immunosuppressants such as hydroxychloroquine for 3 months respectively. Mann-Whitney U test was used for comparison between the two groups, and the difference was statistically significant with ( P<0.05). Results:① The activity of Pin1 in peripheral blood of the newly treated group of RA was [13.62(9.16, 19.42)] higher than that of the healthy control group [8.97(7.62, 11.45)]( Z=42.82 , P<0.05), and Th17 was [18.28(12.76, 24.08)] higher than that of the healthy control group [6.04(4.96, 4.96)]( Z=48.83 , P<0.05). Treg [11.06(5.31, 21.87). It was lower than that of healthy control group [40.41(24.33, 48.52)]( Z=42.21 , P<0.05). ② the activity of Pin1 in peripheral blood of RA patients was positively correlated with CRP, the number of involved joints, DAS28 score, interleukin-6 (IL-6) and tumor necrosis factor-α (TNF-α) ( r=0.396, P<0.05; r=0.683, P<0.05; r=0.466, P<0.05; r=0.315, P<0.05; r=0.416, P<0.05). ③ Compared with the newly treated RA group, the activity of Pin1 [6.94(5.96, 8.77), Z=42.82 , P<0.05] and Th17 7.38 decreased [7.38(3.85, 11.21), Z=48.83 , P<0.05], while Treg [40.41 (17.77, 33.47)] increased ( Z=42.21 , P<0.05). ④ Compared with the traditional medicine group, Treg [28.9(21.73, 37.36)] was higher in the retinoic acid group, and the difference was statistically significant ( Z=-2.683 , P<0.05). The activity of Pin1 was [6.23(5.58, 8.75)], but there was no statistical significance ( Z=-1.622 , P=0.104). Conclusion:Pin1 in peripheral blood of RA patients is over-expressed. Th17 is increased and Treg is decreased. ATRA combined with other traditional drugs can reduce Pin1 activity, promote Treg growth and improve disease activity of RApatients to a certain extent.

Objective To measure serum surfactant protein (SP) A and D levels in patients with interstitial lung disease associated with rheumatoid arthritis (RA).Methods Serum SP-A and SP-D levels of RA,RA-ILD patients and healthy controls were assessed using a sensitive enzyme-linked immunosorbent assay (ELISA).The relationship between SP-A and SP-D and RA-ILD was analyzed.The serum SP-A and SP-Dpositive rate was calculated for the three groups.The correlation between SP-A and SP-D with RF,anti-CCP,antinuclear antibody,antikeratin antibody,anti-perinuclear factor,C-reactive protein,erythrocyte sedimentation rate,were analyzed.Mean value of groups were compared with variance analysis,Spearmam rank correlation test was used for correlation analysis.Results The levels of serum SP-A in RA-ILD patients and RA patients as well as in healthy controls were [ (51.2±9.2),(25.9±2.6),( 15A±0.3 ) μg/L] respectively.The level of serum SP-D of the three groups was [ ( 42.5 ±8.1 ),(20.8 ± 1.5 ),( 16.6±0.8 ) μg/L ] respectively.The levels of serum SP-A and SP-D in patients with RA complicated with ILD were higher than those simple RA patients and healthy controls (P＜0.05).The levels of serum SP-A and SP-D in patients with RA were not significantly higher than those in healthy controls (P＞0.05).The positive rate of serum SP-A and SP-D in RA-ILD patients were significantly higher than those in simple RA patients and healthy controls.The positive rate of serum SP-D of RA-ILD patients was higher than that of SP-A.The levels of serum SP-A and SP-D in patients with RA complicated with ILD were correlated positively with age,C-reactive protein.The level of serum SP-D was correlated positively with RF,anti-CCP,antikeratin antibody.There was no correlation between the level of serum SP-A and SP-D with RA-ILD and antinuclear antibody,antiperinuclear factor,erythrocyte sedimentation rate.Conclusion The levels of serum SP-A and SP-D are correlated with RA-ILD and may be useful markers for ILD in patients with RA.These two paramenters may be helpful to early diagnosis of RA-ILD.The Serum SP-D levels are more sensitive in predicting the development of RA-ILD than other parameters and can help in assessing the severity of lung damage.