Objective:To determine ochratoxin A ( OTA ) in Chinese herbs by high performance liquid chromatography-tandem mass spectrometry method ( HPLC-MS/MS) . Methods: The samples were extracted by 80% methanol water solution and purified by immunoaffinity column. The chromatographic separation was carried out on a C18 column, the mobile phase was methanol-acetonitrile-0. 01 mol·L-1 ammonium acetate, and then OTA was detected by MS/MS in an ESI(-)-MRM mode. Results:The limit of detection was 0. 1 μg·kg-1 , the average recoveries ranged from 84. 8% to 91. 2%, and the relative standard deviations ( RSD) ranged from 3. 6% to 8. 1%(n=9). Conclusion:The method is accurate,sensitive and simple, and suitable for the determination of ochratoxin A in Chinese herbs.

OBJECTIVE To screen a mouse anxiety model with good reproducibility, little stimulation and user-friendliness by comparing the uncertain empty bottle drinking water stimulation with restraint stress to induce anxiety-like emotion in mice in order to find a suitable pharmacodynamic evaluation model for anti-anxiety drugs which provides an experimental basis. METHODS A mouse anxiety model was established using the uncertain empty bottle drinking water stimulation method. The mice were divided into freely drinking water group, regular drinking water group, physiological stress group and empty bottle stimulation group, with eight mice in each group. The freely drinking water group was free to eat and drink all day, the regular drinking water group mice were given free water for 2 periods per day, while the regular drinking water group mice were given water for 10 min at 2 fixed time with a 12 h interval every day. At the same time, physiological stress group mice had access to drinking water for 10 min once a day, but at another time they were given neither water nor empty bottles. The empty bottle stimulation group mice were given empty bottle stimulation randomly once or twice every day so that they drank water 12 times and empty bottle stimulation 16 times. The experiment lasted 14 d. The mouse anxiety model was established with restraint stress method. The experimental mice were divided into normal control group and restraint stress group, with eight mice in each group. The normal control group was normally reared, while the restraint stress group mice were confined to the tube at 9:00 every morning, headed towards the bottom of the centrifuge tube and were restrained for 2 h on the first day, and then the daily binding strength was extended for 2 h until 8 h every day for 21 d. The changes of body mass in mice after 7 and 14 d of empty bottle stimulation and after 7, 14, and 21 d of restraint stress were observed. At the end of the modeling, the mice were tested for autonomous activities and exploration behaviors through the open field test. The elevated plus maze test was used to detect the number of times they entered the open arms and the time spent in the open arms. The plasma corticosterone (CORT) content and the content of serotonin (5-HT) in the hippocampus of mice were determined by ELISA after 21 d of restraint stress. RESULTS Uncertain empty bottle drinking water stimulation method: compared with the freely drinking water group, the regular drinking water group showed a significant decrease in the residence time in the open arm (P<0.05). Compared with the regular drinking water group, the body mass of the mice decreased significantly after 7 d of empty bottle stimulation (P<0.05). There was no significant change in the behavioral indicators. Restraint stress method: compared with the normal control group, the body mass of mice was significantly decreased after 14 d of restraint stress (P<0.05). There was no significant change in the autonomous activity or exploration behavior of mice. The number of times the mice entered the open arms and the residence time in the open arms were significantly decreased (P<0.05). After 21 d of restraint stress, the body mass of the mice decreased significantly (P<0.05), and plasma CORT content (P<0.01) and 5-HT content in hippocampus (PO.01, P<0.05) of mice were significantly reduced. CONCLUSION The mouse anxiety model established with the uncertain empty bottle drinking water stimulation method results in poor stability and sensitivity, with more uncontrollable factors. The mouse anxiety model was not successfully established until 14 d of stimulation. The restraint stress for 14 d could successfully a establish mouse anxiety model, and the mechanism may be related to HPA axis dysfunction and 5-HT metabolic abnormalities.

Objective: To describe the differences of plasma folate concentration and prevalence of folate deficiency in genders and ages in Chinese adults aged 35 to 64 years. Methods: By cross-sectional analysis, plasma folate concentration was measured in 2 545 adults including men and women by microbiological assay. Results: (1) Men tended to have lower plasma folate concentration and higher prevalence of folate deficiency(9.70 nmol/L, 31.0%) than did women(14.2 nmol/L, 12.5%, P=0.001). (2) Men in South have significantly higher plasma folate concentration and lower prevalence of folate deficiency than in North. The difference of plasma folate concentration was not evident between urban and rural area, but evident in North between in winter and spring. There were no significant differences of prevalence of folate deficiency for men between urban and rural areas in both regions. For women, the differences of plasma folate concentration in areas were significant, which were higher in South and urban, and lower in North and rural areas. (3) Plasma folate concentration increased with age in Southern men. A similar trend for others was not significant. Conclusion: Men have lower plasma folate concentration and higher prevalence of plasma folate deficiency than do women. The distribution of plasma folate concentrations in urban and rural areas seems different between men and women.

Objective To evaluate the value of high-frequency echocardiography in assessing Bax gene transfer influence on survival of heterotopic cardiac allograft in rats.Methods Thirty rat models of heterotopic heart transplantation were established.Group A received heart transplantation only; Group B received cyclosporin (CsA) after operatiom Group C received ultrasound targeted microbubble destruction (UTMD) combined with Bax-shRNA.All rats were tested by high-frequency echocardiography at day 1,3,6 after transplantation.The ultrasound parameters included left ventricular internal dimension diastole (LVIDd),left ventricular internal dimension systole(LVIDs),left ventricular ejection fraction(LVEF),left ventricular thickness (LVT),left ventricular thickening (LVTR) and so on.The pathologic examinations were carried out on five rats at day 6 after echocardiography exam.The other rats in each group were observed for the survival time of cardiac allograft.Results ①The left ventricular long axis view and the apical four chamber view could be displayed clearly by high-frequency echocardiography.②The survival time of allografts in group C was (16.21 ± 5.01)d,which was longer than that in group B [(11.14 ± 1.72)d,P ＜ 0.05] and group A [(7.26 ± 1.50)d,P ＜0.01].③LVT index got higher after transfection.At day 6,LVEF of group A was lower than group B and C markedly(P ＜0.05) while no significant difference was found between group B and C(P ＞0.05).At day 3 and 6,LVT of group A and B were higher than group C (P ＜0.05) while LVTR was lower(P ＜0.05).Conclusions The Bax gene transfer influence on survival of heterotopic cardiac allograft in rats could be evaluated accurately by high-frequency echocardiography which could assess cardiac structure and function.LVT and LVTR can be considered as early evaluation indexes for its high sensitivity over LVEF.

Objective To evaluate the causes,prevention and treatment ot laparoscopic comphca- tions in urologic surgery.Methods From May 2000 to May 2004,135 urologic laparoscopies were per- formed,including 17 transperitoneal laparoscopies and 118 retroperitoneal laparoscopies.Of the135 cases,26 had adrenal adenoma extirpation;31 had roof-removal and decompression of renal cyst;25 had radical ne- phrectomy;6 had pyeloplasty;2 had partial resection of renal capsula and encapsulation of the great omen- tum;2 had partial nephrectomy;4 had pelviureteral resection;13 had ligation of renal pedicle lymphatic ves- sel;2 had ureterolysis;7 had ureterolithotomy;12 had ligation of spermophlebectasis;5 had exploration,re- duction and fixation of undescended testis.The intra-and post-operative complications were retrospectively reviewed and clinically analyzed.Results Overall,14 cases(10.4%)had complications.Intraoperative complications occurred in 10 cases,including 6 cases of peritoneal injuries,which were clamped during oper- ation;4 of vascular injuries,which resulted in conversion to open surgery.Postoperative complications oc- curred in 4 cases,including 2 of subcutaneous emphysema,which was spontaneously absorbed at 5 and 7 d after operation;1 of incision seepage,which was drained for 40 d and then disappeared;and 1 of urine reten- tion,for which urethral catheterization was kept for 3 d and then the patient had voluntary voiding.No death occurred in this series.Conclusions Understanding the characteristics of laparoscopic complications in u- rologic surgery and systematic training of the surgeons can reduce the occurrence of complications.

Objective To search a new,safe and effective method to obtain semen using rectal Drobe electroejaculation(RPE).Methods RPE procedures were performed in 8 men with psychological ejaculatory failure,used the electrical stimulate instrument(CGS model,made in Italy).The average age was 26 years.Seminal vesicle was normal by B Uhrasonography or CT examination.Except for 1 case of diabetes,the other 7 cases found no hormones,blood,biochemical and nervous system abnormalities.Three cases had nocturnal emission,but no sexual intercourse ejaculation.Results Adequate sperm was successfully taken out from the 8 patients.Mean seminal fluid volume was 2.3 ml,mean total sperm count was 67×106/ml and mean total motile sperm was 21.3%.Three patients felt abdominal distension during the RPE course.Conclusion RPE is a safe,reliable,non-in-vasive,repeatable method to obtain semen.

ObjectiveAnalyze preoperative clinical relevanted factors of acute type A aortic dissection with hypoxemia according to a group clinical data.MethodsFrom January 2011 to June 2011,we have collected 54 preoperative cases of acute type A aortic dissection,including 42 males,12 females,aged 28-73 years old,onset to treatment time is 0.4-14.0 days.General information:age,gender,time of onset,body mass index,hypertension,diabetes mellitus,smoking,drinking,heart ejection fraction,prothrombin time,quantitative fibrinogen,fibrinogen degradation products,D-dimer,C-reactive protein,procalcitonin,ICU time,length of hospital stay.According to the blood gas analysis of quiet state case without oxygen,with PaO2 ＜ ( 100-age ×0.33 ±5) mm Hg is for the hypoxemia group,equal or higher than this is no-hypoxemia group.ResultsNo-hypoxemia group has 14 cases,11 males,3 females,average aged (51.14 ± 14.24) years old,including 12 operation patients ( no death) and 2 no-operation patients(2 cases death).Hypoxemia group has 40 cases,31 males,9 females,average aged (50.53 ± 9.73 ) years old,including 33 operation patients(2 cases death) and 7 no-operation patients(7 cases death).There is no significant difference in age,gender,time of onset,hypertension,diabetes mellitus,smoking,drinking,cardiac ejection fraction,prothrombin time and fibrinogen.There is statistically significant on body mass index,fibrinogen degradation products,D-dimer,C-reactive protein,procalcitonin,ICU time and length of hospital stay time ( P ＜ 0.05 ).ConclusionPreoperative hypoxemia with acute type A aortic dissection is associated with obesity,excessive inflammation and activation of coagulation and fibrinclytic system,and hypoxemia may prolong the time of operative patients with acute type A aortic dissection in ICU and hospital.

Aim To construct 3 D structure model of cardiac Cav1.2 channel and check its accuracy and re-liability.Methods Homology model of Cav1.2 chan-nel α1 subunit was constructed using SWISS-MODEL server.The model was submitted to an online testing server built by University of California and scored by it.The binding of Cav1.2 channel with blocker or drug was simulated by MOE software molecular docking pro-gram to check the model′s accuracy and reliability.Re-sults Both the target sequence Cav1.2 α1 C and the template sequence Cav1.1 α1 S searched by SWISS-MODEL server belonged to L-type Ca2+channel.Since the homology was 7 1.5% revealed by sequence align-ment,homology modeling was performed using automa-ted mode.L-type Ca2+ channel blockers Verapamil, Nifedipine and Diltiazem could bind to the 3 D structure model of Cav1.2 channel,while sodium channel bloc-ker TTX could not.Furthermore,active ingredient of traditional Chinese drug Praeruptorin A and Berberine could also bind to the 3D structure model of Cav1.2 channel.Conclusion The 3 D structure model of Cav1.2 channel was constructed successfully,which provides reliable materials for further studies and estab-lishes the foundation for the application of homology modeling in the study of 3 D structure prediction of ion channels.

OBJECTIVE To explore the effect and mechanisms of LW-AFC,a new formula derived from Liuwei Dihuang decoction,on chronic unpredictable mild stress(CUMS)-induced mood and cogni-tion impairment in mice. METHODS C57BL/6J mice were randomly placed into seven groups (n=10):normal control group,CUMS group,Fluoxetine(10 mg·kg-1,once per day)group,Liuwei Dihuang de-coction group(LW,10 g·kg-1,once per day),and LW-AFC(0.8 g·kg-1,1.6 g·kg-1,3.2 g·kg-1,once per day) group. The stressed group was given CUMS for 4 weeks to set up a chronic multiple-stressed model.LW and LW-AFC was oral administered a week prior to CUMS and until the end of the study(a total of 35 d),while fluoxetine was administrated orally for 4 weeks.The anxiety behavior was analyzed using the open field test(OFT)and elevated plus maze test(EPM).The depression behavior was ana-lyzed using the sucrose preference test (SPT) and forced swimming test (FST). Spatial cognition was evaluated using Morris water maze (MWM) test and working memory was evaluated using new object recognition test(NORT). RESULTS CUMS for 28 d increased depressive-and anxiety-like behaviors. LW-AFC (1.6 g·kg-1) significantly increased the numbers of entries into the open arm and time in the open arm of CUMS mice (P<0.05). LW-AFC (3.2 g·kg-1) increased sucrose consumption and de-creased the immobility time of FST (P<0.01) of CUMS mice. The MWM test showed that spatial learning andmemory in CUMS mice were remarkably affected relative to controls,whereas LW-AFC(3.2 g·kg-1)im-proves cognitive functions(P<0.05).CONCLUSION The mood and theability of learning and memory of thestressed group can be affected after exposure to CUS.Oral administration of LW-AFC significant-ly improved CUMS-induced impairments of mood and cognition in mice.

Objective To investigate the clinical value of high resolution ultrasound combined with radiological imaging in the diagnosis of Meckel′s diverticulitis (MD),and to analyze the causes of missed diagnosis as well as misdiagnosis by ultrasound.Methods The image data of 61 patients with MD were retrospectively analyzed,the essentials and the causes of missed diagnosis as well as misdiagnosis of ultrasound were summarized and compared with the results of operation,CT,gastrointestinal contrast,as well as radionuclide.Results The patients were divided into simple type(18,29.5%) and complex type(43,70.5%) according to ultrasonography.The simple type diagnostic accuracy rate was 100%,the ultrasonography showed the right lower abdominal tubular bowel shadow or cystic echo mass,and the structure of diverticulum could clearly showed by ultrasound.In another side,the complex type diagnostic accuracy rate was 55.8%,the ultrasonography showed the right lower abdominal mixed echo mass or concomitant with other alimentary tract malformation,while the diverticulum structure was difficult to be distinguished.The diagnostic accuracy rate of radionuclide imaging was 83.6%,CT was 37.0%,while gastrointestinal contrast was 30.0%.Among 19 cases of misdiagnosis by ultrasound,13 cases of MD were confirmed by radionuclide and CT examination,the diagnostic accuracy rate of ultrasound combined with radiological imaging diagnosis was 90.2%.Conclusions The sonographic features of MD are lack of specificity,especially in the complex type,while the patients with recurrent hemafecia or acute abdomen should be alert to the presence of MD.Patients who are suspected of MD by ultrasound diagnosis should also be examined by radiological imaging in order to make an early diagnosis and surgical treatment to improve the prognosis of patients.