Objective To develop a sensitive liquid chromatography-tandem mass spectrometric (LC-MS/MS) method for the determination of forsklin in rat plasma.Methods After extraction with methyl tert-butyl ether,chromatographic separation was performed on a C18 column with the mobile phase consisting of water ( 0.1％ formic acid)-acetonitrile in a gradient elution mode.A tandem mass spectrometer equipped with electrospray ionization (ESI) source was used as detector in the positive ion mode.Quantification was performed using multiple reaction monitoring (MRM) with the precursor product combination ions of m/z 411→375.3 and 285→193 for forsklin and diazepam.Results Good linearity was obtained in the 0.5-1000 ng/ml range for the analyte and the analytical method was validated in terms of specificity,precision,accuracy,recovery,stability and matrix effect.These assays gave RSD values always lower than 14.4％ and RE values between -3.5 ％ and 3.8％.In addition,the specificity,extraction recovery,stability and matrix effect were satisfactory.Conclusion Due to its high sensitivity,specificity and simplicity,the method could be used for pharmacokinetic studies of forsklin.

Objective To explore the clinical effect of magnetic resonance imaging (MRI) and CT scanning in the diagnosis of acute craniocerebral injury.Methods From October 2012 to November 2016,100 patients with acute craniocerebral injury in Zhoushan Hospital were selected.According to the order of patients admitted to hospital,they were randomly divided into observation group and control group,with 50 cases in each group.The observation group received MRI inspection,the control group received CT scanning.The clinical examination results of the two groups were compared.Results The positive rate and missed diagnosis rate of the observation group were 98.0％,2.0％,respectively,which of the control group were 72.0％,24.0％,respectively,the differences were statistically significant (x2 =26.958,17.468,all P ＜ 0.05).The detection rate of craniocerebral injury position lesion in the observation group was significantly higher than control group,the differences between the two groups was statistically significant (x2 =5.030,5.110,11.250,5.430,all P ＜ 0.05).The clinical total effective rate of the observation group was 94.0％,which was higher than 78.0％ of the control group,the difference was statistically significant (x2 =15.432,P ＜ 0.05).Conclusion The clinical value of MRI in the diagnosis of acute craniocerebral injury issuperior to CT scanning.

To facilitate noninvasive diagnosis of anemia, high-performance and portable near infrared (NIR) spectrometer for human blood constituents was designed and fabricated based on linear variable filter (LVF).Meanwhile, the performance of support vector regression (SVR) model for quantitative analysis of human hemoglobin (Hb) was investigated.Spectral data were collected noninvasively from 100 volunteers by self-designed LVF-NIR spectrometer, then divided into calibration set, validation set 1 and 2.To establish a robust SVR model, grid search method was applied to optimize the penalty parameter and kernel function parameter c=5.28, g=0.33.Then, Hb levels in the validation 1 and 2 sets were quantitatively analyzed.The results showed that the root mean square error of prediction (RMSEP) were 10.20 g/L and 10.85 g/L, respectively, and the relative RMSEP (R-RMSEP) were 6.85% and 7.48%, respectively.The results indicated that the SVR model had high prediction accuracy to Hb level and adaptability to different samples, and could satisfy the requirements of clinical measurement.Based on the SVR algorithm, the self-designed LVF-NIR spectrometer has a wide application prospect in the field of non-invasive anemia diagnosis.

Objective: To evaluate the clinical value of CT combined with MRI in the qualitative diagnosis of hepatic lesions. Methods: From August 2015 to March 2018, 100 patients with liver diseases were selected from Zhoushan Hospital.CT, MRI and combined examination were performed respectively.The results of diagnosis of liver diseases in three different ways were compared and analyzed. Results: A total of 96 lesions were detected by CT, the detectable rate was 88.89%, 97 lesions were detected by MRI, and the detectable rate was 89.81%.A total of 105 lesions were detected by CT and MRI, and the detectable rate was 97.22%.The detection rate and number of detected lesions of combined test were significantly higher than those of single test (χ²=8.25, 6.22, all P<0.05), but there were no statistically significant differences in the detection rate and the number of lesions between CT and MRI (all P>0.05). Conclusion The diagnosis of benign and malignant lesions by the combination of CT and MRI is very good.The diagnostic effect is relatively small and the safety factor is high.It is worth widely used in clinical diagnosis of patients.

In order to explore tick proteins as potential targets for further developing vaccine against ticks, the total proteins of unfed female Dermacentor silvarum were screened with anti-D. silvarum serum produced from rabbits. The results of western blot showed that 3 antigenic proteins of about 100, 68, and 52 kDa were detected by polyclonal antibodies, which means that they probably have immunogenicity. Then, unfed female tick proteins were separated by 12% SDS-PAGE, and target proteins (100, 68, and 52 kDa) were cut and analyzed by LC-MS/MS, respectively. The comparative results of peptide sequences showed that they might be vitellogenin (Vg), heat shock protein 60 (Hsp60), and fructose-1, 6-bisphosphate aldolase (FBA), respectively. These data will lay the foundation for the further validation of antigenic proteins to prevent infestation and diseases transmitted by D. silvarum.
Animals ; Antigens/*chemistry/immunology ; Arthropod Proteins/*chemistry/immunology ; Electrophoresis, Polyacrylamide Gel ; Female ; Ixodidae/*chemistry/immunology ; Molecular Weight ; Rabbits ; Tandem Mass Spectrometry

In order to establish the isolation,culture and identification method of cow bone marrow-derived macrophages,three different media(RPMI-1640,DMEM,DMEM/F12)were added with 20％fetal bovine serum(FBS),2.4％chlorine-streptomycin,1.2％glutamine(Gln),and M-CSF(20 ng/mL),respectively,to induce the monocytes extracted from the bone marrow of dairy cows to become macrophages.The induced M0 macrophages were polarized into M1-type macrophages by adding lipopolysaccharide(LPS).The morphology of macrophages was observed by optical mi-croscope at day 1,4 and 7,and the differences of differentiated macrophages between the three media were compared.The effects of prostaglandin D2(PGD2)-DP2 receptor pathway on the secre-tion of cytokines(IL-6,TNF-α)induced by Escherichia coli and phagocytosis of macrophages were also investigated.The results showed that the morphological changes of cells cultured in the medium of RPMI-1640 were the most obvious and the number was large.A large number of char-acteristic markers of mononuclear macrophages were detected(M0 markers:CD1 1b,CD14;M1 markers:CD11b,CD80)expression,M0 and M1 macrophage purity were 79.9％and 93.5％,re-spectively.COX-2 and H-PGDS gene expressions were significantly increased in E.coli group com-pared with the blank control group.The secretion of PGD2also increased significantly(P＜0.000 1).DP2 receptor inhibitors(CAY10471,CAY10595)could significantly inhibit the secretion of E.coli in-duced pro-inflammatory cytokines(IL-6,TNF-α)and significantly enhance the killing effect of macrophages on E.coli.The above results showed that the induced cells had the characteristic mor-phology and immunophenotype of macrophages.E.coli can induce the production of PGD2 in mac-rophages,and the PGD2-DP2 pathway regulates the secretion of cytokines in E.coli infected macro-phages.