Objective:To analyze the correlation between intra-extracranial arterial stenosis and silent cerebral infarction(SCI) in the elder with essential hypertension(EH). Methods:The results of color duplex ultrasonography(Duplex) and digital subtraction angiography(DSA) examination of 86 patients with essential hypertension complicated silent cerebral infarction were analyzed.The incidence of intra-and extracranial arterial stenosis and arthrosclerosis plaque were calculated.The comparative value of Duplex and DSA for evaluating atherosclerosis of extracranial carotid artery was analyzed using statistic methods. Results:The incidence of arthrosclerosis plaque was higher in SCI group than that in normal control group(62.79%,24%,P

Drug metabolism will change significantly during inflammation, including the reduction of expression and activity of many drug metabolizing enzymes and transporters. Body would release a series of inflammatory cytokines which can regulate drug metabolizing enzymes. Recent studies have revealed that drug transporters are also regulated by the cytokines with obvious species difference. Mechanism studies show that several transcription factors play important roles during the signal pathways of regulation. This review focuses on the progress in the regulation of drug transporters during inflammation.

The study aims to identify the related substances in fasudil hydrochloride by hyphenated techniques. A WondaSil C18 (250 mm x 4.6 mm, 5 microm) column was used for the separation of the related substances with a mixture of methanol and ammonium acetate buffer solution as the mobile phase by gradient elution. The structures of the related substances were speculated by electrospray positive ionization LC-TOF/MS accurate ion mass and MS/MS determination and elucidation, and verified further through synthesis and spectroscopic analysis. Fasudil hydrochloride and the related substances were separated under the established HPLC condition. Three related substances in fasudil hydrochloride were characterized by hyphenated techniques. The hyphenated LC-MS method is useful for the identification of related substances in fasudil hydrochloride and the results obtained are valuable for its manufacturing process and quality control.

A sensitive and selective liquid chromatography–tandem mass spectrometric (LC ? MS/MS) method was established to determine 2-oxo-clopidogrel, a crucial intermediate metabolite in human plasma. A chromatographic separation was performed on a Sapphire C18 column following a liquid–liquid extraction sample preparation with methyl t-butyl ether. Detection was carried out on a triple quadrupole mass spectrometer operated in multiple reaction monitoring (MRM) with an electrospray ionization (ESI) mode. The method was validated in terms of specificity, accuracy, precision and limit of quantification. The calibration curves ranged from 0.50 to 50.0 ng/mL with good linearity. The stability was fully validated with addition of 1,4-dithio-DL-threitol (DTT) into the plasma sample prior to and in the preparation procedure. The validated method was proved to be suitable for use in pharmacokinetic study after single oral administration of 75 mg clopidogrel tablets in human subjects, which could make contribution to intensive study of the clinical drug–drug interactions of clopidogrel and individual treatment.

Objective To investigate the application value of three-dimensional (3D) laparoscopic pancreaticoduodenectomy (LPD) and compare the clinical outcomes between 3D-LPD and open pancreaticoduodenectomy (OPD).Methods The retrospective cohort study was adopted.The clinicopathological data of 349 patients who underwent pancreaticoduodenectomy at the Affiliated Tongji Hospital of Tongji Medical College of Huazhong University of Science and Technology between July 2014 and March 2016 were collected.Of 349 patients,146 undergoing 3D-LPD were allocated into the 3D group and 203 undergoing OPD were allocated into the OPD group.Observation indicators:(1) surgical situations:operation time,volume of intraoperative blood loss,cases of blood transfusion,number of lymph node dissected,resection margin and vascular resection and reconstruction,(2) postoperative situations:time of gastric tube removal,duration of intensive care unit (ICU) stay,duration of hospital stay,(3) complications:pancreatic fistula,delayed gastric emptying,intra-abdominal infection or abscess,bile leakage,hemorrhage,pulmonary infection or wound infection,(4) follow-up.All the patients were followed up by telephone interview to detect the tumor-free survival rate up to June 2016.Measurement data with normal distribution were presented as (x) ± s and comparison between groups was analyzed using the t test.Count data were analyzed using the chi-square test.Results (1) Surgical situations:all the patients underwent successful pancreaticoduodenectomy.Operation time,volume of intraoperative blood loss,cases of blood transfusion,number of lymph node dissected and positive resection margin were (334 ± 175)minutes,(254 ± 107)mL,29,13 ±8,1 in the 3D group and(320 ±91)minutes,(290 ± 101) mL,35,14 ±9,5 in the OPD group,respectively,with no statistically significant difference between the 2 groups (t =0.975,1.383,x2=0.390,t =12.155,x2=1.589,P ＞ 0.05).Vascular resection and reconstruction were respectively applied to 0 patient in the 3D group and 14 patients in the OPD group,with a statistically significant difference between the 2 groups (x2 =10.490,P ＜ 0.05).(2) Postoperative situations:time of gastric tube removal,duration of ICU stay and duration of hospital stay were (2.9 ± 1.9) days,(6.9 ± 2.1) days,(12.9 ± 7.2) days in the 3D group and (5.1 ± 1.7) days,(7.4 ± 1.2) days,(19.8 ± 7.1) days in the OPD group,respectively,with statistically significant differences between the 2 groups (t =11.350,2.814,8.903,P ＜ 0.05).(3) Complications:of 146 patients in the 3D group,40 had postoperative complications with incidence of complications of 27.40％ (40/146).Twenty-nine patients with pancreatic fistula (20 in grade A,9 in grade B and C) were improved by conservative treatment.Thirteen patients with delayed gastric emptying were cured by gastrointestinal decompression and enhancing gastric motility.Of 5 patients with postoperative hemorrhage,3 were improved by conservative treatment,and 2 were improved by small vein hemostasis behind the head of pancreas.One patient died of systemic inflammatory response syndrome.Partial patients were combined with multiple complications.Of 203 patients in the OPD group,60 had postoperative complications with incidence of complications of 29.56％ (60/203),including 39 patients with pancreatic fistula (31 in grade A,8 in grade B and C),25 with delayed gastric emptying,15 with intra-abdominal infection and 13 with systemic inflammatory response syndrome,and they were improved by conservative treatment.Of 8 patients with postoperative hemorrhage,4 were improved by conservative treatment,and 4 were cured by hemostatic therapy after ineffectual blood transfusion and interventional treatment.Two patients died of cardiopulmonary complication.Partial patients were combined with multiple complications.There was no statistically significant difference in the incidence of postoperative complication between the 2 groups (x2 =10.490,P ＞ 0.05).(4) Follow-up:all the patients were followed up at postoperative month 6.Tumor-free survival rate was 90.41％ (132/146) in the 3D group and 85.22％ (173/203) in the OPD group,with no statistically significant difference between the 2 groups (x2 =2.076,P ＞ 0.05).Conclusion Compared with OPD,3D-LPD can provide the more realistic visual effects and refinement of surgical procedures,with a good short-term outcome.

Objective To establish a stable and reliable HPLC method and fingerprint reference substance for the measurement of the fingerprint, the practical quality control, and assay of the water-soluble components of Salvia miltiorrhiza. Methods The HPLC was run on C 18 columns with methanol-1.0 % glacial acetic acid solution as mobile phase in gradient elution mode at a flow rate of 1.0 mL/min. The chromatographic system suitability, the gradient elution mode, mobile phase acidity, and the effect of column type on fingerprint repeatability were tested. Results The HPLC fingerprints of the water-soluble extract of reference S. miltiorrhiza were obtained with very good resolution under the established chromatographic system. Fifteen peaks in the chromatograms were selected for the fingerprint identification and quality control of S. miltiorrhiza. The quality of ten batches of S. miltiorrhiza samples from different hibitats were assessed by comparing their chromatographic fingerprints with the reference fingerprints obtained at the same time, and the similarity showed no difference, eventhough the column filler was changed. Conclusion Because the inherent complexity of medicinal material components has been reflected by chromatographic fingerprints, there are many factors affecting the fingerprint repeatability for the changes of column type. The results of the quality assessment of S. miltiorrhiza, using fingerprints from different columns, are not all coincidence. In order to obtain the comparable and repeatable results in different laboratories, it is much practical with both a defined chromatographic system suitability and a fingerprint reference substance.

Objective T o infer the frequency of dosage and m edication history investigate of the victim s in drug facilitated cases by the segm ental analysis of clonazepam in hair. Methods Freezing m illing un-der liquid nitrogen environm ent com bined w ith ultrasonic bath w as used as sam ple pretreatm ent in this study, and liquid chrom atography-tandem m ass spectrom etry w as used for the segm ental analysis of the hair sam ples collected from 6 victim s in different cases. T he concentrations of clonazepam and 7-am in-oclonazepam w ere detected in each hair section. Results C lonazepam and its m etabolite 7-am inoclon-azepam w ere detected in parts of hair sections from the 6 victim s. T he occurrence tim e of drug peak concentration w as consistent w ith the intake tim ing provided by victim s. Conclusion Segm ental analysis of hair can provide the inform ation of frequency of dosage and intake tim ing, w hich show s an unique evidential value in drug facilitated crim es.

Objective:To construct I-Ad/IgG2b Fc baculovirus expression vector and express I-Ad/IgG2b Fc dimer fusion protein in Sf9 insect cells.Methods:I-Ad α,I-Ad β and IgG2b Fc gene sequences were amplified from BALB/c mouse lymphocytes by RT-PCR.I-Ad α and I-Ad β were connected with the leucine zipper sequence Fos and Jun respectively by overlapping PCR to form I-Ad α-Fos and I-Ad β-Jun.I-Ad α-Fos and IgG2b Fc fragments were ligated by restriction sites Xba I to form I-Ad α-Fos-IgG2b Fc recombination sequence.I-Ad α-Fos-IgG2b Fc and I-Ad β-Jun fragments were inserted to PPH and PP10,which were the downstream of the promoters in the plasmid pFastBacTMDual,to form pFastBacTMDual+[I-Ad/IgG2b Fc] recombinant plasmids.The constructed vector was identified by PCR,restriction endonuclease and sequencing.The recombinant plasmids pFastBacTMDual+[I-Ad/IgG2b Fc] was transferred into the DH10Bac competent cell to form recombinant baculovirus Bacmid+[I-Ad/IgG2b Fc].The recombinant baculovirus was transfected into Sf9 insect cells by liposome transfection reagent.After infected with Sf9 insect cells,the supernatant was collected and concentrated by PEG20000 to obtain I-Ad/IgG2b Fc dimer fusion protein.The fusion protein was detected by double-antibody sandwich ELISA and Western blot.Results:PCR,restriction enzyme digestion and sequencing confirmed that the recombinant vector pFastBacTMDual+[I-Ad/IgG2b Fc] had the correct sequence.The double antibody sandwich ELISA and Western blot showed that recombinant bacmid could successfully infect Sf9 insect cells,and the expressed fusion protein had the correct conformation.Conclusion:The pFastBacTMDual+[I-Ad/IgG2b Fc] baculovirus expression vector was successfully constructed and expressed in Sf9 insect cells,laying a foundation for the study of I-Ad-restricted T cells.

Objective T o establish a gas chrom atography-m ass spectrom etry (G C-M S ) m ethod for the determ ination of sulfide ion in blood and apply it to the practical cases. Methods T he 1, 3, 5-tribro-m obenzene w as selected as an internal standard, and 0.2 m L blood sam ple w as collected and analyzed using G C-M S after α-B rom o-2, 3, 4, 5, 6-pentafluorobenzyl brom ide derivatization. Results T he m ass concentration of sulfide ion in blood had good linearity in the range of 0.2-40μg/m L w ith a lim it of detection (L O D ) of 0.05μg/m L . T he m ass concentration of sulfide ion w as less than 0.05μg/m L in blank blood from different sources such as healthy subjects and dead cases. In 3 sulfide poisoning cases, sul-fide ion w as detected in the blood sam ples of 6 victim s, and the m ass concentration range w as 1.02-3.13μg/m L . Conclusion T his study establishes a m ethod for investigation of sulfide ion in blood w hich has been applied successfully to the cases of fatal sulfide poisonings.

After oral administration of Salvia miltiorrhiza (Danshen in Chinese), Panax notoginseng (Sanqi in Chinese) and Danshen Sanqi combination suspensions to Beagle dogs, the plasma concentration-time profiles of danshensu, tanshinone II(A), cryptotanshinone, notoginsenoside R1, ginsenoside Rg1 and Rb1 were analyzed by LC-MS/MS. Pharmacokinetic parameters were calculated and analyzed with BAPP 2.0 software. The results showed that the Cmax and AUC of danshensu, notoginsenoside R1, ginsenoside Rg1 and Rb1 in Danshen Sanqi combination group all decreased in comparison with those of Danshen or Sanqi given alone, while the CLz/F and Vz/F increased to some extent. No significant differences of the pharmacokinetics of tanshinone II(A) and cryptotanshinone were observed between groups.