Objective:To establish an ultra performance liquid chromatography-tandem quadrupole mass spectrometry method for the determination of warfarin and its metabolite 7-hydroxywarfarin in human plasma. Methods: An ACQUITY UPLC? BEH C18 (50 mm × 2. 1 mm, 1. 7 μm) column was used as the stationary phase at 40℃. The mobile phase consisted of acetonitrile and water (con-taining 0. 1% formic acid) with gradient elution at a flow rate of 0. 4 ml·min-1 . Warfarin-d5 was used as the internal standard. The analytes were detected on a triple-quadrupole mass spectrometer equipped with an ESI interface in a positive mode. Results:The reten-tion time of warfarin and 7-hydroxywarfarin was 1. 8 min and 1. 5 min, respectively. Excellent linear calibration curve of warfarin and 7-hydroxywarfarin was obtained within the concentration range of 25-2 000 ng · ml-1 ( r =0. 999 3 ) and 5-500 ng · ml-1 ( r =0. 999 6), respectively. The lower limit of quantification of warfarin and 7-hydroxywarfarin was 5 ng·ml-1 and 2. 5 ng·ml-1 with the average recovery of 96. 9%-105. 3% and 97. 1% -103. 3%, respectively. The intra-and inter-day standard deviations were both less than 10%. Conclusion: The method is accurate and simple, and suitable for the determination of warfarin and its metabolite 7-hydroxywarfarin in human plasma.

The adolescence group is a unique group that deserves being paid close attention to.In terms of public crisis events,they are lack of essential sense of crisis and proper evaluation of crisis information,thus unable to protect and adjust their mental status.Correct responses to public crisis events for adolescence include a re-recognition of the real world,a positive adjustment of mental emotions,effective solutions to problems,and exploration for social support.In terms of intervention of adolescence mental crisis,it is important to implement early stage evaluation and intensification therapy,crisis counseling and treatment,crisis evaluation and intervention,and the usage of crisis intervention hotline.

OBJECTIVE:To establish the method for pharmacokinetic study of luteolin and cynaroside in rats and to determine pharmacokinetic parameters. METHODS:16 SD rats were randomly divided into luteolin group (sublingual iv,1.34 mg/kg) and cynaroside group(sublingual iv,0.64 mg/kg). 0.5 ml blood were collected before administration and 0,15,30 min and 1,2,3,4,6, 8,12,24,48 h after administration respectively to prepare plasma. UPLC-TQ-MS was adopted to determine plasma concentration, and pharmacokinetic parameters were calculated. A CORTECSTM UPLC? C18(100 mm×2.1 mm,1.6 μm)column was used with mobile phase consisted of acetonitrile-water (containing 0.1% formic acid) at a flow rate of 0.4 ml/min,the column temperature was set at 40 ℃,and quercetin was used as internal standard. RESULTS:The linear range of luteolin and cynaroside were 2.5-500 ng/ml (r=0.998 2) and 10-2 500 ng/ml (r=0.993 5). The lowest quantitation limits were 1 and 2.5 ng/ml,and extraction were 70.75%-87.72% and 75.40%-91.18%(n=6);RSD of inter-day and intra-day were all lower than 10%(n=3). Pharmacokinetic parameters as t1/2 were (1.88 ± 0.32) and (1.57 ± 0.08) h;CL were (0.77 ± 0.18) and (0.06 ± 0.01) L/(h·kg);AUC0-6 h were (189.60±40.04)and(1 093.14±187.36)ng·h/ml;AUC0-∞ were(195.18±38.37)and(1 097.11±188.07)ng·h/ml. CONCLU-SIONS:The method can be used for pharmacokinetic study of luteolin and cynaroside in rats,and the pharmacokinetics of them in rats are in line with two-compartment model.

OBJECTIVE:To observe therapeutic efficacy and safety of Yinxing damo injection for neural function recovery af-ter hypertensive intracerebral hemorrhage (HICH) minimally invasive surgery. METHODS:84 HICH patients were randomly di-vided into control group and observation group with 42 patients in each group. Both groups received CT guiding minimally inva-sive aspiration. Control group was given western medicine baseline therapy,such as dehydration and intracranial decompression, controlling blood pressure,preventing infection,alimenting never,symptomatic treatment. Observation group was additionally giv-en Yinxing damo injection 20 ml,ivgtt,bid. Treatment course lasted for 14 d. NIHSS score,GCS score and Fugl-Meyer motor function assessment scale score were comducted before and after treatment. The serum levels of NSE,serum C3,C4 and hs-CRP were determined in 2 groups before and after treatment. RESULTS:After treatment,the effective rate of observation group (85.71%)was better than that of control group(66.67%),with statistical significance(P<0.05);after treatment,NIHSS score, Fugl-Meyer score and GCS score of 2 groups were all better than before,the observation group was better than the control group, the levels of C3 and C4 in observation group were lower than in control group,with statistical significance(P<0.01). There was one case of allergic reaction that the patient can tolerate. Magnesium sulfate for external use was given,which did not affect the treatment. CONCLUSIONS:Yinxing damo injection could improve neurologic impairment,promote the recovery of patients and have good safety.

Objective:To establish an ultra performance liquid chromatography-tandem quadruple mass spectrometry ( UPLC-MS/MS) method to determine CYP2C9 activity in vitro. Methods:An ACQUITY UPLC? BEH C18 (100 mm × 2. 1 mm, 1. 7 μm) column was used as the stationary phase at 30℃. The mobile phase consisted of acetonitrile-water ( containing 0. 1% formic acid and 0. 5%ammonia water) (40∶60, v/v). The flow rate was 0. 2 ml·min-1. Chlorpropamide was used as the internal standard. The MS condi-tions were as follows:ESI with positive ion detection mode. Self-prepared CYP2C9?1, ?2, ?3 and ?13 protein were incubated with tolbutamide at 37℃ and 800μl ethyl acetate was added to stop the reaction. After centrifuged at 10 000g, the organic layer was then dried using nitrogen, the residue was re-dissolved in 200μl mobile phase and determined by UPLC-MS/MS. Results: The reten-tion time of 4-hydroxytolbutamide was 1. 21 min. An excellent linear calibration curve of 4-hydroxytolbutamide was obtained within the concentration range of 0. 05-5 ng·μl-1(r=0. 999 8). The lower limit of quantification of 4-hydroxytolbutamide was 0. 01 ng·μl-1 with the average recovery of 99. 3%-100. 3%. The intra- and inter-day RSDs were all less than 5%. There was no interference from the endogenous substances existing in the incubation system. The catalytic activity of the variants CYP2C9?2,?3 and?13 after tol-butamide was incubated with CYP2C9?1,?2,?3 and?13 was 47. 3%, 11% and 0. 3% of wild type CYP2C9?1. Conclusion:The method is simple and stable, and suitable for the fast evaluation of cytochrome CYP2C9 activity in vitro and relevant studies on the inhibitors.

Objective To explore the efficacy and safety of arterial embolectomy with Solitaire AB stent in treatment of acute intracranial large artery occlusion.Methods The clinical data of 11 patients with acute intracranial large artery occlusion receiving arterial embolectomy with Solitaire AB stent from March to December 2013 were analyzed retrospectively.Among them,5 cases were with simple middle cerebral artery occlusion,1 case combined with occlusion in the intracranial part of the internal carotid artery,4 cases had simple basilar artery occlusion,and 1 case had occlusion in the intracranial portion of the vertebral arteries extending to intracranial portion of the basilar artery.Revascularization and postoperative hemorrhage were analyzed,and nosocomial as well as clinical outcomes after 3 months were evaluated.Results Ten patients showed successful revascularization,of which 8 cases reached the standard of perfect revascularization.After embolectomy,there was obvious stenosis in offending vessel of 4 cases,who were given balloon dilatation then.In 3 cases,the vessels were still narrow,so Solitaire AB stent was placed in the stenosis area to correct stenosis with the residual stenosis rate of less than 40％.Thus electrolytic interruption was given.There was no symptomatic intracranial hemorrhage.Three months after the operation,5 cases recovered,among whom 2 cases had obvious curative effect instantly,1 case had complete remissions in 1 week,and 1 case remained hemiplegia.Five cases died,and 4 cases discharged from the hospital because their family gave up the subsequent treatment.Conclusions Arterial embolectomy with Solitaire AB stent to treat patients with acute intracranial large artery occlusion has a relatively higher revascularization rate.The clinical outcome after 3 months can be significantly improved.Thus,it provides a safe and effective endovascular treatment for patients with acute intracranial large artery occlusion.

Objective: To establish an accurate and selective UPLC-MS/MS) method for the determination of afatinib in rat plas-ma. Methods: Protein precipitating by acetonitrile was used to prepare the samples. A CORTECS BEH C18column ( 50 mm × 2. 1 mm, 1. 6 μm) was used to separate the analytes at 40℃. The mobile phase consisted of acetonitrile and water (0. 1% formic acid) with the flow rate of 0. 4 ml·min-1. The analytes were quantified by multiple reaction monitoring ( MRM) mode with positive electrospray ionization, while the target fragment ions were m/z 486. 19→112. 1 for afatinib and m/z 557. 3→112. 15 for neratinib (IS). Results: The calibration curve obtained good linearity for afatinib within the range of 1–200 ng·ml-1(r=0. 998 1), and the LLOQ in rat plasma was 1. 0 ng/ml. The intra-and inter-day precisions were both≤9. 51% . The recovery of afatinib from plasma was above 77. 1% . After intragastric administration and intravenous administration of afatinib in rats, the t1/2was 7. 19 h and 2. 69 h, Cmax was 97. 78 ng·ml-1and 123. 37 ng·ml-1,and AUC(0-∞)was 1 505. 4 ng·ml-1·h and 405. 55 ng·ml-1·h, respectively. Con-clusion: The validated method can be applied in the pharmacokinetic study of afatinib at the intragastric and intravenous dosage of 10 and 2 mg·kg-1, respectively.

Objective To explore the changes and significance of serum vitamin A levels in children with community acquired pneumonia. Methods A total of 80 children with community-acquired pneumonia (pneumonia group) were selected from October 2015 to March 2016 and were divided into Mycoplasma pneumoniae (MP) infection group, bacteria infection group, MP and bacteria mixed infection group (mixed infection group) according to different pathogens. Thirty healthy children in the same period were selected as the control group. The serum vitamin A concentration was detected by ultra-high-performance liquid chromatography-tandem mass spectrometry. Results The level of serum vitamin A was (0.567±0.163) μmol/L in pneumonia group, (0.578±0.162) μmol/L in MP infection group, (0.557±0.153) μmol/L in bacteria infection group and (0.554±0.186) μmol/L in mixed infection group, and all of them were lower than that in control group (0.759±0.160) μmol/L, and there were significant differences (P<0.05). There was no difference in serum vitamin A level among MP infection group, bacteria infection group and mixed infection group (P>0.05). There was a significant difference in the distribution of vitamin A deficiency between pneumonia group and control group ( P<0.001). The proportion of suspected subclinical vitamin A deficiency in control group was higher, while vitamin A deficiency and subclinical vitamin A deficiency in pneumonia group were higher. Conclusions The serum vitamin A level decreased in children with community-acquired pneumonia, But there was no significant differences in serum vitamin A levels among the children with pneumonia caused by different pathogens.

Purpose: Neoadjuvant therapy modality can increase the operability rate and mitigate pathological risks in locally advanced cervical cancer, but treatment response varies widely. It remains unclear whether genetic alterations correlate with the response to neoadjuvant therapy and disease-free survival (DFS) in locally advanced cervical cancer. Materials and Methods: A total of 62 locally advanced cervical cancer (stage IB-IIA) patients who received neoadjuvant chemoradiation plus radical hysterectomy were retrospectively analyzed. Patients’ tumor biopsy samples were comprehensively profiled using targeted next generation sequencing. Pathologic response to neoadjuvant treatment and DFS were evaluated against the association with genomic traits. Results: Genetic alterations of PIK3CA were most frequent (37%), comparable to that of Caucasian populations from The Cancer Genome Atlas. The mutation frequency of genes including TERT, POLD1, NOS2, and FGFR3 was significantly higher in Chinese patients whereas RPTOR, EGFR, and TP53 were underrepresented in comparison to Caucasians. Germline mutations were identified in 21% (13/62) of the cohort and more than half (57%) had mutations in DNA damage repair genes, including BRCA1/2, TP53 and PALB2. Importantly, high tumor mutation burden, TP53 polymorphism (rs1042522), and KEAP1 mutations were found to be associated with poor pathologic response to neoadjuvant chemoradiation treatment. KEAP1 mutations, PIK3CA-SOX2 co-amplification, TERC copy number gain, and TYMS polymorphism correlated with an increased risk of disease relapse. Conclusion We report the genomic profile of locally advanced cervical cancer patients and the distinction between Asian and Caucasian cohorts. Our findings highlight genomic traits associated with unfavorable neoadjuvant chemoradiation response and a higher risk of early disease recurrence.

Objective To explore the efficacy and safety of Solitaire stent arterial embolectomy in the treatment of acute cardiogenic cerebral embolism.Methods The data of 24 patients who underwent Solitaire stent arterial embolectomy,either alone or in combination with reorganization type tissue plasminogen activator (rt-PA) intravenous thrombolysis,to treat acute cardiogenic cerebral embolism in our hospital were collected.These results between October 2012 and March 2016 were compared with 21 control patients who were treated using only rtPA Ⅳ thrombolysis.The short-term efficacy,long-term clinical outcomes,complications,and mortality rate of these two groups were compared,and the clinical outcomes of these patients with posterior circulation infarction in the two groups were further assessed.Results The time between onset and rt-PA administration in the study group (median time:3.17 h) was significantly shorter than that in the control group (4.00 h,P＜0.05);rt-PA dose used in both two groups was 50 mg.NIHSS scores and Glasgow Coma scale scores at discharge,and modified Rankin scale (mRs) scores 3 and 6 months after treatment in the study group were significantly higher than those in the control group (P＜0.05).There were no significant differences in symptomatic intra-cerebral hemorrhage,high perfusion encephalopathy,incidence of hernia,or mortality between the two groups (P＞0.05).The mRS scores 3 months after treatment in patients with posterior circulation infarction (2) were significantly lower than those in the control group (3,P＜0.05).Conclusions As compared with simple venous thrombolytic therapy,Solitaire stent embolectomy can significantly improve short-term neurological function and long-term prognosis in patients with acute cardiogenic cerebral embolism,enhance their life quality,without increasing the complication incidence and mortality rate.It is safe and effective,and patients with posterior circulation infarction can also be treated by arterial embolectomy.