Objective To compare the difference in pharmacokinetics characteristics of vancomycin in cerebrospinal fluid between administration by continuous infusion and interim infusion.Methods Twenty postoperative patients in the Department of Neurosurgery of Beijing Tiantan Hospital, Capital Medical University admitted into intensive care unit (ICU) to receive vancomycin for prophylaxis of intracranial infection were enrolled, and they were randomly distributed to a continuous intravenous infusion group and a interim intravenous infusion group, each group 10 cases. In continuous intravenous infusion group, the patients received a loading dose of vancomycin (15 mg/kg) by continuous intravenous pump infusion for 1 - 2 hours followed by 30 mg/kg vancomycin in a constant pump infusion rate for 24 hours; while in interim intravenous infusion group, the patients received 15 mg/kg vancomycin administered by intravenous pump infusion for 1 - 2 hours, once every 12 hours. The concentration of vancomycin in the cerebrospinal fluid at different time points was measured by two-dimensional liquid chromatography (2D-LC) method, the parameters of pharmacokinetics were calculated in the two groups, and the adverse reaction was observed.Results The comparison between the ratio of areas under the concentration-time curves (AUC) and minimum inhibitory concentration (MIC) of the continuous and interim groups showed no significant difference (19.7±14.0 vs. 16.1±6.4,P > 0.05). However, in the continuous intravenous infusion group, the drug concentration reached the peak value (0.96± 0.77)μg/mL at 12 hours, and later revealed a plateau concentration 0.91-0.93μg/mL for 12 hours; while in the intravenous infusion interim group, the drug concentration reached the peak value (0.92±0.47)μg/mL at 16 hours, in the later 2 hours declined to (0.84±0.45)μg/mL, and afterwards still had a tendency of persistent declination. In all the patients, no any adverse reaction related to the drug occurred.Conclusion Continuous intravenous infusion and interim intravenous infusion of vancomycin for the postoperative neurosurgical patients without intracranial infection have the similar efficacy of medication, but the former can achieve the peak concentration faster and later the fluctuation of drug concentration in cerebrospinal fluid is smaller than those in the latter.

Objective To evaluate the effects of different positions on cerebral blood flow in patients undergoing laparoscopic surgery.Methods Thirty patients of both sexes,aged 23-64 yr,of American Society of Anesthesiologists physical status Ⅰ or Ⅱ,scheduled for elective laparoscopic surgery,were included.Patients' position was changed using a random number table after induction of anesthesia.Patients were placed in the supine position and tilted 15° head-up or 15° head-down,and the parameters of the middle cerebral artery were monitored using the transcranial Doppler ultrasound.After admission to the operating room (baseline),after induction of anesthesia and before pneumoperitoneum,in the supine position,at head-up tilt and at head-down tilt,the mean blood flow velocity,pulsatility index,resistance index,mean arterial blood pressure (MAP) and heart rate were recorded.Results Compared with the baseline in the supine position,the MAP and bilateral mean blood flow velocity were significantly decreased at head-up tilt,and the MAP and bilateral pulsatility and resistance indices were significantly increased at head-down tilt (P＜0.05).Conclusion During laparoscopic surgery,head-up tilt can lead to a decrease in cerebral blood flow,and head-down tilt exerts no effect on cerebral blood flow.

OBJECTIVE:To establish the method for the determination of tigecycline (TGC) in human plasma. METHODS:After precipitated by acetonitrile,the plasma sample was determined by LC-MS/MS. Using d9-TGC as internal standard,Kromasil C18 column was used with mobile phase consisted of water (containing 0.05% TFA)-acetonitrile (gradient elution) at flow rate of 0.6 ml/min,column temperature of 40 ℃. The ion transitions were performed under ESI positive MRM model at m/z 586.3→513.2 and m/z 595.3→514.3 for TGC and internal standard,respectively. RESULTS:The linear range of TGC was 25-2 000 ng/ml (r=0.999 8),and lowest quantification limit was 25 ng/ml;intra-day and inter-day RSD was 3.15%-7.23%,and relative error was-4.53%-10.48%. Plasma sample kept stable after 3 times of freezing and thawing cycle,at room temperature for 24 h,in automat-ic sample injector for 24 h and freezing for 42 d (RSD<15%). Plasma concentration of TGC was 0-438.0 ng/ml in one patient with pan-drug resistant bacteria infection(0-12 h after administration). CONCLUSIONS:The developed method is accurate,sensi-tive and specific,and can be used for plasma concentration determination of TGC and pharmacokinetic study.

OBJECTIVETo investigate pre- and postoperative changes of regional cerebral cortical blood flow in patients with cerebral arteriovenous malformation.

METHODTwenty-two adult patients with arteriovenous malformation (AVM) were recruited into this study at Beijing Tiantan Hospital from September 2001 to May 2002. Eight patients had giant cerebral AVM and the other 14 had a small one. Cortical cerebral blood flow (CBF) was measured by laser Doppler flowmetry (LDF) before and after AVM resections. After surgery, the probe of LDF was implanted adjacent to the area of AVM and monitored for 24 hours.

RESULTSCBF increased significantly after the resection in all patients regardless of AVM size. In patients with small AVM, CBF returned to the baseline level within 4 hours, but in patients with giant AVM, CBF remained high even after 24 hours.

CONCLUSIONSMonitoring CBF is helpful to understand pre- and postoperative changes of regional cortical CBF in patients with cerebral AVM.

Adult ; Cerebrovascular Circulation ; physiology ; Female ; Humans ; Intracranial Arteriovenous Malformations ; physiopathology ; surgery ; Laser-Doppler Flowmetry ; Male ; Middle Aged ; Regional Blood Flow ; physiology

To investigate the effects of sanguinarine (Sang) combined with cisplatin (Cis) in accelerating the apoptosis of bladder cancer EJ cells, CCK-8 method was used to detect the proliferation of bladder cancer EJ cells treated with different concentrations of Sang with the IC50 values calculated. Annexin V FITC/PI method was used to detect cell apoptosis in the control group, Sang group, Cis group and the combination group. Flow cytometer was used to detect cell cycle arrested. Western blot was used to detect the influence of Bcl-2 expression in the control group, Sang group, Cis group and the combination group. Nude mouse subcutaneous tumor model was constructed to verify that the combination group could accelerate the apoptosis of bladder cancer EJ cells and reduce the side-effects on mice. The safety of the Sang was evaluated by HE staining of vital organs in mice. In vitro, Sang could significantly inhibit the proliferation of EJ cells. Compared with the control group, the number of apoptosis EJ cells in the combination group was significantly increased (P < 0.05), and more cells were arrested in G2/M phase. The expression of Bcl-2 was significantly down-regulated in the combination group (P <0.001). In vivo, compared with the control group, the tumor growth was significantly slower, and a large number of apoptotic cells were inspected (P < 0.05) of the combination group. The side effects of cisplatin were reduced in the combination group. Sang has high biosafety and little side effect. Combined Sang and Cis can increase cell cycle G2/M block, down-regulate Bcl-2 expression, promote cell apoptosis and inhibit tumor growth.

Objective To explore the value of mammogram texture analysis in prediction of metastasis of axillary lymph nodes for breast carcinoma.Methods Mammograms and clinical data of 171 patients with breast carcinoma confirmed by pathology were retrospectively analyzed,and all patients underwent axillary lymph node dissection (ALND).Then the patients were divided into axillary lymph node metastasis group and non-metastasis group according to the result of ALND.The texture features of these lesions were statistically analyzed,including gray-level histogram texture parameters (mean value,standard deviation,skewness,kurtosis and variance) and gray-level co-occurrence matrix texture parameters (energy,entropy,correlation,inertia,inverse difference moment and contrast).Results In all of 171 breast cancer patients,96patients had axillary lymph node metastasis,while 75 patients had no metastasis.Mammograms showed negative axillary lymph nodes in 119 patients and positive axillary lymph nodes in 52 patients,and the sensitivity and specificity of mammograms in the diagnosis of positive axillary lymph nodes was 48.96％ (47/96) and 93.33％ (70/75),respectively.Mammogram texture analysis showed the values of energy,entropy,inverse difference moment and correlation in axillary lymph node metastasis group were higher than those in non-metastasis group,while the values of inertia and contrast in the axillary lymph node metastasis group were lower than those in non-metastasis group (all P＜0.05).The rest texture parameters had no significant differences between two groups (all P＞0.05).Area under curve (AUC) for texture parameters of energy,entropy,inertia,inverse difference moment,correlation and contrast was 0.610,0.610,0.374,0.599,0.612 and 0.421 (all P＜0.05),respectively.AUC of mammography,mammogram texture features,and the combination of mammography and texture features was 0.711,0.676 and 0.787 (all P＜0.05),respectively.The sensitivity and specificity of mammogram texture features,the combination of mammography and texture features in diagnosis of axillary lymph nodes metastasis was 62.5％ and 64.6％,66.7％ and 82.7％,respectively.Conclusion Mammogram texture parameters are helpful for predicting axillary lymph node metastasis,and the combination of mammography and texture features can improve diagnostic efficiency of axillary lymph node metastasis.

The aim of this study was to construct the complete genome of Marek's disease virus serotype 814 strain as an infectious bacterial artificial chromosome (BAC). Using self-designed selection marker Eco-gpt (1.3 kb) and BAC vector pBeloBAC11 (7.5 kb), we constructed the transfer plasmid pUAB-gpt-BAC11. The plasmid pUAB-gpt-BAC11 and MDV total-DNA were cotransfected into secondary CEFs; we put the virus-containing cells in selection medium for eight rounds and obtained purified recombinant viruses. Recombinant viral genomes were extracted and electroporated into E. coli, BAC clones were identified by restriction enzyme digestion and PCR analysis. Finally, we obtained 38 BAC clones, DNA from various MDV-1 BACs was transfected into CEFs, and recombinant virus was reconstituted by transfection of MDV-BAC2 DNA. We successfully cloned the complete genome of MDV-1814 strain as an infectious bacterial artificial chromosome. With these cloned genomes, a revolutionary MDV-DNA engineering platform utilizing RED/ET recombination system was constructed successfully, which can help the understanding of MDV gene functions and promote the using of MDV as a vector for expressing foreign genes. In addition, it opens the possibility to generate novel MDV-1 vaccines based on the BACs.
Animals ; Chickens ; immunology ; virology ; Chromosomes, Artificial, Bacterial ; genetics ; Cloning, Molecular ; DNA, Recombinant ; genetics ; DNA, Viral ; genetics ; Fibroblasts ; metabolism ; Genetic Engineering ; methods ; Mardivirus ; classification ; genetics ; physiology ; Serotyping ; Transfection ; Viral Proteins ; genetics ; physiology ; Virus Replication

Improving expression of antigen is critical to the immunogenicity of DNA vaccines. To achieve this goal, we modified the NDV F48E9 strain HN gene by optimizing the condon usage and inserting the secretary leader sequence [A/Goose/Guangdong/1/96 (H5N1) HA gene, Accession No. AF144305]. The HN gene modified and knocked the signal peptide off were named SoptiHN and optiHN. The three sequence: SoptiHN, optiHN and the NDV F48E9 strain HN gene were inserted into the vector pVAX1 and vector pVAX1-CpG including CpG-ODN sequence respectively. Then we got six recombinant plasmids: pV-SoptiHN, pVC-SoptiHN, pV-optiHN, pVC-optiHN, pV-HN and pVC-HN. By optimizing condon usage in transiently transfected 293T cells, expression levels of HN gene were higher from the codon-optimized gene than the counterpart. Moreover, both optimization of condon usage and addition of signal peptide could improve expression of HN gene in vitro.
Animals ; Chickens ; Codon ; HN Protein ; genetics ; Hemagglutinin Glycoproteins, Influenza Virus ; genetics ; Influenza A Virus, H5N1 Subtype ; genetics ; Newcastle Disease ; immunology ; prevention & control ; Newcastle disease virus ; classification ; genetics ; Vaccines, DNA ; genetics ; immunology ; Viral Vaccines ; genetics ; immunology

Objective To explore the application value of short latency somatosensory evoked potentials (SLSEP) as a tool for preoperative assessment of surgical or interventional treatment in patients with severe aneurysmal subarachnoid hemorrhage (aSAH). Methods A prospective observational cohort study was conducted. The patients with severe aSAH with a WFNS grade of Ⅳ or Ⅴ admitted to intensive care unit (ICU) of Beijing Tiantan Hospital of Capital Medical University from November 2016 to April 2017 were enrolled. The patients received SLSEP monitoring within 12 hours after onset, and the monitoring results were classified according to the Judson scale. Meanwhile, the findings on cerebral CT scans at admission were evaluated by the modified Fisher classification. The follow-up was performed at 3 months after aSAH ictus based on the modified Rankin scale (mRS), and a mRS score 0-3 was defined as favorable outcome, 4-6 was defined as unfavorable outcome. For statistical evaluation, demographic, clinical, neuroimaging and SLSEP data were evaluated by univariate analysis to identify the risk factors associated with prognosis;afterwards, those factors were analyzed by multivariate Logistic regression; also the validity was assessed by calculating the respective sensitivity, specificity, positive predictive value (PPV) and negative predictive value (NPV). Results A total of 41 patients with aSAH were selected, of which 7 were excluded because of the interference of the SLSEP monitoring results, 34 patients with aSAH were enrolled finally. Among them, 21 were classified in the unfavorable outcome group, while the rest (n = 13) were allocated into the favorable outcome group. No significant difference was found in gender, age, body mass index (BMI), time delay from ictus to treatment or the options for therapeutic methods between the two groups. The findings of univariate analysis, however, showed statistically differences in WFNS grade, the modified Fisher scale and Judson scale of SLSEP between the two groups. Yet, the further validity evaluation for these predictors demonstrated that the sensitivity, specificity, PPV and NPV of WFNS grade of Ⅴ and modified Fisher scale of Ⅳ were all less than 85%, whereas the results for SLSEP Judson scale of Ⅲ were much better (sensitivity: 90.5% vs. 71.4% and 71.4%, specificity: 84.6% vs. 69.2% and 76.9%, PPV: 90.5% vs. 79.0% and 83.3%). In the following multivariate Logistic analysis, only Judson scale of Ⅲ was identified to be the independent risk factor for poor outcome [odds ratio (OR) = 45.73, 95% confidence interval (95%CI) = 4.25-499.31, P = 0.002], while the WFNS grade of Ⅴ (OR = 1.14, 95%CI = 0.12-13.06, P = 0.912) and the modified Fisher scale of Ⅳ (OR = 7.22, 95%CI = 0.51-113.20, P = 0.160) were merely associated with poor outcomes without significant independence. Conclusion In comparison with WFNS grade and the modified Fisher scale, SLSEP seems more accurate in the prediction of long-term outcome of severe aSAH prior to surgical or interventional treatment, and thus may be applied as an effective aid in preoperative assessment.

Objective To evaluate the diagnostic validity of application of brainstem auditory evoked potential (BAEP) as an ancillary test in patients with brain death.Methods A prospective observational study was conducted. Adult patients (≥ 18 years) with brain death were consecutively admitted to the Department of Intensive Care Unit (ICU) of Beijing Tiantan Hospital Affiliated to Capital Medical University from July 2015 to June 2017, and they further underwent BAEP monitoring. The following findings: absence of all waveforms at bilateral sides, absence of all waveforms except wave-Ⅰonly present at bilateral orunilateral side were set as the positive criteria of BAEP for the diagnosis of brain death, and its diagnostic sensitivity (SEN), positive predictive value (PPV) and false negative rate (FNR) were further assessed.Results A total of 47 patients were enrolled in the study, and the 43 complete BAEP results were analyzed. The SEN, PPV and FNR of the above 3 types of diagnostic criteria (absence of all waveforms, and unilateral or bilateral presence of wave-Ⅰ only) were 100% [95% confidence interval (95%CI) 90-100], 100% (95%CI 89-100), and 0% (95%CI 0-9), respectively.Conclusion As an ancillary test, BAEP has extremely high validity for the diagnosis of patients with brain death, thus it should be used more extensively in clinical practice.