The attentional guidance and facilitating effects of group behavioral cues on individual college pedestrians' jaywalking decisions.

OBJECTIVE: Jaywalking is an important cause of pedestrian-related automobile accidents. Exploring the factors that influence jaywalking behavior and suggesting appropriate improvement measures are critical for reducing automobile accidents involving pedestrians. METHODS: This study divided traffic situations into high-risk and low-risk situations. Each situation contained three visual attention cues: vehicle, traffic light, and group behavior. Based on this, the role of visual cues in guiding pedestrians' attention and influencing their decisions during jaywalking was examined. Sixty participants, with an average age of 19, were recruited. They were shown 84 crosswalk videos randomly while their crossing decisions and eye movement data were recorded. RESULTS: In low-risk situations, pedestrians spent more attention on group behavioral cues when making jaywalking decisions. The rate of jaywalking increased with the number of other jaywalking pedestrians. In high-risk situations, the pedestrians' total fixation duration at vehicle hazard cues was longer when making jaywalking decisions, and the jaywalking rate decreased. CONCLUSIONS: The results indicate that pedestrians' jaywalking decisions were based on other pedestrians' illegal crossing cues and automatic associative processes in low-risk situations. The higher the number of people crossing the street, the higher the number of pedestrians illegally crossing the road. In high-risk situations, pedestrians paid more attention to vehicle hazard cues before making jaywalking decisions, and fewer illegal crossings. The jaywalking decisions were based on a risk assessment, a controlled analytical process. The results verify the effect of visual cues on pedestrians' attentional guidance and decision-making in different traffic situations, as well as the effectiveness of visual attention in predicting decision intention. The findings provide a theoretical basis and data reference for pedestrian safety education and constructing an intelligent driving pedestrian trajectory prediction model.

The role of anger rumination as a mediator in the relationship between driver moral disengagement and driving angry.

Background: Road rage is a common phenomenon during driving, which not only affects the psychological health of drivers but also may increase the risk of traffic accidents. This article explores the impact of moral disengagement and anger rumination on road rage through two studies. Method: This research combined experimental studies with survey questionnaires. Study one used a driving simulator to investigate whether moral disengagement and anger rumination are psychological triggers of road rage in real-time driving, and whether there are differences in the main psychological triggers of road rage under different road scenarios. Building on the first study, study two employed a survey questionnaire to analyze the relationship between moral disengagement, anger rumination, and road rage. Participants in both studies were drivers with certain driving ages and experience. Data were processed and analyzed using descriptive statistics, factor analysis, reliability and validity tests, and multiple regression analysis. Results: The findings indicated: (1) There were significant differences in the anger induction rate across different road scenarios, χ2 = 35.73, p < 0.01, effect size = 0.29. Significant differences in average anger levels were observed in scenarios involving oncoming vehicles, lane-cutting, sudden stops by the vehicle ahead, pedestrians crossing the road, and traffic congestion (F = 20.41, p < 0.01, ηp2 = 0.36), with anger rumination playing a major role in the formation of road rage; (2) Moral disengagement significantly predicted road rage (ß = 0.25, t = 3.85, p < 0.01). The predictive effect of moral disengagement on anger rumination was significant (ß = 0.39, t = 6.17, p < 0.01), as was the predictive effect of anger rumination on road rage (ß = 0.43, t = 6.3, p < 0.01). The direct effect of moral disengagement on road rage included 0 in the bootstrap 95% confidence interval, while the mediating effect of anger rumination did not include 0 in the bootstrap 95% confidence interval, indicating that anger rumination fully mediated the relationship between moral disengagement and road rage.

Developmental differences in children's adaptation to vehicle distance and speed in street-crossing decision-making.

INTRODUCTION: Young children cannot effectively adapt their behaviors to vehicles at varied distances and speeds, which is a critical cause of road accidents. However, the impact of this crucial ability on children's street-crossing decision-making and the age at which they acquire it remain unclear. METHOD: This study examined the crossing decision-making behavior of children at 6, 8, and 11 years of age in facing 51 different videotaped traffic scenarios with varying vehicle distances and speeds. Sixty Chinese elementary school students, with 20 children evenly distributed into each of the three age groups (6 years, 8 years, and 11 years old), participated in a simulated street-crossing task using video projections. Hierarchical logistic regression models were used to analyze how age moderated the effects of vehicular motion factors (vehicle-pedestrian distance, vehicle speed) on children's crossing safety, including dangerous crossing and crossing decision-making. RESULTS: The results showed that when either vehicle-pedestrian distance decreased or vehicle speed increased all age groups tended to cross less frequently but probability of dangerous crossing increased. Compared to 8-year-old and 11-year-old children, 6-year-old children showed a less pronounced tendency toward both of these crossing decision-making behaviors, and had more dangerous crossing outcomes. CONCLUSIONS: These findings suggest that inadequate adaptation to vehicle-pedestrian distance and vehicle speed may partly contribute to the inferior safety of street-crossing behavior in 6-year-olds compared to 8-year-olds. No significant differences were observed between 8- and 11-year-old children, suggesting the turning point for this ability might occur between 6 and 8 years of age. PRACTICAL APPLICATIONS: Preventive measures aimed to reduce crossing risks for children should consider children's developmental stages.

The small nucleolar RNA SNORA51 enhances breast cancer stem cell-like properties via the RPL3/NPM1/c-MYC pathway.

Breast cancer stem cells (BCSCs) are key players in carcinogenesis and development. Small nucleolar RNAs (snoRNAs) seem to have a crucial influence on regulating stem cell-like properties in various cancers, but the underlying mechanism in breast cancer has not been determined. In this study, we first found that the expression of SNORA51 might be strongly and positively related to BCSCs-like properties. SNORA51 expression was assessed in breast cancer tissues (n = 158 patients) by in situ hybridization. Colony formation, cell counting kit-8, and sphere formation assays were used to detect cell proliferation and self-renewal, respectively. Wound healing and transwell assays were used to detect cell migration. Coimmunoprecipitation and molecular docking were used to determine the underlying mechanism through which SNORA51 regulates BCSCs-like properties. High SNORA51 expression was associated with a worse prognosis, overall survival, and disease-free survival, in 158 breast cancer patients and was also closely related to lymph node status, ER status, the Ki-67 index, histological grade, and TNM stage. Further analysis proved that SNORA51 could enhance and maintain stem cell-like properties, including cell proliferation, self-renewal, and migration, in breast cancer. Moreover, high SNORA51 expression could reduce nucleolar RPL3 expression, induce changes in the expression of NPM1 in the nucleolus and nucleoplasm, and ultimately increase c-MYC expression. Taken together, our findings demonstrated that SNORA51 could enhance BCSCs-like properties via the RPL3/NPM1/c-MYC pathway both in vitro and in vivo. Therefore, SNORA51 might be a significant biomarker and potential therapeutic target and might even provide a new viewpoint on the regulatory mechanism of snoRNAs in breast cancer or other malignant tumors.

Sex difference in driving speed management: The mediation effect of impulse control.

Previous studies have shown that male drivers drive faster than female drivers, but there is no agreement on whether impulsivity could induce this sex difference, nor is there a cross-sectional comparison of the effects of different road environments. The purpose of this study was to verify whether impulsivity and impulse control could explain the sex differences in driving speed. A driving simulator study (study 1, N = 41) was performed to investigate whether there were sex differences in driving speeds in two road sections of different complexity, and a questionnaire survey (study 2, N = 163) was conducted to investigate the relationship between sex, impulsivity, impulse control and driving behavior of the participants. The results showed that male drivers drove faster on simple roads, but this difference did not show on complex roads. There were no sex differences in impulsivity traits, but male participants had significant lower levels of impulse control. The results also reveal a partial mediating role of impulse control in the relationship between sex and driving speed. These results suggest that impulse control can predict dangerous driving behaviors and is an important factor in explaining sex differences in driving speed.

Mental workload accumulation effect of mobile phone distraction in L2 autopilot mode.

As automated vehicles become more common, there is a need for precise measurement and definition of when and in what ways a driver can use a mobile phone in L2 autonomous driving mode, for how long it can be used, the complexity of the call content, and the accumulated mental workload. This study uses a 2 (driving mode) × 2 (call content complexity) × 6 (driving stage) three-factor mixed experimental design to investigate the effect of these factors on the driver's mental workload by measuring the driver's performance on Detection response tasks, pupil diameter, and EEG components in various brain regions in the alpha band. The results showed that drivers' mental workload levels converge between manual and automatic driving modes as the duration of driving increases, regardless of the level of complexity of the mobile phone conversation. This suggests that mobile phone conversations can also disrupt the driver's cognitive resource balance in L2 automatic driving mode, as it increases mental workload while also impairing the normal functioning of brain functions such as cognitive control, problem solving, and judgment, thereby compromising driving safety.

Comparison Between Heated Eye Mask and Intense Pulsed Light Treatment for Contact Lens-Related Dry Eye.

Background: Contact lens-related dry eye (CLDE) is common in contact lens (CL) wearers and can lead to ocular pain, decreased visual quality, and reduced quality of life. Objective: The purpose of this prospective, randomized, examiner masked study was to compare the effectiveness of intense pulsed light (IPL) and heated eye mask (HEM) for relieving CLDE. Methods: The final analysis included 60 patients (30 in the IPL group and 30 in the HEM group) who were using CL and had been suffering contact lens discomfort for more than a year. The IPL treatment group had two IPL treatments spaced 3 weeks apart, whereas the HEM group received HEM daily for 6 weeks (42 days). Non-invasive tear break-up time (NITBUT), tear-film lipid layer (TFLL), conjunctival fluorescein staining, meibum gland quality and expression, conjunctival redness score, ocular surface disease index (OSDI), best-corrected visual acuity, endothelial cell count, and intraocular pressure were assessed at baseline: first visit (V1), second visit (V2), and third visit (V3). Results: The mean age of the IPL group was 28.6 ± 4.3 years (16 females, 53%) and that of the HEM group was 28.6 ± 4.2 years (16 females, 53%). Mean NITBUT for the IPL group at V2 was 5.53 ± 0.77 sec (p < 0.001) and at V3 was 7.72 ± 0.88 sec (p < 0.001); the IPL group demonstrated a clinically and statistically significant improvement in mean NITBUT. In addition, the HEM group showed a clinically and statistically significant improvement in mean NITBUT at V3 (5.86 ± 0.76 sec, p < 0.001). At V3, the C-OSDI, TFLL score, and meibum gland quality and expressibility all increased considerably (p < 0.05) in both groups. Conclusions: Our results imply that IPL or HEM treatment of CLDE can be safely used to relieve symptoms of CLDE. In addition, IPL treatment is more effective in improving the general stability of the tear-film and decreasing the need for artificial tears in CLDE.

Management of Post-LASIK Dry Eye with Intense Pulsed Light in Combination with 0.1% Sodium Hyaluronate and Heated Eye Mask.

INTRODUCTION: Laser in situ keratomelieusis (LASIK) is one of the most frequently performed refractive treatments. Dry eye (DE) is common in patients after LASIK and can be bothersome postoperatively. Therapies such as intense pulsed light (IPL), sodium hyaluronate (SH) and heated eye mask (HEM) have been reported to improve signs and symptoms of DE . AIM: The purpose of this prospective study was to evaluate and compare the effects of IPL and 0.1% SH (IPL group, 50 eyes) and IPL in combination with 0.1% SH and HEM (IPL + group, 50 eyes) in participants with persistent post-LASIK DE. METHODS: The final analysis included 100 patients (100 eyes) who had LASIK for myopic correction and had been experiencing moderate to severe DE following LASIK for over a year. Participants were randomly assigned to either the IPL group (2 IPL sessions) or IPL + group (2 IPL sessions and daily HEM for 4 weeks), and both groups continued the use of daily 0.1% SH (HYLO-COMOD®) preservative-free eye drops. Non-invasive tear break-up time (NITBUT), tear film lipid layer (TFLL), lower tear meniscus height (LTMH), meibomian gland quality (MGQ), meibomian gland expressibility (MGEx), corneal fluorescein staining (CFS), ocular surface disease index (OSDI) and artificial tear usage (ATU) survey were assessed at baseline (BL) and follow-up at 2 (F1) and 4 weeks (F2). RESULTS: Following the treatment protocol, all dry eye (DE) parameters assessed in this study improved significantly (P < 0.05) in both groups at F2 compared with their respective BL measurements. Inter-group comparison at F2 found significant differences in their NITBUT (IPL: 6.06 ± 0.59 vs. IPL +: 6.67 ± 0.86, P < 0.001), TFLL (IPL: 1.90 ± 0.65 vs. IPL +: 1.60 ± 0.64, P = 0.021), LTMH (IPL: 0.186 ± 0.053 vs. 0.204 ± 0.034, P = 0.003), MGQ (IPL: 1.48 ± 0.54 vs. IPL +: 1.26 ± 0.56, P = 0.026), MGEx (IPL: 1.62 ± 0.53 vs. IPL +: 1.44 ± 0.50, P = 0.038) and OSDI (IPL: 32.54 ± 6.85 vs. IPL +: 29.76 ± 4.74, P = 0.001), while CFS score (IPL: 4.02 ± 0.65 vs. IPL +: 3.96 ± 0.73, P = 0.652) and ATU (IPL: 1.88 ± 0.63 vs. IPL +: 1.72 ± 0.50, P = 0.159) showed no significant difference. CONCLUSION: Post-LASIK DE signs and symptoms can be improved by combining therapies such as IPL, HEM and 0.1% SH. Increased TFLL due to a combination of IPL, 0.1% SH and HEM had a greater positive impact on the subjective and objective DE measurements in participants with persistent post-LASIK DE.

Electrophysiological frequency domain analysis of driver passive fatigue under automated driving conditions.

With the continuous improvement of automated vehicles, researchers have found that automated driving is more likely to cause passive fatigue. To explore the impact of automation and scenario complexity on the passive fatigue of a driver, we collected electroencephalography (EEG), detection-response task (DRT) performance, and the subjective report scores of 48 drivers. We found that in automated driving under monotonic conditions, after 40 min, the alpha power of the driver's EEG indicators increased significantly, the accuracy of the detection reaction task decreased, and the reaction time became slower. The receiver characteristic curve was used to calculate the critical threshold of the alpha power during passive fatigue. The determination of the threshold further clarifies the occurrence time and physiological characteristics of passive fatigue and improves the passive fatigue theory.

Effects of automation trust in drivers' visual distraction during automation.

With ongoing improvements in vehicle automation, research on automation trust has attracted considerable attention. In order to explore effects of automation trust on drivers' visual distraction, we designed a three-factor 2 (trust type: high trust group, low trust group) × 2 (video entertainment: variety-show videos, news videos) × 3 (measurement stage: 1-3) experiment. 48 drivers were recruited in Dalian, China for the experiment. With a driving simulator, we used detection-response tasks (DRT) to measure each driver's performance. Their eye movements were recorded, and automation-trust scale was used to divide participants into high trust group and low trust group. The results show that: (1) drivers in the high trust group has lower mental workload and paid more attention to visual non-driving-related tasks; (2) video entertainment also has an impact on distraction behavior, variety-show videos catch more attention than news videos. The findings of the present study indicate that drivers with high automation trust are more likely to be involved in non-driving-related visual tasks.

DEAD-box helicase 27 enhances stem cell-like properties with poor prognosis in breast cancer.

BACKGROUND: Although the rapid development of diagnosis and treatment has improved prognosis in early breast cancer, challenges from different therapeutic response remain due to breast cancer heterogeneity. DEAD-box helicase 27 (DDX27) had been proved to influence ribosome biogenesis and identified as a promoter in gastric and colorectal cancer associated with stem cell-like properties, while the impact of DDX27 on breast cancer prognosis and biological functions is unclear. We aimed to explore the influence of DDX27 on stem cell-like properties and prognosis in breast cancer. METHODS: The expression of DDX27 was evaluated in 24 pairs of fresh breast cancer and normal tissue by western blot. We conducted Immunohistochemical (IHC) staining in paraffin sections of 165 breast cancer patients to analyze the expression of DDX27 and its correlation to stemness biomarker. The Cancer Genome Atlas-Breast Cancer (TCGA-BRCA) database and the Clinical Proteomic Tumor Analysis Consortium (CPTAC) database were used to analyze the expression of DDX27 in breast cancer. Kaplan-Meier survival analysis were used to investigate the implication of DDX27 on breast cancer prognosis. Western blot, CCK-8 assay, Transwell assay and wound-healing assay were carried out to clarify the regulation of DDX27 on stem cell-like properties in breast cancer cells. Gene Set Enrichment Analysis (GSEA) was performed to analyze the potential molecular mechanisms of DDX27 in breast cancer. RESULTS: DDX27 was significantly high expressed in breast cancer compared with normal tissue. High expression of DDX27 was related to larger tumor size (p = 0.0005), positive lymph nodes (p = 0.0008), higher histological grade (p = 0.0040), higher ki-67 (p = 0.0063) and later TNM stage (p < 0.0001). Patients with high DDX27 expression turned out a worse prognosis on overall survival (OS, p = 0.0087) and disease-free survival (DFS, p = 0.0235). Overexpression of DDX27 could enhance the expression of biomarkers related to stemness and promote stem cell-like activities such as proliferation and migration in breast cancer cells. CONCLUSION: DDX27 can enhance stem cell-like properties and cause poor prognosis in breast cancer, also may be expected to become a potential biomarker for breast cancer therapy.

Comparative Analysis of Paper-Based and Web-Based Versions of the National Comprehensive Cancer Network-Functional Assessment of Cancer Therapy-Breast Cancer Symptom Index (NFBSI-16) Questionnaire in Breast Cancer Patients: Randomized Crossover Study.

BACKGROUND: Breast cancer remains the most common neoplasm diagnosed among women in China and globally. Health-related questionnaire assessments in research and clinical oncology settings have gained prominence. The National Comprehensive Cancer Network-Functional Assessment of Cancer Therapy-Breast Cancer Symptom Index (NFBSI-16) is a rapid and powerful tool to help evaluate disease- or treatment-related symptoms, both physical and emotional, in patients with breast cancer for clinical and research purposes. Prevalence of individual smartphones provides a potential web-based approach to administrating the questionnaire; however, the reliability of the NFBSI-16 in electronic format has not been assessed. OBJECTIVE: This study aimed to assess the reliability of a web-based NFBSI-16 questionnaire in breast cancer patients undergoing systematic treatment with a prospective open-label randomized crossover study design. METHODS: We recruited random patients with breast cancer under systematic treatment from the central hospital registry to complete both paper- and web-based versions of the questionnaires. Both versions of the questionnaires were self-assessed. Patients were randomly assigned to group A (paper-based first and web-based second) or group B (web-based first and paper-based second). A total of 354 patients were included in the analysis (group A: n=177, group B: n=177). Descriptive sociodemographic characteristics, reliability and agreement rates for single items, subscales, and total score were analyzed using the Wilcoxon test. The Lin concordance correlation coefficient (CCC) and Spearman and Kendall τ rank correlations were used to assess test-retest reliability. RESULTS: Test-retest reliability measured with CCCs was 0.94 for the total NFBSI-16 score. Significant correlations (Spearman ρ) were documented for all 4 subscales-Disease-Related Symptoms Subscale-Physical (ρ=0.93), Disease-Related Symptoms Subscale-Emotional (ρ=0.85), Treatment Side Effects Subscale (ρ=0.95), and Function and Well-Being Subscale (ρ=0.91)-and total NFBSI-16 score (ρ=0.94). Mean differences of the test and retest were all close to zero (≤0.06). The parallel test-retest reliability of subscales with the Wilcoxon test comparing individual items found GP3 (item 5) to be significantly different (P=.02). A majority of the participants in this study (255/354, 72.0%) preferred the web-based over the paper-based version. CONCLUSIONS: The web-based version of the NFBSI-16 questionnaire is an excellent tool for monitoring individual breast cancer patients under treatment, with the majority of participants preferring it over the paper-based version.

Decoding Analysis of Alpha Oscillation Networks on Maintaining Driver Alertness.

The countermeasure of driver fatigue is valuable for reducing the risk of accidents caused by vigilance failure during prolonged driving. Listening to the radio (RADIO) has been proven to be a relatively effective "in-car" countermeasure. However, the connectivity analysis, which can be used to investigate its alerting effect, is subject to the issue of signal mixing. In this study, we propose a novel framework based on clustering and entropy to improve the performance of the connectivity analysis to reveal the effect of RADIO to maintain driver alertness. Regardless of reducing signal mixing, we introduce clustering algorithm to classify the functional connections with their nodes into different categories to mine the effective information of the alerting effect. Differential entropy (DE) is employed to measure the information content in different brain regions after clustering. Compared with the Louvain-based community detection method, the proposed method shows more superior ability to present RADIO effectin confused functional connection matrices. Our experimental results reveal that the active connection clusters distinguished by the proposed method gradually move from frontal region to parieto-occipital regionwith the progress of fatigue, consistent with the alpha energy changes in these two brain areas. The active class of the clusters in parieto-occipital region significantly decreases and the most active clusters remain in the frontal region when RADIO is taken. The estimation results of DE confirm the significant change (p < 0.05) of information content due to the cluster movements. Hence, preventing the movement of the active clusters from frontal region to parieto-occipital region may correlate with maintaining driver alertness. The revelation of alerting effect is helpful for the targeted upgrade of fatigue countermeasures.

Prevalence of symptomatic dry eye in breast cancer patients undergoing systemic adjuvant treatment: A cross-sectional study.

OBJECTIVES: To investigate the prevalence of symptomatic dry eye (SDE) on women undergoing systemic adjuvant therapy for breast cancer and its association with treatment settings. METHODS: Woman undergoing breast cancer systemic adjuvant therapy were included in exposure group. An age-matched non-treatment control group was recruited. This cross-sectional questionnaire-based study utilised validated Ocular Surface Disease Index (OSDI) and NCCN-FACT-Breast Cancer Symptom Index (NFBSI-16) questionnaires to determine the presence of SDE and investigate other breast cancer treatment complications. Additionally, demographic data and medical histories were collected. RESULTS: Of 423 eligible participants, 200 in each of the control group and the exposure group were included in the final analysis. The prevalence of SDE was 59.0% in breast cancer patients with adjuvant treatment, statistically significantly higher than 25.5% in the control group (P < 0.01). Additionally, exposure group experienced higher prevalence of moderate and severe SDE, which were 20.0% and 19.5% respectively compared with 9.0% and 4.0% in the control group (P = 0.002, P < 0.001). There was a significantly high prevalence of SDE among patients who had received over four cycles of systemic therapy (71.0%, P < 0.001) and the application of targeted therapy (71.2%, P = 0.014). The severity of SDE positively correlated with the cycles of treatment administered. CONCLUSION: SDE was significantly predominant in women with breast cancer undergoing systemic adjuvant treatment. Our findings suggest dry eye assessments among patients receiving more than four cycles of chemotherapy or targeted therapy, thus early revealing possible dry eye conditions to both patients and clinicians for further specialized examination and treatment.

Improving Education About Breast Cancer for Medical Students in China.

In traditional medical school curriculum of cancer education in China, there is a very limited amount of teaching about breast cancer. The current situation may result in indifference to breast cancer education among medical students. Case-based learning (CBL) is a popular teaching method based on clinical cases. To date, there are few research reports about the application and research of CBL in breast cancer education. The aim of this study is to explore the teaching effect about CBL combined with lecture-based learning (LBL) in breast cancer education. Questions of breast cancer in National Medical Licensing Examination (NMLE) from 2011 to 2018 were analyzed. The questions about breast cancer were used as the evaluation criteria for this study. In this pilot study, a total of 140 students were randomly divided into a lecture only group (control group) and a lecture plus CBL group (observation group). The students in the observation group had better academic performances and abilities of memory, understanding, and application. They also had higher favorable impressions of the learning experience. In conclusion, more active approaches yield more learning and are viewed more favorably. CBL plus lecture can significantly improve education about breast cancer among medical students, which may be an important message for the evolution of curriculum in Chinese medical schools.

The Relationship Between Drivers' Cognitive Fatigue and Speed Variability During Monotonous Daytime Driving.

A lack of task workload can lead to drivers' cognitive fatigue and vigilance decrement during a prolonged drive. This study examined the effects of speed variability on driving fatigue in a monotonous drive. Twenty-one participants participated in a 60-min simulated driving task. All participants' cognitive fatigue was assessed using psychological and physiological measurements. Results showed that among all participants, variability of vehicle speed was negatively correlated with sleepiness and hypo-vigilance during the driving task. Further, drivers in the large variability group reported less sleepiness, less fatigue, and more vigilance than those in the small variability group did during the driving task. These drivers also presented a smaller electroencephalogram spectral index (𝜃+α)/ß during the task, where 𝜃, α, and ß are the power spectra of three different frequency bands: theta (𝜃, 4∼8 Hz), alpha (α, 8∼13 Hz), and beta (ß, 13∼30 Hz). Our findings suggested that the larger variability of speed within the speed limit may have a deterrent effect on drivers' cognitive fatigue.

[Development of a single-tube PCR-pyrosequencing method for simultaneous and rapid detection of the genetic polymorphism of warfarin metabolizing enzymes].

The purpose of this article is to develop a new high throughput method for detecting genetic polymorphism of warfarin metabolism-related genes rapidly in a single tube. Genomic DNA from human peripheral blood was extracted, and amplified with biotinylated primer to obtain single-stranded templates for pyrosequencing. Then, the single-stranded tem-plates were subjected to Pyrosequencing analysis using PyroMark ID instrument. Simultaneously, Sanger sequencing was also applied to sequence the products as a control to check the reliability of the pyrosequencing result.. The results dis-played that three variants of the warfarin metabolism-related genetic polymorphism (CYP2C9*2, CYP2C9*3, and VKORC1(-1693)) could be simultaneously detected using three different sequencing primers in a single-tube (one test), and 96 tests could be carried out each time. Repeat test and reliability test indicated that the agreement between the pyrosequencing and the Sanger sequencing methods was 100%. . All of these demonstrated that pyrosequencing could accurately and rapidly detect the genetic polymorphism of the warfarin drug metabolism-related genes with high throughput. Compar-ing with simplex pyrosequencing, the method established in the present study was much more economical and timesaving. It has a great value in personalized medical treatment and could be extended to the other genetic diseases.

Simultaneous determination of 6R-leucovorin, 6S-leucovorin and 5-methyltetrahydrofolate in human plasma using solid phase extraction and chiral liquid chromatography-tandem mass spectrometry.

A selective and rapid method was developed and validated for determination of 6R-leucovorin (LV), 6S-leucovorin and 5-methyltetrahydrofolate (5-MeTHF) in human plasma using stereoselective liquid chromatography-tandem mass spectrometry. All analytes and the internal standard were extracted from plasma by solid phase extraction using Oasis HLB C(18) cartridges. A macrocyclic glycopeptide teicoplanin column was used for chiral separation of LV and 5-MeTHF isomers with NH(4)TFA or NH(4)OAc in methanol as mobile phase. Detection was performed on an API 4000 tandem mass spectrometer with positive electrospray ionization in multiple reaction monitoring mode. The calibration curves were linear in the range of 0.050-20.0microg/mL for 6R-LV and 6S-LV, and 0.025-10.0microg/mL for 5-MeTHF. The intra- and inter-assay precision was 3.6-13.2%, 3.4-12.9% and 5.3-9.3% for 6R-LV, 6S-LV and 5-MeTHF, respectively. The accuracy was 99.4-102.4%, 95.3-96.8% and 93.0-110% for 6R-LV, 6S-LV and 5-MeTHF, respectively. The lower limit of quantification (LLOQ) was 0.050microg/mL for each LV isomer and 0.025microg/mL for 5-MeTHF. The method was successfully applied to a comparative pharmacokinetic study between leucovorin calcium and levoleucovorin calcium in 10 volunteers. No significant differences between levoleucovorin and leucovorin in pharmacokinetic parameters of 6S-LV and 5-MeTHF were found in volunteers.

Validated LC-MS/MS method for quantitative determination of rasagiline in human plasma and its application to a pharmacokinetic study.

A highly sensitive liquid chromatography-tandem mass spectrometric (LC-MS/MS) method has been developed to determine rasagiline in human plasma. The analytical method utilized liquid-liquid extraction of plasma with n-hexane-dichloromethane-isopropanol (20:10:1, v/v/v). Separation of analyte and the internal standard (IS) pseudoephedrine was performed on a Zorbax Extend C(18) column (150 mm x 4.6 mm, 5 microm) with a mobile phase consisting of acetonitrile-5 mM ammonium acetate-acetic acid (40:60:0.05, v/v/v) at a flow rate of 0.5 mL/min. The API 4000 triple quadrupole mass spectrometer was operated in multiple reaction monitoring mode via positive electrospray ionization interface using the transitions m/z 172.1-->m/z (117.1+115.1) for rasagiline, and m/z 166.0-->m/z 148.1 for the internal standard. The method was linear over the concentration range of 0.020-50.0 ng/mL. The intra- and inter-day precisions were less than 11.2% in terms of relative standard deviation (R.S.D.), and the accuracy was within +/-6.4% in terms of relative error (RE). The lower limit of quantification (LLOQ) was identifiable and reproducible at 0.020 ng/mL with acceptable precision and accuracy. The mean extraction-efficiency at three concentrations was 95.6+/-7.0%, 97.9+/-3.0% and 95.3+/-8.3%. The validated method offered increased sensitivity (10 times higher than those reported) and wide linear concentration range. This method was successfully applied for the evaluation of pharmacokinetics of rasagiline after single oral doses of 1, 2 and 5mg rasagiline to 12 Chinese healthy volunteers.