Improving analytical throughput is the focus of many quantitative workflows being developed for early drug discovery. For drug candidate screening, it is common practice to use ultra-high performance liquid chromatography (U-HPLC) coupled with triple quadrupole mass spectrometry. This approach certainly results in short analytical run time; however, in assessing the true throughput, all aspects of the workflow needs to be considered, including instrument optimization and the necessity to re-run samples when information is missed. Here we describe a high-throughput metabolic stability assay with a simplified instrument set-up which significantly improves the overall assay efficiency. In addition, as the data is acquired in a non-biased manner, high information content of both the parent compound and metabolites is gathered at the same time to facilitate the decision of which compounds to proceed through the drug discovery pipeline.
Chromatography, High Pressure Liquid ; methods ; Data Mining ; Humans ; Mass Spectrometry ; methods ; Microsomes, Liver ; metabolism

INTRODUCTIONSuicide is one of the most tragic problems medical schools are facing today. It is an issue that has not escaped medical schools in either developing or developed nations. To combat this trend, medical educators require efficient and effective strategies for the immediate identification of students who are at an elevated risk of harming themselves.

MATERIALS AND METHODSNational Yang Ming University medical students were surveyed on various demographic, academic, personal, and extracurricular subjects as well as assessed for suicidal ideation. In addition, students completed the Chinese Health Questionnaire (CHQ, a translated and modified version of the General Health Questionnaire, GHQ), and the Taiwanese Depression Questionnaire (TDQ, a translated and modified version of the Center for Epidemiologic Studies' Depression Scale, CES-D).

RESULTSThe rate of suicidal ideation was significantly higher in second year students as opposed to fi rst year students (P <0.01). Students of lower socioeconomic status (P = 0.04), with non-inflammatory joint pain (P = 0.02), with headache (P = 0.047), with sleep disorders (P = 0.04), who scored as depressed on the TDQ (P <0.01), and/or who scored abnormally on the CHQ (P <0.01) were all significantly more likely to have experienced suicidal ideation.

CONCLUSIONA number of groups at high risk for suicidal ideation, and thus in greater need of support, were identified. Suicide intervention programmes and depression counselling should target older students and students of lower socioeconomic status. Students presenting to university clinics with non-inflammatory joint pain, headache, and/or sleep disorders should be evaluated for suicidal tendencies. The TDQ and CHQ are potentially valuable screening tests for early detection of potential suicidal students.

Adaptation, Psychological ; Adult ; Depression ; epidemiology ; psychology ; Female ; Humans ; Male ; Psychometrics ; Risk Assessment ; methods ; Social Class ; Statistics as Topic ; Stress, Psychological ; epidemiology ; psychology ; Students, Medical ; psychology ; Suicidal Ideation ; Surveys and Questionnaires ; Young Adult

Acute limb ischemia (ALI) represents one of the most common emergent scenarios encountered by a vascular specialist. Despite expedient revascularization, high rates of limb loss are reported along with high mortality rates which are second only to ruptured abdominal aortic aneurysms. Surgical revascularization is standard of care. Endovascular techniques as an alternative have emerged to be appropriate first line therapy when addressing the threatened limb. We review the etiology and classification of ALI and current endovascular techniques and evidence to date in the management of patients acutely presenting with extremity ischemia.
Aortic Aneurysm, Abdominal ; Classification ; Embolism and Thrombosis ; Endovascular Procedures ; Extremities ; Humans ; Ischemia ; Mortality ; Peripheral Arterial Disease ; Specialization ; Standard of Care

PURPOSE: This study aimed to identify potential epidermal growth factor receptor (EGFR) gene mutations in non-small cell lung cancer that went undetected by amplification refractory mutation system-Scorpion real-time PCR (ARMS-PCR). MATERIALS AND METHODS: A total of 200 specimens were obtained from the First Affiliated Hospital of Guangzhou Medical University from August 2014 to August 2015. In total, 100 ARMS-negative and 100 ARMS-positive specimens were evaluated for EGFR gene mutations by Sanger sequencing. The methodology and sensitivity of each method and the outcomes of EGFR-tyrosine kinase inhibitor (TKI) therapy were analyzed. RESULTS: Among the 100 ARMS-PCR-positive samples, 90 were positive by Sanger sequencing, while 10 cases were considered negative, because the mutation abundance was less than 10%. Among the 100 negative cases, three were positive for a rare EGFR mutation by Sanger sequencing. In the curative effect analysis of EGFR-TKIs, the progression-free survival (PFS) analysis based on ARMS and Sanger sequencing results showed no difference. However, the PFS of patients with a high abundance of EGFR mutation was 12.4 months [95% confidence interval (CI), 11.6−12.4 months], which was significantly higher than that of patients with a low abundance of mutations detected by Sanger sequencing (95% CI, 10.7−11.3 months) (p < 0.001). CONCLUSION: The ARMS method demonstrated higher sensitivity than Sanger sequencing, but was prone to missing mutations due to primer design. Sanger sequencing was able to detect rare EGFR mutations and deemed applicable for confirming EGFR status. A clinical trial evaluating the efficacy of EGFR-TKIs in patients with rare EGFR mutations is needed.
Adenocarcinoma/genetics ; Adenocarcinoma/pathology ; Aged ; Aged, 80 and over ; Animals ; Base Sequence ; Disease-Free Survival ; Female ; Humans ; Lung Neoplasms/genetics ; Lung Neoplasms/pathology ; Male ; Middle Aged ; Mutation/genetics ; Mutation Rate ; Real-Time Polymerase Chain Reaction/methods ; Receptor, Epidermal Growth Factor/genetics ; Sequence Analysis, DNA/methods ; Treatment Outcome

INTRODUCTIONRetinitis pigmentosa (RP) is the most prevalent group of inherited retinopathies and demonstrates considerable clinical and genetic heterogeneity, with wide variations in disease severity, progression, and gene involvement. We studied a large family with RP to determine the pattern of inheritance and to identify the disease-causing gene/locus.

MATERIALS AND METHODSOphthalmic examination was performed on 35 family members to identify affected individuals and carriers and to characterise the disease phenotype. Genetic linkage analysis was performed using short tandem repeat (STR) polymorphic markers encompassing the known loci for Xlinked RP (xlRP) including RP2, RP3, RP6, RP23, and RP24. Mutation screening was performed by direct sequencing of PCR-amplified genomic DNA of the RP2 and RPGR genes of the affected individuals.

RESULTSA highly penetrant, X-linked form of RP was observed in this family. Age of onset was from 5 to 8 years and visual acuity ranged from 20/25 in children to light perception in older adults. Linkage analysis and direct sequencing showed that no known loci/genes were associated with the phenotype in this kindred.

CONCLUSIONA novel disease gene locus/loci is responsible for the xlRP phenotype in this family.

Adolescent ; Adult ; Age of Onset ; Child ; Child, Preschool ; Chromosome Mapping ; DNA Mutational Analysis ; Eye Proteins ; genetics ; Female ; Genetic Diseases, X-Linked ; genetics ; Humans ; Intracellular Signaling Peptides and Proteins ; genetics ; Lod Score ; Male ; Membrane Proteins ; genetics ; Pedigree ; Phenotype ; Retinitis Pigmentosa ; genetics

INTRODUCTIONAntenatal major depression is a relatively common and potentially debilitating illness, but knowledge of its treatment outcomes and strategies is still lacking. This study aimed to explore the clinical profiles and treatment outcomes of patients with antenatal major depression, to look for patterns and associations that could guide subsequent research and clinical applications.

METHODSFrom May 2006 to November 2010, 118 consecutive patients with antenatal major depression were naturalistically assessed over eight months of individualised therapy, and their characteristics were assessed as potential predictors of treatment outcome.

RESULTSAll participants accepted supportive counselling and case management, although only 51 (43.2%) participants accepted low-dose antidepressant therapy. Overall, 95 (80.5%) of them were successfully discharged, while 12 (10.2%) required extended treatment into the postnatal period. An equation for prognosticating the need for extended treatment was obtained using multiple logistic regression analysis, which incorporated three predictors: previous depression (odds ratio [OR] 12.4, 95% confidence interval [CI] 1.40-110; p = 0.024); maternal age < 26 years or > 35 years (OR 6.88, 95% CI 1.67-28.4; p = 0.008); and no use of antidepressant (OR 6.94, 95% CI 0.79-60.9; p = 0.080). Among participants with previous depression and at either extreme of maternal age, the number needed to treat with antidepressants to avert extended treatment was three.

CONCLUSIONThe majority of women with antenatal major depression recovered after receiving short-term treatment. Those with previous depression and who were of relative extreme maternal age were most likely to benefit from antidepressant treatment to expedite recovery.

Objective: To determine the prevalence and determinants of folic acid (FA) supplementation in Chinese couples planning for pregnancy and in women during early pregnancy. Methods: This was a cross-sectional study based on the Shanghai PreConception Cohort (SPCC) study. Data on FA supplementation and socio-demographic features were collected using questionnaires. Couples visiting clinics for pre-pregnancy examination and pregnant women at < 14 gestational weeks were recruited in Shanghai, China, between March 2016 and September 2018. Results: Among the pregnancy planners, 42.4% (4,710/11,099) women and 17.1% (1,377/8,045) men used FA supplements, while 93.4% (14,585/15,615) of the pregnant women used FA supplements. FA supplement use was higher in female pregnancy planners who were older ( : 1.13, 95% : 1.08-1.18), had higher education ( : 1.71, 95% : 1.53-1.92), and were residing in urban districts ( : 1.06, 95% : 1.01-1.11) of FA supplementation; female pregnancy planners with alcohol consumption ( : 0.95, 95% : 0.90-0.99) had lower odds of FA supplementation. In early pregnancy, women with higher educational level ( : 1.04, 95% : 1.03-1.06), who underwent pre-pregnancy examination ( : 1.02, 95% : 1.01-1.03) had higher odds of using an FA supplement; older aged ( : 0.99, 95% : 0.98-0.99), and multigravida ( : 0.97, 95% : 0.96-0.98) had lower odds of FA supplementation. Conclusion Although the majority of pregnant women took FA supplements, more than half of the women planning for pregnancy did not. Urgent strategies are needed to improve pre-conception FA supplementation.
Adult ; China ; Cohort Studies ; Cross-Sectional Studies ; Diet ; Dietary Supplements ; analysis ; Female ; Folic Acid ; administration & dosage ; Humans ; Male ; Pregnancy ; Surveys and Questionnaires ; Vitamin B Complex ; administration & dosage ; Young Adult