Determination of ITS1 haplotypes of Fritillariae Cirrhosae Bulbus by amplicon sequencing.

BACKGROUND: Fritillariae Cirrhosae Bulbus is an antitussive and expectorant Chinese medicinal material derived from the dried bulbs of six Fritillaria species. In the 2015 edition of the Chinese Pharmacopoeia, the polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) is the officially listed method for their authenfication. Specifically, the ~ 300-bp ITS1 amplicon of only Fritillariae Cirrhosae Bulbus but not other Fritillaria species can be cleaved into two smaller fragments with restriction enzyme SmaI. Considering repeated reported cases of incomplete digestion of ITS1 amplicon, this study aims to investigate the possibility of heterogeneous ITS1 sequences contained in the Fritillariae Cirrhosae Bulbus. METHODS: In this study, ITS1 amplicons of Fritillaria Cirrhosae Bulbus and four other Fritillaria species were sequenced on Illumina platform. We utilised high-throughout amplicon sequencing to determine ITS1 haplotypes and their frequencies in Fritillaria genomes. RESULTS: Our results showed that all six botanical sources of Fritillariae Cirrhosae Bulbus indeed possess ITS1 haplotypes with no SmaI restriction site, and the average percentages of ITS1 reads containing SmaI restriction site ranged from 63.60% to 91.81%. CONCLUSION: Our findings suggest that the incomplete digestion in PCR-RFLP analysis of Fritillariae Cirrhosae Bulbus is caused by the presence of ITS1 haplotypes without SmaI restriction site due to intragenomic heterogeneity.

Enhancing Testing Laboratory Engagement in Plant DNA Barcoding through a Routine Workflow-A Case Study on Chinese Materia Medica (CMM).

Introduction of DNA standards into Pharmacopoeia in different parts of the world enables identification of herbal materials in a complementary manner. However, little has been discussed about the quality requirements for a testing laboratory to implement DNA barcoding methods for herbal materials, which has limited the test method to be developed as a routine service. To encourage the engagement of testing laboratory in application of DNA barcode, a practical workflow including the components of analytical run and the corresponding quality control plan was suggested and employed to address a real-life challenge faced by the differentiation of plant-derived Chinese Materia Medica (CMM), Herba Potentillae Chinensis (Wei ling Cai), Herba Potentillae Discoloris (Fan Bai Cai), Radix Pulsatillae (Bai Tou Weng), and Radix Arnebiae (Zi Cao), which share similar morphological characteristics and multiple species involved. The ITS2 barcode results indicated that there are significant differences among the four CMM, together with quality control plan data to ensure the measurement traceability and validity of test results.

Resilience and well-being of university nursing students in Hong Kong: a cross-sectional study.

BACKGROUND: University nursing students experience higher levels of academic stress than those of other disciplines. Academic stress leads to psychological distress and has detrimental effects on well-being. The ability to overcome such adversity and learn to be stronger from the experience is regarded as resilience. Resilience is found to have an impact on learning experience, academic performance, course completion and, in the longer term, professional practice. Resilience and positive coping strategies can resist stress and improve personal well-being. However, the relationship between resilience and well-being remains unexplored in nursing students, which are significant attributes to their academic success and future career persistence. METHODS: The study was a cross-sectional descriptive correlational design. Inclusion criteria for recruitment was students studying pre-registration nursing programmes at both undergraduate and postgraduate levels. The 10-item Connor-Davidson Resilience Scale (CD-RISC-10) and World Health Organisation-5 Well-Being Index (WHO-5) were used to measure resilience and psychological well-being respectively. RESULTS: A convenience sample of 678 university nursing students was recruited from a university. The mean score of CD-RISC-10 was 24.0. When comparing the resilience levels of undergraduate and postgraduate students, the total scores were found to be 23.8 and 24.9 respectively. There was a statistically significant difference between the groups (p = .020). With regard to perceived well-being, the mean score of WHO-5 was 15.5. There was no significant difference between undergraduates and postgraduates (p = .131). Bivariate analysis showed that self-reported resilience had a medium, positive correlation with perceived well-being (r = .378, p = .000), and senior students had significantly higher level of perceived well-being than junior students (16.0 vs 15.1, p = .003). Multivariable regression analysis on perceived well-being indicated that self-reported resilience emerged as a significant predictor of perceived well-being (regression coefficient B = 0.259; p < .001). CONCLUSIONS: The results demonstrate that nursing students with a high level of resilience have better perceived well-being, and the level of resilience of postgraduates was significantly higher than that of undergraduates. Therefore, educational strategies should be developed in the nursing curriculum and a supportive learning environment should be created to foster resilience in the students.

Identification of 5-fluorouracil response proteins in colorectal carcinoma cell line SW480 by two-dimensional electrophoresis and MALDI-TOF mass spectrometry.

Colorectal cancer (CRC) is the second most prevalent cause of cancer-related deaths in the Western world. 5-Fluorouracil (5-FU) is a standard chemotherapeutic drug to treat CRC. However, the response rate is less than 20% and patients who have responded to 5-FU may become resistant. Therefore there is an urgent need to examine the 5-FU response proteins so that patients with no response to 5-FU can change to other treatment strategies promptly. In this study, the proteomic expression profile in a CRC cell line SW480 before and after 5-FU treatment was examined using 2-dimensional electrophoresis technology. Fourteen proteins with differential expression were identified using mass spectrometry and 7 of them were validated using immunocytochemical (ICC) staining. Protein identification indicated that cyclophilin A, cytokeratin 19 (CK19), cytokeratin 8 (CK8), ras-related nuclear protein, heat shock protein 27 (hsp27) and peroxiredoxin 6 (Prx 6) were upregulated whereas heat shock protein 60 (hsp60), cytokeratin 18 (CK18), cytokeratin 9 (CK9), carbamoylphosphate synthetase I, alpha-enolase, heat shock protein 70 (hsp70), nm23 and beta-actin were down-regulated. Seven of the 14 proteins detected were validated by ICC staining, which showed that the expression of hsp27, Prx 6 and hsp70 correlated with that from proteomics profiling. Our results suggest that hsp27, Prx 6 and hsp70 are potential 5-FU response proteins and they may represent potential targets for further evaluation in other 5-FU-sensitive and -resistant CRC cell lines.