RT-nPCR Assays for Amplification and Sequencing of VP1 Genes in Human Enterovirus A–D from Clinical Specimens

Objective To develop RT-nPCR assays for amplifying partial and complete VP1 genes of human enteroviruses(HEVs)from clinical samples and to contribute to etiological surveillance of HEV-related diseases.Methods A panel of RT-nPCR assays,consisting of published combined primer pairs for VP1 genes of HEV A–C and in-house designed primers for HEV-D,was established in this study.The sensitivity of each RT-nPCR assay was evaluated with serially diluted virus stocks of five serotypes expressed as CCID50 perμL and copies perμL,and the newly established methods were tested in clinical specimens collected in recent years.Results The sensitivity of RT-nPCR assays for amplifying partial VP1 gene of HEVs was 0.1 CCID50 perμL and 10 virus copies perμL,and for the complete VP1 gene was 1 CCID50 perμL and 100 virus copies perμL,using serially-diluted virus stocks of five serotypes.As a proof-of-concept,25 serotypes were identified and complete VP1 sequences of 23 serotypes were obtained by this system among 858 clinical specimens positive for HEVs during the past eight surveillance seasons.Conclusion This RT-nPCR system is capable of amplifying the partial and complete VP1 gene of HEV A–D,providing rapid,sensitive,and reliable options for molecular typing and molecular epidemiology of HEVs in clinical specimens.

Nanomorphology in A–D–A type small molecular acceptors-based bulk heterojunction polymer solar cells

Recent developments in acceptor–donor–acceptor(A–D–A) type non-fullerene acceptors have led to substantial improvements in bulk-heterojunction polymer solar cells efficiency. The device performance strongly depends on photoactive layer morphology, as the molecular packing, donor–acceptor interface and phase separation significantly affect the charge-transfer states and charge carrier dynamics. In this review, we start with a brief introduction of the techniques most effectively utilized to characterize multiphase morphology. Then, we summarize recent progress in A–D–A type acceptors, with the emphasis on understanding the molecular structure–morphology–performance relationships. Finally, an outlook on correlating morphological characteristics with photovoltage losses is presented for further improving device performance.

Unprecedented C_(19)-diterpenoid alkaloid glycosides from an aqueous extract of“fu zi”:Neoline 14-O-L-arabinosides with four isomeric L-anabinosyls

Four structurally unprecedented aconitane-type C_（19）-diterpenoid alkaloid glycosides with isomeric arabinosyls, named aconicarmichosides A–D（1–4）, were isolated from an aqueous extract of ＂fu zi＂, the lateral roots of Aconitum carmichaelii. Their structures were determined as neoline 14-O-a-and 14-O-b-L-arabinopyranosides（1 and 2） and 14-O-a-and 14-O-b-L-arabinofuranosides（3 and 4）, by spectroscopic and chemical methods including 2D NMR experiments and acid hydrolysis. Compounds 1–4 represent the first examples of glycosidic diterpenoid alkaloids.