Asset-light operation strategy for car-sharing model with vertical shareholding: financial leasing or instalment factoring

Supply chain finance is an important way of realising the transformation of the asset-heavy operation mode in car-sharing services to an asset-light operation mode, by transforming the ownership cost into a variable cost. This study considers a service supply chain with vertical shareholding comprising a vehicle manufacturer and a car-sharing operator; further, it establishes financing decision-making models under financial leasing (FL) and instalment factoring finance (IFF) and explores the impact of profitability and shareholding ratios on FL and IFF. The results how that service profitability has a fundamental influence on financing decisions. The pooling effect and service satisfaction rate affect ownership costs and rental income, respectively. In turn, these factors jointly affect the choice of financing strategy. Moreover, there is a threshold of financing strategy choice related to the shareholding ratio: a large shareholding ratio means IFF is optimal, while when the ratio is low, FL is optimal. Finally, under certain conditions, a composite contract with revenue sharing and linear transfer payment for the optimal financing strategies can be adopted to improve supply chain performance. This study thus provides effective strategies for realising asset-light operation in car sharing by transforming ownership cost into variable cost.</p

Facile Synthesis of Semiconductor and Noble Metal Nanocrystals in High-Boiling Two-Phase Liquid/Liquid Systems

Novel high-boiling liquid/liquid systems were introduced for the synthesis of semiconductor and noble metal nanocrystals. Polyols and long-chain hydrocarbons were utilized to form such two-phase systems. Because of the high-boiling nature, these systems may be conveniently employed for nanocrystal synthesis at relatively high temperatures (under the normal pressure) and thus are useful alternatives to toluene/water. CdS and Ag nanocrystals were successfully prepared in an octadecene (ODE)/glycerol system via interfacial processes, demonstrating the effectiveness of this class of two-phase systems. As-prepared nanocrystals had hydrophobic surfaces and were dispersed in the ODE phase of the system. Furthermore, a congeneric two-phase system of liquid paraffin/glycerol was also found to be effective for the synthesis of CdS and Ag nanocrystals

Additional file 2 of Dynamic co-expression modular network analysis in nonalcoholic fatty liver disease

Additional file 2: Supplementary table 1. DEGs in different profiles

Additional file 1 of Dynamic co-expression modular network analysis in nonalcoholic fatty liver disease

Additional file 1: Supplementary figure 1. The flow diagram of dynamic network analysis

Schematic diagram of yeast-based ZFNs screening and validation system.

<p>(<b>A</b>) <b>Schematic representation of simultaneous screening and validation of ZFNs in yeast.</b> Host strain AH109 harbored ZFNs expression and reporter plasmids and co-expressing ZFNs that could cut target sites on <i>Gal4</i> to generate double strand breaks (DSBs). DSBs were repaired via cellular single strand annealing (SSA) and <i>Gal4</i> was restored by removing the target site and one homology region. Thus, functional Gal4 started to drive expression of reporter genes in AH109, and yeast colonies survived on selective medium. By comparison, ZFNs had no abilities to cut target sites and dysfunctional <i>Gal4</i> could not drive reporter genes expression either. Therefore, yeast colonies could not survive on selective medium lacking histidine and adenine. Gal4HL: Gal4 homology left arm, Gal4HR: Gal4 homology right arm, Gal4AD: Gal4 active domain, Gal4DBD: Gal4 DNA binding domain. (<b>B</b>) <b>The procedure of screening efficient ZFNs via three-step selection.</b> The first step aimed at enriching efficient ZFNs target 3-bp subsites from randomized ZFNs libraries. 3 enriched single fingers target 9-bp half sites were amplified and assembled by overlap PCR and cloned into vector pLeu-FokI between <i>Xba</i>I/<i>Bam</i>HI sites to generate re-constructed ZFNs libraries. The second step screening aimed at screening for efficient ZFNs target 9-bp half sites from re-constructed ZFNs libraries. Finally, pairs of efficient ZFNs were obtained after the third step screening. RZF1: Enriched randomized-zinc finger 1, RZF2: Enriched randomized-zinc finger 2, RZF3: Enriched randomized-zinc finger 3.</p

Primers used for construction of reporter plasmids.

<p>Primers used for construction of reporter plasmids.</p

Primers used for construction of randomized ZFNs libraries.

<p>Primers used for construction of randomized ZFNs libraries.</p

Detection of ZFNs activities targeting endogenous locus in GME cells.

<p>(<b>A</b>) <b>Illustration of puromycin-based system for enrichment of genome modification-positive cells.</b> The reporter gene of <i>puromycin resistance</i> (<i>PuroR</i>) was divided into two fragments carrying 300-bp direct repeats and a ZFN target site. The target sites in reporter vectors as well as the genome could be cut by introducing ZFNs in GME cells. Thus, restored <i>PuroR</i> conferred GME cells resistance to high concentration of puromycin pressure. Meanwhile, genomes of surviving clones were also potentially targeted by ZFNs. Pcmv: CMV promoter, Puro-L: <i>PuroR</i> left homology, Puro-R: <i>PuroR</i> right homology, PAsv40: SV40 polyA. (<b>B</b>) <b>Schematic representation of T7 endonuclease I assay.</b> Genomic DNA was isolated from surviving clones and negative control. PCR reaction was used to amplify target sequences. Then, 200 ng PCR products were re-denatured and re-annealed to generate heteroduplexes. T7 EI enzyme specifically recognizes and cleaves mismatches, and cleaved fragments were isolated by 3% agarose gel. (<b>C</b>) <b>Levels of endogenous locus modification mediated by 3 pairs of ZFNs.</b> Goat mammalian epithelial (GME) cells were electroporated with reporter and ZFNs expression vectors or control plasmids. 5 µg/ml puromycin was added to culture 2 days post electroporation, and genomic DNA was isolated 5 days after treatment with puromycin. The T7 EI assay demonstrated that 3 pairs of ZFNs Z1, Z2 and Z3 generated gene disruption frequencies at 9.4%, 15.9% and 4.1%, respectively. No gene modification was detected in GME cells electroporated with control plasmids of empty expression vectors. (<b>D</b>) <b>Sequences of small deletions and insertions in target site of </b><b><i>α s1-casein</i></b><b> induced by ZFNs.</b></p

Construction of reporter vectors and randomized ZFN libraries.

<p>(<b>A</b>) <b>Schematic of constructing reporter plasmids.</b> Target sites with <i>Not</i>I/<i>Bam</i>HI sticky ends were generated via two oligonucleotides direct annealing (90°C 5 min, 70°C 10 min, cool down to room temperature), and inserted into plasmid pADH-Gal-MCS, resulting in reporter vector pADH-Gal4-BS. Reporter plasmids containing palindromic sequences of target sites for screening three 3-bp subsites of left (right) were substituted by BCR subsites, respectively. 9-bp half sites took place of BCR target sequence completely in LF (left half) and RF (right half) target sites. (<b>B</b>) <b>Illustration of constructing 3 randomized ZFN libraries.</b> The randomized finger 1 fragments were amplified with primers F1/F1R, finger 2 and finger 3 were amplified with primers F1D/FR and all 3 fingers were fused together by overlap PCR. PCR products were cloned in yeast expression plasmids pLeu-FokI between <i>Xba</i>I/<i>Bam</i>HI sites resulting in randomized ZFN1 library. The construction of randomized ZFN2 and ZFN3 libraries was similar to that of ZFN1 library. (<b>C</b>) <b>Sequence alignment results of randomized ZFN libraries.</b> The regions of red cube represent sequences of randomized zinc fingers in each library in which DNA sequences of key amino acids were mutated by PCR with randomized primers and the other two zinc fingers were fixed to the corresponding fingers of BCR-ZFP.</p

Forest plot showing the association between <i>PDE4B</i> SNPs and schizophrenia under recessive model.

<p>Forest plot showing the association between <i>PDE4B</i> SNPs and schizophrenia under recessive model.</p