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The expression of stage-specific genes during spermatogenesis was determined by isolating two segments of rat seminiferous tubule at different stages of the germinal epithelium cycle delineated by transillumination-delineated microdissection, combined with differential display polymerase chain reaction to identify the differential transcripts formed. A total of 22 cDNAs were identified and accepted by GenBank as new expressed sequence tags. One of the expressed sequence tags was radiolabeled and used as a probe to screen a rat testis cDNA library. A novel full-length cDNA composed of 2228 bp, designated as RSD-3 (rat sperm DNA no.3, GenBank accession no. AF094609) was isolated and characterized. The reading frame encodes a polypeptide consisting of 526 amino acid residues, containing a number of DNA binding motifs and phosphorylation sites for PKC, CK-II, and p34cdc2. Northern blot of mRNA prepared from various tissues of adult rats showed that RSD-3 is expressed only in the testis. The initial expression of the RSD-3 gene was detected in the testis on the 30th postnatal day and attained adult level on the 60th postnatal day. Immunolocalization of RSD-3 in germ cells of rat testis showed that its expression is restricted to primary spermatocytes, undergoing meiosis division I. A human testis homologue of RSD-3 cDNA, designated as HSD-3.1 (GenBank accession no. AF144487) was isolated by screening the Human Testis Rapid-Screen arrayed cDNA library panels by RT-PCR. The exon-intron boundaries of HSD-3.1 gene were determined by aligning the cDNA sequence with the corresponding genome sequence. The cDNA consisted of 12 exons that span approximately 52.8 kb of the genome sequence and was mapped to chromosome 14q31.3.

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A cDNA, designated as rtSH3p13, was isolated from a rat testis cDNA library. It consists of 1463 bp nuclear acids, which encodes a protein of 312 amino acids and was assigned the GenBank accession number AF227439. The deduced rtSH3p13 protein is a truncated isoform of SH3p13 as a result of mRNA alternative splicing. It is mainly expressed in the rat testis, detected in spermatid at the steps 8-19 of spermiogenesis, and found around the acrosome. During postnatal development, rtSH3p13 appears on day 18 and reaches maximum on day 60. Further experimental results suggested that rtSH3p13 forms a complex with activated epidermal growth factor receptor (EGFR) and interacts with synaptojanin I. Surprisingly, similar to SH3 domain, the V region of rtSH3p13 also inhibits endocytosis in CHO cells. Our results reveal a link between an rtSH3p13-synaptojanin-clathrin complex-mediated formation of pits and the process of spermiogenesis.

Under the challenge of climate change, fuel-based vehicles have been receiving increasingly harsh criticism. To promote the use of battery electric vehicles (BEVs) as an alternative, many researchers have studied the deployment of BEVs. This paper proposes a new method to choose locations for new BEV charging stations considering drivers’ perceived time cost and the existing infrastructure. We construct probability equations to estimate drivers’ demanding time for charging (and waiting to charge), use the Voronoi diagram to separate the study area (i.e., Shanghai) into service areas, and apply an optimization algorithm to deploy the charging stations in the right locations. The results show that (1) the probability of charging at public charging stations is 39.6%, indicating BEV drivers prefer to charge at home; (2) Shanghai’s central area and two airports have the busiest charging stations, but drivers’ time costs are relatively low; and (3) our optimization algorithm successfully located two new charging stations surrounding the central area, matching with our expectations. This study provides a time-efficient way to decide where to build new charging stations to improve the existing infrastructure.

YWK-II protein is a sperm membrane component, structurally related to human placenta amyloid precursor protein homolog (APPH) and rat amyloid precursor-like protein 2 (APLP2). Its transmembrane-cytoplasmic domain has high homology (70.6%) with that of betaA4-amyloid precursor protein (APP) found in brain plaques of subjects with Alzheimer's disease. The gene encoding the YWK-II protein is expressed in various mammalian cells and tissues. In the present study, splicing patterns of YWK-II mRNA and the content of YWK-II mRNA in mouse testes, eggs, and cumulus cells were determined. Three different YWK-II transcripts were found in testes and eggs, while cumulus cells contained an additional transcript. In mouse eggs, the content of YWK-II transcript exceeded that of APP. An alternative splicing region was located in the vicinity of the kunitz protease inhibitor (KPI) domain, which may be the basis for the formation of multiple transcripts. YWK-II protein was immunolocated in male and female gametes. It was localized in the plasma membrane of mouse eggs and spermatozoa. In the male reproductive system of the mouse, the YWK-II gene was expressed in germ cells at various stages of differentiation. In mature spermatozoa, the YWK-II protein occurred in the plasma membrane enveloping the acrosome. Triggering the acrosome reaction incited a release of the YWK-II protein attached to the liberated membrane vesicles. The occurrence of the YWK-II protein in the plasma membranes of mouse gametes suggests its involvement in sperm-egg interaction.

Conventional enhanced oil recovery (EOR) approaches are inefficient in the unconventional reservoirs. This paper provides a novel approach to enhance oil recovery from unconventional oil reservoirs through synchronous inter-fracture injection and production (SiFIP) and asynchronous inter-fracture injection and production (AiFIP). The compartmental embedded discrete fracture model (cEDFM) is introduced to simulate complex fracture geometries to quantitatively evaluate the performance of SiFIP and AiFIP. EOR performances using multiple producing methods are investigated (i.e., depletion, fluid flood, fluid Huff and Puff, SiFIP, AiFIP. Higher cumulative oil production rates can be achieved by AiFIP and SiFIP. AiFIP yields the highest oil recovery factor, two times higher than depletion. Compared with SiFIP, AiFIP may be a preferred method when CO2/water resources are short. The impacts of fracture and injection parameters on oil production are discussed. The feasible well completions for AiFIP and SiFIP are provided. AiFIP (CO2) achieves the best EOR performance among different producing methods. This paper demonstrates the feasibility of SiFIP and AiFIP to improve oil recovery. The proposed methods improve flooding performance by transforming fluid injection among wells to among hydraulic fractures from the same Multi-fractured horizontal well (MFHW), which is a promising EOR approach in unconventional oil reservoirs. The proposed EOR method (AiFIP-CO2) can improve the oil recovery, and mitigate the emission of CO2 as well as reduce the waste of water resources.