Potent anti-tumor effects of a dual specific oncolytic adenovirus expressing apoptin in vitro and in vivo

<p>Abstract</p> <p>Background</p> <p>Oncolytic virotherapy is an attractive drug platform of cancer gene therapy, but efficacy and specificity are important prerequisites for success of such strategies. Previous studies determined that Apoptin is a p53 independent, bcl-2 insensitive apoptotic protein with the ability to specifically induce apoptosis in tumor cells. Here, we generated a conditional replication-competent adenovirus (CRCA), designated Ad-hTERT-E1a-Apoptin, and investigated the effectiveness of the CRCA a gene therapy agent for further clinical trials.</p> <p>Results</p> <p>The observation that infection with Ad-hTERT-E1a-Apoptin significantly inhibited growth of the melanoma cells, protecting normal human epidermal melanocytes from growth inhibition confirmed cancer cell selective adenoviral replication, growth inhibition, and apoptosis induction of this therapeutic approach. The <it>in vivo </it>assays performed by using C57BL/6 mice containing established primary or metastatic tumors expanded the <it>in vitro </it>studies. When treated with Ad-hTERT-E1a-Apoptin, the subcutaneous primary tumor volume reduction was not only observed in intratumoral injection group but in systemic delivery mice. In the lung metastasis model, Ad-hTERT-E1a-Apoptin effectively suppressed pulmonary metastatic lesions. Furthermore, treatment of primary and metastatic models with Ad-hTERT-E1a-Apoptin increased mice survival.</p> <p>Conclusions</p> <p>These data further reinforce the previously research showing that an adenovirus expressing Apoptin is more effective and advocate the potential applications of Ad-hTERT-E1a-Apoptin in the treatment of neoplastic diseases in future clinical trials.</p

Comprehensive Mapping of Common Immunodominant Epitopes in the West Nile Virus Nonstructural Protein 1 Recognized by Avian Antibody Responses

West Nile virus (WNV) is a mosquito-borne flavivirus that primarily infects birds but occasionally infects humans and horses. Certain species of birds, including crows, house sparrows, geese, blue jays and ravens, are considered highly susceptible hosts to WNV. The nonstructural protein 1 (NS1) of WNV can elicit protective immune responses, including NS1-reactive antibodies, during infection of animals. The antigenicity of NS1 suggests that NS1-reactive antibodies could provide a basis for serological diagnostic reagents. To further define serological reagents for diagnostic use, the antigenic sites in NS1 that are targeted by host immune responses need to be identified and the potential diagnostic value of individual antigenic sites also needs to be defined. The present study describes comprehensive mapping of common immunodominant linear B-cell epitopes in the WNV NS1 using avian WNV NS1 antisera. We screened antisera from chickens, ducks and geese immunized with purified NS1 for reactivity against 35 partially overlapping peptides covering the entire WNV NS1. This study identified twelve, nine and six peptide epitopes recognized by chicken, duck and goose antibody responses, respectively. Three epitopes (NS1-3, 14 and 24) were recognized by antibodies elicited by immunization in all three avian species tested. We also found that NS1-3 and 24 were WNV-specific epitopes, whereas the NS1-14 epitope was conserved among the Japanese encephalitis virus (JEV) serocomplex viruses based on the reactivity of avian WNV NS1 antisera against polypeptides derived from the NS1 sequences of viruses of the JEV serocomplex. Further analysis showed that the three common polypeptide epitopes were not recognized by antibodies in Avian Influenza Virus (AIV), Newcastle Disease Virus (NDV), Duck Plague Virus (DPV) and Goose Parvovirus (GPV) antisera. The knowledge and reagents generated in this study have potential applications in differential diagnostic approaches and subunit vaccines development for WNV and other viruses of the JEV serocomplex

A New Relative Permeability Characterization Method Considering High Waterflooding Pore Volume

In the process of waterflooding development, high waterflooding PVs will make the fluid percolation in the reservoir more complicated, resulting in lower efficiency of waterflooding. High waterflooding PVs will affect the relative permeability and change the seepage law of oil&ndash;water two-phase flow in a high water-cut period. In this study, we performed high waterflooding PVs relative permeability experiments using nine natural cores. The unsteady measurement method is used to test the relative permeability curve. The results show that: (1) the relative permeability is affected by the waterflooding PVs, the recovery efficiency of 2000 waterflooding PVs is 10.72% higher than that of 50 waterflooding PVs on the core scale; (2) it makes water mobility increase sharply, while oil phase flow capacity remains low and decreases at high water cut stage. A new relative permeability characterization method considering high waterflooding PVs is established, which is applied to the numerical simulator. It shows that the remaining oil saturation of the high-permeability belt is higher than the calculation results of the traditional numerical simulator. It means that the injected water does not diffuse much into the low-permeability zone of the formation. The modified simulator is validated with the actual China offshore oilfield model. The numerical saturation of the key section of the passing well is in good agreement with the actual logging interpretation results, and the water cut curve fits better in the whole area. The modified simulator could predict oil production accurately after high waterflooding PVs treatment

Ovarian tissue transplantation ameliorates osteoporosis and dyslipidaemia in ovariectomised mice

Abstract Background Ovarian insufficiency frequently renders postmenopausal women susceptible to osteoporosis and dyslipidaemia. Postmenopausal transplant women are at a higher risk developing osteoporosis and dyslipidaemia due to the concomitant application of glucocorticoids and immunosuppressants after solid organ transplantation. Thus, this study aimed to explore the feasibility of ovarian tissue transplantation (OTT) as an alternative to Hormone replacement therapy (HRT) for postmenopausal women with solid organ transplant needs. Results Sixty mice were randomly divided into four groups: sham operation, ovariectomised (OVX group), ovariectomy plus oestrogen (E2 group), and ovariectomy plus OTT (OTT group). The inhibin levels in the OTT group were increased and the follicle stimulating hormone and luteinizing hormone were suppressed to normal levels, which could not be achieved in the E2 group. The femoral bone mineral density in the OTT group was significantly increased than the E2 group (P < 0.05), and the probability of fracture was reduced by 1.4–2.6 times. Additionally, the high-density lipoprotein cholesterol levels were higher in the OTT group than in the E2 group and the triglyceride levels were lower in the OTT group than in the E2 group (P < 0.05). Conclusion OTT not only achieves certain endocrine effects by participating in the regulation of the hypothalamic-pituitary-ovarian feedback control loop, but also ameliorates osteoporosis and dyslipidaemia, which may be an alternative to traditional HRT for postmenopausal women with solid organ transplant needs

A New Relative Permeability Characterization Method Considering High Waterflooding Pore Volume

In the process of waterflooding development, high waterflooding PVs will make the fluid percolation in the reservoir more complicated, resulting in lower efficiency of waterflooding. High waterflooding PVs will affect the relative permeability and change the seepage law of oil–water two-phase flow in a high water-cut period. In this study, we performed high waterflooding PVs relative permeability experiments using nine natural cores. The unsteady measurement method is used to test the relative permeability curve. The results show that: (1) the relative permeability is affected by the waterflooding PVs, the recovery efficiency of 2000 waterflooding PVs is 10.72% higher than that of 50 waterflooding PVs on the core scale; (2) it makes water mobility increase sharply, while oil phase flow capacity remains low and decreases at high water cut stage. A new relative permeability characterization method considering high waterflooding PVs is established, which is applied to the numerical simulator. It shows that the remaining oil saturation of the high-permeability belt is higher than the calculation results of the traditional numerical simulator. It means that the injected water does not diffuse much into the low-permeability zone of the formation. The modified simulator is validated with the actual China offshore oilfield model. The numerical saturation of the key section of the passing well is in good agreement with the actual logging interpretation results, and the water cut curve fits better in the whole area. The modified simulator could predict oil production accurately after high waterflooding PVs treatment

Development and optimization of a DNA-based reverse genetics systems for epizootic hemorrhagic disease virus

Epizootic hemorrhagic disease virus (EHDV) is a member of the genus Orbivirus, family Reoviridae, and has a genome consisting of 10 linear double-stranded (ds) RNA segments. The current reverse genetics system (RGS) for engineering the EHDV genome relies on the use of in vitro-synthesized capped viral RNA transcripts. To obtain more-efficient and simpler RGSs for EHDV, we developed an entirely DNA (plasmid or PCR amplicon)-based RGS for viral rescue. This RGS enabled the rescue of infectious EHDV from BSR-T7 cells following co-transfection with seven helper viral protein expression plasmids and 10 cDNA rescue plasmids or PCR amplicons representing the EHDV genome. Furthermore, we optimized the DNA-based systems and confirmed that some of the helper expression plasmids were not essential for the recovery of infectious EHDV. Thus, DNA-based RGSs may offer a more efficient method of recombinant virus recovery and accelerate the study of the biological characteristics of EHDV and the development of novel vaccines.The National Key R&D Program of China and the Central Public-Interest Scientific Institution Basal Fund.http://link.springer.com/journal/705hj2021BiochemistryGeneticsMicrobiology and Plant Patholog

More but Correct: Generating Diversified and Entity-revised Medical Response

Medical Dialogue Generation (MDG) is intended to build a medical dialogue system for intelligent consultation, which can communicate with patients in real-time, thereby improving the efficiency of clinical diagnosis with broad application prospects. This paper presents our proposed framework for the Chinese MDG organized by the 2021 China conference on knowledge graph and semantic computing (CCKS) competition, which requires generating context-consistent and medically meaningful responses conditioned on the dialogue history. In our framework, we propose a pipeline system composed of entity prediction and entity-aware dialogue generation, by adding predicted entities to the dialogue model with a fusion mechanism, thereby utilizing information from different sources. At the decoding stage, we propose a new decoding mechanism named Entity-revised Diverse Beam Search (EDBS) to improve entity correctness and promote the length and quality of the final response. The proposed method wins both the CCKS and the International Conference on Learning Representations (ICLR) 2021 Workshop Machine Learning for Preventing and Combating Pandemics (MLPCP) Track 1 Entity-aware MED competitions, which demonstrate the practicality and effectiveness of our method.Comment: 12 pages, 4 figures, 7 table

Thickness dependent properties of ultrathin perovskite nanosheets with Ruddlesden–Popper-like atomic stackings

Ruddlesden-Popper perovskites possess a rich variety of multiple phases due to their mixed organic cations and variable layer numbers. However, the direct observation of these phases and their optical performance in ultrathin nanosheets, have rarely been reported. Here we demonstrate, through a one-pot CVD synthesis method to incorporate MA(+) and NMA(+) cations into PbI2 simultaneously, that the stackings of Ruddlesden-Popper phases with a distribution of a number of layers n can be produced within a single perovskite nanosheet. As featured by the micro-, time-resolved and temperature-dependent photoluminescence measurements, the optical properties are highly dependent on the nanosheet thickness.The work was financially supported by the National Key R&D Program of China (Grant No. 2020YFA0308900), the National Natural Science Foundation of China (Grant No. 92064010, 61801210, 91833302), the Natural Science Foundation of Jiangsu Province (Grant No. BK20180686), the China postdoctoral Science Foundation (2020M683555), the funding for "Distinguished professors" and "High-level talents in six industries" of Jiangsu Province (Grant No. XYDXX-021), the Fundamental Research Funds for the Central Universities, the Key Research and Development Program of Shaanxi Province (2020GXLH-Z-020, 2020GXLH-Z-027) and the start-up foundation of Northwestern Polytechnical University and Nanjing Tech University

Autophagy Activated by Bluetongue Virus Infection Plays a Positive Role in Its Replication

Bluetongue virus (BTV) is an important pathogen of wild and domestic ruminants. Despite extensive study in recent decades, the interplay between BTV and host cells is not clearly understood. Autophagy as a cellular adaptive response plays a part in many viral infections. In our study, we found that BTV1 infection triggers the complete autophagic process in host cells, as demonstrated by the appearance of obvious double-membrane autophagosome-like vesicles, GFP-LC3 dots accumulation, the conversion of LC3-I to LC3-II and increased levels of autophagic flux in BSR cells (baby hamster kidney cell clones) and primary lamb lingual epithelial cells upon BTV1 infection. Moreover, the results of a UV-inactivated BTV1 infection assay suggested that the induction of autophagy was dependent on BTV1 replication. Therefore, we investigated the role of autophagy in BTV1 replication. The inhibition of autophagy by pharmacological inhibitors (3-MA, CQ) and RNA interference (siBeclin1) significantly decreased viral protein synthesis and virus yields. In contrast, treating BSR cells with rapamycin, an inducer of autophagy, promoted viral protein expression and the production of infectious BTV1. These findings lead us to conclude that autophagy is activated by BTV1 and contributes to its replication, and provide novel insights into BTV-host interactions

Comprehensive Mapping of Common Immunodominant Epitopes in the Eastern Equine Encephalitis Virus E2 Protein Recognized by Avian Antibody Responses

<div><p>Eastern equine encephalitis virus (EEEV) is a mosquito-borne virus that can cause both human and equine encephalitis with high case fatality rates. EEEV can also be widespread among birds, including pheasants, ostriches, emu, turkeys, whooping cranes and chickens. The E2 protein of EEEV and other <i>Alphaviruses</i> is an important immunogenic protein that elicits antibodies of diagnostic value. While many therapeutic and diagnostic applications of E2 protein-specific antibodies have been reported, the specific epitopes on E2 protein recognized by the antibody responses of different susceptible hosts, including avian species, remain poorly defined. In the present study, the avian E2-reactive polyclonal antibody (PAb) response was mapped to linear peptide epitopes using PAbs elicited in chickens and ducks following immunization with recombinant EEEV E2 protein and a series of 42 partially overlapping peptides covering the entire EEEV E2 protein. We identified 12 and 13 peptides recognized by the chicken and duck PAb response, respectively. Six of these linear peptides were commonly recognized by PAbs elicited in both avian species. Among them five epitopes recognized by both avian, the epitopes located at amino acids 211–226 and 331–352 were conserved among the EEEV antigenic complex, but not other associated alphaviruses, whereas the epitopes at amino acids 11–26, 30–45 and 151–166 were specific to EEEV subtype I. The five common peptide epitopes were not recognized by avian PAbs against Avian Influenza Virus (AIV) and Duck Plague Virus (DPV). The identification and characterization of EEEV E2 antibody epitopes may be aid the development of diagnostic tools and facilitate the design of epitope-based vaccines for EEEV. These results also offer information with which to study the structure of EEEV E2 protein.</p></div