Targeting the ribosome to treat multiple myeloma.

The high rates of protein synthesis and processing render multiple myeloma (MM) cells vulnerable to perturbations in protein homeostasis. The induction of proteotoxic stress by targeting protein degradation with proteasome inhibitors (PIs) has revolutionized the treatment of MM. However, resistance to PIs is inevitable and represents an ongoing clinical challenge. Our first-in-human study of the selective inhibitor of RNA polymerase I transcription of ribosomal RNA genes, CX-5461, has demonstrated a potential signal for anti-tumor activity in three of six heavily pre-treated MM patients. Here, we show that CX-5461 has potent anti-myeloma activity in PI-resistant MM preclinical models in vitro and in vivo. In addition to inhibiting ribosome biogenesis, CX-5461 causes topoisomerase II trapping and replication-dependent DNA damage, leading to G2/M cell-cycle arrest and apoptotic cell death. Combining CX-5461 with PI does not further enhance the anti-myeloma activity of CX-5461 in vivo. In contrast, CX-5461 shows synergistic interaction with the histone deacetylase inhibitor panobinostat in both the Vk∗MYC and the 5T33-KaLwRij mouse models of MM by targeting ribosome biogenesis and protein synthesis through distinct mechanisms. Our findings thus provide strong evidence to facilitate the clinical development of targeting the ribosome to treat relapsed and refractory MM.

A Computational Model of Cn2 Profile Inversion for Atmospheric Laser Communication in the Vertical Path.

In this paper, an atmospheric structure constant Cn2 model is proposed for evaluating the channel turbulence degree of atmospheric laser communication. First, we derive a mathematical model for the correlation between the atmospheric coherence length r0, the isoplanatic angle θ0 and Cn2 using the Hufnagel-Valley (HV) turbulence model. Then, we calculate the seven parameters of the HV model with the actual measured r0 and θ0 data as input quantities, so as to draw the Cn2 profile and the θ0 profile. The experimental results show that the fitted average Cn2 contours and single-day Cn2 contours have superior fitting performance compared with our historical data, and the daily correlation coefficient between the single-day computed θ0 contours and the measured θ0 contours is up to 87%. This result verifies the feasibility of the proposed method. The results validate the feasibility of the proposed method and provide a new technical tool for the inversion of turbulence Cn2 profiles.

Discovery of Novel, Selective Prostaglandin EP4 Receptor Antagonists with Efficacy in Cancer Models.

Prostaglandin E2 (PGE2) receptor 4 (EP4) is one of four EP receptors commonly upregulated in the tumor microenvironment and plays vital roles in stimulating cell proliferation, invasion, and metastasis. Biochemical blockade of the PGE2-EP4 signaling pathway is a promising strategy for controlling inflammatory and immune related disorders. Recently combination therapies of EP4 antagonists with anti-PD-1 or chemotherapy agents have emerged in clinical studies for lung, breast, colon, and pancreatic cancers. Herein, a novel series of indole-2-carboxamide derivatives were identified as selective EP4 antagonists, and SAR studies led to the discovery of the potent compound 36. Due to favorable pharmacokinetics properties and good oral bioavailability (F = 76%), compound 36 was chosen for in vivo efficacy studies. Compound 36 inhibited tumor growth in a CT-26 colon cancer xenograft better than E7046 and a combination of 36 with capecitabine significantly suppressed tumor growth (TGI up to 94.26%) in mouse models.

Molecular Characterization of HN1304M, a Cat Que Virus Isolated from Midges in China.

The Cat Que orthobunyavirus has been found in mosquitoes, birds, pigs, and humans, suggesting its wide range of hosts and potential public health implications. During arbovirus surveillance in 2013, the HN1304M virus was isolated from naturally occurring Culicoides biting midges in Hunan Province, southern China. The virus was cytopathic to BHK-21 cells and showed stable passage, but was not cytopathic to C6/36 cells. Determination and analysis of the viral genome sequence revealed that HN1304M is an RNA virus with three gene segments, namely, L, M, and S. The nucleotide and amino acid sequence homologies of HN1304M to Cat Que viruses in the Manzanilla species complex were 90.3-99.4%, and 95-100%, respectively, while the homologies to other viruses in this species complex were 74-86.6% and 78.1-96.1%, respectively. A phylogenetic analysis of the viral genes revealed that HN1304M formed an evolutionary branch with other Cat Que viruses isolated from mosquitoes, pigs, birds, and humans, which was completely independent of the other viruses in this complex. The fact that the Cat Que virus was isolated from Culicoides suggests that biting midges may participate in the natural circulation of Cat Que viruses.

Teacher Online Informal Learning as a Means to Innovative Teaching During Home Quarantine in the COVID-19 Pandemic.

The home quarantine in the COVID-19 pandemic has created challenges for teaching across the world and called for innovative teaching, as well as teachers' learning. Given the rapid development of teachers' online learning and teaching, identifying effective ways to facilitate innovative teaching under such challenging conditions is a critical issue. Although researchers have realized that workplace informal learning (IL) increasingly reveals its potential value to individual development, the relationship between IL and innovation has been under-explored. The purpose of this study was to evaluate the impact of IL on innovative teaching, through the mediating roles of three types of teaching-related efficacy, with a particular focus on college teachers and online context. A sample of 479 Chinese college teachers was randomly selected to participate in the survey. The results showed that teachers' online IL in pandemic improved their personal teaching efficacy and ICT efficacy (information and communication technology efficacy), and then facilitated their innovative teaching without differences of gender and teaching-age effect. Whereas, general teaching efficacy was not a mediator between online IL and innovative teaching. Hence, we proposed a can-do motivating model of teacher efficacy in fostering innovative teaching through informal learning. It implies three properties of teachers' online IL: social interaction, autonomous learning and novelty-seeking. It also revealed that innovative teaching can be driven in COVID-19 pandemic, mainly by learning domain-specific knowledge and skills, thus enhancing personal teaching efficacy and ICT efficacy in online teaching context.

Comparison of bayberry fermented wine aroma from different cultivars by GC-MS combined with electronic nose analysis.

Four bayberry cultivars (Biqi, Dongkui, Wandao, and Dingao) in eastern China were selected to produce the fermented bayberry wine. The volatile flavor compounds in different bayberry wine were compared by gas chromatography-mass spectrometry (GC-MS) and electronic nose. The results showed that 46 volatile flavor compounds were found in bayberry wine, including 19 esters, 7 alcohols, 6 acids, 2 aldehydes, 2 ketones, 3 terpenes, and 7 others compounds. The most important contribution to the aroma of bayberry wine was esters and alcohols, respectively. Differentiation of four kinds of bayberry wine was conducted analysis by E-nose. Sensory evaluation showed that Biqi bayberry wine was highly evaluated for its highest score in color, floral aroma, overall acceptability, and fruity aroma. Our results suggest that there were differences in the flavor characteristics of bayberry wine brewed from different varieties of bayberry. The results of this study will provide valuable information for bayberry wine makers to select raw materials.

Development of Chiral Spiro Phosphoramidites for Rhodium-Catalyzed Enantioselective Reactions.

A series of 1,1'-spirobiindane-7,7'-diol (SPINOL) analogues bearing a 2,2'-dimethyl-, cyclopentyl-, or cyclohexyl-fused ring were synthesized, and their distinct structural features were elucidated by X-ray crystallography. On the basis of these scaffolds, chiral monophosphoramidite ligands 6 a-m were synthesized, which demonstrated excellent enantioselectivity in RhI -catalyzed asymmetric hydrogenation of a dehydro amino acid methyl ester. Ligands 6 a-m were also successfully applied in the RhI -catalyzed enantioselective [4+2] cycloaddition of α,ß-unsaturated imines with isocyanates, which afforded the corresponding pyrimidinones in good yields (60-92 %) with high enantioselectivities (75-92 % ee).

Detection of West Nile Virus Infection in Viral Encephalitis Cases, China.

This study detected West Nile virus (WNV) infection in serum samples of patients clinically diagnosed with viral encephalitis in the Japanese encephalitis virus (JEV) endemic area (seven provinces) and JEV nonendemic area (Xinjiang province) in China from 2011 to 2012. In JEV endemic areas, there were 22 positive cases of WNV immunoglobulin M (IgM) antibody in serum specimens of 65 JEV patients (JEV IgM antibody positive) in the acute phase, whereas WNV IgM antibodies were not detected in serum specimens of 63 non-JEV patients (JEV IgM antibody negative). However, the titer of JEV-neutralizing antibody was four times higher than that of WNV-neutralizing antibody in WNV-IgM-positive serum specimens. Detection was also conducted in serum specimens collected from 12 patients clinically diagnosed as viral encephalitis in Xinjiang; five serum specimens were WNV IgM antibody positive, and there were fourfold differences in WNV-neutralizing antibody titers between convalescent and acute serum. Meanwhile JEV-neutralizing antibody titer was negative or significantly lower than that of WNV-neutralizing antibody in the same specimens. WNV IgM antibodies positive were detected in acute serum specimens of patients clinically diagnosed with JEV infection in JEV-endemic areas, but no WNV neutralization antibodies were detected fourfold greater than that of the corresponding JEV antibodies. Clinical cases of WNV infection were detected in patients clinically diagnosed with viral encephalitis in Xinjiang.

Changing Geographic Distribution of Japanese Encephalitis Virus Genotypes, 1935-2017.

Japanese encephalitis virus (JEV) is a representative virus of the JEV serogroup in genus Flavivirus, family Flaviviridae. JEV is a mosquito-borne virus that causes Japanese encephalitis (JE), one of the most severe viral encephalitis diseases in the world. JEV is divided into five genotypes (G1-G5), and each genotype has its own distribution pattern. However, the distribution of different JEV genotypes has changed markedly in recent years. JEV G1 has replaced G3 as the dominant genotype in the traditional epidemic areas in Asia, while G3 has spread from Asia to Europe and Africa and caused domestic JE cases in Africa. G2 and G5, which were endemic in Malaysia, exhibited great geographical changes as well. G2 migrated southward and led to prevalence of JE in Australia, while G5 emerged in China and South Korea after decades of silence. Along with these changes, JE occurred in some non-traditional epidemic regions as an emerging infectious disease. The regional changes in JEV pose a great threat to human health, leading to huge disease burdens. Therefore, it is of great importance to strengthen the monitoring of JEV as well as virus genotypes, especially in non-traditional epidemic areas.

Isolation and Genome Phylogenetic Analysis of Arthropod-Borne Viruses, Including Akabane Virus, from Mosquitoes Collected in Hunan Province, China.

This study investigated the abundance of mosquitoes and circulation of mosquito-borne arboviruses from 16 villages in 8 cities of Hunan Province, China, in July-August of 2010 and in August of 2011. In total, 16,076 mosquitoes consisting of seven species from four genera were collected by ultraviolet-light trap. Culex quinquefasciatus was the most common species, accounting for 50.63% (8140/16,076) of the total. Anopheles sinensis (24.26%, 3900/16,076) made up the second most common species, followed by Culex tritaeniorhynchus (9.76%, 1569/16,076). The proportions of Culex pipiens pallens, Armigeres subalbatus, and Culex modestus were 6.7%, 5.2%, and 3.31%, respectively. Fourteen Aedes albopictus were detected. The mosquitoes were identified by morphologic characteristics and frozen in liquid nitrogen. The mosquitoes were pooled, triturated, and centrifuged. The clarified supernatant was used to inoculate monolayers of C6/36 and baby hamster kidney-21 cells. We obtained six virus isolates that caused cytopathic effects. Phylogenetic analysis revealed that two isolates were Akabane virus (AKAV, from A. sinensis and C. quinquefasciatus), two isolates were Japanese encephalitis virus (from C. pipiens pallens and C. quinquefasciatus), and two isolates were Tibet orbivirus (from C. quinquefasciatus and C. tritaeniorhynchus). This is the first report of AKAV isolated from A. sinensis and C. quinquefasciatus in nature in China. The detection of AKAV in these species confirms circulation of AKAV in Hunan province and suggests potential challenges to the prevention and control of arthropod-borne animal viruses in mainland China.

Chiral Cyclohexyl-Fused Spirobiindanes: Practical Synthesis, Ligand Development, and Asymmetric Catalysis.

1,1'-Spirobiindane has been one type of privileged skeleton for chiral ligand design, and 1,1'-spirobiindane-based chiral ligands have demonstrated outstanding performance in various asymmetric catalysis. However, the access to enantiopure spirobiindane is quite tedious, which obstructs its practical application. In the present article, a facile enantioselective synthesis of cyclohexyl-fused chiral spirobiindanes has been accomplished, in high yields and excellent stereoselectivities (up to >99% ee), via a sequence of Ir-catalyzed asymmetric hydrogenation of α,α'-bis(arylidene)ketones and TiCl4 promoted asymmetric spiroannulation of the hydrogenated chiral ketones. The protocol can be performed in one pot and is readily scalable, and has been utilized in a 25 g scale asymmetric synthesis of cyclohexyl-fused spirobiindanediol (1 S,2 S,2' S)-5, in >99% ee and 67% overall yield for four steps without chromatographic purification. Facile derivations of (1 S,2 S,2' S)-5 provided straightforward access to chiral monodentate phosphoramidites 6a-c and a tridentate phosphorus-amidopyridine 11, which were evaluated as chiral ligands in several benchmark enantioselective reactions (hydrogenation, hydroacylation, and [2 + 2] reaction) catalyzed by transition metal (Rh, Au, or Ir). Preliminary results from comparative studies showcased the excellent catalytic performances of these ligands, with a competency essentially equal to the corresponding well-established privileged ligands bearing a regular spirobiindane backbone. X-ray crystallography revealed a close resemblance between the structures of the precatalysts 20 and 21 and their analogues, which ultimately help to rationalize the almost identical stereochemical outcomes of reactions catalyzed by metal complexes of spirobiindane-derived ligands with or without a fused cyclohexyl ring on the backbone. This work is expected to stimulate further applications of this type of readily accessible skeletons in development of chiral ligands and functional molecules.

Arboviruses and their related infections in China: A comprehensive field and laboratory investigation over the last 3 decades.

Since the 1980s, a comprehensive field and laboratory investigation has been conducted throughout China, and a total of 29 virus species belonging to 7 families and 13 genera were identified through virological, morphological, and immunological methods, as well as whole-genome sequencing and molecular genetic analyses. Most of the virus isolates belong to 9 genera in the families Flaviviridae, Bunyaviridae, Togaviridae, and Reoviridae. Among them, 4 genera (Orthobunyavirus, Bunyavirus, Phlebovirus, and Nairovirus) belong to the family Bunyaviridae and 3 genera (Seadonavirus, Orbivirus, and Cypovirus) belong to the family Reoviridae. Analyses of the relationships between viruses and human/animal diseases indicated that Japanese encephalitis virus, dengue virus, severe fever with thrombocytopenia syndrome virus, tick-borne encephalitis virus, Crimean-Congo hemorrhagic fever virus, West Nile virus, and Tahyna virus can cause human and animal infections and disease epidemics in China. This review systematically introduces the current status of the diversity and geographical distribution of arboviruses and vectors in China. In addition, our results provide strong technical support for the prevention and control of arboviral diseases, the treatment of epidemics, and the early warning and prediction of diseases, and so they are significant for the control and prevention of arboviral diseases in Asia and around the world.

Rapid deployment of a mobile biosafety level-3 laboratory in Sierra Leone during the 2014 Ebola virus epidemic.

BACKGROUND: Ebola virus emerged in West Africa in December 2013. The high population mobility and poor public health infrastructure in this region led to the development of the largest Ebola virus disease (EVD) outbreak to date. METHODOLOGY/PRINCIPAL FINDINGS: On September 26, 2014, China dispatched a Mobile Biosafety Level-3 Laboratory (MBSL-3 Lab) and a well-trained diagnostic team to Sierra Leone to assist in EVD diagnosis using quantitative real-time PCR, which allowed the diagnosis of suspected EVD cases in less than 4 hours from the time of sample receiving. This laboratory was composed of three container vehicles equipped with advanced ventilation system, communication system, electricity and gas supply system. We strictly applied multiple safety precautions to reduce exposure risks. Personnel, materials, water and air flow management were the key elements of the biosafety measures in the MBSL-3 Lab. Air samples were regularly collected from the MBSL-3 Lab, but no evidence of Ebola virus infectious aerosols was detected. Potentially contaminated objects were also tested by collecting swabs. On one occasion, a pipette tested positive for EVD. A total of 1,635 suspected EVD cases (824 positive [50.4%]) were tested from September 28 to November 11, 2014, and no member of the diagnostic team was infected with Ebola virus or other pathogens, including Lassa fever. The specimens tested included blood (69.2%) and oral swabs (30.8%) with positivity rates of 54.2% and 41.9%, respectively. The China mobile laboratory was thus instrumental in the EVD outbreak response by providing timely and reliable diagnostics. CONCLUSIONS/SIGNIFICANCE: The MBSL-3 Lab significantly contributed to establishing a suitable laboratory response capacity during the emergence of EVD in Sierra Leone.

Discovery and genetic analysis of novel coronaviruses in least horseshoe bats in southwestern China.

To investigate bat coronaviruses (CoVs), we collected 132 rectal swabs and urine samples from five bat species in three countries in southwestern China. Seven CoVs belonging to distinct groups of severe acute respiratory syndrome (SARS)-like CoVs and α-CoVs were detected in samples from least horseshoe bats. Samples from other bat species were negative for these viruses, indicating that the least horseshoe bat represents one of the natural reservoirs and mixers for strains of CoVs and has a pivotal role in the evolution and dissemination of these viruses. The genetic and evolutionary characteristics of these strains were described. Whole-genome sequencing of a new isolate (F46) from a rectal swab from a least horseshoe bat showed that it contained 29 699 nucleotides, excluding the poly (A) tail, with 13 open reading frames (ORFs). Phylogenetic and recombination analyses of F46 provided evidence of natural recombination between bat SARS-like CoVs (Rs3367 and LYRa11) or SARS-CoV (BJ01), suggesting that F46 could be a new recombinant virus from SARS-like CoVs or SARS-CoVs.

Molecular evolution of emerging Banna virus.

Banna virus (BAV) is an emerging pathogen that causes human viral encephalitis and has been isolated from types of blood-sucking insects and mammals in Asia. However, there are no reported systematic studies that describe the origin and evolution of BAV. Here, a phylogenetic analysis of BAVs isolated from a variety of potential vectors and vertebrate hosts worldwide revealed that BAVs emerged in the beginning of the 20th century and do not exhibit a species barrier. The mean substitution rate of BAVs was 2.467×10-2substitution/site/year (95% HPD, 1.093×10-3 to 5.628×10-2). The lineage is mainly composed of BAVs from high-latitude regions, which are the most recently emerged viruses with significantly higher substitution rates compared with the lineage comprised of the isolates from middle or low-latitude regions. The genetic differences between BAV strains are positively correlated with the geographic distribution. Strains from the same latitude regions are almost 100% identical, whereas the differences between strains from long distance regions with different latitudes could be >60%. Our results demonstrate that BAV is an emerging virus at a stage that involves rapid evolution and has great potential for introduction into non-endemic areas. Thus, enhanced surveillance of BAV is highly recommended worldwide.

Clinical and Virological Characteristics of Ebola Virus Disease Patients Treated With Favipiravir (T-705)-Sierra Leone, 2014.

BACKGROUND: During 2014-2015, an outbreak of Ebola virus disease (EVD) swept across parts of West Africa. No approved antiviral drugs are available for Ebola treatment currently. METHODS: A retrospective clinical case series was performed for EVD patients in Sierra Leone-China Friendship Hospital. Patients with confirmed EVD were sequentially enrolled and treated with either World Health Organization (WHO)-recommended supportive therapy (control group) from 10 to 30 October, or treated with WHO-recommended therapy plus favipiravir (T-705) from 1 to 10 November 2014. Survival and virological characteristics were observed for 85 patients in the control group and 39 in the T-705 treatment group. RESULTS: The overall survival rate in the T-705 treatment group was higher than that of the control group (56.4% [22/39] vs 35.3% [30/85]; P = .027). Among the 35 patients who finished all designed endpoint observations, the survival rate in the T-705 treatment group (64.8% [11/17]) was higher than that of the control group (27.8% [5/18]). Furthermore, the average survival time of the treatment group (46.9 ± 5.6 days) was longer than that of the control group (28.9 ± 4.7 days). Most symptoms of patients in the treatment group improved significantly. Additionally, 52.9% of patients who received T-705 had a >100-fold viral load reduction, compared with only 16.7% of patients in the control group. CONCLUSIONS: Treatment of EVD with T-705 was associated with prolonged survival and markedly reduced viral load, which makes a compelling case for further randomized controlled trials of T-705 for treating EVD.

[Eukaryotic Expression and Immunogenic Research of Recombination Ebola Virus Membrane Protein Gp-Fc].

We used 293 cells to express the recombinant membrane protein of the Ebola virus. Then, the immunogenicity of the recombinant protein was studied by immunized BALB/c mice. According to the codon use frequency of humans, the gene encoding the extracellular domain of the Ebola virus membrane protein was optimized, synthesized, and inserted into the eukaryotic expression plasmid pXG-Fc to construct the human IgG Fc and Ebola GP fusion protein expression plasmid pXG-modGP-Fc. To achieve expression, the fusion protein expression vector was transfected into high-density 293 cells using transient transfection technology. The recombinant protein was purified by protein A affinity chromatography. BALB/c mice were immunized with the purified fusion protein, and serum antibody titers evaluated by an indirect enzyme-linked immunosorbent assay (ELISA). Purification and analyses of the protein revealed that the eukaryotic expression vector could express the recombinant protein GP-Fc effectively, and that the recombinant protein in the supernatant of the cell culture was present as a dimer. After immunization with the purified recombinant protein, a high titer of antigen-specific IgG could be detected in the serum of immunized mice by indirect ELISA, showing that the recombinant protein had good immunogenicity. These data suggest that we obtained a recombinant protein with good immunogenicity. Our study is the basis for development of a vaccine against the Ebola virus and for screening of monoclonal antibodies.

Low Protective Efficacy of the Current Japanese Encephalitis Vaccine against the Emerging Genotype 5 Japanese Encephalitis Virus.

BACKGROUND: The current Japanese encephalitis (JE) vaccine derived from G3 JE virus (JEV) can induce protective immunity against G1-G4 JEV genotypes. However, protective efficacy against the emerging G5 genotype has not been reported. METHODS/PRINCIPAL FINDINGS: Using in vitro and in vivo tests, biological phenotype and cross-immunoreactions were compared between G3 JEV and G5 JEV (wild strains). The PRNT90 method was used to detect neutralizing antibodies against different genotypes of JEV in JE vaccine-immunized subjects and JE patients. In JE vaccine-immunized mice, the lethal challenge protection rates against G3 and G5 JEV wild strains were 100% and 50%, respectively. The seroconversion rates (SCRs) of virus antibodies against G3 and G5 JEV among vaccinated healthy subjects were 100% and 35%, respectively. All clinically identified JE patients showed high levels of G3 JEV neutralizing antibodies (≥1:10-1280) with positive serum geometric mean titers (GMTs) of 43.2, while for G5 JEV, neutralizing antibody conversion rates were only 64% with positive serum GMTs of 11.14. Moreover, the positive rate of JEV neutralizing antibodies against G5 JEV in pediatric patients was lower than in adults. CONCLUSIONS/SIGNIFICANCE: Low levels of neutralizing/protective antibodies induced by the current JE vaccine, based on the G3 genotype, were observed against the emerging G5 JEV genotype. Our results demonstrate the need for more detailed studies to reevaluate whether or not the apparent emergence of G5 JEV can be attributed to failure of the current vaccine to induce appropriate immune protectivity against this genotype of JEV.

Ebola Virus Outbreak Investigation, Sierra Leone, September 28-November 11, 2014.

During 2014-2015, an outbreak of Ebola virus disease (EVD) swept across parts of West Africa. The China Mobile Laboratory Testing Team was dispatched to support response efforts; during September 28-November 11, 2014, they conducted PCR testing on samples from 1,635 suspected EVD patients. Of those patients, 50.4% were positive, of whom 84.6% lived within a 3-km zone along main roads connecting rural towns and densely populated cities. The median time from symptom onset to testing was 5 days. At testing, 75.7% of the confirmed patients had fever, and 94.1% reported at least 1 gastrointestinal symptom; all symptoms, except rash and hemorrhage, were more frequent in confirmed than nonconfirmed patients. Virus loads were significantly higher in EVD patients with fever, diarrhea, fatigue, or headache. The case-fatality rate was lower among patients 15-44 years of age and with virus loads of <100,000 RNA copies/mL. These findings are key for optimizing EVD control and treatment measures.