Dipterocarpoidae genomics reveal their demography and adaptations to Asian rainforests.

Dipterocarpoideae species form the emergent layer of Asian rainforests. They are the indicator species for Asian rainforest distribution, but they are severely threatened. Here, to understand their adaptation and population decline, we assemble high-quality genomes of seven Dipterocarpoideae species including two autotetraploid species. We estimate the divergence time between Dipterocarpoideae and Malvaceae and within Dipterocarpoideae to be 108.2 (97.8‒118.2) and 88.4 (77.7‒102.9) million years ago, and we identify a whole genome duplication event preceding dipterocarp lineage diversification. We find several genes that showed a signature of selection, likely associated with the adaptation to Asian rainforests. By resequencing of two endangered species, we detect an expansion of effective population size after the last glacial period and a recent sharp decline coinciding with the history of local human activities. Our findings contribute to understanding the diversification and adaptation of dipterocarps and highlight anthropogenic disturbances as a major factor in their endangered status.

Apoptosis-resistant megakaryocytes produce large and hyperreactive platelets in response to radiation injury.

BACKGROUND: The essential roles of platelets in thrombosis have been well recognized. Unexpectedly, thrombosis is prevalent during thrombocytopenia induced by cytotoxicity of biological, physical and chemical origins, which could be suffered by military personnel and civilians during chemical, biological, radioactive, and nuclear events. Especially, thrombosis is considered a major cause of mortality from radiation injury-induced thrombocytopenia, while the underlying pathogenic mechanism remains elusive. METHODS: A mouse model of radiation injury-induced thrombocytopenia was built by exposing mice to a sublethal dose of ionizing radiation (IR). The phenotypic and functional changes of platelets and megakaryocytes (MKs) were determined by a comprehensive set of in vitro and in vivo assays, including flow cytometry, flow chamber, histopathology, Western blotting, and chromatin immunoprecipitation, in combination with transcriptomic analysis. The molecular mechanism was investigated both in vitro and in vivo, and was consolidated using MK-specific knockout mice. The translational potential was evaluated using a human MK cell line and several pharmacological inhibitors. RESULTS: In contrast to primitive MKs, mature MKs (mMKs) are intrinsically programmed to be apoptosis-resistant through reprogramming the Bcl-xL-BAX/BAK axis. Interestingly, mMKs undergo minority mitochondrial outer membrane permeabilization (MOMP) post IR, resulting in the activation of the cyclic GMP-AMP synthase-stimulator of IFN genes (cGAS-STING) pathway via the release of mitochondrial DNA. The subsequent interferon-ß (IFN-ß) response in mMKs upregulates a GTPase guanylate-binding protein 2 (GBP2) to produce large and hyperreactive platelets that favor thrombosis. Further, we unmask that autophagy restrains minority MOMP in mMKs post IR. CONCLUSIONS: Our study identifies that megakaryocytic mitochondria-cGAS/STING-IFN-ß-GBP2 axis serves as a fundamental checkpoint that instructs the size and function of platelets upon radiation injury and can be harnessed to treat platelet pathologies.

Effects of Reticuloendotheliosis virus on TLR-3/IFN-Β pathway in specific pathogen-free chickens.

Birds infected by Reticuloendotheliosis virus (REV) are vulnerable to other microorganisms. This immunosuppression is related to the immune organs (thymus, bursa of Fabricius, and spleen) damaged by REV. The regulation of IFN-ß greatly depends on pattern recognition receptor TLR-3 and nuclear factors IRF-7, NF-κB. To address if and how the TLR-3/IFN-ß pathway is disturbed by REV, 60 one-day-old specific-pathogen-free chickens were intraperitoneally injected with RE virus dilution (n = 30) or stroke-physiological saline solution (n = 30). At 1, 3, 7, 21, and 28 days post-infection, after collecting thymuses, bursas, and spleens, we monitor the kinetics of TLR-3, IFN-ß, NF-κB p65, and IRF-7 at transcriptional and translational levels using qPCR, Western blotting, and ELISA separately. As a result, compared with control chickens, the mRNA levels of TLR-3, IRF-7, and NF-κB p65 showed increasingly differences in the early period of REV infection. Synchronal changes occurred at translation levels. In the latter infection period, a decrease of NF-κB p65 was contemporaneous with a fall in IFN-ß at both transcriptional and translational levels in the thymuses and bursas. These data suggest that the changes of IFN-ß content are closely related to NF-κB p65 when REV invades chicken central immune organs. That reveals new insights into the immunosuppression mechanism of REV in avian.

A Triple-Heterozygous ß-Thalassemia Patient Demonstrated an Unusual Electrophoresis Pattern Due to a Novel ß0 Mutation [an IVS-II-654 (C>T) mutation with a Hb Zürich-Langstrasse (HBB: c.151A>T) mutation in cis].

ß-Thalassemia (ß-thal) is caused by mutations on the ß-globin genes, causing reduced (ß+) or absent (ß0) synthesis of the ß chains of hemoglobin (Hb). In this report, a 28-year-old male patient with anemia and jaundice, was diagnosed with triple-heterozygous ß-thal [an IVS-II-654 (C>T) mutation, a Hb Zürich-Langstrasse (HBB: c.151A>T) mutation and a Hb G-Siriraj (HBB: c.22G>A) mutation] by gene sequencing. However, his electrophoresis pattern was unusual: 90.8% Hb G-Siriraj, 5.9% Hb A2, 3.3% Hb F, no Hb A, no Hb Zürich-Langstrasse. His mother carried a ß-thal trait (ßA/ßIVS-II-654) having mild anemia, with a classical electrophoresis pattern (95.1% Hb A, 4.4% Hb A2, 0.5% Hb F). His father was heterozygous for Hb G-Siriraj (ßA/ßG-Siriraj) but asymptomatic, with a corresponding electrophoresis pattern (63.9% Hb A, 3.5% Hb A2, 32.6% Hb G-Siriraj). In view of the family study results, the Hb Zürich-Langstrasse mutation in the proband was considered a de novo mutation occurring on the ßIVS-II-654 allele that he inherited from his mother, resulting in a ßIVS-II-654/Hb Zürich-Langstrasse genotype, which should be interpreted as a novel ß0 mutation. This report illustrates that mutations in cis can confound genotype-phenotype correlations, therefore, just as DNA testing and Hb analysis, family study is also indispensable to the accurate identification of ß-thal mutations.

Interpretation of clot-based lupus anticoagulant assays-Normalizing clotting time against different denominators.

OBJECTIVES: The endpoint for all lupus anticoagulant (LA) assays is a clotting time in seconds. This study aimed to clarify the use of normalizing clotting time to ratio and how the use of different denominators is relevant. METHODS: Whether normalization could reduce reagent variability and possess better diagnostic performances was assessed; denominators included reference interval (RI) mean, local-obtained pooled plasma clotting time, standard plasma clotting time, and control plasma clotting time (CNPPct). Moreover, whether day-to-day variation in CNPPct would impact its application was studied. RESULTS: If not normalized, significant difference existed among different reagent batches; if normalized (against any denominators), no statistically significant difference existed anymore. The validation of in-house RIs achieved a 100% success rate. Normalization against different denominators had different RIs, but the same diagnostic efficacies (when a prolonged LA1 is used to suggest further LA-related testings, normalized LA1 demonstrated a better sensitivity: 1.0 vs. 0.95). Normalization against a "daily" CNPPct (obtained alongside test plasmas day to day) demonstrated low inter-day variations (LA1: ~1%, LA2: ~1%), and it could employ the RI for normalization against a "fixed" CNPPct (obtained alongside normal plasmas when the RI was established). CONCLUSIONS: Normalizing clotting time reduces reagent-batch variability and promotes the adoption of common RIs, and therefore reduces the necessity of establishing RI for new reagent batches. Normalized LA1 is more sensitive when used to suggest further LA-related testings, and therefore reduces the rate of missed LA diagnosis. All denominators are of the same application value. Day-to-day variation in CNPPct did not impact its application as a reliable denominator.

Peak urea level, leukocyte count and use of invasive ventilation as risk factors of mortality in acute pancreatitis: A retrospective study.

BACKGROUND: Acute pancreatitis (AP) is associated with high complications. Early, reliable prediction of mortality may improve patient management. METHODS: We retrospectively reviewed medical records of 1,599 patients with AP treated at a single large hospital in southwest China. Models to predict mortality were derived from a subset of 1,062 patients (development dataset), and the models were then validated in the remaining 537 patients (validation dataset). Independent risk factors and prediction models for mortality were identified using logistic regression. RESULTS: A total of 33 patients in the development dataset and 13 in the validation dataset died during hospitalization. Independent risk factors for mortality were found to be plasma urea levels, glucose levels and platelet counts at admission; as well as peak urea levels, leukocyte counts and use of invasive ventilation during hospitalization. Based on the development dataset, a mortality prediction model based only on urea level at admission gave an area under the curve (AUC) of 0.81, which did not significantly improve by incorporating glucose level or platelet count at admission. Significantly better was a model taking into account three in-hospital parameters: peak urea level, leukocyte count and use of invasive ventilation (AUC 0.97). CONCLUSIONS: While mortality of AP patients can be predicted reasonably well based only on urea values at admission, predictions are more reliable when they take into account in-hospital data on peak urea level, leukocyte count and use of invasive ventilation.

[Adjusting Platelet Counts for Platelet Aggregation Tests].

OBJECTIVE: To explore a better method to adjust platelet counts for light transmission aggregometry (LTA). METHODS: Blood samples from 36 healthy participants aged from 18 to 50 yr. were collected.Platelet-rich plasma (PRP) was diluted using platelet-poor plasma (PPP) and physiological saline (PS),respectively,in a ratio of 1.5,2,2.5 and 3 times. Platelet aggregation was induced by adenosine diphosphate (ADP),arachidonic acid (ARA),collagen (COL), epinephrine (EPI),or ristocetin (RIS). The maximal aggregation rates (MAs) of different approaches were compared. We also compared the MAs induced by RIS between PRP-obtained-PPP and whole blood-obtained-PPP (2 100×g, 5 min). RESULTS: Compared with the original PRP,the MAs induced by ADP,ARA,and EPI decreased in PPP-adjusted PRP (significant at 2-3 times dilution ratio,P<0.05),but not in PS-adjusted PRP (P>0.05). The MA induced by RIS decreased in PS-adjusted PRP (significant at all dilution ratios,P<0.05),but not in PPP-adjusted PRP (P>0.05). No changes in the MA induced by COL were found in PS-adjusted PRP and PPP-adjusted PRP (P>0.05). Whole blood-obtained-PPP (2 100×g, 5 min) had the same MA induced by ristocetin compared with PRP-obtained-PPP (P>0.05). CONCLUSION: PS is recommended for adjusting platelets counts for platelet aggregation induced by ADP,ARA,COL and EPI. Whole blood-obtained-PPP (2 100 ×g, 5 min) is recommended for RIS-induced aggregation as a matter of convenience.

A Phenotypically Unusual Hemoglobin H Disease Resulting from a Rare α-Globin Genotype (-SEA/-α27.6).

Hemoglobin H (Hb H) disease is usually characterized by the existence of Hb H, which influences the degree of functional anemia. We here report a patient with a rare Hb H disease genotype (-SEA/-α27.6), who was observed to paradoxically have no detectable Hb H fraction on electrophoresis. To date, the reason why the quantity of Hb H component and the clinical presentation in Hb H disease vary widely is still incompletely understood. Our report demonstrates a possible explanation - the different degradation ability of excess ß-globin chains, which might be regulated by the 27.6 kb sequence of α-globin gene.

Biochemical traits and proteomic changes in postharvest flowers of medicinal chrysanthemum exposed to enhanced UV-B radiation.

The article studied UV-B effects on biochemical traits and proteomic changes in postharvest flowers of medicinal chrysanthemum. The experiment about UV-B effects on biochemical traits in flowers included six levels of UV-B treatments (0 (UV0), 50 (UV50), 200 (UV200), 400 (UV400), 600 (UV600) and 800 (UV800) µWcm(-2)). UV400, UV600 and UV800 treatments significantly increased the contents of hydrogen peroxide, malondialdehyde and UV-B absorbing compounds, and the activity of phenylalanine ammonia lyase enzyme over the control. The contents of chlorogenic acid and flavone in flowers were significantly increased by UV-B treatments (except for UV50 and UV800). Two-dimensional gel electrophoresis was utilized to analyze proteomic changes in flowers with or without UV-B radiation. Results indicated that 43 protein spots (>1.5-fold difference in volume) were detected, including 19 spots with a decreasing trend and 24 spots with an increasing trend, and 19 differentially expressed protein spots were successfully indentified by MALDI-TOF MS. The indentified proteins were classified based on functions, the most of which were involved in photosynthesis, respiration, protein biosynthesis and degradation and defence. An overall assessment using biochemical and differential proteomic data revealed that UV-B radiation could affect biochemical reaction and promote secondary metabolism processes in postharvest flowers.

[Generation high yield vaccine strain wholly derived from avian influenza viruses by reverse genetics].

Highly pathogenic avian influenza A (HPAI) viruses of the H5N1 subtypes caused enormous economical loss to poultry farms in China and Southeastern Asian countries. The vaccination program is a reliable strategy in controlling the prevalence of these disastrous diseases. The six internal genes of the high-yield influenza virus A/Goose/Dalian/3/01 (H9N2), the hemagglutinin (HA) gene of A/Goose/HLJ/QFY/04 (H5N1) strain, and the neuraminidase gene from A/Duck/Germany/1215/73 (H2N3) reference strain were amplified by RT-PCR technique. The HA gene was modified by the deletion of four basic amino acids of the connecting peptide between HA1 and HA2. Eight gene expressing plasmids were constructed, and the recombinant virus rH5N3 was generated by cells transfection. The infection of chicken embryos and the challenge tests involving chickens demonstrated that the recombinant H5N3 (rH5N3) influenza virus is avirulent. The allantoic fluids of rH5N3-infected eggs contain high-titer influenza viruses with hemagglutination unit of 1:2048, which are eight times those of the parental H5N1 virus. The rH5N3 oil-emulsified vaccine could induce hemagglutination inhibition (HI) antibodies in chickens in 2 weeks post-vaccination, and maximum geometric mean HI-titer were observed 4 approximately 5 weeks post-vaccination and were kept under observation for 18 weeks. The rH5N3-vaccinated chickens were fully protected against morbidity and mortality of the lethal challenge of the H5N1 HPAI viruses, A/Goose/Guangdong/1/96 and A/Goose/HLJ/QFY/04, which had 8 years expansion and differences among multiple amino acids in HA protein. The N3 neuraminidase protein marker makes it possible to distinguish between H5N1 infected- and H5N3 vaccinated animals.