Mitogenome phylogeny reveals Indochina Peninsula origin and spatiotemporal diversification of freshwater crabs (Potamidae: Potamiscinae) in China.

The Asian potamid subfamily Potamiscinae is the most species-rich group of primary freshwater crabs in the world, and a large number of these (318 species in 52 genera) are found in China where they exhibit a remarkably high rate of endemism. However, the evolutionary history and phylogenetic relationships of this subfamily is still poorly understood and the hypothesis that the Chinese potamids originated in the Indochina Peninsula has yet to be tested using a modern phylogeographic reconstruction and molecular dating methods. Here we analyze the phylogenetic relationships and biogeographic history of the Chinese potamiscines based on whole mitogenomes of 72 species of Asian freshwater crabs representing 65% of the potamiscine genera found in China, including new mitogenome sequences for 57 species. The mitogenomic phylogeny generated four separate clades: Southwest China (SWC), Indochina-Southwest China (ISWC), Central China (CC), and South China-adjacent Islands (SCI). Our results suggest that the Chinese potamiscine fauna most likely originated in Indochina during the Eocene. The establishment of the East Asian monsoon during the Eocene/Oligocene resulted in major climate changes that coincide with the local diversification of the CC and SCI clades. In addition, diversification of two other clades (ISWC and SWC) may have been driven by past large scale environmental changes taking place at that time such as increased local precipitation and expanding freshwater habitats associated with the uplifting of the Hengduan Mountains. In addition, our phylogenies did not support monophyly for six currently recognized potamiscine genera, highlighting the need for further taxonomic revisions of this fauna. The broad taxonomic sampling and well-supported molecular phylogenies used here allow for the updating of our understanding of the evolutionary origins and diversification of the potamiscine freshwater crabs in China.

Phylogenetic implications of mitogenome rearrangements in East Asian potamiscine freshwater crabs (Brachyura: Potamidae).

We present here a combined mitogenome gene order dataset totalling 62% of the known genera of East Asian potamiscine freshwater crabs that includes first-time mitogenome data for 14 species and gene order data from 37 other species of potamiscines. A surprisingly high number of mitogenome gene order rearrangements were found in the taxa studied (comprising nine different rearrangements and seven major patterns, one of which has two sub-arrangements). Our phylogenetic reconstructions indicate that the mitogenome gene order rearrangements are associated with the evolutionary history of potamiscine lineages. We also used a new Event-based Maximum Parsimony method to reconstruct ancestral gene orders, which takes into consideration gene duplication, pseudogeneticization, and tandem duplication random loss. Furthermore, shared mitogenome gene order patterns were used to inform the taxonomic placement of Sinopotamon parvum, and the cryptic diversity in Potamiscus. The remarkably frequent mitogenome rearrangements in potamiscine freshwater crabs have great potential to contribute to our understanding of the evolutionary history of these highly diverse decapods in East Asia.

Phylomitogenomics reconfirm the phylogenetic position of the genus Metaplax inferred from the two grapsid crabs (Decapoda: Brachyura: Grapsoidea).

Two new complete mitogenomes of the grapsids, Metaplax longipes Stimpson, 1858 and Nanosesarma minutum (De Man, 1887) were sequenced using next-generation sequencing (NGS). By analyzing a combination of 75 Brachyura taxa, our phylomitogenomic inferences suggested that Metaplax crab seperated earlier from the sesarmid crabs and closely related to the varunids with respect to Nanosesarma crab. It reconfirmed that the Metaplax should be removed from the Sesarmidae and assinged to the Varunidae. Additional mitogenomic comparisons including gene rearrangement and genomic organization were conducted among the 33 taxa of Grapsoidea and Ocypodoidea, and a shared rearrangement pattern between Metaplax longipes and the varunids were recovered, which also strongly supported the inference for the phylogenetic position of the Metaplax.

PD-1 blockade enhances radio-immunotherapy efficacy in murine tumor models.

PURPOSE: It has become increasingly clear in cancer treatment that radiotherapy can be enhanced by immunotherapy. In the present study, we evaluated a novel triple combination therapy consisting of local radiotherapy, intratumoral CpG, and systemic PD-1 blockade in lung cancer models. METHODS: The efficacy of a novel triple therapy was examined by recording tumor volume and survival time. The immunologic effects of this novel triple therapy were evaluated by the frequency and percentage of immune cells and cytokines using flow cytometry. RESULTS: This triple combination proved more effective than its subcomponents and its positive antitumor effects included reducing tumor growth and improving host survival. The antitumor effect was not only observed in directly irradiated tumors but also in at distant tumor sites in a CD8+ T-cell-dependent fashion. Phenotypic analyses of CD8+ T cells revealed that the triple combination therapy increased the percentage of effector memory T cells in the spleen. Furthermore, the combination therapy significantly increased the frequency of IFN-γ and TNF-α-positive-CD8+ tumor-infiltrating lymphocytes (TIL) and mature-activated dendritic cells (DCs) within treated tumors, indicating that the antitumor effects likely depend on the activation of a DC subset specialized in antigen crosspriming to induce cytotoxic lymphocyte (CTLs). In addition, the triple therapy reduced immunosuppressive factors, like regulatory T cells (Tregs) in the spleen and tumor microenvironment while inducing the robust systemic antitumor effect. Finally, the triple treatment was, indeed, well tolerated and had a little effect on the hemogram and lung. CONCLUSIONS: These results suggest that this triple therapy promotes a local antitumor immune response with systemic consequences. The efficacy and limited toxicity of this strategy are attractive for clinical translation.

Characterization of four new mitogenomes from Ocypodoidea & Grapsoidea, and phylomitogenomic insights into thoracotreme evolution.

Four new complete mitochondrial genomes (mitogenomes) from the two superfamilies Ocypodoidea and Grapsoidea were sequenced, which represented Uca (Gelasimus) borealis (Ocypodidae: Ucinae), Dotilla wichmani (Dotillidae), Metopograpsus quadridentatus (Grapsidae: Grapsinae), and Gaetice depressus (Varunidae: Gaeticinae). All of the mitogenomes shared the complete set of 37 mitochondrial genes. Mitogenome lengths were 15,659, 15,600, 15,517, and 16,288 bp, respectively, with A + T contents of 69.41%, 68.46%, 70.30%, and 72.96%, respectively. Comparative genomic analyses suggested that they exhibited different genomic rearrangements. In particular, G. depressus shared a major rearrangement pattern present in Eriocheir crabs, while the remainder shared the brachyuran ground genomic rearrangement patterns. Phylomitogenomic inferences provided new evidence for the strongly supported nesting of Thoracotremata within Heterotremata clades. A close phylogenetic relationship was observed between Varunidae and Macrophthalmidae crabs, and between Dotillidae and Grapsidae crabs, which was consistent with mitochondrial genomic rearrangement similarities. Altogether, these results suggest the presence of reciprocal paraphyly for Ocypodoidea and Grapsoidea.

The complete mitochondrial genome of the semiterrestrial crab, Chiromantes neglectum (Eubrachyura: Grapsoidea: Sesarmidae).

The complete mitogenome of the semiterrestrial crab Chiromantes neglectum was sequenced. It contained the entire set of 37 genes. The gene order was basically identical to pancrustacean ground pattern, except for thetrnH and trnQ genes. Phylogenetic inferences based on protein-coding genes (PCGs) provide strong evidence that places C. neglectum within an intermingled 'Grapsoidea & Ocypodoidea' clade.