Plastid phylogenomics provides new insights into the systematics,diversification,and biogeography of Cymbidium(Orchidaceae)

Cymbidium(Orchidaceae:Epidendroideae),with around 60 species,is widely-distributed across Southeast Asia,providing a nice system for studying the processes that underlie patterns of biodiversity in the region.However,phylogenetic relationships of Cymbidium have not been well resolved,hampering investigations of species diversification and the biogeographical history of this genus.In this study,we construct a plastome phylogeny of 56 Cymbidium species,with four well-resolved major clades,which provides a framework for biogeographical and diversification rate analyses.Molecular dating and biogeographical analyses show that Cymbidium likely originated in the region spanning northern IndoBurma to the eastern Himalayas during the early Miocene(~21.10 Ma).It then rapidly diversified into four major clades in East Asia within approximately a million years during the middle Miocene.Cymbidium spp.migration to the adjacent regions(Borneo,Philippines,and Sulawesi)primarily occurred during the Pliocene-Pleistocene period.Our analyses indicate that the net diversification rate of Cymbidium has decreased since its origin,and is positively associated with changes in temperature and monsoon intensity.Favorable hydrothermal conditions brought by monsoon intensification in the early Miocene possibly contributed to the initial rapid diversification,after which the net diversification rate was reduced with the cooling climate after the middle Miocene.The transition from epiphytic to terrestrial habits may have enabled adaptation to cooler environments and colonization of northern niches,yet without a significant effect on diversification rates.This study provides new insights into how monsoon activity and temperature changes affected the diversification dynamics of plants in Southeast Asia.

Comparative analyses of mitogenomes in the social bees with insights into evolution of long inverted repeats in the Meliponini

The insect mitogenome is typically a compact circular molecule with highly conserved gene contents.Nonetheless,mitogenome structural variations have been reported in specific taxa,and gene rearrangements,usually the tRNAs,occur in different lineages.Because synapomorphies of mitogenome organizations can provide information for phylogenetic inferences,comparative analyses of mitogenomes have been given increasing attention.However,most studies use a very few species to represent the whole genus,tribe,family,or even order,overlooking potential variations at lower taxonomic levels,which might lead to some incorrect inferences.To provide new insights into mitogenome organizations and their implications for phylogenetic inference,this study conducted comparative analyses for mitogenomes of three social bee tribes(Meliponini,Bombini,and Apini)based on the phylogenetic framework with denser taxonomic sampling at the species and population levels.Comparative analyses revealed that mitogenomes of Apini and Bombini are the typical type,while those of Meliponini show diverse variations in mitogenome sizes and organizations.Large inverted repeats(IRs)cause significant gene rearrangements of protein coding genes(PCGs)and rRNAs in Indo-Malay/Australian stingless bee species.Molecular evolution analyses showed that the lineage with IRs have lower dN/dS ratios for PCGs than lineages without IRs,indicating potential effects of IRs on the evolution of mitochondrial genes.The finding of IRs and different patterns of gene rearrangements suggested that Meliponini is a hotspot in mitogenome evolution.Unlike conserved PCGs and rRNAs whose rearrangements were found only in the mentioned lineages within Meliponini,tRNA rearrangements are common across all three tribes of social bees,and are significant even at the species level,indicating that comprehensive sampling is needed to fully understand the patterns of tRNA rearrangements,and their implications for phylogenetic inference.

Decoding the evolution and transmissions of the novel pneumonia coronavirus(SARS-CoV-2/HCoV-19)using whole genomic data

The outbreak of COVID-19 started in mid-December2019 in Wuhan, China. Up to 29 February 2020,SARS-CoV-2(HCoV-19/2019-nCoV) had infected more than 85 000 people in the world. In this study,we used 93 complete genomes of SARS-CoV-2 from the GISAID EpiFlu TM database to investigate the evolution and human-to-human transmissions of SARS-CoV-2 in the first two months of the outbreak.We constructed haplotypes of the SARS-CoV-2 genomes, performed phylogenomic analyses and estimated the potential population size changes of the virus. The date of population expansion was calculated based on the expansion parameter tau(τ)using the formula t=τ/2 u. A total of 120 substitution sites with 119 codons, including 79 non-synonymous and 40 synonymous substitutions, were found in eight coding-regions in the SARS-CoV-2 genomes.Forty non-synonymous substitutions are potentially associated with virus adaptation. No combinations were detected. The 58 haplotypes(31 found in samples from China and 31 from outside China)were identified in 93 viral genomes under study and could be classified into five groups. By applying the reported bat coronavirus genome(bat-RaTG13-CoV)as the outgroup, we found that haplotypes H13 and H38 might be considered as ancestral haplotypes,and later H1 was derived from the intermediate haplotype H3. The population size of the SARS-CoV-2 was estimated to have undergone a recent expansion on 06 January 2020, and an early expansion on 08 December 2019. Furthermore,phyloepidemiologic approaches have recovered specific directions of human-to-human transmissions and the potential sources for international infected cases.

Influence of CXCR4/SDF-1 axis on E-cadherin/β-catenin complex expression in HT29 colon cancer cells

AIM： To study the influence of CXCR4/stromal cell- derived factor-1 （SDF-1） axis on E-cadherin/β-catenin complex expression in HT29 colon cancer ceils and its underlying mechanisms. METHODS： Effect of SDF-1 on E-cadherin/β-catenin expression was detected by immunocytochemistry. E-cadherin and/3-catenin mRNA expression levels were measured by reverse transcriptase-polymerase chain reaction. SDF-l-induced phosphorylation of phosphati- dylinositol 3-kinase （PI3K）/AKT and β-catenin was detected by Western blotting. RESULTS： The E-cadherin and β-catenin mRNA ex-pression levels in HT29 cells were lower 48 h after incubated with SDF-1 at the concentrations of 20 and 40 ng/mL （P 〈 0.05）. SDF-l-induced significant phosphorylation of PI3K/AKT and β-catenin. AMD3100 and LY294002 inhibited the phosphorylation of PI3K/AKT and β-catenin. CONCLUSION： SDF-1 down-regulates the E-cadherin/ β-catenin complex expression in HT29 cells by decreasing mRNA synthesis and increasing β-catenin phosphorylation.

Low-pressure pneumoperitoneum with abdominal wall lift in laparoscopic total mesorectal excision for rectal cancer:initial experience

AIM To evaluate the safety and feasibility of a new technology combining low-pressure pneumoperitoneum(LPP) and abdominal wall lift(AWL) in laparoscopic total mesorectal excision(TME) for rectal cancer.METHODS From November 2015 to July 2017,26 patients underwent laparoscopic TME for rectal cancer using LPP(6-8 mm Hg) with subcutaneous AWL in Qilu Hospital of Shandong University,Jinan,China.Clinical data regarding patients' demographics,intraoperative monitoring indices,operation-related indices andpathological outcomes were prospectively collected.RESULTS Laparoscopic TME was performed in 26 cases(14 anterior resection and 12 abdominoperineal resection) successfully,without conversion to open or laparoscopic surgery with standard-pressure pneumoperitoneum.Intraoperative monitoring showed stable heart rate,blood pressure and paw airway pressure.The mean operative time was 194.29 ± 41.27 min(range:125-270 min) and 200.41 ± 20.56 min(range:170-230 min) for anterior resection and abdominoperineal resection,respectively.The mean number of lymph nodes harvested was 16.71 ± 5.06(range:7-27).There was no positive circumferential or distal resection margin.No local recurrence was observed during a median follow-up period of 11.96 ± 5.55 mo(range:5-23 mo).CONCLUSION LPP combined with AWL is safe and feasible for laparoscopic TME.The technique can provide satisfactory exposure of the operative field and stable operative monitoring indices.

Effects of ytterbium addition and heat treatment on the mechanical properties and biocorrosion behaviors of Mg-Zn-Zr alloy

Mechanical properties and biocorrosion behaviors in simulated body fluid(SBF)of newly developed Mg-5.8 Zn-0.5 Zr-x Yb(ZK60-X Yb,x=0,1.0,2.0wt%)magnesium alloys in the solution-treated(T4)and artificially-aged(T6)conditions were investigated.The results of mechanical properties show that with Yb addition,the microhardness and the ultimate tensile strength(UTS)of the tested alloys are significantly increased despite a slight decrease in tensile elongation in both T4 and T6 conditions.Especially,after the T6 treatment,the microhardness and the UTS of the samples were further improved,which was mainly attributed to the precipitation strengthening.The biocorrosion behaviors of the tested alloys were studied using electrochemical examinations and immersion tests.The results indicate that the biocorrosion resistance of the tested alloys is significantly improved by Yb addition in both T4 and T6 conditions.Although the corrosion resistance was slightly deteriorated after T6 treatment,the aged ZK60-2.0 Yb alloy still exhibited a favorable corrosion behavior,which was mainly ascribed to the corrosion barrier effect of a more compact and uniform protective film induced by the dispersed nano-scale precipitates.Electrochemical measurements also confirmed these observations.Given the favorable comprehensive performance in mechanical and biocorrosion behaviors,the T6 treated ZK60-2.0 Yb alloy may be considered as a promising candidate for biomedical applications.

Four new species and a new record of Orchidinae(Orchidaceae:Orchideae)from China

Four new species of Orchidaceae from China,Heminium lijiangense,Peristylus fasciculatus,Platanthera milinensis,and Ponerorchis gongshanensis,together with a new country record,Peristylus tenuicallus,are described and illustrated based on morphological and/or phylogenetic analyses.Heminium lijiangense is closely related to H.elisabethae but differs from it by having the dorsal sepal ovate-orbicular and lip midlobe distinctly shorter than lateral lobes.P.fasciculatus is close to Peristylus tradescantifolius but is distinguished from it by having several fascicled and straight,root-like tubers(vs.one or two oblongoid tubers),old stems usually persistent,middle lobe of lip narrowly ligulate-lanceolate and half as long as the lateral lobes(vs.middle lobe deltoid,about a third as long as the lateral lobes or less),a raised callus at the base of each lateral lobe(vs.callus absent),spur gradually attenuate toward the apex(vs.spur clavate).Platanthera milinensis is similar to P.stenochila by sharing small green flowers and lip without a spur,but differs in having a creeping rhizome,a corymbose inflorescence,and a broadly ovate and slightly 3-lobed lip.Ponerochis gongshanensis is similar to P.faberi in its small flowers,but differs in having a linear leaf c.3 mm wide(vs.leaf 5-13 mm wide),in the lip having collar-like raised margins on the sides of the spur entrance,and a mid-lobe which is notched at the apex but not divided into two divergent lobules that are nearly as large as the lateral lobes,as in P.faberi.All the proposed species obtained high support in phylogenetic analysis as new species.The recently described genus Apetalanthe is reduced to synonymy of Ponerorchis and a new combination is made.

Three new species of Liparis s.l.(Orchidaceae:Malaxideae)from Southwest China based on morphological characters and phylogenetic evidence

Liparis aureolabella and L mengziensis,two new species from the karst region of southwestern China,and L bingzhongluoensis,a new species from montane region in Yunnan,are described and illustrated.L aureolabella is easily distinguished from its relatives by having abaxially purple leave with purple reticulate veins prominent adaxially,a lip auriculate at base,and falcate-lanceolate pollinia.Liparis mengziensis is closely related to L.petiolata and L.auriculata,but differs from them by having an ovate to broadly ovate leaf,purple lip and apex connate along the margins.Liparis bingzhongluoensis is similar to Liparis nanlingensis,but the new species is characterized by having a lip with two transparent ridges on its disc,longitudinally concave basal callus and triangular column wings.Phylogenetic analyses based on nuclear ribosomal ITS and plastid matK sequences showed that L aureolabella and L mengziensis are nested with L petiolata or L auriculata in a monophyletic clade.L bingzhongluoensis is sister to a clade formed by L.nanlingensis,L.tsii,L sasakii and L.krameri.Moreover,morphological comparisons strongly support that the three species as separated species newly to science.

Modification effect of Yb and Na_3PO_4 on microstructure of Mg_2Si/Mg-4Si alloy and mechanism

The modification effects of ytterbium(Yb), Na_3PO_4 and Yb + Na_3PO_4 on primary Mg_2Si phase in Mg-4Si alloys were investigated by means of X-ray diffraction(XRD), optical microscopy(OM), scanning electron microscopy(SEM) and energy dispersive spectroscopy(EDS) analysis in this work. The results indicate that the morphology of the primary Mg_2Si phase apparently changes from coarse dendrites to fine dispersive polygonal particles and the mean size decreases from 276.6 μm to 7.1 μm, with combined modification of 0.8wt.% Yb and 2.64 wt.% Na_3PO_4. Such a morphological evolution results in improvement in the ultimate tensile strength and elongation of the alloys as compared to the base alloy. This may be attributed to the formation of the YbP particles that acted as the heterogeneous nucleation substrates for the primary Mg_2Si particles, resulting in a refined distribution of these precipitates. The results of XRD examination show that there was no reaction between Si and Yb or Na_3PO_4. Solo addition of Yb or Na_3PO_4 into the melt has no real modification effect on the microstructure, but the primary Mg_2Si particles and α-Mg phases become coarser than that in the unmodified alloy.

Duodenal-jejunal bypass increases intraduodenal bile acids and upregulates duodenal SIRT1 expression in high-fat diet and streptozotocin-induced diabetic rats

BACKGROUND The mechanisms underlying diabetes remission after duodenal-jejunal bypass(DJB) remain elusive. In DJB surgery, the duodenum is excluded. However, the duodenum has emerged as an important regulator of glucose homeostasis, and elevated duodenal SIRT1 leads to improved hepatic insulin sensitivity. After DJB, bile acids(BAs) in the duodenum are not mixed and diluted by the ingested food. And activation of BA receptors promotes SIRT1 expression in many tissues. We hypothesized that BA-mediated upregulation of SIRT1 may contribute to diabetic control after DJB.AIM To investigate the surgical effects of DJB on duodenal SIRT1 expression and uncover the potential crosslinks between BAs and SIRT1.METHODS Twenty diabetic rats were randomly allocated to the sham(n = 10) and DJB(n = 10) groups. Body weight, food intake, fasting blood glucose(FBG), serum and intraduodenal total BA(TBA) levels were measured accordingly. Oral glucose tolerance test(OGTT) and intraperitoneal pyruvate tolerance test(ip PTT) were performed to evaluate the effects of surgeries on systemic glucose disposal and hepatic gluconeogenesis. The key genes of BA signaling pathway in the duodenal mucosa, including farnesoid X receptor(FXR), small heterodimer partner(SHP), and Takeda G-protein-coupled receptor 5(TGR5) were evaluated by real-time quantitative polymerase chain reaction 8 wk postoperatively. The duodenal SIRT1, AMPK, and phosphorylated AMPK(p-AMPK) levels were evaluated by western blotting. Rat small intestine epithelial IEC-6 cells were treated with GW4064 and INT-777 to verify the effects of BAs on SIRT1 expression in enterocytes.RESULTS The DJB group exhibited body weight and food intake comparable to those of the sham group at all postoperative time points. The FBG level and area under the curve for the OGTT and ip PTT were significantly lower in the DJB group. The DJB group exhibited higher fasting and postprandial serum TBA levels than the sham group at both 2 and 8 wk postoperatively. At 8 wk after surgery, the DJB group showed higher intraluminal TBA concentration, upregulated m RNA expression of FXR and SHP, and elevated protein expression of SIRT1 and p-AMPK in the descending and horizontal segments of the duodenum. Activation of FXR and TGR5 receptors by GW4064 and INT-777 increased the m RNA and protein expression of SIRT1 and promoted the phosphorylation of AMPK in IEC-6 cells.CONCLUSION DJB elevates intraduodenal BA levels and activates the duodenal BA signaling pathway, which may upregulate duodenal SIRT1 and further contribute to improved glucose homeostasis after DJB.

Fibroblast growth factor 15,induced by elevated bile acids,mediates the improvement of hepatic glucose metabolism after sleeve gastrectomy

BACKGROUND Fibroblast growth factor(FGF)15/19,which is expressed in and secreted from the distal ileum,can regulate hepatic glucose metabolism in an endocrine manner.The levels of both bile acids(BAs)and FGF15/19 are elevated after bariatric surgery.However,it is unclear whether the increase in FGF15/19 is induced by BAs.Moreover,it remains to be understood whether FGF15/19 elevations contribute to improvements in hepatic glucose metabolism after bariatric surgery.AIM To investigate the mechanism of improvement of hepatic glucose metabolism by elevated BAs after sleeve gastrectomy(SG).METHODS By calculating and comparing the changes of body weight after SG with SHAM group,we examined the weight-loss effect of SG.The oral glucose tolerance test(OGTT)test and area under the curve of OGTT curves were used to assess the anti-diabetic effects of SG.By detecting the glycogen content,expression and activity of glycogen synthase as well as the glucose-6-phosphatase(G6Pase)and phosphoenolpyruvate carboxykinase(Pepck),we evaluated the hepatic glycogen content and gluconeogenesis activity.We examined the levels of total BA(TBA)together with the farnesoid X receptor(FXR)-agonistic BA subspecies in systemic serum and portal vein at week 12 post-surgery.Then the histological expression of ileal FXR and FGF15 and hepatic FGF receptor 4(FGFR4)with its corresponding signal pathways involved in glucose metabolism were detected.RESULTS After surgery,food intake and body weight gain of SG group was decreased compare with the SHAM group.The hepatic glycogen content and glycogen synthase activity was significantly stimulated after SG,while the expression of the key enzyme for hepatic gluconeogenesis:G6Pase and Pepck,were depressed.TBA levels in serum and portal vein were both elevated after SG,the FXR-agonistic BA subspecies:Chenodeoxycholic acid(CDCA),lithocholic acid(LCA)in serum and CDCA,DCA,LCA in portal vein were all higher in SG group than that in SHAM group.Consequently,the ileal expression of FXR and FGF15 were also advanced in SG group.Moreover,the hepatic expression of FGFR4 was stimulated in SG-operated rats.As a result,the activity of its corresponding pathway for glycogen synthesis:FGFR4-Ras-extracellular signal regulated kinase pathway was stimulated,while the corresponding pathway for hepatic gluconeogenesis:FGFR4-cAMP regulatory element-binding protein-peroxisome proliferator-activated receptorγcoactivator-1αpathway was suppressed.CONCLUSION Elevated BAs after SG induced FGF15 expression in distal ileum by activating their receptor FXR.Furthermore,the promoted FGF15 partly mediated the improving effects on hepatic glucose metabolism of SG.

Pollen size strongly correlates with stigma depth among Pedicularis species

Summary Darwin proposed that pollen size should be positively correlated with stigma depth rather than style length among species given that pollen tubes first enter the stigma autotrophically, then grow through the style heterotrophically. However, studies often show a posi-tive relationship between pollen size and style length. Five floral traits were observed to be correlated among 42 bumblebee-pollinated Pedicularis species （Orobancha- ceae） in which stigmas are distinct from styles. The phylogenetic independent contrast analysis revealed that pollen grain volume was more strongly correlated with stigma depth than with style length, consistent with Darwin＇s functional hypothesis between pollen size and stigma depth.

Pollen Morphology of Pedicularis sect.Cyathophora,a Group Endemic to the Eastern Himalaya-Hengduan Mountains Region

Pedicularis sect. Cyathophora is a distinctive group endemic to the eastern Himalaya-Hengduan Mountains region. It was regarded as a ＇grex＇ or section and included all four general corolla types of Pedicularis. A unique feature of it is that the leaf and bract bases are fused together to form a cup-like structure around the stem at each node. Pollen morphology of seven species in sect. Cyathophora was investigated by scanning electron microscopy （SEM） and light microscopy （LM）. Two different pollen apertures could be distinguished （i.e. trisyncolpate and bisyncolpate） using LM, while examination with SEM made it possible to recognize three types of exine ornamentation （i.e. microscabrate, microfoveolate and microreticulate）. The microfoveolate exine ornamentation was found in trisyncolpate pollen grains for the first time. Possible relationships between pollen data and the corolla types were discussed. Comparisons of floral and phyllotaxy characters of the genus Pedicularis, together with the pollen characters of sect. Cyathophora, could help us to better understand the evolutionary trends in Pedicularis.

Al–9.00 %Si–0.25 %Mg alloys modified by ytterbium

The effects of ytterbium（Yb） on microstructure and solidification behavior of Al-9.00%Si-0.25%Mg alloys were investigated.By optical microscope（OM） and scanning electron microscopy（SEM）,it is found that the morphology of eutectic silicon changes from coarse plates to fine fibers by the addition of 0.7%wt Yb.In addition,the grains of α-Al matrix are refined by Yb addition.Phase constitution of the alloy was analyzed by X-ray diffractometer（XRD） and energy dispersive spectroscopy（EDS）attached with SEM and mechanical properties were measured by hardness test.It is concluded that the Yb atoms are incorporated into the silicon by the adsorption at the solid-liquid growth front to cause the modification of eutectic silicon.Furthermore,the results of XRD and EDS analysis reveal that the Yb-containing phase forming in the alloys is Al_3Yb.