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Abstract Background Evidence shows that full mycoheterotrophs and holoparasites often have reduced plastid genomes with rampant gene loss, elevated substitution rates, and deeply altered to conventional evolution in mitochondrial genomes, but mechanisms of cytonuclear evolution is unknown. Endoparasitic Sapria himalayana and mycoheterotrophic Gastrodia and Platanthera guangdongensis represent different heterotrophic types, providing a basis to illustrate cytonuclear evolution. Here, we focused on nuclear-encoded plastid / mitochondrial (N-pt / mt) -targeting protein complexes, including caseinolytic protease (ClpP), ribulose-1,5-bisphosphate carboxylase/oxygenase (RuBisCo), oxidative phosphorylation system (OXPHOS), DNA recombination, replication, and repair (DNA-RRR) system, and pentatricopeptide repeat (PPR) proteins, to identify evolutionary drivers for cytonuclear interaction. Results The severity of gene loss of N-pt PPR and pt-RRR genes was positively associated with increased degree of heterotrophy in full mycoheterotrophs and S. himalayana, while N-mt PPR and mt-RRR genes were retained. Substitution rates of organellar and nuclear genes encoding N-pt/mt subunits in protein complexes were evaluated, cytonuclear coevolution was identified in S. himalayana, whereas disproportionate rates of evolution were observed in the OXPHOS complex in full mycoheterotrophs, only slight accelerations in substitution rates were identified in N-mt genes of full mycoheterotrophs. Conclusions Nuclear compensatory evolution was identified in protein complexes encoded by plastid and N-pt genes. Selection shaping codon preferences, functional constraint, mt-RRR gene regulation, and post-transcriptional regulation of PPR genes all facilitate mito-nuclear evolution. Our study enriches our understanding of genomic coevolution scenarios in fully heterotrophic plants.

Abstract Echinoderm microtubule-associated protein 4 (EML4) - anaplastic lymphoma kinase (ALK) fusion gene detection is of great significance in personalized tumor treatment. With the development of EML4-ALK fusion variants detection, it is necessary to establish traceability to ensure the consistency and comparability of its detection results in clinical practice. The establishment of traceability relies on SI traceable reference materials (RMs) and potential reference measurement procedures (RMPs). In this study, a potential RMP for the quantitative detection of V1 and V3 fusion mutations and the reference type (ALK-ref, including wild type, V1 and V3 mutant type) based on reverse transcription dPCR (RT-dPCR) and EML4-ALK fusion gene RMs were established. The proposed potential RMP has high specificity, good inter-laboratory reproducibility (CV

Orchidaceae are one of the largest families of angiosperms in terms of species richness. In the last decade, numerous studies have delved into reconstructing the phylogenetic framework of Orchidaceae, leveraging data from plastid, mitochondrial and nuclear sources. These studies have provided new insights into the systematics, diversification and biogeography of Orchidaceae, establishing a robust foundation for future research. Nevertheless, pronounced controversies persist regarding the precise placement of certain lineages within these phylogenetic frameworks. To address these discrepancies and deepen our understanding of the phylogenetic structure of Orchidaceae, we provide a comprehensive overview and analysis of phylogenetic studies focusing on contentious groups within Orchidaceae since 2015, delving into discussions on the underlying reasons for observed topological conflicts. We also provide a novel phylogenetic framework at the subtribal level. Furthermore, we examine the tempo and mode underlying orchid species diversity from the perspective of historical biogeography, highlighting factors contributing to extensive speciation. Ultimately, we delineate avenues for future research aimed at enhancing our understanding of Orchidaceae phylogeny and diversity.

IntroductionOur objective was to explore the efficacy of hyperbaric oxygen in the treatment of sudden sensorineural hearing loss by conducting an umbrella review of all existing evidence.MethodsWe conducted an umbrella review, searching for related articles in the PubMed, Web of Science, Embase, and Scopus databases. The search period covered from the inception of each database until April 2024. We extracted authors, country of publication, time of publication, number of included studies and participants, interventions, summary of results, P-values, I2, relative risk (95% CI), and outcome measures. The methodological quality, evidence quality, and overlap rate of the included articles were assessed using AMSTAR 2, GRADE, and OVErviews (GROOVE).ResultsMethodological quality was assessed using AMSTAR 2. Of the nine included articles, two were assessed as “high,” three as “moderate,” two as “low,” and the remaining two as “very low.” The quality of evidence was assessed using the GRADE system. It was found that the quality of evidence in most of the studies was unsatisfactory. It was found that there was a slight overlap among the included articles. Six studies reported positive results (OR 1.37; 95% CI, 1.17–1.61; P = 0.04), with high heterogeneity observed (I2 = 63%). Egger's test indicated bias (P = 0.000101). Three studies reported negative results (MD 1.49; 95% CI, −0.32 to 3.29; P = 0.43; I2 = 0%), with no significant bias detected (P = 0.106) according to Egger's test.ConclusionHBO therapy is shown to be an effective treatment for SSNHL with fewer side effects. However, the methodological quality and evidence of the systematic reviews and meta-analysis included in this study were generally low. Therefore, more high-quality, large-scale, multi-center randomized controlled trials are needed in the future to verify the efficacy of HBO therapy for SSNHL.Systematic review registrationhttps://www.crd.york.ac.uk/prospero, identifier [CRD42024523651].

This research aimed to study the impact of different altitudes on the flue gas temperature and thermal stratification in the highway tunnel with a vertical shaft, by establishing a full-sized tunnel model using the numerical simulation software Fire Dynamics Simulator (FDS). Simulation experimental results were analyzed and it was found: first, with the same heat release power of fire source, the vertical, transverse and roof temperatures of both fire source and non-fire source sections in the tunnel all increased to a certain extent along with the increase of altitude; second, the vertical, transverse and roof temperatures of the tunnel all increased with the increased heat release rate of fire source at the same altitude, and the temperature of the fire source section was higher than that of the non-fire source section as a whole; third, the average temperature and stratification intensity of vertical flue gas in both sections showed an increasing trend with the increase of altitude, indicating that the intensity of flue gas thermal stratification at high altitude was greater than that at low altitude, because a large amount of flue gas accumulated in the upper tunnel and led to the increase of the roof flue gas temperature at high altitude.

A new species of Orchidaceae, Phalaenopsis zhanhuoensis, from Xichou County, Yunnan, China, is described and illustrated. The novelty is close to P. taenialis, P. wilsonii, and P. stobartiana, but differs from them by having a distinct, fleshy anterior callus with a deeply lobed apex at the base of the labellum and lateral lobes of labellum reflexed and facing outward.

Economic recession from the coronavirus outbreak continues to have negative rippling effects on local and international financial investments in public–private partnership (PPP) projects in Ghana, a developing country. Together with poor reports on operating cash-inflows for PPP projects such as those covering schools, hospitals, railways, fishing harbors, cocoa warehouses, recreational parks and affordable housing, there is an urgent need to develop and institutionalise sustainable and robust financial risk management measures. These measures are meant to mitigate financial losses, promote sustainability practices and prolong the longevity of infrastructure developments within PPP pacts. Therefore, this study aims to assess the mitigation measures of PPP infrastructure management in Ghana. Primary data from survey questionnaires were utilised in this study, with the data obtained from PPP practitioners and experts. Data were grouped into project type, sectors and practitioners of PPP projects to aid the analysis using tools such as the Kruskal–Wallis test, the Mann–Whitney U test and factor analysis. The results demonstrate sustainable and green finance, the innovative skills and competencies of project teams, green financial risk models and inclusive cost reduction strategies as crucial to minimising financial risks in PPP project delivery. These findings have significant implications for PPP practitioners and researchers in Ghana and similar developing countries to understand and develop measures to respond to financial risks for sustainable PPP project development and future research studies.

Abstract Background Sapria himalayana (Rafflesiaceae) is an endoparasitic plant characterized by a greatly reduced vegetative body and giant flowers; however, the mechanisms underlying its special lifestyle and greatly altered plant form remain unknown. To illustrate the evolution and adaptation of S. himalayasna, we report its de novo assembled genome and key insights into the molecular basis of its floral development, flowering time, fatty acid biosynthesis, and defense responses. Results The genome of S. himalayana is ~ 1.92 Gb with 13,670 protein-coding genes, indicating remarkable gene loss (~ 54%), especially genes involved in photosynthesis, plant body, nutrients, and defense response. Genes specifying floral organ identity and controlling organ size were identified in S. himalayana and Rafflesia cantleyi, and showed analogous spatiotemporal expression patterns in both plant species. Although the plastid genome had been lost, plastids likely biosynthesize essential fatty acids and amino acids (aromatic amino acids and lysine). A set of credible and functional horizontal gene transfer (HGT) events (involving genes and mRNAs) were identified in the nuclear and mitochondrial genomes of S. himalayana, most of which were under purifying selection. Convergent HGTs in Cuscuta, Orobanchaceae, and S. himalayana were mainly expressed at the parasite–host interface. Together, these results suggest that HGTs act as a bridge between the parasite and host, assisting the parasite in acquiring nutrients from the host. Conclusions Our results provide new insights into the flower development process and endoparasitic lifestyle of Rafflesiaceae plants. The amount of gene loss in S. himalayana is consistent with the degree of reduction in its body plan. HGT events are common among endoparasites and play an important role in their lifestyle adaptation.

Public–private partnership (PPP) is a prominent tool for sustainable infrastructure development. However, the positive contributions of PPPs toward the attainment of sustainable, climate resilience and zero-carbon infrastructure projects are hampered by poor financial risk management. This problem is more prevalent in developing countries like Ghana where private investment inflow has plummeted due to the COVID-19 recession and poor project performance. Thus, this study aims to assess the key financial risk management strategies in ensuring sustainable PPP infrastructure projects in Ghana. The study utilised primary data from PPP practitioners in Ghana solicited through survey questionnaires. Factor analysis, mean scores and fuzzy synthetic analysis are the data analysis techniques for this study. The results revealed that sustainable and green funding models, effective cost-reduction initiatives, a competent team with committed leadership and emerging technologies and regulations constitute the key strategies for managing the financial risks of sustainable PPP infrastructure projects. Although future studies must expand the scope of data gathering, the findings of the study enrich the theoretical understanding of financial risks in sustainable investments in PPP infrastructures. Relevant remedies that will aid the development of practical financial risk management guidelines are also provided in this study for PPP practitioners.

The linear economy model that is generally practised in the construction industry is one of the main reasons hindering the achievement of sustainability in construction. Alternatively, the Circular Economy (CE) model is becoming a promising approach to achieve sustainable construction, in which materials and products are circulated at their highest economic value and, thereby, contribute towards mitigating the negative economic, social, and environmental impacts of the construction industry. For a successful transition towards CE in the construction industry, it is important not only to understand CE practices that could be implemented across different stages of the life cycle of construction projects, but also to understand how a particular practice relates to another at those stages. Having limited studies identifying such relationships, this study aims to improve the successful implementation of CE practices in the construction industry by comprehending the practices, along with their relationships, throughout the project life cycle. A qualitative research method consisting of a systematic literature review was adopted using the Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) framework to identify CE practices. Amongst the identified highly cited CE practices applicable to construction projects, design for disassembly, design for adaptability and flexibility, and design for manufacture and assembly ended up being the most significant ones. Consequently, a relationship matrix was developed, highlighting the relationships among the highly cited practices. The matrix emphasises that practices applied in the design stage relate frequently to ones at the following stages, necessitating projects to start with proper planning. This study ultimately assists construction industry stakeholders in successfully and rationally implementing CE practices throughout the project life cycle while enhancing the value addition on the economy, society, and environment.

Trollius, a genus in the Ranunculaceae family, has significant medicinal and ornamental value. It is widely distributed in China with 16 different species accepted. However, due to the lack of enough samples and information sites, the molecular phylogenetic relationships of Trollius have been unresolved till now. Here we sequenced, assembled and annotated the plastomes of 16 Trollius species to investigate their genomic characteristics, inverted repeat (IR) boundaries, sequence repeats, and hypervariable loci. In addition, the phylogenetic relationships of this genus was reconstructed based on the whole plastomes and the protein-coding sequences data-sets. The plastomes of Trollius ranged between 159,597 bp and 160,202 bp in length, and contained 113 unique genes, including 79 protein coding, 30 tRNA, and 4 rRNA. The IR boundaries were relatively conserved within the genus Trollius. 959 simple sequence repeats and 657 long sequence repeats were detected in the Trollius plastomes. We identified 12 highly polymorphic loci (Pi > 0.0115) that can be used as plastid markers in molecular identification and phylogenetic investigation of the genus. Besides, Trollius was a monophyletic group with the earliest divergence clade being Trollius lilacinus Bunge, and the remaining species were divided into two strongly-supported clades. The phylogeny in our study supported the traditional classification systems based on the color of sepal, but not the previous classification system based on the types and relative lengths of the nectaries, and distribution. The genomic resources provided in our study can be used in the taxonomy of the genus Trollius, promoting the development and utilization of this genus.

A non-integrated induced pluripotent stem cell (iPSC) line was generated derived from a healthy 20-day-old male using peripheral blood mononuclear cells. The established induced pluripotent stem cell line possesses a normal karyotype, expresses pluripotency markers and able to differentiate into all three germ layers in vitro. This cell line could serve as a health control or a basis for disease modeling, to explore molecular pathogenesis.

This paper aims to study the effects of altitude and the size of smoke outlet on fire smoke diffusion and discharge in semi-lateral smoke exhaust highway tunnels. At first, simulations of semi-lateral smoke exhaust highway tunnels were carried out in FDS (Fire Dynamics Simulator), then the distribution laws of temperature, CO concentration, smoke mass flow, and visibility in the tunnel under the conditions of different altitudes and smoke outlet areas were analyzed to figure out the effects of altitude and size of smoke outlet on fire smoke diffusion and discharge in the said tunnels. The results suggest that, in case of the same fire source power, the velocity of smoke diffusion increases with the altitude; for curves of different altitudes, the tunnel roof temperature features are basically the same, that is, the higher the altitude, the higher the temperature at the tunnel roof. When the fire source power is 20 MW, the smoke mass flow at the smoke outlet decreases with the increase of altitude, but the CO concentration grows with it, indicating that the smoke exhaust efficiency is higher in high-altitude areas. When the altitude reaches 4200 m and the fire source power is 20 MW, with the increase of smoke outlet area, the smoke discharge effect of the tunnel shows an upward trend, taking both the smoke discharge effect and economy into consideration; the smoke outlet should take a size of 4 x 3 m.

Abstract Background Plastomes of heterotrophic plants have been greatly altered in structure and gene content, owing to the relaxation of selection on photosynthesis-related genes. The orchid tribe Gastrodieae is the largest and probably the oldest mycoheterotrophic clade of the extant family Orchidaceae. To characterize plastome evolution across members of this key important mycoheterotrophic lineage, we sequenced and analyzed the plastomes of eleven Gastrodieae members, including representative species of two genera, as well as members of the sister group Nervilieae. Results The plastomes of Gastrodieae members contain 20 protein-coding, four rRNA and five tRNA genes. Evolutionary analysis indicated that all rrn genes were transferred laterally and together, forming an rrn block in the plastomes of Gastrodieae. The plastome GC content of Gastrodia species ranged from 23.10% (G. flexistyla) to 25.79% (G. javanica). The plastome of Didymoplexis pallens contains two copies each of ycf1 and ycf2. The synonymous and nonsynonymous substitution rates were very high in the plastomes of Gastrodieae among mycoheterotrophic species in Orchidaceae and varied between genes. Conclusions The plastomes of Gastrodieae are greatly reduced and characterized by low GC content, rrn block formation, lineage-specific reconfiguration and gene content, which might be positively selected. Overall, the plastomes of Gastrodieae not only serve as an excellent model for illustrating the evolution of plastomes but also provide new insights into plastome evolution in parasitic plants.

Summary: National key protected wild plants (NKPWPs) are considered flagship species for plant diversity conservation in China. Using data for 1101 species, we characterized NKPWPs distribution patterns in China and assessed conservation effectiveness and conservation gaps. In total, 4880 grid cells at a 20 × 20 km resolution were filled with occurrence records for NKPWPs. We identified 444 hotspot grid cells and 27 diversity hotspot regions, containing 92.37% of NKPWPs. However, 43.24% of these hotspot grid cells were fully or partially covered by national nature reserves (NNRs), where 70.21% of species were distributed. Approximately 61.49% of the NKPWPs species were protected by NNRs, but the populations or habitats of 963 species were partially or fully outside of NNRs. With global warming, the overall change in the extent of suitable habitats for NKPWPs is expected to be small, however, habitat quality in some areas with a high habitat suitability index will decrease.

Pathogenic variants in Jagged-1 (JAG1), which encodes the ligand of the Notch receptor, had been demonstrated to cause Alagille syndrome. However, there is no evidence to support any genotype-phenotype correlations. Here, we generated a gene-edited human embryonic stem cell (hESC) line (H9) carrying the c.1615C > T mutation in JAG1 that was identified in a patient with Alagille syndrome (ALGS). This modified cell line was accomplished by using cytosine base editor (CBE), and may serve as a valuable model for JAG1 mutaion related disease, and facilitate to gain more insight into the biological function of JAG1.

BackgroundThe role of matrix metalloproteinase 9 (MMP-9) in the pathophysiology of chronic kidney disease (CKD), which is associated with a nearly two-fold greater risk for urinary calculi compared to people without CKD, has been demonstrated. The aim of the research is to evaluate the association between MMP-9-1562C>T polymorphism, MMP-9 serum levels and nephrolithiasis risk.MethodsA hospital-based case-control study involving 302 kidney stone patients and 408 controls without kidney stone from southern China was conducted. Sanger sequencing was used to genotype the MMP-9-1562C>T polymorphism. The serum MMP-9 was measured in 105 kidney stone patients and 77 controls by enzyme-linked immunosorbent assay.ResultsCompared to the control group, the CT genotype was more frequent in nephrolithiasis patients (adjusted OR = 1.60, 95% CI = 1.09–2.37: the risk of developing nephrolithiasis in individuals with CT genotype compared to CC genotype). Moreover, there was also a higher frequency of CT/TT genotypes among patients with nephrolithiasis (adjusted OR = 1.49, 95% CI = 1.02–2.19: the risk of developing nephrolithiasis in individuals with CT/TT genotypes compared to CC genotype). The risk remained for the subgroups of patients aged >53, smokers with pack-years of smoking >20, non-drinkers, non-diabetic patients, patients with hypertension, recurrent episodes and calcium oxalate stones (OR = 2.26, 95% CI = 1.31–3.91; OR = 5.47, 95% CI = 1.10–27.30; OR = 1.76, 95% CI = 1.14–2.72; OR = 1.54, 95% CI = 1.03–2.30; OR = 1.97, 95% CI = 1.01–3.82; OR = 1.67, 95% CI = 1.06–2.62; OR = 1.54, 95% CI = 1.02–2.32, respectively). Biochemical parameters did not differ between genotypes. Compared to controls (18.57 ± 5.80 ng/mL), nephrolithiasis patients had significantly higher serum MMP-9 levels (30.17 ± 6.78 ng/mL, p T were significantly higher than those with CC genotype (32.00 ± 6.33 vs. 29.13 ± 6.85 ng/mL, p = 0.037).ConclusionThe MMP-9-1562C>T polymorphism in association with its soluble protein increased the risk of kidney stone, thus suggesting it could be used as a susceptibility biomarker for nephrolithiasis. Further functional studies and larger studies that include environmental exposure data are needed to confirm the findings.

About 47% of pathogenic point mutations could be corrected by ABE-induced A·T-to-G·C conversions. However, the applications of ABEs are still hindered by undesired editing efficiency, limited editing scopes, and off-targeting effects. Here, we develop a new adenine base editor, by embedding TadA-8e monomer into SpRY-nCas9, named as CE-8e-SpRY, which exhibits higher activity at NRN than NYN PAMs favored by SpRY nuclease. CE-8e-SpRY could target nearly all genomic sites in principle and induces the highest targeting efficiency among tested SpRY-based ABEs. In addition, CE-8e-SpRY also shows reduced RNA and DNA off-targeting activities. With optimized sgRNAs, CE-8e-SpRY induces efficient or desired target editing at some disease-relevant loci where conventional ABEs were unable to induce precise and satisfied editing. Taken together, our CE-8e-SpRY could broaden the applicability of ABEs in correcting or introducing pathogenic point mutations.

Abstract Cyclin-dependent kinase 5 (Cdk5) /p35 is involved in many developmental processes of the central nervous system. Cdk5/p35 is also implicated in synaptic plasticity, learning and memory. Several lines of conditional Cdk5 knockout mice (KO) have been generated and have shown different outcomes for learning and memory. Here, we present our analysis of p35 conditional KO mice (p35cKO) in hippocampal pyramidal neurons or forebrain GABAergic neurons using electrophysiological and behavioral methods. In the fear conditioning task, CamKII-p35cKO mice showed impaired memory retention. Furthermore, NMDAR-dependent long-term depression (LTD) induction by low-frequency stimuli in hippocampal slices from CamkII-p35cKO mice was impaired compared to that in control mice. In contrast, Dlx-p35cKO mice showed no abnormalities in behavioral tasks and electrophysiological analysis in their hippocampal slices. These results indicated that Cdk5/p35 in excitatory neurons is important for the hippocampal synaptic plasticity and associative memory retention.

Abstract Background To illustrate the molecular mechanism of mycoheterotrophic interactions between orchids and fungi, we assembled chromosome-level reference genome of Gastrodia menghaiensis (Orchidaceae) and analyzed the genomes of two species of Gastrodia. Results Our analyses indicated that the genomes of Gastrodia are globally diminished in comparison to autotrophic orchids, even compared to Cuscuta (a plant parasite). Genes involved in arbuscular mycorrhizae colonization were found in genomes of Gastrodia, and many of the genes involved biological interaction between Gatrodia and symbiotic microbionts are more numerous than in photosynthetic orchids. The highly expressed genes for fatty acid and ammonium root transporters suggest that fungi receive material from orchids, although most raw materials flow from the fungi. Many nuclear genes (e.g. biosynthesis of aromatic amino acid L-tryptophan) supporting plastid functions are expanded compared to photosynthetic orchids, an indication of the importance of plastids even in totally mycoheterotrophic species. Conclusion Gastrodia menghaiensis has the smallest proteome thus far among angiosperms. Many of the genes involved biological interaction between Gatrodia and symbiotic microbionts are more numerous than in photosynthetic orchids.

This paper discusses four kinds of hole sealing technologies for gas drainage in soft coal seam, which are polyurethane hole sealing, cement mortar hole sealing, full-length screen pipe hole sealing and pipe direct connection technology. Through the observation of negative pressure loss, the sealing effect of four sealing technologies is analyzed, the polyurethane sealing effect is poor, the cement mortar, the full-length screen pipe combined with cement mortar and the direct connection technology of pipe are good; through the observation of gas concentration, the gas drainage effect of four sealing technologies is analyzed, the polyurethane sealing technology is the worst, and the direct connection technology of pipe is the worst. The gas drainage effect is the best; according to the sealing effect and gas drainage effect, the traditional polyurethane and cement mortar sealing technology cannot meet the gas drainage demand of the soft coal seam. In the case of slow drilling deformation, the full-length screen pipe sealing technology can be used. In the case of rapid drilling deformation, the direct connection of pipe sealing technology should be used.

Abstract Background Hearing loss (HL) is the most frequent sensory deficit in humans, HL has strong genetic heterogeneity. The genetic diagnosis of HL is very important to aid treatment decisions and to provide prognostic information and genetic counseling for the patient’s family. Methods We undertook pedigree analysis in 92 Chinese non-syndromic HL patients by targeted next-generation sequencing and Sanger sequencing. Results Among the 92 HL patients, 18 were assigned a molecular diagnosis with 33 different variants in 14 deafness genes. Eighteen of the variants in 12 deafness genes were novel. Variants in TMC1, CDH23, LOXHD1 and USH2A were each detected in two probands, and variants in POU3F4, OTOA, GPR98, GJB6, TRIOBP, SLC26A4, MYO15A, TNC, STRC and TMPRSS3 were each detected in one proband. Conclusion Our findings expand the spectrum of deafness gene variation, which will inform genetic diagnosis of deafness and add to the theoretical basis for the prevention of deafness.

We review achievements in the conservation of orchid diversity in China over the last 21 years. We provide updated information on orchid biodiversity and suggestions for orchid conservation in China. We outline national policies of biodiversity conservation, especially of orchid conservation, which provide general guidelines for orchid conservation in China. There are now approximately 1708 known species of Orchidaceae in 181 genera in China, including five new genera and 365 new species described over the last 21 years. The assessment of risk of extinction of all 1502 known native orchid species in China in 2013 indicated that 653 species were identified as threatened, 132 species were treated as data-deficient, and four species endemic to China were classified as extinct. Approximately 1100 species (ca. 65%) are protected in national nature reserves, and another ~66 species in provincial nature reserves. About 800 native orchid species have living collections in major botanical gardens. The pollination biology of 74 native orchid species and the genetic diversity and spatial genetic structure of 29 orchid species have been investigated at a local scale and/or across species distributions. The mycorrhizal fungal community composition has been investigated in many genera, such as Bletilla, Coelogyne, Cymbidium, Cypripedium, and Dendrobium. Approximately 292 species will be included in the list of national key protected wild plants this year. Two major tasks for near future include in situ conservation and monitoring population dynamics of endangered species.

ObjectivePeripheral atherosclerosis that accumulates in the extracranial carotid and lower extremity arteries can lead to significant morbidity and mortality. However, atherosclerotic disease progression is often not homogenous and is accelerated by diabetes. We previously observed increased phospholipid content in minimally (Min)-diseased arterial segments compared to maximally (Max)-diseased segments. Since Peroxisome Proliferator-Activated Receptor alpha (PPARα) is a key regulator of lipid metabolism, we hypothesized that it may have differential expression and signaling in Min vs. Max-diseased peripheral arterial segments.MethodsEighteen patients who underwent carotid endarterectomy (CEA), and 34 patients who underwent major lower extremity amputation were prospectively enrolled into a vascular tissue biobank. Min and Max-diseased segments were obtained in real-time from CEA plaque and amputated lower extremity arterial segments. mRNA and protein were isolated from specimens and the relative expression of ppara, and its downstream genes Acyl-CoA Oxidase 1 (acox1) and Carnitine Palmitoyltransferase 1A (cpt1a) were also evaluated. We evaluated gene expression and protein content relative to atherosclerotic disease severity and clinical diabetes status. Gene expression was also evaluated relative to Hemoglobin A1c and serum lipid profiles.ResultsIn CEA segments of patients with diabetes, we observed significantly higher ppara and acox1 gene expression (p < 0.01 and p < 0.001 respectively), and higher PPARα protein content (p < 0.05). Hemoglobin A1c significantly correlated with expression of ppara (R2 = 0.66, p < 0.001), acox1 (R2 = 0.31, p < 0.05), and cpt1a (R2 = 0.4, p < 0.05). There was no significant difference in gene expression between Min vs. Max-diseased CEA plaque segments. Conversely, in lower extremity arterial segments of patients with diabetes, we observed significantly lower ppara, acox1, and cpt1a expression (p < 0.05, p < 0.001, and p < 0.0001 respectively). Interestingly, CPT1A content was lower in arterial segments of patients with diabetes (p < 0.05). Hemoglobin A1c and HDL-cholesterol had negative correlations with ppara (R2 = 0.44, p < 0.05; R2 = 0.42, p < 0.05; respectively).ConclusionThis study demonstrates the significant differential expression of ppara and its immediate downstream genes in human carotid and lower extremity arteries relative to disease severity and diabetes. These findings highlight that mechanisms that influence atheroprogression in the carotid and lower extremities peripheral arteries are not homogenous and can be impacted by patient diabetes status and serum cholesterol profiles. Further elucidating these differential molecular mechanisms can help improve targeted therapy of atherosclerosis in different peripheral arterial beds.

Background and Aims: The Cancer Genome Atlas (TCGA) project has identified HER2 mutations or amplification in 7% of colon cancers. In addition to HER2 mutations, colon cancer patients also possess co-occurring mutations in genes such as APC. Here, we investigated the role of HER2 and APC mutations on the crypt-villus architecture of the intestinal epithelium, localization of secretory cells, and expression of intestinal stem cell markers. Methods: We generated a HER2 transgenic mouse (HER2V777L Tg) possessing an activating mutation commonly found in colorectal cancer patients, HER2V777L, using transcription activator-like effector nucleases–based gene editing technology. We expressed the HER2V777L transgene in mouse small intestine and colon using Lgr5-Cre and Villin-Cre recombinases. In addition, we analyzed Lgr5-Cre; APCmin; HER2V777L Tg mice by morphologic and gene expression assays on intestinal sections and organoids derived from the epithelium. Results: HER2V777L expression resulted in hypertrophic crypt formation with expanded zones of proliferation. Proximal intestinal villi showed increased abundance of multiple differentiated lineages including extensive intermediate cell differentiation, as evidenced by MUC2/MMP7 co-immunofluorescence and transmission electron microscopy. HER2V777L expression in the context of APC loss resulted in further enhancement and expansion of the proliferative crypt compartment. Conclusions: We established an epithelial intrinsic role for HER2V777L on enhanced cellular proliferation. Additionally, we determined that HER2 and APC mutations, when combined, promote enhanced proliferation of intestinal crypts.

Fagales are an order of woody plants and comprise more than 1,100 species, most of which produce economically important timbers, nuts, and fruits. Their nuclear and plastid genomes are well-sequenced and provided valuable resources to study their phylogeny, breeding, resistance, etc. However, little is known about the mitochondrial genomes (mitogenomes), which hinder a full understanding of their genome evolution. In this study, we assembled complete mitogenomes of 23 species, covering five of the seven families of Fagales. These mitogenomes had similar gene sets but varied 2.4 times in size. The mitochondrial genes were highly conserved, and their capacity in phylogeny was challenging. The mitogenomic structure was extremely dynamic, and synteny among species was poor. Further analyses of the Fagales mitogenomes revealed extremely mosaic characteristics, with horizontal transfer (HGT)-like sequences from almost all seed plant taxa and even mitoviruses. The largest mitogenome, Carpinus cordata, did not have large amounts of specific sequences but instead contained a high proportion of sequences homologous to other Fagales. Independent and unequal transfers of third-party DNA, including nuclear genome and other resources, may partially account for the HGT-like fragments and unbalanced size expansions observed in Fagales mitogenomes. Supporting this, a mitochondrial plasmid-like of nuclear origin was found in Carpinus. Overall, we deciphered the last genetic materials of Fagales, and our large-scale analyses provide new insights into plant mitogenome evolution and size variation.

Background: To investigate the expression of miR-451a in prostate cancer tissues and its effect on prognosis. Methods: Each of 78 specimens of prostate cancer tissues and corresponding adjacent normal tissues were collected from patients in Changshu Hospital Affiliated to Soochow University, Changshu, China from Apr 2014 to Jun 2015. Real-time quantitative RT-PCR (qRT-PCR) was used to detect the expression of miR-451a in tissues. The relationship between the expression of miR-451a and clinical pathological parameters was analyzed. The median expression of miR-451a in the experimental group was used to distinguish the high and low expressions of miR-451a in the experimental group. Kaplan-Meier was used to analyze the survival of miR-451a high and low expression groups. Results: The expressions of miR-451a in the patient's tissues and serum were decreased, and the correlation analysis found that they were positively correlated. ROC curve analysis showed that miR-451a had a high clinical value in the diagnosis of prostate cancer and the area under the curve was 0.921. The incidence of stage III+IV lymph node metastasis, Gleason score of >7 points and a serum Prostate-specific antigen (PSA) level of >20 ng/ml in patients of the low expression group increased significantly. The 5-yr survival rate of patients with low expression was significantly lower than that of those with high expression (P=0.005). MiR-451a was an independent factor affecting the prognosis of patients. Conclusion: miR-451a is lowly expressed in prostate cancer, and patients with low expression have a poor prognosis.

The COVID-19 pandemic has brought massive online activities and increased cybersecurity incidents and cybercrime. As a result of this, the cyber reputation of organisations has also received increased scrutiny and global attention. Due to increased cybercrime, reputation displaying a more important role within risk management frameworks both within public and private institutions is vital. This study identifies key factors in determining reputational damage to public and private sector institutions through cyberattacks. Researchers conducted an extensive review of the literature, which addresses factors relating to risk management of reputation post-cyber breach. The study identified 42 potential factors, which were then classified using the STAR model. This model is an organisational design framework and was suitable due to its alignment with organisations. A qualitative study using semi-structured and structured questions was conducted with purposively selected cybersecurity experts in both public and private sector institutions. Data obtained from the expert forum were analysed using thematic analysis, which revealed that a commonly accepted definition for cyber reputation was lacking despite the growing use of the term “online reputation”. In addition, the structured questions data were analysed using relative importance index rankings. The analysis results revealed significant factors in determining reputational damage due to cyberattacks, as well as highlighting reputation factor discrepancies between private and public institutions. Theoretically, this study contributes to the body of knowledge relating to cybersecurity of organisations. Practically, this research is expected to aid organisations to properly position themselves to meet cyber incidents and become more competitive in the post-COVID-19 era.

East Asia is one of the plant diversity and endemism centers in the world, and the temporal and spatial patterns and processes of vascular plants attracted ecologists’ and biogeographers’ attention. However, the biogeographic patterns of endemic epiphytic plants in East Asia are still unclear. Here, we investigated the historical biogeography of an East Asian endemic epiphytic genus Holcoglossum Schltr. (Orchidaceae). Using DNA sequences of eleven chloroplast genes and one nuclear gene, we reconstructed a robust phylogenetic framework for Holcoglossum and used a relaxed-clock method to estimate divergent times for the genus. We inferred the ancestral range of lineages under the statistical dispersal-extinction cladogenesis (S-DEC) and statistical dispersal-vicariance analysis (S-DIVA), respectively. Biogeographical analysis suggested that the most recent common ancestor of Holcoglossum occurred in the Palaeotropical region in the late Miocene (6.33 Ma). Four dispersal events were inferred to explain the Holcoglossum expansion to Sino-Himalayan, Sino-Japanese, and Taiwan regions from the latest Miocene to Quaternary. The episodes of these events were associated with intensification of East Asian monsoon around 3.6–2.6 Ma and global cooling since the latest Pliocene. The disjunct distribution between mainland China and Taiwan was attributed by the sea-level fluctuations and climate changes during the late Pliocene. This study shed light on the biogeographic processes of endemic epiphytic plants in East Asia.

Mycoheterotrophic plants are highly specialized species able to acquire organic carbon from symbiotic fungi, with relaxed dependence on photosynthesis for carbon fixation. The relaxation of the functional constraint of photosynthesis and thereby the relaxed selective pressure on functional photosynthetic genes usually lead to substantial gene loss and a highly degraded plastid genome in heterotrophs. In this study, we sequenced and analyzed the plastome of the eudicot Exacum paucisquama, providing the first plastid genome of a mycoheterotroph in the family Gentianaceae to date. The E. paucisquama plastome was 44,028 bp in length, which is much smaller than the plastomes of autotrophic eudicots. Although the E. paucisquama plastome had a quadripartite structure, a distinct boundary shift was observed in comparison with the plastomes of other eudicots. We detected extensive gene loss and only 21 putative functional genes (15 protein-coding genes, four rRNA genes and two tRNA genes). Our results provide valuable information for comparative evolutionary analyses of plastomes of heterotrophic species belonging to different phylogenetic groups.

Odontochilus putaoensis, a new species of Orchidaceae, is described and illustrated from Putao Township, Kachin State, Myanmar. Odontochilus putaoensis is close to O. duplex, but can be easily distinguished from the latter by having a light yellow lip, a bisaccate hypochile with a small, erect, blade-like and emarginate callus within each sac, a mesochile with a pair of dentate-pectinate flanges and a bilobed epichile with a pair of widely diverging lobes that are erect and concave. An identification key to the Southeast Asian species of Odontochilus and colour photographs of O. putaoensis are provided. A preliminary conservation assessment according to the IUCN Red List Categories and Criteria is given for the new species.

Gastrodia elata is an obligate mycoheterotrophic plant with highly reduced leaves and bracts in scape. Here, Yuan et al sequence and analyze its 1.06 Gb genome which provides insights in adaptation to a lifestyle of heterotrophy.

The determination of the geographic distributions of orchid species and their relationships with environmental factors are considered fundamental to their conservation. Paphiopedilum subgenus Brachypetalum is one of the most primitive, ornamental, and threatened groups of Orchidaceae. However, little is known about the distribution of Brachypetalum orchids and how they are influenced by environmental factors. In this study, we developed a database on the geographical distribution of Brachypetalum orchids based on a large-scale field investigation in the Guangxi, Guizhou, and Yunnan provinces of southwest China (2019–2020). Using this database, we first adopted the nonparametric Mann–Whitney U test to analyze the differences in the geographical distributions and growth environments of Brachypetalum orchids. In addition, we also used the method of principal component analysis (PCA) to explore distribution patterns of Brachypetalum orchids in relation to environmental factors (topography, climate, anthropogenic disturbance, productivity, and soil) in southwest China. Our results indicated that Brachypetalum orchid species were mainly distributed in the karst limestone habitats of southwest China. In general, there were 194 existing localities with the occurrence of seven target orchids in the investigated area. Of the discovered species in our study, 176 locations (~90.7%) were distributed primarily in the karst habitat. Among them, the range of 780–1267 m was the most concentrated elevation of Brachypetalum orchids. In addition, the findings also suggested that the distribution of Brachypetalum orchids in southwest China was relatively scattered in geographical space. However, the density of the distribution of Brachypetalum orchids was high, between 104° and 108° E and between 25° and 26° N. The results of the Mann–Whitney U test revealed that there are obviously different geographical distributions and growth environments of Brachypetalum in southwest China. More specifically, we found some extremely significant differences (p < 0.001) in elevation, mean diurnal range, precipitation of coldest quarter, solar radiation, and exchangeable Ca2+ between the provinces of southwest China. The PCA analysis revealed that elevation, solar radiation, temperature (mean diurnal range, annual temperature range) and precipitation (precipitation seasonality, precipitation of the warmest quarter) were found to be the most significant factors in determining Brachypetalum orchids’ distribution. These findings have implications in assessing conservation effectiveness and determining niche breadth to better protect the populations of these Brachypetalum orchid species in the future.

The growing interest in Knowledge Management (KM) has led to increased attention to Social Network Analysis (SNA) as a tool to map the relationships in networks. SNA can be used to evaluate knowledge flows between project teams, contributing to collaborative working and improved performance. Similarly, it has the potential to be used for construction projects and organisations. This paper aims at identifying current trends and future research directions related to using SNA for KM in construction. A systematic review and thematic analysis were used to critically review the existing studies and identify potential research areas in construction specifically related to research approaches and explore the possibilities for extension of SNA in KM. The findings revealed that there are knowledge gaps in research approaches with case study-based research involving external stakeholders, collaborations, development of communication protocols, which are priority areas identified for future research. SNA in KM related to construction could be extended to develop models that capture both formal and informal relationships as well as the KM process in pre-construction, construction, and post-construction stages to improve the performance of projects. Similarly, SNA can be integrated with methodological concepts, such as Analytic Hierarchy Process (AHP), knowledge broker, and so forth, to improve KM processes in construction. This study identifies potential research areas that provide the basis for stakeholders and academia to resolve current issues in the use of SNA for KM in construction.

Selecting a better performing contractor at the procurement stage is crucial in achieving a successful outcome for a construction project. The construction industry lacks a systematic and purpose driven method to assess performance of contractors using objective metrics. There are many approaches to measuring construction performance, but most are complicated and have high dependency on data that is difficult to attain. This paper aims to create a model for evaluating construction contractors’ performance based on directly attributable measures that are quantitative and easy to gather. This makes such a model more attractive and easier to use. Initially, a detailed literature review revealed different categories of measures of performance (MoP) and corresponding critical measures of performance (CMoP). Through a series of Delphi-based expert forums, the set of measures were fine-tuned and shortlisted. Fuzzy analytic hierarchy process-based comparisons were then used for developing a contractors’ performance model to quantify their level of performance based on a limited set of organisation-specific and project-specific measures. The results indicate a shift from traditional measures and a higher preference towards non-price measures. The performance model can be further developed to systematically rank the prospective contractors at the procurement stage based on seven non-price measures.

Coelogyne putaoensis, a new species of section Ocellatae from Putao, Kachin State, Myanmar, is described and illustrated. It is morphologically similar to C. taronensis and C. weixiensis, presumably its nearest relatives. An identification key and colour photographs are provided. A preliminary risk-of-extinction assessment according to the IUCN Red List categories and criteria is given for the new species.

This paper aims to clarify the effect of water on the desorption and release of coal seam gas (CSG). To this end, lab simulations were combined with field tests to disclose the changes in the amount and rate of gas desorption under different water saturation conditions. The results show that water had an obvious inhibitory effect on gas desorption. Specifically, when the equilibrium pressure and desorption time remained constant, the gas desorption is negatively correlated with the water content of the coal samples; when the water content reached a certain level, the desorption rate no longer decreased with the increase of water. Meanwhile, several water injection tests were carried out on the initial velocity q and desorption rate K1 of the CSG, trying to effectively prevent and control coal and gas outbursts. The tests results reveal that the values of both indices varied greatly with the increase of the water content, when the latter was between 4.5% and 5%. This means the two indices were highly sensitive to water content in this phase. The decrease of the values slowed down as the water content fell between 6% and 6.5%, and stabilized when the latter increased to 9%~10%.To sum up, water injection of coal seam, i.e. using the q value and K1 value as the indicators of outburst hazards, only partially mitigate rather than eliminate the potential threat of gas outburst.

Carbon emissions are categorised as Embodied Carbon (EC) occurring in the production phase and Operational Carbon (OC) occurring in the operational phase of buildings. The current focus on producing zero-carbon buildings, emphasises reducing OC and ignores the importance of reducing EC emissions. This study focuses on EC. Methods available in EC estimating currently produce estimates that often do not complement each other. This makes it important to develop a robust and accurate methodology for estimating EC. Blockchain is an emerging technology that has significant potential for transaction processing in supply chains. The construction industry being the second least digitalised industry, the adoption of innovative technologies is predominantly important. This paper explores the potential application of blockchain for accurate estimation of EC in construction supply chains. A detailed literature review and expert interviews revealed that, compared to traditional information systems, blockchain systems could eliminate issues in EC estimating highlighting its potential credible application for EC estimating. Scalability was identified as a feature that was lacking in a blockchain system, however, for EC estimating, its impact was identified as minimal. It will be difficult to generalise the findings of the study due to interview based qualitative methodology adopted in this study along with the fact that blockchain is an emerging and fairly new technology. However, a similar process could be followed by other studies to compare blockchain with traditional information systems, to evaluate the suitability of blockchain technology to develop prototype systems.

This paper aims to draw the whole picture of serious and extraordinarily serious (SES) work safety accidents in China. To this end, relevant data were collected from authentic sources like the accident query system of the State Administration of Coal Mine Safety and employed for statistical analysis of the national SES work safety accidents in China from 2005 to 2015. For better presentation, the occurrences and death tolls of SES work safety accidents were plotted for every province in Chinese mainland, and the accidents were classified into different categories by different standards. Then, the author discussed the regional, temporal and classification features of such accidents in great details. The results show that the SES work safety accidents in China during the study period are mainly coal mine accidents and traffic accidents; the leading type of coal mine accidents is gas explosion, and that of traffic accidents is highway accidents. In terms of regional distribution, Shanxi, Guizhou, Hunan, Henan and Yunnan had extremely high occurrence of SES work safety accidents. For coal mine accidents, Shanxi, Guizhou and Henan are the high-risk areas, and March, April, May and October are the peak months; For traffic accidents, Yunnan, Huizhou and Hunan are the high-risk areas, and February, March, April and August are the peak months; For fire accidents, Guangdong, Zhejiang and Jilin are the high-risk areas, and May, November and December are the peak months. The research findings provide a valuable guidance for the prediction and prevention of SES work safety accidents.

During the process of coal mining, the CH4 produced by the coal will be released in the workspace. With the movement of the working face, the overlying strata will be moved or destroyed, resulting in the continuous development of mining cracks and thus the forming a methane enriched area. The study of methane enrichment law is helpful for methane extraction and utilization, and at the same time reduces the risk of workspace. The development of the mining fracture field in overlying strata is determined by the process of overlying strata movement. Since there are many factors affecting the migration of overlying strata, the migration process of the rock strata is complex and the field is limited by many conditions, it is difficult to detect the distribution of the fractured field in the overlying strata. In this paper, the evolution law of mining fracture field will be studied by similar material model test. Based on the geological conditions of the 12# coal seam in Nanzhuang coal mine, Yangquan, the paper uses the similar model test to analyze and discuss the evolution law of the mining fracture field in the overlying strata of the coal seam, which provides a theoretical basis for the design of the mining scheme for the mining in the adjacent layer of the coal seam of the 12# coal seam.

Proper density and morphology of dendritic spines are important for higher brain functions such as learning and memory. However, our knowledge about molecular mechanisms that regulate the development and maintenance of dendritic spines is limited. We recently reported that cyclin-dependent kinase 5 (Cdk5) is required for the development and maintenance of dendritic spines of cortical neurons in the mouse brain. Previous in vitro studies have suggested the involvement of Cdk5 substrates in the formation of dendritic spines; however, their role in spine development has not been tested in vivo. Here, we demonstrate that Cdk5 phosphorylates collapsin response mediator protein 2 (CRMP2) in the dendritic spines of cultured hippocampal neurons and in vivo in the mouse brain. When we eliminated CRMP2 phosphorylation in CRMP2KI/KI mice, the densities of dendritic spines significantly decreased in hippocampal CA1 pyramidal neurons in the mouse brain. These results indicate that phosphorylation of CRMP2 by Cdk5 is important for dendritic spine development in cortical neurons in the mouse hippocampus.

DNA barcoding has been proposed to be one of the most promising tools for accurate and rapid identification of taxa. However, few publications have evaluated the efficiency of DNA barcoding for the large genera of flowering plants. Dendrobium, one of the largest genera of flowering plants, contains many species that are important in horticulture, medicine and biodiversity conservation. Besides, Dendrobium is a notoriously difficult group to identify. DNA barcoding was expected to be a supplementary means for species identification, conservation and future studies in Dendrobium. We assessed the power of 11 candidate barcodes on the basis of 1,698 accessions of 184 Dendrobium species obtained primarily from mainland Asia. Our results indicated that five single barcodes, i.e., ITS, ITS2, matK, rbcL and trnH-psbA, can be easily amplified and sequenced with the currently established primers. Four barcodes, ITS, ITS2, ITS+matK, and ITS2+matK, have distinct barcoding gaps. ITS+matK was the optimal barcode based on all evaluation methods. Furthermore, the efficiency of ITS+matK was verified in four other large genera including Ficus, Lysimachia, Paphiopedilum, and Pedicularis in this study. Therefore, we tentatively recommend the combination of ITS+matK as a core DNA barcode for large flowering plant genera.

Determining the generic delimitations within Aeridinae has been a significant issue in the taxonomy of Orchidaceae, and Holcoglossum is a typical case. We investigated the phylogenetic utility of the morphological traits of leaf epidermis in the taxonomy of Holcoglossum s.l. by using light and scanning electron microscopy to analyze 38 samples representing 12 species of Holcoglossum, with five species from five closely related genera, such as Ascocentrum, Luisia, Papilionanthe, Rhynchostylis and Vanda. Our results indicated that Holcoglossum can be distinguished from the related genera based on cuticular wax characteristics, and the inclusion of Holcoglossum himalaicum in Holcoglossum is supported by the epidermis characteristics found by LM and SEM. The percentage of the tetracytic, brachyparacytic, and laterocytic stomata types as well as the stomata index and certain combinations of special wax types support infrageneric clades and phylogenetic relationships that have been inferred from molecular data. Laterocytic and polarcytic stomata are perhaps ecological adaptations to the strong winds and ample rains in the alpine region of the Hengduanshan Mountains.

Interleukin-10 (IL-10) is a multifunctional cytokine which participates in the development and progression of various malignant tumors. To date, a number of case-control studies were conducted to detect the association between IL-10-592C>A polymorphism and cancer risk in humans. However, the results of these studies on the association remain conflicting. In an effort to solve this controversy, we performed a meta-analysis based on 70 case-control studies from 65 articles, including 16 785 cancer cases and 19 713 controls. We used odds ratios (ORs) with 95% confidence intervals (CIs) to assess the strength of the association. The overall results suggested that the variant homozygote genotype AA of the IL-10-592C>A polymorphism was associated with a moderately decreased risk of all cancer types (OR = 0.90, 95% CI = 0.83-0.98 for homozygote comparison, OR = 0.92, 95% CI = 0.86-0.98 for recessive model). In the stratified analyses, the risk remained for studies of smoking-related cancer, Asian populations and hospital-based studies. These results suggested that the IL-10-592C>A polymorphism might contribute to the cancer susceptibility, especially in smoking-related cancer, Asians and hospital-based studies. Further studies are needed to confirm the relationship.

Recently, there have been a lot of intense debates about the acceptance/rejection of paraphyletic groups in biological classification. On the one hand, evolutionary classification states that similarity and common descent are two criteria for biological classification and paraphyletic groups are natural units of biological classification. On the other hand, cladistic classification considers that common descent is the only criterion in biological classification and monophyly should be strictly adhered to. Holcoglossum is used herein as a case to illustrate this problem. Although Holcoglossum is a small orchid genus of less than 20 species, there is little consensus about its generic circumscription since it was established, which leads to confusion in taxonomic treatments in the Aerides-Vanda group. Based on the analyses of molecular and morphological evidence, our results suggest that the clade comprising Holcoglossum s.s., Ascolabium, Penkimia and Ascocentrum himalaicum is strongly supported as a monophyly, and that the three taxa are nested within different subclades of Holcoglossum s.s. Thus, it is reasonable to recognize a monophyletic circumscription of Holcoglossum, which is also well supported by some vegetative and floral characters. The Holcoglossum s.l. would facilitate a better understanding of pollinator-driven floral divergence and vegetative stasis than a paraphyletic and narrowly defined genus.