Research progress in the use of leeches for medical purposes

Leeches are invertebrates that have a long history of application in the development of human medicine in both the East and the West.This paper comprehensively analyzes and evaluates current research and the latest progress with regard to the application of leeches,their medical value,and their application prospects from various perspectives,so as to provide a reference for new viewpoints and directions for research on leeches.Modern research has revealed that leeches contain various bioactive components,which have pharmacological effects such as anticoagulation,antithrombosis,blood viscosity reduction,and anti-atherosclerosis.Leech therapy is an important treatment approach for venous congestion after microsurgery and is also an effective adjuvant treatment for diabetic feet,chronic pain,and tumors.Therefore,leeches are of importance for the research and development of new drugs,the restoration of blood supply after surgery,and the adjuvant treatment of diseases accompanied by blood blocking.In addition,leeches can also be used as model organisms for research in evolutionary biology and invertebrate neurophysiology as well as in neurophysiological,behavioral,and functional studies.

Forest characteristics and population structure of Glyptostrobus pensilis, a globally endangered relict species of southeastern China

The Chinese water pine Glyptostrobus pensilis is the sole surviving species of the genus Glyptostrobus. It is endemic to southern China, central Vietnam, and eastern Laos, and today it is nearly extinct in the wild. Forest community characteristics and population structure of G. pensilis in China have remained un-known up to now. We investigated six swamp forest stands and analyzed their forest community characteristics (i.e. vertical stratification, species composition, and diversity) and population structure, including the frequency distribution of DBH (diameter at breast height) and age-classes as found in Fujian Province, southeastern China. The vertical stratifications of all the forest stands were rather simple. The remaining wild specimens ranged from roughly 15 to some 357 years for an average of ca. 85 years, with only a few individuals less than 20 years old. Compared with the stands and populations of G. pensilis in Vietnam, the taxonomic compositions of the stands in the two regions were different, except for the dominant species-G. pensilis. The Shannon-Wiener index showed the overstory of each stand had much lower diversity (0.26 on average) in Fujian Province than that (1.97 on average) in Vietnam, whereas the diversity indices were about the same (around 2.41) for the understories in the two regions. Furthermore, we discovered 18 G. pensilis seedlings at the study sites in Fujian Province. This discovery demonstrates that G. pensilis regeneration is extremely poor and its populations are declining, although these populations are rela-tively healthier than those in Vietnam.

Photosynthetic traits and antioxidative defense responses of Pinus yunnanensis after joint attack by bark beetles Tomicus yunnanensis and T.minor

Bark beetles Tomicus yunnanensis and T.minor are two important pests of Pinus yunnanensis and can cause massive death of pine trees.In this study,we examined several traits related to photosynthesis in P.yunnanensis and their relationship with antibiotic defense responses after joint attack by the two bark beetles at the shoot and the trunk stages.When shoots were attacked by the beetles,the abundance of chlorophylls,carotenoids,and the rates of net photosynthesis(Pn)and transpiration(E)decreased in needles,while the levels of superoxide dismutase and malondialdehyde remained unchanged in both needles and phloem.The activity of peroxidases also remained unchanged in needles,but increased in phloem.The activity of catalases increased in both needles and phloem.When trunks were attacked by the bark beetles,chlorophyll abundance,Pn,E,and antioxidative enzyme activities all declined,and the declines were more pronounced than in the attacked shoots.A decrease in protein concentrations was also observed in needles and phloem from the attacked pines.Attack on shoots by the bark beetles suppressed host defense and provided a favorable environment for larval growth and development,resulting in long-term decline of pine growth potential.The results suggest that attacks on trunks by beetles caused more severe damage to host trees than attacks on shoots.

Trees as hotspots:Using forests,trees,and agroforestry to foster diverse sustainable landscapes

Forests,trees,and agroforestry(FTA)are ecosystem hotspots.They exemplify the contributions of biodiversity to sustainable and resilient landscapes,green circular economy and to sustainable agriculture and food systems for healthy diets.However,most research on these topics have been performed separately and lack comparison.The International FTA-Kunming Conference'Forests,trees and agroforestry for diverse sustainable landscapes'22nd–24th June 2021,focused on these contributions,brought together scientists NGOs,and policy makers to further the understanding of tree diversity;provided a communication platform for scientists to share their research results;evaluated the role of tree diversity in agroecology and circular agriculture;assessed benefits of landscape restoration;and explored applied research in mountain ecosystems and food security.The goals were to gather evidence that ground the design of solutions that can contribute to the implementation of the post 2020 Global Biodiversity Framework and towards the UN Food Systems Summit,and the overall implementation of the SDGs.This paper summarizes the outcomes of the international FTA Conference in Kunming 2021 and points out the highlights of research involved in six major themes.

Distribution and development strategy for Jatropha curcas L. in Yunnan Province, Southwest China

Yunnan Province is the main distributing area ofJatropha curcas L. This plant is abundant in several drainage areas of the dry-hot, dry-warm and sub-humid valleys in the south subtropical area of Yunnan Province. The seeds that were picked from trees blossoming between April and May and fructifying between September and October will have large seed yield and fine quality. For developing bio-diesel stock forest ofJ. curcas in areas with adaptive climate, seeding measures for afforestation should be taken and techniques on breeding, fast-growing, and high-yielding plantation cultivation are very important.

Epitypification:should we epitypify?

Epitypification can solve many taxonomic problems and stabilize the understanding of species, genera, families or orders. The aim of this paper is to illustrate how to epitypify. A few examples where taxa have been epitypified are considered and the benefits and disadvantages of epitypification are discussed. We also outline some examples of taxa which need to be epitypifled with reasons.

Protostropharia luteonitens(Basidiomycota,Agaricales):new to China

Protostropharia luteonitens is characterized by its pointedly papillate pileus,the large dark basidiospores,2-spored basidia and absence of chrysocystidia.It is known from North America,Europe and Asia(Japan).In this paper,P.luteonitens is reported for the first time from China.This species is presented here with a full description,colour photographs,and line drawings.

Edible insects in China： Utilization and prospects

The use of edible insects has a long history in China, where they have been consumed for more than 2000 years. In general, the level of acceptance is high for the consumption of insects in China. Many studies on edible insects have been conducted in the last 20 years, and the scope of the research includes the culture ofentomophagy and the identification, nutritional value, farming and breeding of edible insects, in addition to food production and safety. Currently, 324 species of insects from 11 orders are documented that are either edible or associated with entomophagy in China, which include the common edible species, some less commonly consumed species and some medicinal insects. How- ever, only approximately 10 to 20 types of insects are regularly consumed. The nutritional values for 174 species are available in China, including edible, feed and medicinal species. Although the nutritional values vary among species, all the insects examined contain pro- tein, fat, vitamins and minerals at levels that meet human nutritional requirements. Edible insects were, and continue to be, consumed by different ethnic groups in many parts of China. People directly consume insects or food products made from insects. The processing of products from insect protein powder, oil and chitin, and the development of healthcare foods has been studied in China. People also consume insects indirectly by eating livestock that were fed insects, which may be a more acceptable pathway to use insects in human diets. Although limited, the data on the food safety of insects indicate that insects are safe for food or feed. Incidences of allergic reactions after consuming silkworm pupae, cicadas and crickets have been reported in China. Insect farming is a unique breeding industry in rural China and is a source of income for local people. Insects are reared and bred for human food, medicine and animal feed using two approaches in China： the insects are either fully domesticated and reared completely in captivity or are partially raised in cap- tivity, and the insect habitat is manipulated to increase production. Depending on the type of relationship the insect has with humans, plants and the environment, different farming strategies are used. The social and scientific communities must work together to promote the use of insects as food and feed.

Complete sequence and gene organization of the mitochondrial genome of Batocera lineolata Chevrolat(Coleoptera:Cerambycidae)

Batocera lineolata Chevrolat,1852 is an important forest pest that is found mainly in China,Vietnam,India,and Japan.The complete mitochondrial genome(mitogenome) sequence of B.lineolata was determined by long polymerase chain reaction(PCR) and conserved primer walking approaches.The results showed that the entire mitogenome is 15418 bp long with 74.48% A+T content.The positions and arrangement of the 37 genes encoded by the genome are identical to the mitogenomes of two other longhorn beetles for which the complete gene content and arrangement are publicly available.All protein-coding genes start with the ATN codon that is a typical initiation codon in insects.All transfer RNAs(tRNAs) were predicted to form the standard clover-leaf structure,except for tRNA Ser(AGN),which lacks the dihydrouridine(DHU) arm.The most unusual feature that was found was the use of TCT as the tRNA Ser(AGN) anticodon instead of the GCT that is used in most other arthropods.The lack of tandem repeat motif in the 735 bp long A+T-rich region was another unusual feature of the B.lineolata mitogenome.The short,highly conserved polythymidine stretch that was previously described in the Orthoptera and Diptera orders was also present in the A+T-rich region of the B.lineolata(order Coleoptera) mitogenome.The sequence and annotation of the mitogenome of B.lineolata will provide further insights into the diversity and evolution of the Cerambycidae family of long-horned beetles.The B.lineolata mitogenome sequence has been deposited in GenBank(http://www.ncbi.nlm.nih.gov/Genbank/) under accession number JN986793.

Towards a natural classification of Botryosphaeriales

The type specimens of Auerswaldia,Auerswaldiella,Barriopsis,Botryosphaeria,Leptoguignardia,Melanops,Neodeightonia,Phaeobotryon,Phaeobotryosphaeria,Phyllachorella,Pyrenostigme,Saccharata,Sivanesania,Spencermartinsia and Vestergrenia were examined and fresh specimens of Botryosphaeriales were collected from Thailand.This material is used to provide a systematic treatment of Botryosphaeriales based on morphology and phylogeny.Two new genera,Botryobambusa and Cophinforma are introduced and comparedwith existing genera.Four species newto science,Auerswaldia dothiorella,A.lignicola,Botryosphaeria fusispora and Phaeobotryosphaeria eucalypti,are also described and justified.We accept 29 genera in Botryosphaeriales,with Macrovalsaria being newly placed.In the phylogenetic tree,the 114 strains of Botyrosphaeriales included in the analysis cluster into two major clades with 80%,96%and 1.00(MP,ML and BY)support,with Clade A containing the family type of Botryosphaeriaceae,and Clade B containing Phyllosticta,Saccharata and Melanops species.This group may represent Phyllostictaceae.In Clade A the taxa analyzed cluster in eight sub-clades(Clades A1-8).Clade A1 comprises three distinct subclusters corresponding to the genera Diplodia(Diplodia Clade),Neodeightonia(Neodeightonia Clade)and Lasiodiplodia(Lasiodiplodia Clade).Clade A2 clusters into three groups representing Phaeobotryosphaeria(100%),Phaeobotryon(100%)and Barriopsis(94%).Clade A3 incorporates 17 strains that cluster into three well-supported genera(Dothiorella(86%),Spencermartinsia(100%)and Auerswaldia(63%);the position of Macrophomina is not stable.Clade A4 is a single lineage(100%)representing the new genus Botryobambusa.Clade A5 is a wellsupported subclade incorporating Neofussicoccum.Clade A6 represents the type species of Botryosphaeria,three other Botryosphaeria species and two other genera,Neoscytalidium and Cophinforma gen.nov.Clade A7 comprises two Pseudofusicoccum species and Clade A8 has two Aplosporella species.These sub-clades may eventually require separate families but this requires analysis of a much larger dataset.Our data advances the understanding of Botryosphaeriales,there is,however,still much research to be carried out with resolution of families and genera,linkage of sexual and asexual morphs and differentiation of cryptic species.

Trichomeriaceae, a new sooty mould family of Chaetothyriales

Trichomerium is a genus of foliar epiphytes with the appearance of sooty moulds,mostly occurring on the surface of living leaves and apparently gaining their nutrients from insect exudates.Species have ascostromata with setae and develop on a loosely interwoven mycelial mass of dark brown hyphae,while asci have a bitunicate appearance with hyaline ascospores.In this study,we made 16 collections of Trichomerium from Thailand.All were isolated,and the LSU and ITS rDNA gene regions sequenced.Phylogenetic analysis indicated that the Trichomerium species form a monophyletic clade within Chaetothyriales and warrant the introduction of a new family Trichomeriaceae.Bootstrap support for the Chaetothyriales is 100%and clearly separates Trichomeriaceae from Capnodiales which are morphologically very similar.A detailed account of Trichomerium is provided and we describe and illustrate three new species based on morphological and molecular data.We propose that T.foliicola is adopted as the generic type of Trichomerium because it has been impossible to obtain the holotype specimen of T.coffeicola and also no molecular data exists in worldwide databases for this species or genus.

Taxonomy and the evolutionary history of Micropeltidaceae

Micropeltidaceae species are flyspeck fungi which have been subjected to few systematic studies.We re-examined 27 genera which were accepted in the Micropeltidaceae and re-described them based on herbaria materials and protologues.Based on morphology and phylogenetic investigations,we transfer Micropeltidaceae to a new order,Micropeltidales(Lecanoromycetes).Genera with bluish or greenish upper walls(Dictyopeltella,Dictyothyriella,Dictyothyrina,Dictyothyrium,Haplopeltheca,Micropeltis,Scolecopeltidium and Stomiopeltopsis)are accepted in the new taxonomic concept for Micropeltidaceae.A molecular clock approach estimated the divergence time of the Micropeltidaceae crown group at 130(165–104)Mya,which also supports its rank as an order(diverging from 220–100 Mya).The evolutionary histories between Micropeltidaceae species and host plants are interpreted by cophylogenetic analyses calibrated by their divergence times.The result indicates that the diversification of Angiospermae(130–80 Mya)fosters the formation of genera of Micropeltidaceae mainly via cospeciation events,and this codivergent period would be an important reference when establishing generic boundaries of epifoliar fungi.

Families of Dothideomycetes

Dothideomycetes comprise a highly diverse range of fungi characterized mainly by asci with two wall layers(bitunicate asci)and often with fissitunicate dehiscence.Many species are saprobes,with many asexual states comprising important plant pathogens.They are also endophytes,epiphytes,fungicolous,lichenized,or lichenicolous fungi.They occur in terrestrial,freshwater and marine habitats in almost every part of the world.We accept 105 families in Dothideomycetes with the new families Anteagloniaceae,Bambusicolaceae,Biatriosporaceae,Lichenoconiaceae,Muyocopronaceae,Paranectriellaceae,Roussoellaceae,Salsugineaceae,Seynesiopeltidaceae and Thyridariaceae introduced in this paper.Each family is provided with a description and notes,including asexual and asexual states,and if more than one genus is included,the type genus is also characterized.Each family is provided with at least one figure-plate,usually illustrating the type genus,a list of accepted genera,including asexual genera,and a key to these genera.A phylogenetic tree based on four gene combined analysis add support for 64 of the families and 22 orders,including the novel orders,Dyfrolomycetales,Lichenoconiales,Lichenotheliales,Monoblastiales,Natipusillales,Phaeotrichales and Strigulales.The paper is expected to provide a working document on Dothideomycetes which can be modified as new data comes to light.It is hoped that by illustrating types we provide stimulation and interest so that more work is carried out in this remarkable group of fungi.

Foldable Units and Wing Expansion of the Oakleaf Butterfly During Eclosion

Eclosion is a rapid process of morphological changes in insects,especially for the wings of butterflies.The orange oakleaf butterfly(Kallima inachus)transits from pupae to adults with a 9.3 fold instant increase in the surface area of their wings.To explore the mechanism for the rapid morphological changes in butterfly wings,we analyzed changes in microstructures in the wings of K.inachus.We found that there were lots of micron-sized foldable units in the wings at the pupal stage.The foldable units could provide as much as 31.35 times of increase in wing surface area.During eclosion,foldable units were flattened sequentially and resulted in a rapid increase in wing surface areas.The unfolding process was regulated by the structures and layouts of wing veins.Based on our observation,foldable units play important roles in both deformation and stretching of wings.The foldable units of microstructures may provide mimics for simulating entities of large-deformational bionic structures with practical application.

Refined families of Dothideomycetes:orders and families incertae sedis in Dothideomycetes

Numerous new taxa and classifications of Dothideomycetes have been published following the last monograph of families of Dothideomycetes in 2013.A recent publication by Honsanan et al.in 2020 expanded information of families in Dothideo-mycetidae and Pleosporomycetidae with modern classifications.In this paper,we provide a refined updated document on orders and families incertae sedis of Dothideomycetes.Each family is provided with an updated description,notes,including figures to represent the morphology,a list of accepted genera,and economic and ecological significances.We also provide phylogenetic trees for each order.In this study,31 orders which consist 50 families are assigned as orders incertae sedis in Dothideomycetes,and 41 families are treated as families incertae sedis due to lack of molecular or morphological evidence.The new order,Catinellales,and four new families,Catinellaceae,Morenoinaceae Neobuelliellaceae and Thyrinulaceae are introduced.Seven genera(Neobuelliella,Pseudomicrothyrium,Flagellostrigula,Swinscowia,Macroconstrictolumina,Pseudobogoriella,and Schummia)are introduced.Seven new species(Acrospermum urticae,Bogoriella complexoluminata,Dothiorella ostryae,Dyfrolomyces distoseptatus,Macroconstrictolumina megalateralis,Patellaria microspora,and Pseu-domicrothyrium thailandicum)are introduced base on morphology and phylogeny,together with two new records/reports and five new collections from different families.Ninety new combinations are also provided in this paper.

Effects of Land Uses on Soil Organic Carbon and Carbon Pool Management Index

Carbon management index（ICM） is used to evaluate the scientificity of soil management.Soil organic carbon（SOC） and readily oxidized carbon （ROC） contents under Leucaena leucocephala stand, Acacia glauca stand,Acacia auriculiformis stand, Azadirachta indica stand,wasteland and dry cropland were determined and their ICM were calculated.The results showed that the SOC and ROC contents under the 6 land use types were 4.22-5.92 g·kg-1 and 1.34-2.33 g·kg-1,respectively.No significant differences in SOC contents among these land uses were observed.The ROC contents under the 4 types of woodland,however,were significantly higher than these under dry cropland or wasteland（P【0.05）. The ROC were high significantly（P【0.001） correlated（R2 is 66.3%） with SOC.Compared with wasteland,the ICM ranged from 1.77 to 2.36 under these woodlands,and 0.99 under dry cropland.It is revealed that the litter quantity and land management under these land uses may be the key factors resulting in the variation of ROC.At ecological fragile region of Dry-hot Valley,closure management is useful to sequestrate C in woodland soils.However,the management systems for soil carbon pool under dry cropland are not in a sound way.

Relationship between Geographical Distribution of Pinus yunnanensis and Climate

A distribution map of Pinus yunnanensis was made according to the data on geographical distribution of P.yunnanensis by means of applying geographic information systems software ArcGIS, and the climatic parameters of the distribution areas for P.yunnanensis were determined by the climate forecast model Climate China.The relationship between the geographical distribution of P.yunnanensis and the climate was studied by adopting the indexes of Kira including the warmth index(WI),coldness index(CI) and aridity index (K),Xu’s humidity index(HI),Holdridge’s life temperature(BT),potential evapotranspiration (PET),potential evapotranspiration ratio(PER) and the single factors as the mean annual temperature, the mean temperature in January,the mean temperature in July,the mean annual precipitation, the】5℃accumulated temperature,the】18℃accumulated temperature,the maximum temperature, the minimum temperature,the temperature variation, the ratio of temperature and precipitation of the year, etc.And the Kira’s water-temperature indexes on distributional upper limit,low limit and north limit were discussed.The major climatic factors effecting the distribution of P.yunnanensis were screened out by statistical analysis software SPSS and the results showed the impact factors are in the order of temperature】humidity】the temperature difference between mean warmest month temperature and mean coldest month temperature.

Prophenoloxidase from Pieris rapae:gene cloning,activity,and transcription in response to venom/calyx fluid from the endoparasitoid wasp Cotesia glomerata

Prophenoloxidase (PPO) plays an important role in melanization, necessary for defense against intruding parasitoids. Parasitoids have evolved to inject maternal virulence factors into the host hemocoel to suppress hemolymph melanization for the successful development of their progeny. In this study, the full-length complementary DNA (cDNA) of a Pieris rapae PPO was cloned. Its cDNA contained a 2 076-base pair (bp) open reading frame (ORF) encoding 691 amino acids (aa). Two putative copper-binding sites, a proteolytic activation site, three conserved hemocyanin domains, and a thiol ester motif were found in the deduced amino acid sequence. According to both multiple alignment and phylogenetic analysis, P. rapae PPO gene cloned here is a member of the lepidopteran PPO-2 family. Injection of Cotesia glomerata venom or calyx fluid resulted in reduction of P. rapae hemolymph phenoloxidase activity, demonstrating the ability to inhibit the host's melanization. Real-time reverse transcriptase polymerase chain reaction (RT-PCR) showed that transcripts of P. rapae PPO-2 in the haemocytes from larvae had not significantly changed following venom injection, suggesting that the regulation of PPO messenger RNA (mRNA) expression by venom was not employed by C. glomerata to cause failure of melanization in parasitized host. While decreased P. rapae PPO-2 gene expression was observed in the haemocytes after calyx fluid injection, no detectable transcriptional change was induced by parasitization, indicating that transcriptional down-regulation of PPO by calyx fluid might play a minor role involved in inhibiting the host's melanization.

The MNS16A polymorphism in the TERT gene in peri-centenarians from the Han Chinese population

MNS16A, a variable number of tandem repeats polymorphism in the TERT gene, has been suggested to regulate telomerase activity. As telomerase activity has been reported to be related to life-span, we hypothesized that this polymorphism might affect human longevity by controlling the length of the telomere. To test this hypothesis, we collected 446 unrelated pericentenarian individuals （age）90, mean 94.45±3.45 years） and 332 normal controls （age 22-53, mean 35.0±12.0 years） from Dujiangyan, Sichuan, China. We typed the MNS16A polymorphism in both groups, and compared the allele and genotype frequencies between the peri-centenarian and control groups using the chi-squared test. There was no significant difference between the peri-centenarian and control groups. Thus, the MNS16A polymorphism in TERT might not influence human life-span, at least in the Hart Chinese population studied here.

Dothideales

The type specimens or representative specimens of the potentially dothidealean genera Bagnisiella,Botryochora,Coccostromella,Columnosphaeria,Delphinel la,Dictyodothis,Discosphaerina,Dothidea,Dothiora,Endodothiora,Jaffuela,Mycoporis,Omphalospora,Pachysacca,Plowrightia,Saccothecium,Stylodothis,Sydowia and Yoshinagaia were examined while,fresh specimens of Aureobasidium pullulans,Dothidea insculpta,Plowrightia ribesia and Saccothecium sepincola were made from Italy and Thailand.An introduction and the history of these genera,their family placement,morphology,and molecular phylogeny are provided.Morphology plus GenBank data are used to provide a systematic treatment of Dothideales.Phylogenetic analysis of LSU,SSU and ITS gene regions was carried out and in the resulting phylogenetic tree the taxa cluster in two clades with high bootstrap support.Clade A comprises Dothideaceae,the family type of Dothideales.The family Dothioraceae is not recognized as a distinct family and is synonymized under Dothideaceae.Neocylindroseptoria is introduced to accommodate Cylindroseptoria pistaciae as it forms a well-supported distinct clade in Dothideaceae.Clade B comprises Aureobasidium,Kabatiella,Pseudoseptoria,Saccothecium and Selenophoma species and Columnosphaeria fagi,for which we propose a new family,Aureobasidiaceae.The recently introduced Sydowia eucalypti also clustered within Clade B and therefore based on morphology and molecular phylogeny a new genus Pseudosydowia is introduced for Sydowia eucalypti.Celosporium laricicola is separated in a distinct clade,and therefore it is placed in Dothideales,genera,incertae sedis.The genera Bagnisiella,Botryochora,Coccostromella,Jaffuela,Lucidascocarpa,Mycoporis,Omphalospora,Pachysacca and Yoshinagaia are excluded from Dothideales and their placements are discussed.