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— Leucostegia has been long recognized as a small fern genus with only two species, L. immersa and L. pallida. Here we elevate a variety of L. immersa to species level and make the new combination Leucostegia amplissima based on morphological and molecular phylogenetic evidence. Morphologically, Leucostegia amplissima differs from L. immersa by having larger evergreen fronds, smaller indusia, and glandular hairs on the basal stipes. The study of Taiwanese populations further suggested an ecological divergence between L. immersa and L. amplissima.

— Two new fern species from Sarawak, Antrophyum hovenkampii and Pteris hovenkampii are described in memory of renowned pteridologist, Peter Hans Hovenkamp. Their relationships are inferred using morphological comparisons and molecular phylogenetic analyses. A description, photographs of key characters, distribution map, and habitat description of both new species are provided. Keys to all known species of Antrophyum and Pteris in Borneo are also included.

Five new combinations are made in Haplopteris C.Presl (Pteridaceae: Vittarioideae), involving four transfers from Vittaria Sm. and one from Monogramma Comm. ex Schkuhr. Lectotypes are designated for V. pluridichotoma Bonap., V. schaeferi Hieron. and V. stenophylla Copel. Additionally, H. stenophylla (Copel.) C.W.Chen & S.Linds. is recognised as distinct from H. mediosora (Hayata) X.C.Zhang based on rhizome scale morphology.

Microsoroideae is the third largest of the six subfamilies of Polypodiaceae, containing over 180 species. These ferns are widely distributed in the tropical and subtropical regions of the Old World and Oceania. We documented the spore ornamentation and integrated these data into the latest phylogenetic hypotheses, including a sampling of 100 taxa representing each of 17 major lineages of microsoroid ferns. This enabled us to reconstruct the ancestral states of the spore morphology. The results show verrucate ornamentation as an ancestral state for Goniophlebieae and Lecanoptereae, globular for Microsoreae, and rugulate surface for Lepisoreae. In addition, spore ornamentation can be used to distinguish certain clades of the microsoroid ferns. Among all five tribes, Lecanoptereae show most diversity in spore surface ornamentation. Electronic supplementary material The online version of this article (10.1007/s10265-020-01238-4) contains supplementary material, which is available to authorized users.

—A new species, Antrophyum nambanense, is described here based on the evidence from morphology and molecular phylogeny. Morphologically, short taeniform soral paraphyses and large fronds are the diagnostic characteristics of A. nambanense. Phylogenetically, it is most closely related to A. annamense, but can be readily distinguished from A. annamense by taeniform soral paraphyses (vs. filiform). We suggest A. nambanense is an allopolyploid of hybrid origin involving A. annamense and A. henryi, based on the evidence of morphology, phylogenetic position, chromosome number, and partially malformed spores. A description, photographs of key characters, distribution, conservation status, and ecology of the new species are presented. In addition, A. nambanense is also the first confirmed species with chlorophyllous spores in Antrophyum. Lastly, an updated key to all known Antrophyum species in Vietnam, including A. henryi as a new record for the country, is provided.

One genus (Chrysogrammitis) and twelve species (Asplenium pifongiae, Chrysogrammitis musgraviana, Crepidomanes makinoi, Ctenitis sinii, Ctenopterella khaoluangensis, Didymoglossum wallii, Dryopteris hendersonii, Hymenophyllum palmatifidum, H. paniculiflorum, Polystichum prionolepis, Pteris latipinna, and Stegnogramma wilfordii) of ferns are reported as new records for Vietnam based on newly collected specimens from Lang Biang Plateau. We provide photographs of key characters, type information, voucher specimens, distribution, ecology, and notes for each newly recorded species.

Four new species of Hymenasplenium (Aspleniaceae) from Africa and Asia are here described based on both morphological and molecular evidence. These four new species include H. guineense and H. kenyense from mainland Africa and H. sabahense and H. wusugongii from Asia. These new species have been erroneously treated as H. unilaterale by earlier pteridologists. The information on their distributions, habitats, and major distinguishing characters of these new species are provided.

—Two poorly known Malesian vittarioid ferns, Haplopteris angustissima and Monogramma capillaris, are noteworthy for their similar gross morphology. In this study, their systematic relationship was clarified by using molecular phylogenetic analysis which resolved the species in two different clades. Diagnostic morphological characters were further revealed by detailed comparison of almost all the known collections. Although overlaps were found in most of the investigated morphological characters, the venation and silica bodies were found to be useful for species delimitation. Type information, field images, illustrations, and taxonomic notes are provided for both species. A new combination, Haplopteris capillaris, is made and its occurrence in Taiwan as independent gametophytes is confirmed.

The infrageneric relationships and taxonomy of the largest fern genus, Asplenium (Aspleniaceae), have remained poorly understood. Previous studies have focused mainly on specific species complexes involving a few or dozens of species only, or have achieved a large taxon sampling but only one plastid marker was used. In the present study, DNA sequences from six plastid markers (atpB, rbcL, rps4, rps4-trnS, trnL and trnL-F) of 1030 accessions (616 of them newly sequenced here) representing c. 420 species of Asplenium (60% of estimated species diversity), 16 species of Hymenasplenium, three Diplaziopsidaceae, and four Rhachidosoraceae were used to produce the largest genus-level phylogeny yet for ferns. Our major results include: (i) Asplenium as broadly circumscribed is monophyletic based on our inclusion of representatives of 32 of 38 named segregate genera; (ii) 11 major clades in Asplenium are identified, and their relationships are mostly well-resolved and strongly supported; (iii) numerous species, unsampled in previous studies, suggest new relationships and numerous cryptic species and species complexes in Asplenium; and (iv) the accrued molecular evidence provides an essential foundation for further investigations of complex patterns of geographical diversification, speciation and reticulate evolution in this family.

In the process of undertaking a comprehensive review of the pteridophytes of the Solomon Islands, multiple unidentified specimens of the fern genus Ptisana Murdock (Marattiaceae) were collected. Morphological and molecular phylogenetic analyses as well as field observations were required to identify the Solomon Islands taxa. Four species and one variety are recognized from the Solomon Islands: Ptisana ambulans Murdock & C.W. Chen, sp. nov., Ptisana decipiens Murdock & C.W. Chen, sp. nov., Ptisana decipiens var. delicata Murdock & C.W. Chen, var. nov., Ptisana papuana (Alderw.) Murdock & C.W. Chen, comb. nov., and Ptisana smithii (Mett. ex Kuhn) Murdock. The complexities in the identification of Solomon Islands collections show the limits of morphology in the genus and illuminate a path forward for untangling the Ptisana taxonomy on a broader scale.

A new species was collected during a fern expedition in the Nuluhon Trusmadi Forest Reserve, Sabah, Borneo. Morphologically, it is similar to both Pteris decrescens Christ and P. parviloba Christ but can be distinguished by frond characters. Specifically, the new species differs from the other two species by having basiscopic secondary pinnae (vs. without basiscopic secondary pinnae), petiolulate pinnae (vs. sessile pinnae), without bristles on the stipes (vs. with bristles on the stipes), a crenate margin of the pinna-segment apex (vs. a serrate apex), and entire pseudoindusia (vs. ciliate pseudoindusia). Furthermore, P. decrescens and P. parviloba differ in the number of lateral pinnae (5 or 6 vs. 8–16 pairs) and the length of the basal pinna-segments being shortest or longest of all lateral pinnae. Two plastid genes, rbcL and matK, were used to decipher their phylogenetic relationships. The new species was resolved in different clades from P. decrescens and P. parviloba, providing further support of its uniqueness. The morphology, illustrations, ecology, and distribution of the new species are provided. We also found that the type materials of P. decrescens included two different species, P. decrescens and P. parviloba.A lectotype of P. decrescens was therefore designated. Based on the results of this study, P. decrescens and P. parviloba are re-circumscribed, and the new species is described as P. borneensis sp. nov.

The fern genus Tectaria (Tectariaceae) is morphologically diverse and difficult in terms of recognizing species and species groups. To infer the systematic positions of some species and identity-unknown collections with special morphological characters, we conducted phylogenetic analyses based on sequences of five plastid regions (atpB, ndhF plus ndhF-trnL, rbcL, rps16-matK plus matK, and trnL-F). Three analysis methods (maximum parsimony, maximum likelihood, and Bayesian inference) were used to reconstruct the phylogeny of Tectaria. The most surprising result is that T. menyanthidis, T. ternata, and T. variabilis are revealed to represent a distinct lineage from Tectaria, which should be called Polydictyum and is supported as sister to Pteridrys. Other accessions of Tectaria are well resolved into four major clades, which is consistent with the results of previous studies. Of the four clades, Clade II (T. subtriphylla group) is unpredictable, with morphologically very diverse species clustered there, and is supposed to be a minor evolutionary line within Tectaria in the Old World. In addition, the position of the climbing genus Arthropteris and the utility of molecular data in recognizing species of Tectaria are briefly discussed. As a conclusion we formally reinstate the genus Polydictyum by providing diagnostic characters, key to species, nomenclature, and information of detailed distribution and habit for the currently known three species.

The Old World fern genus Pyrrosia (Polypodiaceae) offers a rare system in ferns to study morphological evolution because almost all species of this genus are well studied for their morphology, anatomy, and spore features, and various hypotheses have been proposed in terms of the phylogeny and evolution in this genus. However, the molecular phylogeny of the genus lags behind. The monophyly of the genus has been uncertain and a modern phylogenetic study of the genus based on molecular data has been lacking. In the present study, DNA sequences of five plastid markers of 220 accessions of Polypodiaceae representing two species of Drymoglossum, 14 species of Platycerium, 50 species of Pyrrosia, and the only species of Saxiglossum (subfamily Platycerioideae), and 12 species of other Polypodiaceae representing the remaining four subfamilies are used to infer a phylogeny of the genus. Major results and conclusions of this study include: (1) Pyrrosia as currently circumscribed is paraphyletic in relation to Platycerium and can be divided into two genera: Pyrrosia s.s. and Hovenkampia (gen. nov.), with Hovenkampia and Platycerium forming a strongly supported clade sister to Pyrrosia s.s.; (2) Subfamily Platycerioideae should contain three genera only, Hovenkampia, Platycerium, and Pyrrosia s.s.; (3) Based on the molecular phylogeny, macromorphology, anatomical features, and spore morphology, four major clades in the genus are identified and three of the four are further resolved into four, four, and six subclades, respectively; (4) Three species, P. angustissima, P. foveolata, and P. mannii, not assigned to any groups by Hovenkamp (1986) because of their unusual morphology, each form monospecific clades; (5) Drymoglossum is not monophyletic and those species previously assigned to this genus are resolved in two different subclades; (6) Saxiglossum is resolved as the first lineage in the Niphopsis clade; and (7) The evolution of ten major morphological characters in the subfamily is inferred based on the phylogeny and various morphological synapomorphies for various clades and subclades are identified.

The monotypic fern genus Dryopolystichum Copel. combines a unique assortment of characters that obscures its relationship to other ferns. Its thin-walled sporangium with a vertical and interrupted annulus, round sorus with peltate indusium, and petiole with several vascular bundles place it in suborder Polypodiineae, but more precise placement has eluded previous authors. Here we investigate its phylogenetic position using three plastid DNA markers, rbcL, rps4-trnS, and trnL-F, and a broad sampling of Polypodiineae. We also provide new data on Dryopolystichum including spore number counts, reproductive mode, spore SEM images, and chromosome counts. Our maximum-likelihood and Bayesian-inference phylogenetic analyses unambiguously place Dryopolystichum within Lomariopsidaceae, a position not previously suggested. Dryopolystichum was resolved as sister to a clade comprising Dracoglossum and Lomariopsis, with Cyclopeltis as sister to these, but clade support is not robust. All examined sporangia of Dryopolystichum produced 32 spores, and the chromosome number of sporophyte somatic cells is ca. 164. Flow cytometric results indicated that the genome size in the spore nuclei is approximately half the size of those from sporophyte leaf tissues, suggesting that Dryopolystichum reproduces sexually. Our findings render Lomariopsidaceae as one of the most morphologically heterogeneous fern families. A recircumscription is provided for both Lomariopsidaceae and Dryopolystichum, and selected characters are briefly discussed considering the newly generated data.

Davalliaceae have been classified on the basis of morphological data or morphological and molecular data. The family has been divided into various genera, the circumscription of which may differ be ...

The phylogenetic affinities of the fern genus Aenigmopteris have been the subject of considerable disagreement, but until now, no molecular data were available from the genus. Based on the analysis of three chloroplast DNA regions (rbcL, rps16-matK, and trnL-F) we demonstrate that Aenigmopteris dubia (the type species of the genus) and A. elegans are closely related and deeply imbedded in Tectaria. The other three species of genus are morphologically very similar; we therefore transfer all five known species into Tectaria. Detailed morphological comparison further shows that previously proposed diagnostic characters of Aenigmopteris fall within the range of variation of a broadly circumscribed Tectaria.

A vittarioid fern from the Solomon Islands, which has long been treated as Antrophyum semicostatum, is described here as a new species, Antrophyum solomonense. A description, illustration, and distribution of the new species are presented. Boot-shaped paraphysal apical cells, rounded stipe in cross section, and thin rhizome scale cell walls are the diagnostic characteristics of A. solomonense. A molecular phylogeny inferred from four chloroplast markers further supports its systematic uniqueness.

A vittarioid fern (Pteridaceae) distributed in Eastern Asia is described here as a new species, Haplopteris heterophylla, which has long been included in H. hainanensis. A description, illustration, distribution, ecology, and a key to congeneric relatives of the new species are provided. Obovoid-shaped paraphyses and frond dimorphism are the diagnostic characteristics of H. heterophylla. In a trnL-F + matK + ndhF phylogeny of 14 Asian Haplopteris species, including H. hainanensis, both morphological observations and the cpDNA phylogeny support the systematic uniqueness of H. heterophylla.

In kidney collecting duct cells, filamentous actin (F-actin) depolymerization is a critical step in vasopressin-induced trafficking of aquaporin-2 to the apical plasma membrane. However, the molecular components of this response are largely unknown. Using stable isotope-based quantitative protein mass spectrometry and surface biotinylation, we identified 100 proteins that showed significant abundance changes in the apical plasma membrane of mouse cortical collecting duct cells in response to vasopressin. Fourteen of these proteins are involved in actin cytoskeleton regulation, including actin itself, 10 actin-associated proteins, and 3 regulatory proteins. Identified were two integral membrane proteins (Clmn, Nckap1) and one actin-binding protein (Mpp5) that link F-actin to the plasma membrane, five F-actin end-binding proteins (Arpc2, Arpc4, Gsn, Scin, and Capzb) involved in F-actin reorganization, and two actin adaptor proteins (Dbn1, Lasp1) that regulate actin cytoskeleton organization. There were also protease (Capn1), protein kinase (Cdc42bpb), and Rho guanine nucleotide exchange factor 2 (Arhgef2) that mediate signal-induced F-actin changes. Based on these findings, we devised a live-cell imaging method to observe vasopressin-induced F-actin dynamics in polarized mouse cortical collecting duct cells. In response to vasopressin, F-actin gradually disappeared near the center of the apical plasma membrane while consolidating laterally near the tight junction. This F-actin peripheralization was blocked by calcium ion chelation. Vasopressin-induced apical aquaporin-2 trafficking and forskolin-induced water permeability increase were blocked by F-actin disruption. In conclusion, we identified a vasopressin-regulated actin network potentially responsible for vasopressin-induced apical F-actin dynamics that could explain regulation of apical aquaporin-2 trafficking and water permeability increase.

BACKGROUND AND AIMS The gametophyte phase of ferns plays an important role in habitat selection, dispersal, adaptation and evolution. However, ecological studies on fern gametophytes have been impeded due to the difficulty of species identification of free-living gametophytes. DNA barcoding provides an alternative approach to identifying fern gametophytes but is rarely applied to field studies. In this study, an example of field vittarioid gametophyte identification using DNA barcoding, which has not been done before, is given. METHODS A combination of distance-based and tree-based approaches was performed to evaluate the discriminating power of three candidate barcodes (matK, rbcL and trnL-F) on 16 vittarioid sporophytes. Sequences of the trnL-F region were generated from 15 fern gametophyte populations by tissue-direct PCR and were compared against the sporophyte dataset, using BLAST. KEY RESULTS trnL-F earns highest primer universality and discriminatory ability scores, whereas PCR success rates were very low for matK and rbcL regions (10·8 % and 41·3 %, respectively). BLAST analyses showed that all the sampled field gametophytes could be successfully identified to species level. Three gametophyte populations were also discovered to be living beyond the known occurrence of their sporophyte counterparts. CONCLUSIONS This study demonstrates that DNA barcoding (i.e. reference databasing, tissue-direct PCR and molecular analysis), especially the trnL-F region, is an efficient tool to identify field gametophytes, and has considerable potential in exploring the ecology of fern gametophytes.

Independent gametophyte ferns are unique among vascular plants because they are sporophyteless and reproduce asexually to maintain their populations in the gametophyte generation. Such ferns had been primarily discovered in temperate zone, and usually hypothesized with (sub)tropical origins and subsequent extinction of sporophyte due to climate change during glaciations. Presumably, independent fern gametophytes are unlikely to be distributed in tropics and subtropics because of relatively stable climates which are less affected by glaciations. Nonetheless, the current study presents cases of two independent gametophyte fern species in subtropic East Asia. In this study, we applied plastid DNA sequences (trnL-L-F and matK + ndhF + chlL datasets) and comprehensive sampling (~80%) of congeneric species for molecular identification and divergence time estimation of these independent fern gametophytes. The two independent gametophyte ferns were found belonging to genus Haplopteris (vittarioids, Pteridaceae) and no genetic identical sporophyte species in East Asia. For one species, divergence times between its populations imply recent oversea dispersal(s) by spores occurred during Pleistocene. By examining their ex situ and in situ fertility, prezygotic sterility was found in these two Haplopteris, in which gametangia were not or very seldom observed, and this prezygotic sterility might attribute to their lacks of functional sporophytes. Our field observation and survey on their habitats suggest microhabitat conditions might attribute to this prezygotic sterility. These findings point to consideration of whether recent climate change during the Pleistocene glaciation resulted in ecophysiological maladaptation of non-temperate independent gametophyte ferns. In addition, we provided a new definition to classify fern gametophyte independences at the population level. We expect that continued investigations into tropical and subtropical fern gametophyte floras will further illustrate the biogeographic significance of non-temperate fern gametophyte independence.

A new combination, Pyrrosia annamensis (Christ) Li Bing Zhang, X.M.Zhou & C.W.Chen, for a Southeast Asian species formerly treated in Niphobolus is made. We also lectotypify Cyclophorus rhomboidalis Bonaparte.

We examined the global historical biogeography of grammitid ferns (Polypodiaceae) within a phylogenetic context. We inferred phylogenetic relationships of 190 species representing 31 of the 33 currently recognized genera of grammitid ferns by analyzing DNA sequence variation of five plastid DNA regions. We estimated the ages of cladogenetic events on an inferred phylogeny using secondary fossil calibration points. Historical biogeographical patterns were inferred via ancestral area reconstruction. Our results supported four large-scale phylogenetic and biogeographic patterns: (1) a monophyletic grammitid clade that arose among Neotropical polypod ancestors about 31.4 Ma; (2) a paraphyletic assemblage of clades distributed in the Neotropics and the Afro-Malagasy region; (3) a large clade distributed throughout the Asia–Malesia–Pacific region that originated about 23.4 Ma; and, (4) an Australian or New Zealand origin of the circumaustral genus Notogrammitis. Most genera were supported as monophyletic except for Grammitis, Oreogrammitis, Radiogrammitis, and Zygophlebia. Grammitid ferns are a well-supported monophyletic group with two biogeographically distinct lineages: a primarily Neotropical grade exhibiting several independent successful colonizations to the Afro-Malagasy region and a primarily Paleotropical clade exhibiting multiple independent dispersals to remote Pacific islands and temperate, austral regions.

Cheng-Wei Chen,1 Ming-Kung Yeh,2 Chia-Yang Shiau,3 Chiao-Hsi Chiang,4,* Da-Wen Lu5,*1Chengwei Biotechnology Co, Ltd, 2Bureau of Pharmaceutical Affairs, Military of National Defense Medical Affairs Bureau, 3Graduate Institute of Medical Sciences, 4School of Pharmacy, National Defense Medical Center, 5Department of Ophthalmology, Tri-Service General Hospital, Taipei, Taiwan *These authors contributed equally to this workBackground: The purpose of this study was to demonstrate the effectiveness of an integrin peptide ligand-labeled liposomal delivery system loaded with vascular endothelial growth factor (VEGF)-siRNA in a model study of gene therapy for retinopathy using human retinal pigment epithelial cells.Methods: Arg(R)-Gly(G)-Asp(D) motif peptide conjugating polyethylene glycol modified (RGD-PEGylated) liposomes were prepared using a thin-film hydration method and optimized for surface charge, particle size, small interfering RNA (siRNA) load, and entrapment efficiency. Reverse transcriptase-polymerase chain reaction and enzyme-linked immunosorbent assays were used to determine VEGF levels in retinal pigment epithelial cells. Cytotoxicity was determined using the 3-[4, 5-dimethylthiazol-2-yl]-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium (MTS) assay and flow cytometry.Results: Physicochemical properties, including particle size, zeta potential, and siRNA load, of the prepared RGD-PEGylated liposomes and their entrapment efficiency were determined to be within the following ranges: 123.8–234.1 nm, 17.31–40.09 mV, 5.27%–6.33%, and >97%, respectively. RGD-PEGylated liposome-mediated fluorescent-labeled siRNA delivery demonstrated significantly enhanced cellular uptake, and 3 mol% RGD-PEGylated liposomes (having 3β-[N-(N´, N´-dimethylaminoethane) carbamoyl] cholesterol (DC-cholesterol) DSPE and DSPE-PEG(2000)-RGD with molar ratio of 50/47/3) were shown to have better efficacy with regard to specificity for retinal pigment epithelial cells, reduced cytotoxicity, and knockdown of the target molecule.Conclusion: By integrin receptor-mediated endocytosis, 3 mol% RGD-PEGylated liposomes were shown to be a suitable vector when loaded with VEGF-siRNA for efficient downregulation of VEGF in retinal pigment epithelial cells at both the protein and gene levels. This integrin ligand-modified liposomal delivery system has therapeutic potential for ocular gene therapy.Keywords: vascular endothelial growth factor, siRNA delivery, liposome, retinal pigment epithelial cellsA Letter to the Editor has been received and published for this article.

PREMISE OF THE STUDY Primers were designed for amplifying intron 1 of the single-copy nuclear LEAFY gene for species of Davalliaceae. METHODS AND RESULTS New primer sets were designed and successfully amplified for intron 1 of the LEAFY gene in 13 species representing the five genera of Davalliaceae. The orthology of these sequences was further confirmed by phylogenetic analyses. Site variation in LEAFY intron 1 sequences across genera of the Davalliaceae and among accessions of the Humata repens complex were 18% and 8%, respectively. Such variation was greater than that for the cpDNA atpB-rbcL intergenic spacer region across the same taxa and accessions. CONCLUSIONS Using our newly designed primers, intron 1 of the LEAFY gene could be amplified for all species tested. In addition, this single-copy, biparentally inherited, and quickly evolving region showed considerable potential for addressing infraspecific-level questions.

Study on small animal model using multiple imaging is an important procedure in medicine and physiology. Recently, Single Photon Emission Computed Tomography (SPECT) of dopamine transporter with 99cmTc-TRODA-1 has been proved to be a valuable and feasible mean of assessing the integrity of dopamine neurons. In this paper, an image registration algorithm for MRI and SPECT is proposed to evaluate the changes of a Parkinson’s rat in brain organization and function. In addition, an automatic labeling algorithm which highlights striatum is necessary because manually segmenting these tissues from MRI is usually time consuming. As a result, we have built a computer-aid clinical diagnosis system which automates all the processes. This paper provides effective evaluation of Parkinson’s symptoms by calculating the values of binding potentials for dopamine for quantification. Experimental result shows that the performance of the proposed method is better than the previous approach based on automated image registration.

Three new species (Calymmodon concinnus, Oreogrammitis parvula, Xiphopterella parva), and seven new records (Dasygrammitis brevivenosa, Oreogrammitis sinohirtella, Radiogrammitis beddomeana, R. jagoriana, Scleroglossum pyxidatum, S. sulcatum and Themelium halconense) of grammitid ferns (Polypodiaceae) are reported for the flora of Vietnam, mostly from newly collected specimens. We provide detailed morphological descriptions, illustrations, ecology and distribution, and taxonomic notes for each new species. Furthermore, type information, distribution, voucher specimens and taxonomic notes are provided for each newly recorded species. A checklist including synonymy and a key to Vietnamese grammitid ferns are also given.

AIM: To evaluate the efficacy and safety of capsule oxymatrine in the treatment of chronic hepatitis B. METHODS: A randomised double-blind and placebo-controlled multicenter trial was conducted. Injection of oxymatrine was used as positive-control drug. A total of 216 patients with chronic hepatitis B entered the study for 24 wk, of them 108 received capsule oxymatrine, 36 received injection of oxymatrine, and 72 received placebo. After and before the treatment, clinical symptoms, liver function, serum hepatitis B virus markers, and adverse drug reaction were observed. RESULTS: Among the 216 patients, six were dropped off, and 11 inconsistent with the standard were excluded. Therefore, the efficacy and safety of oxymatrine in patients were analysed. In the capsule treated patients, 76.47% became normal in ALT level, 38.61% and 31.91% became negative both in HBV DNA and in HBeAg. In the injection treated patients, 83.33% became normal in ALT level, 43.33% and 39.29% became negative both in HBV DNA and in HBeAg. In the placebo treated patients, 40.00% became normal in ALT level, 7.46% and 6.45% became negative both in HBV DNA and in HBeAg. The rates of complete response and partial response were 24.51% and 57.84% in the capsule treated patients, and 33.33% and 50.00% in the injection treated patients, and 2.99% and 41.79% in the placebo treated patients, respectively. There was no significance between the two groups of patients, but both were significantly higher than the placebo. The adverse drug reaction rates of the capsule, injection and placebo were 7.77%, 6.67% and 8.82%, respectively. There was no statistically significant difference among them. CONCLUSION: Oxymatrine is an effective and safe agent for the treatment of chronic hepatitis B.

A new species of vittarioid ferns (Pteridaceae) that has long been regarded as Haplopteris amboinensis or H. forrestiana (a synonym of H. doniana ), is described here as Haplopteris yakushimensis . Both macro- and micromorphological characteristics were compared with these two species. Chartaceous matured fronds, shallowly grooved adaxial costae and obtuse carinate abaxial costae are the diagnostic characteristics that distinguish H. yakushimensis from the other species in the genus. A phylogeny based on plastid regions chlL , ndhF and matK of 14 East-Asian Haplopteris species, including our new taxon is presented here .

We describe here a strategy for preparing a human membrane and secreted protein (MSP)-enriched cDNA library based on human MSP- and non-MSP-encoding cDNA sequences in the databases. The signal peptide parts of the MSP-encoding cDNA sequences, which currently comprise about half of the estimated total number in humans, were analyzed for common patterns. These patterns form a ‘minimal’ set of polymerase chain reaction primer candidates of length varying from 9 to 21 nt. The products stemming from each primer candidate were determined and the results allowed us to obtain an ‘optimal’ mixed-length primer set. Ninety-six percent of the primers in this set were predicted to yield ≤10% undesired products, and the desired MSP-cDNA products could be easily separated by gel electrophoresis. The present analysis establishes a methodology for preparing a cDNA library that enables the analysis of individual MSPs. This methodology may also help identify new MSPs. As many cell regulatory processes are mediated by secreted proteins and their membrane-bound receptors, the preparation of a MSP-enriched cDNA library should benefit research on MSPs.

Background Human retinal pigment epithelial cells are promising target sites for small interfering RNA (siRNA) that might be used for the prevention and/or treatment of choroidal neovascularization by inhibiting the expression of angiogenic factor; for example, by downregulating expression of the vascular endothelial growth factor gene. Methods A novel functional lipid, DSPE-PEG-RGD, a Arg(R)-Gly(G)-Asp(D) motif peptide conjugated to 1, 2-distearoyl-sn-glycero-3-phosphoethanolamine- N-[maleimide (polyethylene glycol)-2000], was synthesized for the preparation of siRNA-loaded RGD-PEGylated liposomes to enhance uptake of encapsulated siRNA in retinal pigment epithelial cells. Various liposomes, with 1 mol% and 5 mol% PEGylated lipid or 1 mol% and 5 mol% RGD-PEGylated lipid, were fabricated. Results Characterization of the liposomes, including siRNA entrapment efficiency, average particle size and ζ-potential, were determined to be as follows: >96%, 129.7 ± 51 to 230.7 ± 60.7 nm, and 17.3 ± 0.6 to 32 ± 1.3 mV, respectively. For the in vitro retinal pigment epithelial cell studies, the RGD-PEGylated liposomes had high delivery efficiency with siRNA delivery, about a four-fold increase compared with the PEGylated liposomes. Comparison of the various liposomes showed that the 1 mol% RGD-modified liposome had less cytotoxicity and higher siRNA delivery efficiency than the other liposomes. The antibody blocking assay confirmed that uptake of the 1 mol% RGD-PEGylated liposome was via integrin receptor- mediated endocytosis in retinal pigment epithelial cells. Conclusion The results of this study suggest that RGD-PEGylated liposomes might be useful for siRNA delivery into retinal pigment epithelial cells by integrin receptor-medicated endocytosis., Video abstract Video