Genetic diversities in wild and cultivated populations of the two closely-related medical plants species, Tripterygium Wilfordii and T. Hypoglaucum (Celastraceae).

BACKGROUND: The sustainable supply of medicinal plants is important, and cultivating and domesticating them has been suggested as an optimal strategy. However, this can lead to a loss of genetic diversity. Tripterygium wilfordii Hook. f. is a medicinal plant commonly used in traditional Chinese medicine, but its wild populations are dwindling due to excessive harvesting. To protect the species and meet the increasing demand, it is urgent to cultivate it on a large scale. However, distinguishing between T. wilfordii and T. hypoglaucum, two similar species with different medicinal properties, is challenging. Therefore, it is crucial to understand the genetic diversity and population structure of these species for their sustainable utilization. RESULTS: In this study, we investigated the genetic diversity and population structure of the two traditional medicinal semiwoody vines plant species, Tripterygium wilfordii and T. hypoglaucum, including wild and cultivated populations using chloroplast DNA (cpDNA) sequences and microsatellite loci. Our results indicated that the two species maintain a high level of genetic divergence, indicating possible genetic bases for the different contents of bioactive compounds of the two species. T. wilfordii showed lower genetic diversity and less subdivided population structures of both markers than T. hypoglaucum. The potential factors in shaping these interesting differences might be differentiated pollen-to-seed migration rates, interbreeding, and history of population divergence. Analyses of cpDNA and microsatellite loci supported that the two species are genetically distinct entities. In addition, a significant reduction of genetic diversity was observed for cultivated populations of the two species, which mainly resulted from the small initial population size and propagated vegetative practice during their cultivation. CONCLUSION: Our findings indicate significant genetic divergence between T. wilfordii and T. hypoglaucum. The genetic diversity and population structure analyses provide important insights into the sustainable cultivation and utilization of these medicinal plants. Accurate identification and conservation efforts are necessary for both species to ensure the safety and effectiveness of crude drug use. Our study also highlighted the importance of combined analyses of different DNA markers in addressing population genetics of medicinal plants because of the contrasts of inheritance and rates of gene flow. Large-scale cultivation programs should consider preserving genetic diversity to enhance the long-term sustainability of T. wilfordii and T. hypoglaucum. Our study proposed that some populations showed higher genetic diversity and distinctness, which can be considered with priority for conservation and as the sources for future breeding and genetic improvement.

Two new species of Rhizoplaca (Lecanoraceae) from Southwest China.

In this study, two new species, Rhizoplacaadpressa Y. Y. Zhang & Li S. Wang and R.auriculata Y. Y. Zhang, Li S. Wang & Printzen, are described from Southwest China, based on their morphology, phylogeny and chemistry. In phylogeny, the two new species are monophyletic, and sister to each other within Rhizoplacachrysoleuca-complex. Rhizoplacaadpressa is characterized by its placodioid and closely adnate thallus, pale green and heavily pruinose upper surface, narrow (ca. 1 mm) and white free margin on the lower surface of marginal squamules, the absence of a lower cortex, and its basally non-constricted apothecia with orange discs that turn reddish-brown at maturity. Rhizoplacaauriculata is characterized by its squamulose to placodioid thallus, yellowish green and marginally pruinose squamules, wide (1-3 mm) and bluish-black free margin on the lower surface of marginal squamules, the absence of a lower cortex, and its basally constricted apothecia with persistently orange discs. Rhizoplacaadpressa and R.auriculata share the same secondary metabolites of usnic and placodiolic acids.

The phylogeny and taxonomy of Upretia (Caloplacoideae, Teloschistaceae), reveal three new species from Southwestern China.

Several specimens of Upretia from Southwest China are morphologically and phylogenetically distinct from currently recognized species in the genus. These specimens are here accommodated within a new species, Upretiazeorina Li J. Li & Printzen. It is characterized by an areolate to squamulose thallus with brown to blackish brown upper surface, pruinose, zeorine type apothecia, black discs, narrowly bacilliform conidia, and the production of gyrophoric acid. Two other specimens of Upretia from China are distinct from currently accepted species and tentatively referred to as Upretia sp. 1 and Upretia sp. 2. A key to all known species of Upretia is also provided.

The phylogeny and taxonomy of Glypholecia (Acarosporaceae, lichenized Ascomycota), including a new species from northwestern China.

Glypholeciaqinghaiensis An C. Yin, Q. Y. Zhong & Li S. Wang is described as new to science. It is characterized by its squamulose thallus, compound apothecia, ellipsoid ascospores, and the presence of rhizines on the lower surface of the thallus. A phylogenetic tree of Glypholecia species was constructed based on nrITS and mtSSU sequences. Two species G.qinghaiensis and G.scabra are confirmed in China.

The Expression of Parthanatos Markers and miR-7 Mimic Protects Photoreceptors from Parthanatos by Repressing α-Synuclein in Retinal Detachment.

Retinal detachment (RD) refers to the separation between the neuroepithelium and the pigment epithelium layer. It is an important disease leading to irreversible vision damage worldwide, in which photoreceptor cell death plays a major role. α-Synuclein (α-syn) is reportedly involved in numerous mechanisms of neurodegenerative diseases, but the association with photoreceptor damage in RD has not been studied. In this study, elevated transcription levels of α-syn and parthanatos proteins were observed in the vitreous of patients with RD. The expression of α-syn- and parthanatos-related proteins was increased in experimental rat RD, and was involved in the mechanism of photoreceptor damage, which was related to the decreased expression of miR-7a-5p (miR-7). Interestingly, subretinal injection of miR-7 mimic in rats with RD inhibited the expression of retinal α-syn and down-regulated the parthanatos pathway, thereby protecting retinal structure and function. In addition, interference with α-syn in 661W cells decreased the expression of parthanatos death pathway in oxygen and glucose deprivation model. In conclusion, this study demonstrates the presence of parthanatos-related proteins in patients with RD and the role of the miR-7/α-syn/parthanatos pathway in photoreceptor damage in RD.

Mycobiont-specific primers facilitate the amplification of mitochondrial small subunit ribosomal DNA: a focus on the lichenized fungal genus Melanelia (Ascomycota, Parmeliaceae) in Iceland.

The fungal mitochondrial small subunit (mtSSU) ribosomal DNA is one of the most commonly used loci for phylogenetic analysis of lichen-forming fungi, but their primer specificity to mycobionts has not been evaluated. The current study aimed to design mycobiont-specific mtSSU primers and highlights their utility with an example from the saxicolous lichen-forming fungal genus Melanelia Essl. in Iceland. The study found a 12.5% success rate (3 out of 24 specimens with good-quality mycobiont mtSSU sequences) using universal primers (i.e. mrSSU1 and mrSSU3R), not including off-target amplification of environmental fungi, e.g. Cladophialophoracarrionii and Lichenotheliaconvexa. New mycobiont-specific primers (mt-SSU-581-5' and mt-SSU-1345-3') were designed by targeting mycobiont-specific nucleotide sites in comparison with environmental fungal sequences, and assessed for mycobiont primer specificity using in silico PCR. The new mycobiont-specific mtSSU primers had a success rate of 91.7% (22 out of 24 specimens with good-quality mycobiont mtSSU sequences) on the studied Melanelia specimens. Additional testing confirmed the specificity and yielded amplicons from 79 specimens of other Parmeliaceae mycobiont lineages. This study highlights the effectiveness of designing mycobiont-specific primers for studies on lichen identification, barcoding and phylogenetics.

Phylogeographic analyses of an epiphytic foliose lichen show multiple dispersal events westward from the Hengduan Mountains of Yunnan into the Himalayas.

Lobaria pindarensis is an endemic species of the Himalayas and the Hengduan Mountains. Little information is available on the phylogeography genetics and colonization history of this species or how its distribution patterns changed in response to the orographic history of the Himalayas and Hengduan Mountains. Based on samples covering a major part of the species' distribution range, we used 443 newly generated sequences of nine loci for molecular coalescent analyses in order to reconstruct the evolutionary history of L. pindarensis, and to reconstruct the species' ancestral phylogeographic distributions using Bayesian binary MCMC analyses. The results suggest that current populations originated from the Yunnan region of the Hengduan Mountains in the middle Pliocene, and that the Himalayas of Bhutan were colonized by a lineage that diverged from Yunnan ca. 2.72 Ma. The analysis additionally indicates that the Nepal and Xizang areas of the Himalayas were colonized from Yunnan as well, and that there was later a second dispersal event from Yunnan to Bhutan. We conclude that the change in climate and habitat related to the continuous uplift of the Himalayas and the Hengduan Mountains in the late Pliocene and middle Pleistocene influenced the geographic distribution pattern of L. pindarensis.

Baseline Severity of Sacroiliitis and Extensive Fat Metaplasia Predicts the Progression of Backfill at the Sacroiliac Joint in Patients With Axial Spondyloarthritis.

Objectives: Fat metaplasia in an erosion cavity, also known as backfill, is an essential intermediary in new bone formation in axial spondyloarthritis (axSpA) patients; however, the predictors of backfill progression are unknown. This longitudinal study aimed to assess the predictors of backfill progression in axSpA patients on magnetic resonance imaging (MRI). Methods: Clinical and MRI data were collected at baseline and follow-up in 52 axSpA patients. Backfill progression was defined as the new or increased T1 hyperintensity within the SI joint space. Logistic regression analyses were performed to identify the predictors of the backfill progression. Results: A total of 19 patients had "backfill" at baseline and 30 patients exhibited the backfill progression after follow-up. The mean disease duration and SPARCC scores at baseline were significantly different between patients with and without backfill progression (P<0.001, P=0.003, respectively). Patients with backfill progression had a higher frequency of backfill at baseline, a higher SSS score of fat metaplasia, and a higher SSS score of backfill than those without (P=0.001, P<0.001, and P=0.002, respectively). A higher fat fraction value in the fat metaplasia area at the baseline was more frequent in patients with, than without, backfill progression (P=0.019). In the univariate logistic regression analyses, a higher SPARCC score for inflammation and a higher SSS score for fat metaplasia at baseline were associated with backfill progression. Conclusions: Severity of sacroiliitis and extensive fat metaplasia at baseline are predictors of the backfill progression in axSpA patients.

Providing Scale to a Known Taxonomic Unknown-At Least a 70-Fold Increase in Species Diversity in a Cosmopolitan Nominal Taxon of Lichen-Forming Fungi.

Robust species delimitations provide a foundation for investigating speciation, phylogeography, and conservation. Here we attempted to elucidate species boundaries in the cosmopolitan lichen-forming fungal taxon Lecanora polytropa. This nominal taxon is morphologically variable, with distinct populations occurring on all seven continents. To delimit candidate species, we compiled ITS sequence data from populations worldwide. For a subset of the samples, we also generated alignments for 1209 single-copy nuclear genes and an alignment spanning most of the mitochondrial genome to assess concordance among the ITS, nuclear, and mitochondrial inferences. Species partitions were empirically delimited from the ITS alignment using ASAP and bPTP. We also inferred a phylogeny for the L. polytropa clade using a four-marker dataset. ASAP species delimitations revealed up to 103 species in the L. polytropa clade, with 75 corresponding to the nominal taxon L. polytropa. Inferences from phylogenomic alignments generally supported that these represent evolutionarily independent lineages or species. Less than 10% of the candidate species were comprised of specimens from multiple continents. High levels of candidate species were recovered at local scales but generally with limited overlap across regions. Lecanora polytropa likely ranks as one of the largest species complexes of lichen-forming fungi known to date.

LINC00667 regulates MPP+ -induced neuronal injury in Parkinson's disease.

OBJECTIVE: Parkinson's disease (PD), also known as paralysis tremor, is a chronic disease of the central nervous system. It has been reported that hepatocyte nuclear factor 4 alpha (HNF4A) is upregulated in PD, but its specific function has not been well explored. METHODS: We established an in vitro PD model in SH-SY5Y cells stimulated with 1-methyl-4-phenylpyridinium (MPP+ ). Meanwhile, the effect of HNF4A on MPP+ -treated SH-SY5Y cell behavior was monitored by functional assays. Mechanism assays were conducted to verify the relationship among LINC00667/miR-34c-5p/HNF4A. Rescue experiments validated the regulatory mechanism in PD model. RESULTS: The results revealed that depletion of HNF4A suppressed cell cytotoxicity and apoptosis caused by MPP+ . Knockdown of HNF4A recovered MPP+ -stimulated oxidative stress and neuroinflammation. Mechanically, HNF4A was targeted and inhibited by miR-34c-5p. Furthermore, we found that LINC00667 positively modulated HNF4A expression via sequestering miR-34c-5p in MPP+ -stimulated SH-SY5Y cells. Interestingly, the data indicated that HNF4A could transcriptionally activate LINC00667 expression. Rescue experiments presented that miR-34c-5p interference or HNF4A overexpression could mitigate the effects of LINC00667 knockdown on cell viability, cytotoxicity, cell apoptosis, oxidative stress, and neuroinflammation in MPP+ -treated SH-SY5Y cells. CONCLUSION: Our study first proved LINC00667, miR-34c-5p, and HNF4A constructed a positive feedback loop in MPP+ -treated SH-SY5Y cells, enriching our understanding of PD.

Revision of Immersaria and a new lecanorine genus in Lecideaceae (lichenised Ascomycota, Lecanoromycetes).

The species Immersariacupreoatra has been included in Bellemerea. This caused us to reconsider the relationships between Bellemerea and the lecanorine species of Immersaria and to question the monophyly of Immersaria. Amongst 25 genera of the family Lecideaceae, most have lecideine apothecia, the exceptions being Bellemerea and Koerberiella, which have lecanorine apothecia. According to previous classifications, Immersaria included species with both lecanorine and lecideine apothecia. A five-loci phylogenetic tree (nrITS, nrLSU, RPB1, RPB2, and mtSSU) for Lecideaceae showed that Immersaria was split into two clades: firstly, all the lecideine apotheciate species and secondly, all the lecanorine apotheciate species. The latter clade was closely related to the remaining lecanorine apotheciate genera: Bellemerea and Koerberiella. Therefore, the genus concept of Immersaria is revised accordingly and a new lecanorine genus Lecaimmeria is proposed. Furthermore, four new species for Immersaria and seven new species and three new combinations for the new genus Lecaimmeria are proposed. Keys to Immersaria and the new genus Lecaimmeria are provided.

LncRNA THRIL promotes high glucose-induced proliferation and migration of human retina microvascular endothelial cells through enhancing autophagy.

AIMS: Diabetes retinopathy (DR) is associated with retinal microvascular system injury induced by high glucose (HG). This study aims to explore the role and mechanism of long non-coding RNA THRIL in regulating cell proliferation and migration of human retina microvascular endothelial cells (hRMECs) under HG condition. METHOD: The gene and protein expression were detetced by RT-PCR and western blot, respectively. Cell proliferation and migration of hRMECs were examined using MTT assay and Transwell assay, respectively. The interaction between miR-125b-5p and THRIL or autophagy-related gene 4D (ATG4D) was analyzed using luciferase activity assay. RESULTS: THRIL expression was induced by HG in hRMECs. THRIL overexpression enhanced the proliferation and migration of hRMECs induced by HG, whereas THRIL silencing yielded the opposite results. Furthermore, THRIL overexpression induced autophagy activation, and inhibition of autophagy by 3-methyladenine abrogated the promotory effects of THRIL overexpression on cell proliferation and migration of hRMECs. Mechanismly, THRIL inhibited miR-125b-5p to upregulate the expression of ATG4D (an important autophagy-related gene), thereby promoting autophagy. Moreover, miR-125b-5p overexpression or ATG4D silencing alone abolished the promoting effects of THRIL overexpression on HG-induced autophagy, proliferation and migration of hRMECs. CONCLUSIONS: THRIL promotes HG-induced cell proliferation and migration of hRMECs through activation of autophagy via the miR-125b-5p/ATG4D axis. THRIL may serve as a potential therapeutic target for DR.

Nakamurella leprariae sp. nov., isolated from a lichen sample.

A novel actinobacterium, YIM 132084T, was isolated from Lepraria sp. lichen collected from Yunnan province, south-west PR China and identified by a polyphasic taxonomic approach. The strain was Gram-stain-positive, aerobic, catalase-positive, oxidase-negative, non-motile and coccus-shaped. Colonies were round, convex, smooth and light orange yellow in color. It grew at 10-40 °C (optimum 28 °C), at pH 6.0-11.0 (optimum pH 7.0) and in the presence of 0-4% NaCl (optimum 0%). Strain YIM 132084T comprised diphosphatidylglycerol, phosphatidylethanolamine and phosphatidylinositol as the major polar lipids, MK-8(H4) as the predominant menaquinone, and anteiso-C15:0, anteiso-C17:0, iso-C15:0 and iso-C16:0 as major fatty acids. Strain YIM 132084T had meso-diaminopimelic acid as the diagnostic diamino acid in the cell-wall peptidoglycan, and mannose, ribose, glucose and rhamnose as whole-cell sugars. The 16S rRNA gene sequence showed high level of similarity with Nakamurella flavida KCTC 19127T (97.7%) and Nakamurella flava CGMCC 4.7524T (97.7%). The G + C content of the genomic DNA was 72.4 mol%. Based on draft genome sequences, strain YIM 132084T showed an average nucleotide identity value of 76.1% and 74.9%, a digital DNA-DNA hybridization value of 20.9% and 20.6% with the reference strains Nakamurella flavida and Nakamurella flava, respectively. The results of the phenotypic, chemotaxonomic and phylogenetic analyses showed that strain YIM 132084T represents a novel species of the genus Nakamurella, for which the name Nakamurella leprariae sp. nov. is proposed. The type strain is YIM 132084T (= CGMCC 4.7667T = NBRC 114280T = KCTC 49367T).

EphA2 inhibits SRA01/04 cells apoptosis by suppressing autophagy via activating PI3K/Akt/mTOR pathway.

This study attempted to determine the effect of EphA2 on H2O2-treated lens epithelial cells (SRA01/04) and the underlying mechanisms. MTT assay and flow cytometry were performed to assess cell viability and cell apoptosis. Western blot was carried out to examine the levels of proteins associated with apoptosis and autophagy. Our results revealed that EphA2 significantly elevated the reduced cell viability, and inhibited the increased cell apoptosis in H2O2-treated SRA01/04 cells, along with the significant up-regulated Bcl-2 and down-regulated Cleaved-caspase-3 and Bax protein levels, but which were all abolished by Rapa (autophagy activator). We also found that EphA2 significantly suppressed cell autophagy in H2O2-treated SRA01/04 cells. Additionally, EphA2 significantly up-regulated the protein levels of p-Akt and p-mTOR in H2O2-treated SRA01/04 cells, and the inhibition of Akt by MK-2206 and inhibition of mTOR by Rapa both obviously reversed EphA2-mediated the inhibition of autophagy in H2O2-treated SRA01/04 cells. In summary, these data demonstrated that EphA2 inhibited the apoptosis of SRA01/04 cells by inhibiting autophagy via activating PI3K/Akt/mTOR pathway.

Glaciibacter flavus sp. nov., isolated from a lichen sample.

A novel Actinobacterium strain YIM 131861 T, was isolated from lichen collected from the South Bank Forest of the Baltic Sea, Germany. It was Gram-stain-positive, strictly aerobic, catalase positive and oxidase negative, yellow pigmented. Cells were motile with a polar flagellum, irregular rod shaped and did not display spore formation. The strain grew at 15 - 30 °C (optimum 25 °C), at pH 6.0 - 10.0 (optimum pH 7.0) and in the presence of 0 - 1.5% (w/v) NaCl (optimum 1%). Phylogenetic analysis based on 16S rRNA gene sequences revealed that strain YIM 131861 T belonged to the genus Glaciibacter, and exhibited a high sequence similarity (96.4%) with Glaciibacter superstes NBRC 104264 T. The genomic DNA G + C content of strain YIM 131861 T was 68.2 mol%. The average nucleotide identity (ANI) and digital DNA-DNA hybridization (dDDH) values between strain YIM 131861 T and Glaciibacter superstes NBRC 104264 T were 73.2 and 19.9% based on the draft genome sequence. The cell-wall peptidoglycan type was B2γ and contained the 2, 4-diaminobutyric acid as the diagnostic amino acid. Whole cell sugars were galactose, rhamnose, ribose and glucose. It contained MK-12 and MK-13 as the predominant menaquinones. The major cellular fatty acids (> 10%) were identified as anteiso-C15:0, iso-C16:0 and anteiso-C17:0. The polar lipids were diphosphatidylglycerol, phosphatidylglycerol and two unknown glycolipids. Based on the results of phenotypic, chemotaxonomic and phylogenetic analyses, strain YIM 131861 T should belong to the genus Glaciibacter and represents a novel species of the genus Glaciibacter, for which the name Glaciibacter flavus sp. nov. is proposed. The type strain is YIM 131861 T (= CGMCC 1.16588 T = NBRC 113572 T).

Paracoccus lichenicola sp. nov., Isolated from Lichen.

Strain YIM 132242T, isolated from lichen collected from Pu'er, Yunnan Province, China, was short-rod-shaped, Gram-reaction-negative, aerobic, catalase- and oxidase-positive. Growth of the strain was occurred at 10-39 °C (optimum, 28-35 °C), at pH 4.0-10.0 (optimum, pH 7.0-8.0) and at salinities of 0-8% (w/v) NaCl (optimum, 0-2%). Phylogenetic analyses based on 16S rRNA gene sequences indicated that strain YIM 132242T belonged to the genus Paracoccus and had the highest levels of sequence similarity to Paracoccus aerius KCTC 42845T (97.0% similarity), Paracoccus sediminis CMB17T (96.8% similarity), and Paracoccus fontiphilus MVW-1T (96.4% similarity). The major fatty acid was identified as C18:1 ω7c (77.6%). The predominant respiratory quinone was ubiquinone-10 (Q-10). Polar lipid analysis indicated the presence of phosphatidylglycerol (PG), phosphatidylethanolamine (PE), phosphatidylcholine (PC), diphosphatidylglycerol (DPG), an unidentified lipid (L), and three unidentified phospholipids (PL1-PL3). Based on the draft genome sequence, the DNA G + C content of the strain was 67.1 mol%, and the values of average nucleotide identity (ANI) and digital DNA-DNA hybridization (dDDH) of strain YIM 132242T with Paracoccus aerius KCTC 42845T were 85.4% and 29.1%, respectively. On the basis of the data from this polyphasic characterization, strain YIM 132242T represents a novel species of the genus Paracoccus, for which the name Paracoccus lichenicola sp. nov. is proposed. The type strain is YIM 132242T (= KCTC 72463T = CGMCC1.17191T).

Aspergillus fumigatus-Stimulated Human Corneal Epithelial Cells Induce Pyroptosis of THP-1 Macrophages by Secreting TSLP.

Fungal keratitis (FK) is a keratopathy caused by pathogenic fungal infection. The aim of this work is to explore the role of thymic stromal lymphopoietin (TSLP) in FK. Human corneal epithelial cells (HCECs) were treated with Aspergillus fumigatus hyphae, and we found that TSLP was highly expressed and secreted in the hyphae-treated HCECs. Hyphae-treated HCECs or TSLP treatment enhanced the expression of caspase-1 P20, GSDMD-N (p30), IL-1ß, and IL-18 in the human THP-1 macrophages. The influence conferred by hyphae-treated HCECs or TSLP treatment was rescued by TSLP neutralizing antibody or VX-765 (caspase-1 inhibitor) treatment. Moreover, TSLP treatment promoted the expression of NLRP3, ASC, caspase-1 P20, GSDMD-N (p30), IL-1ß, and IL-18 in the THP-1 macrophages, which was abolished by NLRP3 knockdown. Furthermore, TSLPR silencing suppressed the expression of NLRP3, ASC, caspase-1 P20, GSDMD-N (p30), IL-1ß, and IL-18 in the TSLP-treated THP-1 macrophages. In conclusion, our article confirms that Aspergillus fumigatus-stimulated HCECs induce pyroptosis of THP-1 macrophages by secreting TSLP. TSLP/TSLPR induces caspase-1-dependent pyroptosis through activation of NLRP3 inflammasome. Thus, our work suggests that TSLP may be a potential target for FK treatment.