Acute Toxicity of a Novel anti-fouling Material Additive DCOIT to Marine Chlorella sp.

DCOIT (4,5-dichloro-2-n-octyl-4-isothiazolin-3-one) is the main ingredient in SeaNine-211, a new antifouling agent that replaces organotin compounds to prevent the growth of fouling organisms on board. Biocides from antifoulants can cause problems for marine ecosystems by destroying non-target algal species. This study evaluated the potential adverse effects DCOIT using the Marine Chlorella sp. The concentration of DCOIT were set according to the semi-inhibitory concentrations for acute exposure experiments, and relevant oxidative stress indicators were measured to assess the acute toxic effects. The results showed that the inhibition concentrations (IC50) of DCOIT against Marine Chlorella sp was 2.522 mg/L. The genes related to photosynthesis and antioxidant capacity showed the effect of promoting low concentration and inhibiting high concentration. In addition, based on the ultrastructural observation and the expression analysis of photosynthesis related genes, it was found that DCOIT had a significant effect on plant photosynthesis.

Gender- and age-related differences in distinct phenotypes of hypertrophic cardiomyopathy-associated mutation MYBPC3-E334K.

The mutation MYBPC3-E334K is a culprit mutation of hypertrophic cardiomyopathy (HCM). The pathogenicity of MYBPC3-E334K is conflicting in ClinVar because of the limited segregation data and the relatively high frequency in gnomAD (0.03% overall, with 0.3% in East Asians and 0.8% in Japanese). The main aim is to clarify the clinical importance and phenotype-genotype correlations in subjects with or without MYBPC3-E334K alone. The prevalence of MYBPC3-E334K was sequenced in 1017 HCM unrelated probands. The clinical features, morphology phenotypes, and electrical phenotypes were further analyzed according to the phenotype and genotype status in families with single-mutation MYBPC3-E334K. Nine of 1017 (0.88%) unrelated HCM probands were detected harboring MYBPC3-E334K, and three of them harbored a second variant in sarcomere protein gene. Family study and co-segregation analyses indicated that patients with single-mutation MYBPC3-E334K showed autosomal dominant mode of inheritance with incomplete penetrance. The overall disease penetrance was 52.6%, and the disease penetrance was higher in males than in females (100% in men vs 25% in women, p = 0.003). The mean age at diagnosis of males was approximately 25 years younger than females (36.57 ± 18.65 vs 62.33 ± 12.10, p = 0.062). The variant MYBPC3-E334K was classified as a likely pathogenic variant, and a second sarcomere variant did not reveal obvious cumulative effects. The patients harboring single-mutation MYBPC3-E334K had incomplete penetrance, and males demonstrated higher penetrance and early onset HCM than females. A second sarcomere variant did not reveal obvious cumulative effects.

Two new compounds from Streptomyces sp. HS-NF-813.

Two new derivatives of cytotoxic substance BE-52211, designed as BE-52211D (1) and BE-52211E (2), were isolated from the fermentation broth of the strain Streptomyces sp. HS-NF-813. Their structures were determined by 1D and 2D NMR techniques, ESI-MS and comparison with data from the literature. The absolute stereochemistry of 1 was elucidated by NMR data of the Mosher ester derivatives. Compounds 1 and 2 showed moderate cytotoxic activity against three human tumor cell lines.

[Treating lupus nephritis by a drug pair of radix astragali and rehmanniae radix combined with glucocorticoid: a preliminary clinical study].

OBJECTIVE: To observe the therapeutic effect of a drug pair of Radix Astragali and Rehmanniae Radix combined with glucocorticoid (GC) in treating lupus nephritis (LN) patients and its influence on some experimental indices. METHODS: Totally 52 LN patients were randomly assigned to the treatment group (treated by routine Western medicine and a drug pair of Radix Astragali and Rehmanniae Radix, 25 cases) and the control group (treated by Western medicine, mainly by GC and cyclophosphamide, 27 cases). All patients received 6-month therapy. The GC dosage, the withdrawal and reduction dosage of GC, clinical efficacy, systemic lupus erythematosus disease activity index (SLEDAI) score, adverse reactions, and laboratory indicators were recorded. RESULTS: (1) All patients got relieved to some degree with the dosage of GC reduced. The total withdrawal and reduction dosage of GC was slightly higher in the treatment group than in the control group [(50.23 +/- 12.43) mg vs (48.76 +/- 13.61) mg, P > 0.05]. Besides, the prednisone dosage in the treatment group was lower than that in the control group, but without statistical difference (P > 0.05). The ratio of patients in need of adding prednisone for aggravating disease was 24.0%, significantly lower than that in the control group (44.44%, P < 0.05). (2) There was no statistical difference in the SLEDAI score, inflammatory indicators, liver and renal functions, blood electrolytes, blood glucose, blood and urine routines between the two groups (P > 0.05). The 24-h urinary protein count was (1.06 +/- 0.22) g/L in the treatment group, obviously lower than that in the control group (1.43 +/- 0.55 g/L, P < 0.05). (3) There was no statistical difference in the incidence rate of infection, gastrointestinal hemorrhage, psychoneuroses, Cushing's syndrome, cardiovascular anomalies, and femoral head necrosis between the two groups (P > 0.05). But the incidence of adverse reactions such as insomnia, tidal fever, spontaneous sweat, and obesity was less in the treatment group than in the control group (P < 0.05). CONCLUSIONS: Using a drug pair of Radix Astragali and Rehmanniae Radix combined with GC in treating LN could withdraw the dosage of GC and relieve symptoms it induced. It was advantageous in reducing the dosage of GC and stabilizing patients' conditions.

Indigoferajintongpenensis (Fabaceae, Papilionoideae, Indigofereae), a new species from Yunnan, southwest China.

Indigoferajintongpenensis, a new species of the subfamily Papilionoideae of Fabaceae, is described and illustrated from Yunnan, southwest China. The new species is characterised by having a prostrate habit, flexible stems and branches, as well as spreading, sub-basifixed, asymmetrically 2-branched trichomes covering the entire plant, discoid calyx, and racemose inflorescences 6-8-flowered, short, 1-2 (-3.5) cm in length, apparently shorter than the leaf. A distribution map and comparison of morphological diagnostic characters with its morphologically similar species are provided. Additionally, a preliminary conservation assessment of I.jintongpenensis is proposed following IUCN criteria.

Metagenomics analysis reveals the effects of norfloxacin on the gut microbiota of juvenile common carp (Cyprinus carpio).

Norfloxacin (NOR) is an early third-generation quinolone antibiotic that has been widely used in animal husbandry and aquaculture because of its bactericidal properties. As an emerging contaminant, NOR may have toxic effects on fish. This study assessed the chronic toxicity (6 weeks) of 0 (control group), 100 ng/L (environmental concentration), and 1 mg/L NOR to the gut microbiota of juvenile common carp (Cyprinus carpio) based on metagenomic sequencing. Metagenomic analysis revealed that the Proteobacteria, Bacteroidetes, Fusobacteria, Firmicutes, and Actinobacteria were the dominant bacteria in the gut of common carp. The relative abundance of Actinobacteria was highest in the control group. The alpha diversity of the environmental concentration NOR was significantly lower than the control group. Principal coordinates analysis (PCoA) indicated that the bacterial community between the different groups formed clear separate clusters. NOR exposure adversely could affect immune function and some substance metabolic pathways in the gut microbiota of common carp. Furthermore, environmental concentrations of NOR produce antibiotic resistance genes (ARGs) in the gut microbiota, enhancing resistance to drugs. In conclusion, environmental concentrations of NOR could alter the composition, structure, and abundance of ARGs in the gut microbiota, thereby affecting the intestinal health of fish.

Assessing the ecotoxicity of combined exposure to triphenyltin and norfloxacin at environmental levels: A case study of immunotoxicity and metabolic regulation in carp (Cyprinus carpio).

This paper evaluates the coexistence risks of triphenyltin (TPT) and norfloxacin (NOR) to aquatic organisms in the aquatic environment. Carp (Cyprinus carpio) was used as the test organism, the control and exposure groups (1 µg/L TPT), 1 mg/L (NOR), 1 µg/LTPT+1 mg/LNOR (TPT_NOR)) were set up according to the environmental concentration in the severely polluted area for 42 days. The single/combined toxic effects of TPT and NOR on aquatic organisms were evaluated by analyzing carp brain transcriptome sequencing, gut microbiota structure, and detection of biochemical indicators and RT-qPCR. Our results show that TPT and NOR induce lipid metabolism disorder in carp brain tissue, affecting the metabolism of cytochrome P450 to exogenous substances, and NOR also induces immunosuppression in carp. Long-term exposure to TPT combined with NOR amplifies the monotoxicity of TPT or NOR on lipid metabolism and immunosuppression in carp, induces immune dysfunction in brain tissue and changes in gut microbiota structure. However, TPT_NOR has no obvious neurotoxicity on the brain, but it can inhibit the level of intestinal MDA. This highlights that co-exposure of TPT and NOR amplifies metabolic disorders and immunosuppressive functions in carp.

Phylogenomics and plastome evolution of Indigofera (Fabaceae).

Introduction: Indigofera L. is the third largest genus in Fabaceae and includes economically important species that are used for indigo dye-producing, medicinal, ornamental, and soil and water conservation. The genus is taxonomically difficult due to the high level of overlap in morphological characters of interspecies, fewer reliability states for classification, and extensive adaptive evolution. Previous characteristic-based taxonomy and nuclear ITS-based phylogenies have contributed to our understanding of Indigofera taxonomy and evolution. However, the lack of chloroplast genomic resources limits our comprehensive understanding of the phylogenetic relationships and evolutionary processes of Indigofera. Methods: Here, we newly assembled 18 chloroplast genomes of Indigofera. We performed a series of analyses of genome structure, nucleotide diversity, phylogenetic analysis, species pairwise Ka/Ks ratios, and positive selection analysis by combining with allied species in Papilionoideae. Results and discussion: The chloroplast genomes of Indigofera exhibited highly conserved structures and ranged in size from 157,918 to 160,040 bp, containing 83 protein-coding genes, 37 tRNA genes, and eight rRNA genes. Thirteen highly variable regions were identified, of which trnK-rbcL, ndhF-trnL, and ycf1 were considered as candidate DNA barcodes for species identification of Indigofera. Phylogenetic analysis using maximum likelihood (ML) and Bayesian inference (BI) methods based on complete chloroplast genome and protein-coding genes (PCGs) generated a well-resolved phylogeny of Indigofera and allied species. Indigofera monophyly was strongly supported, and four monophyletic lineages (i.e., the Pantropical, East Asian, Tethyan, and Palaeotropical clades) were resolved within the genus. The species pairwise Ka/Ks ratios showed values lower than 1, and 13 genes with significant posterior probabilities for codon sites were identified in the positive selection analysis using the branch-site model, eight of which were associated with photosynthesis. Positive selection of accD suggested that Indigofera species have experienced adaptive evolution to selection pressures imposed by their herbivores and pathogens. Our study provided insight into the structural variation of chloroplast genomes, phylogenetic relationships, and adaptive evolution in Indigofera. These results will facilitate future studies on species identification, interspecific and intraspecific delimitation, adaptive evolution, and the phylogenetic relationships of the genus Indigofera.

Combined effect of microplastic and triphenyltin: Insights from the gut-brain axis.

Microplastics (MPs), an emerging group of pollutants, not only have direct toxic effects on aquatic organisms but also cause combined toxicity by absorbing other pollutants. Triphenyltin (TPT), one of the most widely used organotin compounds, has adverse effects on aquatic organisms. However, little is known about the combined toxicity of MPs and TPT to aquatic organisms. To investigate the individual and combined toxicity of MPs and TPT, we selected the common carp (Cyprinus carpio) for a 42-day exposure experiment. Based on the environmental concentrations in a heavily polluted area, the experimental concentrations of MPs and TPT were set at 0.5 mg L-1 and 1 µg L-1, respectively. The effects of MPs combined with TPT on the carp gut-brain axis were evaluated by detecting gut physiology and biochemical parameters, gut microbial 16S rRNA, and brain transcriptome sequencing. Our results suggest that a single TPT caused lipid metabolism disorder and a single MP induced immunosuppression in carp. When MPs were combined with TPT, the involvement of TPT amplified the immunotoxic effect induced by MPs. In this study, we also explored the gut-brain axis relationship of carp immunosuppression, providing new insights for assessing the combined toxicity of MPs and TPT. At the same time, our study provides a theoretical basis for evaluating the coexistence risk of MPs and TPT in the aquatic environment.

A reversible plasmonic nanoprobe for dynamic imaging of intracellular pH during endocytosis.

Understanding the pH evolution during endocytosis is essential for our comprehension of the fundamental processes of biology as well as effective nanotherapeutic design. Herein, we constructed a plasmonic Au@PANI core-shell nanoprobe, which possessed significantly different scattering properties under acidic and basic conditions. Encouragingly, the scattering signal of Au@PANI nanoprobes displayed a positive linear correlation with the pH value not only in PBS but also in nigericin-treated cells. Ultimately, benefiting from the excellent response properties as well as the excellent biocompatibility and stability, the Au@PANI nanoprobes have successfully enabled a dynamic assessment of the evolving pH in the endosomal package as the endosome matured from early to late, and eventually to the lysosome, by reporting scattering signal changes.

Phytotoxicity of environmental norfloxacin concentrations on the aquatic plant Spirodela polyrrhiza: Evaluation of growth parameters, photosynthetic toxicity and biochemical traits.

As an emerging pollutant, the increasing use of antibiotics in wastewater posed a serious threat to non-target organisms in the environment. Duckweed (Spirodela polyrrhiza) is a common higher aquatic plant broadly used in phytotoxicity tests for xenobiotic substances. The aim of this study was to evaluate the chronic toxicity of norfloxacin (NOR) on Spirodela polyrrhiza during 18 days of exposure. Our study investigated the addition of NOR into the medium with environment-related concentrations (0, 0.1, 10, and 1000 µg L-1). Subsequently, biomarkers of toxicity such as growth, pigment, chlorophyll fluorescence parameters, indicators of oxidative stress, and osmotic regulatory substances content were analyzed in duckweed. In response to NOR exposure, obvious chlorosis, declines in growth and photosynthetic pigment, and photosystem II inhibition were noted in a concentration dependent manner. Reactive oxygen species (ROS) and antioxidant activity content increased in the treated fronds, which indicated that oxidative stress was specifically affected by NOR exposure. A slight increase in osmotic regulatory substances in NOR treated setups than in the control represented the increasing stress resistance. These results suggest NOR exerts its toxic effects on the aquatic plant Spirodela polyrrhiza.

Effects of long-term exposure of norfloxacin on the HPG and HPT axes in juvenile common carp.

Currently, there is a relatively lack of relevant research on the interference effect of quinolone antibiotics on the endocrine of aquatic animals. In this study, the toxicity of norfloxacin (NOR) on the endocrine system of juvenile common carp (Cyprinus carpio) was evaluated, as well as the hematocyte parameters. Specifically, two important endocrine axes were assessed: the hypothalamus-pituitary-thyroid (HPT) axis and hypothalamus-pituitary-gonadal (HPG) axis. Norfloxacin was used as a representative of quinolone antibiotics. According to the concentration of water pollution areas and considering the bad situation that may be caused by wastewater discharge, a control, 100 ng/L NOR, and 1 mg/L NOR treatment groups were set up. The juvenile carp, as the test animal, was subjected to an exposure experiment for 42 days. Thyroid hormones (T3 and T4) and related genes in HPT axis and sex hormones (11-ketotestosterone [11-KT] and progesterone [PROG]) and related genes in HPG axis and blood count are tested. It was found that the T4 iodine level and conversion process were enhanced after NOR treatment, which in turn led to the increase of T3 content and biological activity in the blood. One hundred nanograms per liter NOR can inhibit the level of sex hormones and inhibit the expression of HPG axis-related genes. In the 1 mg/L NOR treatment group, long-term exposure over a certain concentration range may lead to the development of adaptive mechanisms, making the changes in hormones and related genes insignificant. In conclusion, this study provides reference data for the endocrine interference of quinolone antibiotics on aquatic organisms, and has ecological significance for assessing the health of fish populations of quinolone antibiotics. However, the specific sites and mechanisms of action related to the effects of NOR on the endocrine system remain unclear and require further study.

Chronic toxic effects of polystyrene micro-plastics, DCOIT and their combination on marine Chlorella sp.

Polystyrene (PS) is one of the most dangerous polymers, mainly because of the mutagenic or carcinogenic risk of the monomers used to produce it. Sea-Nine 211 is a commercial antifouling agent; its active ingredient is the biocide 4,5-dichloro-2-octyl-4-isothiazolinone-3-one (DCOIT). Micro- and nano-plastics have different synergistic effects on marine organisms together with organic pollutants. To understand the toxic effects of DCOIT and PS alone and in combination, marine Chlorella sp was selected as the experimental organism. The exposure concentration of DCOIT was set at 50 µg/L, and that of PS was set at 10 µg/L. The results show that all exposed groups promoted the growth of marine Chlorella sp in the late stage of exposure, and the recovery time of marine Chlorella sp in the exposed group containing PS was earlier. Changing trend of chlorophyll a was consistent with the growth trend. On the 15th day of exposure, the gene expression of the photosynthesis system in the combined exposed group showed a significant difference, and the cells produced oxidative stress. Scanning electron microscope observation shows the algae adhered to each other. The volume of algae cells in DCOIT and PS exposed groups decreased, and the internal structure of algae cells in each exposed group was damaged.

Delayed improvements in visual memory task performance among chronic schizophrenia patients after high-frequency repetitive transcranial magnetic stimulation.

BACKGROUND: Cognitive impairments are core characteristics of schizophrenia, but are largely resistant to current treatments. Several recent studies have shown that high-frequency repetitive transcranial magnetic stimulation (rTMS) of the left dor-solateral prefrontal cortex (DLPFC) can reduce negative symptoms and improve certain cognitive deficits in schizophrenia patients. However, results are inconsistent across studies. AIM: To examine if high-frequency rTMS of the DLPFC can improve visual memory deficits in patients with schizophrenia. METHODS: Forty-seven chronic schizophrenia patients with severe negative symptoms on stable treatment regimens were randomly assigned to receive active rTMS to the DLPFC (n = 25) or sham stimulation (n = 22) on weekdays for four consecutive weeks. Patients performed the pattern recognition memory (PRM) task from the Cambridge Neuropsychological Test Automated Battery at baseline, at the end of rTMS treatment (week 4), and 4 wk after rTMS treatment (week 8). Clinical symptoms were also measured at these same time points using the Scale for the Assessment of Negative Symptoms (SANS) and the Positive and Negative Syndrome Scale (PANSS). RESULTS: There were no significant differences in PRM performance metrics, SANS total score, SANS subscores, PANSS total score, and PANSS subscores between active and sham rTMS groups at the end of the 4-wk treatment period, but PRM performance metrics (percent correct and number correct) and changes in these metrics from baseline were significantly greater in the active rTMS group at week 8 compared to the sham group (all P < 0.05). Active rTMS treatment also significantly reduced SANS score at week 8 compared to sham treatment. Moreover, the improvement in visual memory was correlated with the reduction in negative symptoms at week 8. In contrast, there were no between-group differences in PANSS total score and subscale scores at either week 4 or week 8 (all P > 0.05). CONCLUSION: High-frequency transcranial magnetic stimulation improves visual memory and reduces negative symptoms in schizophrenia, but these effects are delayed, potentially due to the requirement for extensive neuroplastic changes within DLPFC networks.

Assessing the impact of land surface temperature on urban net primary productivity increment based on geographically weighted regression model.

Urbanization had a huge impact on the regional ecosystem net primary productivity (NPP). Although the urban heat island (UHI) caused by urbanization has been found to have a certain promoting effect on urban vegetation NPP, the factors on the impact still are not identified. In this study, the impact of urbanization on NPP was divided into direct impact (NPPdir) and indirect impact (NPPind), taking Kunming city as a case study area. Then, the spatial heterogeneity impact of land surface temperature (LST) on NPPind was analyzed based on the geographically weighted regression (GWR) model. The results indicated that NPP, LST, NPPdir and NPPind in 2001, 2009 and 2018 had significant spatial autocorrelation in Kunming based on spatial analytical model. LST had a positive impact on NPPind in the central area of Kunming. The positively correlation areas of LST on NPPind increased by 4.56%, and the NPPind caused by the UHI effect increased by an average of 4.423 gC m-2 from 2009 to 2018. GWR model can reveal significant spatial heterogeneity in the impacts of LST on NPPind. Overall, our findings indicated that LST has a certain role in promoting urban NPP.

Effects of environmental norfloxacin concentrations on the intestinal health and function of juvenile common carp and potential risk to humans.

Antibiotics are emerging pollutants in our environment. These treatments have been widely used for their low cost, convenient use, and prominent effects. However, the prolonged or excessive use of such drugs can cause toxicity in aquatic organisms. These effects include genotoxicity, metabolic alteration, delayed development and decreased immunity, which carry further risks for ecological systems. In the present study, juvenile common carp (Cyprinus carpio) were exposed to norfloxacin (NOR) for 42 days, with NOR concentrations ranging from 100 ng/L to 1 mg/L, to assess the effects of environmental concentrations of antibiotics, to investigate the effects of NOR on intestinal morphology, enzymatic activity, and transcriptomic levels of RNA in fish, as well as a risk assessment on human health was carried out. The results demonstrated that oxidative stress was induced, the barrier function of the intestine was damaged, and changes occurred in the expression of immune-related genes in fish chronically exposed to antibiotics. Moreover, NOR could affect the regulation of the NF-κB signaling pathway. Thus, environmental concentrations of antibiotics can influence the intestinal health of fish and potentially posing health risks to humans.

Hepatotoxicity in carp (Cyprinus carpio) exposed to environmental levels of norfloxacin (NOR): Some latest evidences from transcriptomics analysis, biochemical parameters and histopathological changes.

Recently, the residues of quinolones have received widespread attention. However, toxicological studies on aquatic organisms are relatively scarce, especially on the liver metabolism and immune effects of these aquatic organisms. In this study, we investigated the toxic effects of carp exposed to 0, 100 ng/L, and 1 mg/L norfloxacin (NOR) at environmental concentrations for 42 days. In this study, through transcriptomics analysis, we found that some genes involved in lipid metabolism, immune response, and cytochrome P450 metabolism, especially genes accounting for the metabolism-related disorders of glucose and lipid. Defects in these genes and thus their related pathways increase the risk of coming down with nonalcoholic fatty liver disease. Compared to those of the control, results from the biochemical indicators of the treatment group changed significantly, including levels of total cholesterol, triglycerides, glucose, and insulin. Moreover, our results confirmed that NOR at environmental concentrations disordered the metabolism of glucose and lipid in the carp also resulted in hepatocellular and nuclear enlargement. Our results, therefore, confirmed that long-term exposure to NOR can induce carp liver toxicity at histological, biochemical, and transcriptional levels and provided the latest data and theoretical basis for the toxicology study of quinolones in the natural environment.

Living-Cell MicroRNA Imaging with Self-Assembling Fragments of Fluorescent Protein-Mimic RNA Aptamer.

As the cellular roles of RNA abundance continue to increase, there is an urgent need for the corresponding tools to elucidate native RNA functions and dynamics, especially those of short, low-abundance RNAs in live cells. Fluorescent RNA aptamers provide a useful strategy to create the RNA tag and biosensor devices. Corn, which binds with 3,5-difluoro-4-hydroxybenzylidene-imidazolinone-2-oxime (DFHO), is a good candidate for the RNA tag because of its enhanced photostability and red-shifted spectrum. Herein, we report for the first time the utilization of Corn as a split aptamer system, combined with RNA-initiated fluorescence complementation (RIFC), for monitoring RNA self-assembly and sensing microRNA. In this platform, the 28-nt Corn was divided into two nonfunctional halves (named probe I and probe II), and an additional target RNA recognition and stem part was introduced in each probe. The target RNA can trigger the self-assembly reconstitution of the Corn's G-quadruplex scaffold for DFHO binding and turn-on fluorescence. These probes can be transfected stably into mammalian cells and deliver the light-up fluorescent response to microRNA-21 (miR-21). Significantly, the probes have good photostability, with minimal fluorescence loss after continuous irradiation, and can be used for imaging of miR-21 in living mammalian cells. The proposed method is universal and could be applied to the sensing of other tumor-associated RNAs, including messenger RNA and noncoding RNA, as well as for monitoring RNA/RNA interactions. The Corn-based splitting aptamers show promising potential in the real-time visualization and mechanistic analysis of nucleic acids.

Clinical and molecular characterization of seven patients with Danon disease.

Danon disease is an X-linked glycogen storage disease characterized by skeletal myopathy, cardiomyopathy and intellectual impairment. It is caused by a loss-of-function mutation in the lysosome-associated membrane protein-2 (LAMP2) gene. In the present study, exon and boarding intron analysis of 96 cardio disease-associated genes was performed in 770 patients with hypertrophic cardiomyopathy (HCM) using second-generation sequencing. Next, the identified mutations were confirmed in family members of the patients and 300 healthy controls. Detailed clinical, electrocardiographic (ECG) and echocardiographic findings were recorded. A pathogenic mutation in LAMP2 was identified in 7 patients who phenotypically presented with HCM. A total of four patients had a fragmented QRS complex (fQRS) on surface ECG. In addition, two patients presented with ventricular preexcitation with a short PR interval. Compared with the patients with protein kinase AMP-activated non-catalytic subunit γ2 syndrome and Fabry disease, the 7 patients with Danon disease presented at an earlier age, had a smaller left atrial size, a thinner maximal left ventricular wall thickness and a lower probability of pacemaker implantation. Compared with 12 sex- and age-matched patients with sarcomere-protein mutations, the 4 patients with Danon disease had a lower left ventricular outflow tract gradient and worse diastolic function. The present study provided a comprehensive comparison of different pathologies presenting with HCM and reported on features of early-onset Danon disease, including the characteristic preexcitation and fQRS on ECG. This may provide valuable information that may be utilized for the early diagnosis and treatment of patients with Danon disease. The present study was registered as a clinical trial with ClinicalTrials.gov (Sep. 2, 2016; registry no. NCT02888132).

Comparative Genomics and Phylogenetic Analyses of Christia vespertilionis and Urariopsis brevissima in the Tribe Desmodieae (Fabaceae: Papilionoideae) Based on Complete Chloroplast Genomes.

Taxonomic and phylogenetic relationships of Christia, Urariopsis, Uraria and related genera within the tribe Desmodieae (Fabaceae: Papilionoideae) have long been controversial. Here, we report the complete chloroplast (cp) genomes of Christia vespertilionis and Urariopsis brevissima and perform comparative and phylogenetic analyses with Uraria lagopodioides and other relatives in the Desmodieae. The cp genomes of C. vespertilionis and U. brevissima are 149,656 and 149,930 bp long, with 128 unique genes (83 protein-coding genes, 37 tRNA genes and 8 rRNA genes), respectively. Comparative analyses revealed 95-129 simple sequence repeats (SSRs) and eleven highly variable regions (trnK-rbcL, rbcL-atpB, ndhJ-trnF, trnL-trnT, psbD-rpoB, accD-cemA, petA-psbL, psbE-petL, rps11-rps19, ndhF-ccsA, and rps15-ycf1) among six Desmodieae species. Phylogenetic analyses clearly resolved two subtribes (Desmodiinae and Lespedezinae) of Desmodieae as monophyletic, and the newly reported C. vespertilionis and U. brevissima clustered in subtribe Desmodiinae. A sister relationship of C. vespertilionis to U. lagopodioides was supported. Evidence was presented to support the treatment of Urariopsis as a distinct genus rather than in synonymy with Uraria. The results provide valuable information for further studies on species delimitation, phylogenetics, population genetics, and the evolutionary process of speciation in the Desmodieae.