A high-fidelity RNA-targeting Cas13 restores paternal Ube3a expression and improves motor functions in Angelman syndrome mice.

Angelman syndrome (AS) is a rare neurodevelopmental disorder caused by loss of function mutations in maternally expressed UBE3A. No gene-specific treatment is available for patients so far. Although intact and transcriptionally active, paternally inherited UBE3A is silenced by elongation of antisense long noncoding RNA UBE3A-ATS in neurons. Here, we demonstrated that RNA targeting of paternal Ube3a-ATS with a high-fidelity CRISPR-Cas13 (hfCas13x.1) system could restore Ube3a expression to similar levels as that of maternal Ube3a in the cultured mouse neurons. Furthermore, injection into lateral ventricles with neuron-specific hSyn1 promoter-driven hfCas13x.1 packaged in adeno-associated virus (AAV-PHP.eb) could restore paternal Ube3a expression in cortex and hippocampus of neonatal AS mice for up to 4 months after treatment. Behavioral tests showed that expression of paternal Ube3a significantly alleviated AS-related symptoms, including obesity and motor function. Our results suggested that hfCas13x.1-mediated suppression of the Ube3a-ATS lncRNA potentially serves as a promising targeted intervention for AS.

Inhibiting L1CAM Reverses Cisplatin Resistance of Triple Negative Breast Cancer Cells by Blocking AKT Signaling Pathway.

DDP-resistant MDA-MB-231 cells (MDA-MB-231/DDP) cells had higher expression of L1CAM than their parental cells. L1CAM siRNA decreased the IC50 of MDA-MB-231/DDP cells to DDP. L1CAM inhibition down-regulated p-AKT/AKT in MDA-MB-231/DDP cells; meanwhile, it could promote MDA-MB-231/DDP cell apoptosis, inhibit cell EMT, invasion, and migration. Moreover, SC79 (an AKT activator) increased the DDP-resistance of MDA-MB-231/DDP cells, which was reversed by L1CAM inhibition. Furthermore, co-treatment of L1CAM shRNA and cisplatin injection had better anti-tumor effects in vivo than these two single treatments with decreased p-AKT/AKT. Thus, silencing L1CAM reversed the DDP resistance by inhibiting the AKT pathway.

Simultaneous quantification of hepatitis A virus and norovirus genogroup I and II by triplex droplet digital PCR.

The representative enteric viruses responsible for global foodborne outbreaks that have become an essential concern for health authorities are Norovirus (NoV) and Hepatitis A virus (HAV). Droplet digital PCR (ddPCR) has recently emerged as an alternative platform for virus quantification due to its high precision, ultra-sensitivity, and lack of a standard curve need. Using a ratio-based probe-mixing strategy, we established a triplex ddPCR method to detect norovirus genogroup I (GI), genogroup II (GII), and HAV in food, drinking water, and faecal samples. The probe concentration, annealing temperature, and annealing/extension time were all tuned in the PCR amplification program. The detection limit for NoV GI, NoV GII, and HAV was 7.5, 5.0, and 5.0 copies/reaction, respectively. Furthermore, the suggested approach was validated on 114 samples, demonstrating greater sensitivity, accuracy, and anti-interference performance features than RT-qPCR.

[Hereditary vitreous degeneration muddy: report of ten cases].

Hereditary vitreous degeneration muddy is rare in clinic. Here we report ten cases (thirteen eyes) of hereditary vitreous degeneration muddy from two families. All patients presented with vitreous opacity, and the textures appeared tough and tensile. Two cases had concurrent detachment of rhegmatogenous retina. HE staining showed red changeableness, and methyl violet staining appeared purple. All patients received vitrectomy with traditional Chinese medicine treatment, and got satisfactory efficacy.

Phylogenetic analysis of Cyprinus acutidorsalis (Wang, 1979) from the Hainan population using complete mitochondrial genome.

Cyprinus acutidorsalis (Wang, 1979) is an endemic fish in China that is sparsely distributed in the Hainan provinces and Guangxi Zhuang Autonomous Region (GZAR). In this study, the complete mitochondrial genome of C. acutidorsalis from the Hainan population from the Wanquan River was sequenced, and its phylogenetic relationship was analyzed. The circular mtDNA was 16,581 bp in length, and the overall base composition was A (32.0%), C (27.5%), T (24.8%), and G (15.70%), with a slight bias toward A + T. The complete mitogenome encoded 13 protein-coding genes (PCGs), 22 tRNA genes, two rRNA genes, and a control region. Phylogenetic analysis indicated that the most closely related fish to C. acutidorsalis from the Hainan population was C. acutidorsalis from the Guangxi population. These findings offer basic molecular data and a better understanding of the phylogenetic relationships among the Cyprinus species.

Dietary Astragalus polysaccharides improve the growth and innate immune response of giant freshwater prawn Macrobrachium rosenbergii: Insights from the brain-gut axis.

Supplementation with Astragalus polysaccharides (APS) has beneficial effects on aquatic animals. Herein, we aimed to investigate the effects of different doses of APS on the growth, innate immune response, and brain-gut axis of Macrobrachium rosenbergii. The molecular weight and the monosaccharide composition of APS were analyzed. APS were added at concentration of 0 (control), 0.05, 0.10, 0.15, and 0.20 % in practical diets. Growth performance increased significantly under 0.05 to 0.20 % APS, with enhanced lipase and protease activities in intestinal tissues. Prawns receiving APS supplementation had significantly lower amounts of pathogenic intestinal bacteria (Vibrio and Aeromonas) and a markedly different microbial community structure compared with those of the control group. The fecal short chain fatty acid (SCFA) and neurotransmitters γ-aminobutyric acid contents increased in the brains of prawns receiving APS, which was potentially associated with increased Lactobacillus and Bacillus levels. Prawns receiving APS supplementation displayed a significantly enhanced immune function (such as total hemocyte count, total protein concentration, phenoloxidase activity, serum agglutination titer, and lysozyme activity) and improved disease resistance to Vibrio anguillarum compared those in the control group. Thus, dietary APS positively affected the gut-brain axis by altering the microbiota composition, increasing the fecal SCFA content, and enhancing prawn immunity.

The complete mitochondrial genome of a new geographical population of freshwater fish Macropodus hongkongensis (Freyhof & Herder, 2002).

Macropodus hongkongensis (Freyhof & Herder, 2002), is sparsely distributed in Hong Kong and Guangdong provinces. Recently, a new geographical population of M. hongkongensis was discovered in the Wanquan River in the Hainan province. Therefore, this study focused on sequencing the complete mitochondrial genome of the new geographical population of Macropodus hongkongensis from the Wanquan River. The circular mtDNA molecule was 16,492 bp in size, and the overall base compositions were A (30.30%), C (24.90%), T (29.80%), and G (15.00%), with a slight bias toward A + T. The complete mitogenome encoded 13 protein-coding genes (PCGs), 22 tRNA genes, 2 rRNA genes, and a control region. Phylogenetic analysis indicated that M. hongkongensis of the Hainan Wanquan River was most closely related to the M. hongkongensis of the Gongdong population. These results provide useful genetic information for species identification and phylogenetic studies of Macropodus species.

The AKT inhibitor, MK-2206, attenuates ABCG2-mediated drug resistance in lung and colon cancer cells.

Introduction: The overexpression of ATP-binding cassette (ABC) transporters, ABCB1 and ABCG2, are two of the major mediators of multidrug resistance (MDR) in cancers. Although multiple ABCB1 and ABCG2 inhibitors have been developed and some have undergone evaluation in clinical trials, none have been clinically approved. The compound, MK-2206, an inhibitor of the protein kinases AKT1/2/3, is undergoing evaluation in multiple clinical trials for the treatment of certain types of cancers, including those resistant to erlotinib. In this in vitro study, we conducted in vitro experiments to determine if MK-2206 attenuates multidrug resistance in cancer cells overexpressing the ABCB1 or ABCG2 transporter. Methodology: The efficacy of MK-2206 (0.03-1 µM), in combination with the ABCB1 transporter sub-strates doxorubicin and paclitaxel, and ABCG2 transporter substrates mitoxantrone, SN-38 and topotecan, were determined in the cancer cell lines, KB-C2 and SW620/Ad300, which overexpress the ABCB1 transporter or H460/MX20 and S1-M1-80, which overexpress the ABCG2 transporter, respectively. The expression level and the localization of ABCG2 transporter on the cancer cells membranes were determined using western blot and immunofluorescence assays, respectively, following the incubation of cells with MK-2206. Finally, the interaction between MK-2206 and human ABCG2 transporter was predicted using computer-aided molecular modeling. Results: MK-2206 significantly increased the efficacy of anticancer compounds that were substrates for the ABCG2 but not the ABCB1 transporter. MK-2206 alone (0.03-1 µM) did not significantly alter the viability of H460/MX20 and S1-M1-80 cancer cells, which overexpress the ABCG2 transporter, compared to cells incubated with vehicle. However, MK-2206 (0.3 and 1 µM) significantly increased the anticancer efficacy of mitoxantrone, SN-38 and topotecan, in H460/MX20 and S1-M1-80 cancer cells, as indicated by a significant decrease in their IC50 values, compared to cells incubated with vehicle. MK-2206 significantly increased the basal activity of the ABCG2 ATPase (EC50 = 0.46 µM) but did not significantly alter its expression level and sub-localization in the membrane. The molecular modeling results suggested that MK-2206 binds to the active pocket of the ABCG2 transporter, by a hydrogen bond, hydrophobic interactions and π-π stacking. Conclusion: These in vitro data indicated that MK-2206 surmounts resistance to mitoxantrone, SN-38 and topotecan in cancer cells overexpressing the ABCG2 transporter. If these results can be translated to humans, it is possible that MK-2206 could be used to surmount MDR in cancer cells overexpressing the ABCG2 transporter.

Thyroid Cancer and COVID-19: Prospects for Therapeutic Approaches and Drug Development.

Thyroid cancer is the most prevalent endocrine malignancy and the reported incidence of thyroid cancer has continued to increase in recent years. Since 2019, coronavirus disease 2019 (COVID-19) has been spreading worldwide in a global pandemic. COVID-19 aggravates primary illnesses and affects disease management; relevant changes include delayed diagnosis and treatment. The thyroid is an endocrine organ that is susceptible to autoimmune attack; thus, thyroid cancer after COVID-19 has gradually attracted attention. Whether COVID-19 affects the diagnosis and treatment of thyroid cancer has also attracted the attention of many researchers. This review examines the literature regarding the influence of COVID-19 on the pathogenesis, diagnosis, and treatment of thyroid cancer; it also focuses on drug therapies to promote research into strategies for improving therapy and management in thyroid cancer patients with COVID-19.

Factors affecting microplastic accumulation by wild fish: A case study in the Nandu River, South China.

Quantifying microplastic accumulation in aquatic organisms is important for risk assessments in natural habitats. However, studies on the factors influencing microplastic accumulation by fish, particularly in natural water bodies, remain limited. In this study, we monitored the microplastic pollution characteristics in the fish digestive tract and surface water of the Nandu River, the largest river in Hainan, China. Among the 179 fish examined, microplastics were detected in 90.5 %, covering 27 fish species. Moreover, low abundance of microplastics occurred among the different fish, with an average of 3.20 ± 2.60 particles per fish. Fibrous microplastics with a large size (>500 µm) were dominant in both the water column and fish. In this study, fish size, feeding habits, and living habitats did not affect microplastic abundance in fish. Based on the analysis of differences in the abundance and morphological characteristics of microplastics in water column and fish, we suggested that the random intake and differential excretion of microplastics with different characteristics by fish might lead to the homogeneity of microplastic accumulation in different fish. Our study suggest that more attention should be given to the negative effects of microplastics on small fish owing to the higher microplastic exposure per gram weight, and further improvement in microplastic risk assessment methods is required.

Species-specific bioaccumulation and health risk assessment of heavy metal in seaweeds in tropic coasts of South China Sea.

Seaweeds are widely known superfood in coasts where most anthropogenic heavy metal discharges are inputted and stored. The present study analyzed 11 seaweed species and 13 heavy metals to test the hypothesis that the species-specific capacity of heavy metal bioaccumulation had great significance to health risk of human. The seaweeds were collected from tropic coasts of Hainan Island. We comparatively determined the bioaccumulation level of metals in different species. The results revealed that the red algae mainly concentrated V, Se, Mn, Ni, and Ag. The brown algae mainly concentrated Cr, Co, Cu, Cd, As and Fe, while the green algae mainly concentrated Zn and Pb. The cluster analysis, principal component analysis and metal pollution index indicated that Padina crassa, Sargassum thunbergii, Caulerpa racemosa and Asparagopsis taxiformis showed similar metal bioaccumulation behavior. The health risk assessment revealed that the overall hazard index (HI) of seaweeds consumption to adults was less than 1, while the HI of Sargassum oligocystum, Turbinaria ornate, Sargassum polycystum and Sargassum thunbergii consumption to children was greater than 1, suggesting a moderate or high risk to children. Moreover, the exposure amount and the carcinogenic risk parameter indicated that As and Cr were the limiting factor for seaweeds consumption. Overall, our findings here largely supported our hypothesis that the heavy metal bioaccumulation behavior and health risk was highly variable and complex among different species. We thus suggested that the species-specific health risk of heavy metals in seaweeds should be cautiously evaluated in natural environments.

Complete mitochondrial genome of Vanmanenia hainanensis (Cypriniformes: Gastromyzontidae).

Vanmanenia hainanensis Chen & Zheng 1980 is endemic to Hainan Island, China. The complete mitogenome of the species was sequenced in this study. It was 16,555 bp in length, containing 13 protein-coding genes (PCGs), 22 tRNA genes, 2 rRNA genes, and 1 control region. The base composition was 29.5% A, 25.4% T, 16.7% G, and 28.4% C. All genes were encoded on the H-strand except for ND6 and 8 tRNA genes, located on the l-strand. Phylogenetic analysis based on 13 protein-coding genes indicated that the genus Vanmanenia did not form monophyly and it had the closest relationship with Formosania. This study aimed at providing useful genetic information for future studies on taxonomy, phylogeny, and evolution of Vanmanenia species.

The complete mitogenome of Liniparhomaloptera qiongzhongensis (Cypriniformes: Gastromyzontidae) and phylogenetic implications.

Liniparhomaloptera qiongzhongensis Zheng and Chen 1980 is distributed in Hainan Island, China. The complete mitogenome of the species was sequenced in this study. It had a circular molecule of 16,554 bp in length with a total of 56.5% A + T content, containing 13 protein-coding genes (PCGs), 22 tRNA genes, two rRNA genes, and one control region. Five overlapping regions and 14 intergenic sequences were observed with the length of 2-10 bp and 1-31 bp, respectively. The phylogenetic analysis based on 13 protein-coding genes revealed that L. qiongzhongensis had the closest relationship with L. disparis. This study would provide useful genetic information for future studies on taxonomy, phylogeny and evolution of Liniparhomaloptera species.

A new long-legged terrestrial freshwater crab, Calcipotamon puglabrum gen. nov. et sp. nov. (Crustacea: Decapoda: Potamidae), from Hainan Island, China.

A new genus and new species of terrestrial freshwater crab, Calcipotamon puglabrum gen. nov. et sp. nov., is described from the limestone forests of Changjiang, Hainan Island, China, based on morphology and mitochondrial 16S rDNA sequences. The new genus is closest to Neotiwaripotamon Dai Naiyanetr, 1994, and Tiwaripotamon Bott, 1970, but differs in a combination of carapace, third maxilliped, ambulatory leg and male gonopod characters. Molecular analysis shows that the new genus is closely related with but not clustered within other Hainan potamid genera. Notes on the general biology of this new species are also provided.

Complete chloroplast genome sequence of an endangered Ottelia cordata and its phylogenetic analysis.

Ottelia cordata is a typical submerged plant to the family of hydrocharitaceae. In this work, complete chloroplast genome of O. cordata was assembled and characterized from high-throughput Illumina sequencing data. The chloroplasgenome of O. cordatais 157,869bp in length. The O. cordata cp genome encodes 128 functional genes, including 85 protein-coding genes, 35 tRNA genes, and 8 rRNA genes. The GC content of the genome was 36,57%. Its nucleotide composition is asymmetric with an overall A + T content of 63.44%. Phylogenetic analysis indicated that O. cordata is closely related to the Elodea. These results could offer molecular markers and helpful in understanding about the role of O. cordata evolution and protection.

Linc00426 accelerates lung adenocarcinoma progression by regulating miR-455-5p as a molecular sponge.

Increasing lines of evidence indicate the role of long non-coding RNAs (LncRNAs) in gene regulation and tumor development. Hence, it is important to elucidate the mechanisms of LncRNAs underlying the proliferation, metastasis, and invasion of lung adenocarcinoma (LUAD). We employed microarrays to screen LncRNAs in LUAD tissues with and without lymph node metastasis and revealed their effects on LUAD. Among them, Linc00426 was selected for further exploration in its expression, the biological significance, and the underlying molecular mechanisms. Linc00426 exhibits ectopic expression in LUAD tissues and cells. The ectopic expression has been clinically linked to tumor size, lymphatic metastasis, and tumor differentiation of patients with LUAD. The deregulation of Linc00426 contributes to a notable impairment in proliferation, invasion, metastasis, and epithelial-mesenchymal transition (EMT) in vitro and in vivo. Mechanistically, the deregulation of Linc00426 could reduce cytoskeleton rearrangement and matrix metalloproteinase expression. Meanwhile, decreasing the level of Linc00426 or increasing miR-455-5p could down-regulate the level of UBE2V1. Thus, Linc00426 may act as a competing endogenous RNA (ceRNA) to abate miR-455-5p-dependent UBE2V1 reduction. We conclude that Linc00426 accelerates LUAD progression by acting as a molecular sponge to regulate miR-455-5p, and may be a potential novel tumor marker for LUAD.

Alterations of the gut microbiome of largemouth bronze gudgeon (Coreius guichenoti) suffering from furunculosis.

High-throughput sequencing was applied to compare the intestinal microbiota in largemouth bronze gudgeon either healthy or affected by furunculosis. Proteobacteria, Actinobacteria, Tenericutes, Firmicutes and Bacteroidetes were detected as the predominant bacterial phyla in the gut of both diseased and healthy fish. The abundance of Proteobacteria differed significantly between the two groups of fish, mainly due to the overwhelming prevalence of Aeromonas in the diseased fish (81% ± 17%), while the genus was unevenly spread among the apparently healthy fish (33% ± 33%). The bacterial diversity in the intestine of diseased fish was markedly lower than in healthy fish. Analysis revealed the significant dissimilarity between the gut microbiota of diseased and healthy fish. The bacterial profiles in the gut were further characterized with the 28 phylotypes that were shared by the two groups. In diseased fish, two shared OTUs (OTU0001 and OTU0013) were closely related to Aeromonas salmonicida, their total proportion exceeding 70% of the sequences in diseased fish, while averaging 5.2% ± 4.6% in the healthy fish. This result suggested the presence of healthy carriers of pathogenic A. salmonicida among the farmed fish, and the gut appeared as a probable infection source for furunculosis in largemouth bronze gudgeon.

Transcriptomic and proteomic analyses of splenic immune mechanisms of rainbow trout (Oncorhynchus mykiss) infected by Aeromonas salmonicida subsp. salmonicida.

Furunculosis caused by Aeromonas salmonicida subsp. salmonicida is an epidemic disease among salmonids, including rainbow trout (Oncorhynchus mykiss). However, the immune mechanisms that are elicited in rainbow trout against the invasion of A. salmonicida are not yet fully understood. In this study, we examined the spleen to investigate the immune response of rainbow trout at 3days post-infection by A. salmonicida at the transcriptome and proteome levels by using Illumina-seq and iTRAQ methods, respectively. A total of 1036 genes and 133 proteins were found to undergo differential expression during the immune response of the spleen against A. salmonicida infection. Gene ontology and KEGG analysis were conducted among the differentially expressed genes and proteins, revealing that immune system process and response to stimulus were the top two biological processes, and immune system, signaling molecules and interaction, and immune diseases were the differential pathways activated. Correlation analysis of transcriptomic and proteomic results showed 17 proteins (11 upregulated and 6 downregulated) having consistent expression at RNA and protein levels. Moreover, protein-protein interaction analysis showed that diseases, proteasome, aminoacyl-tRNA biosynthesis, and nucleotide metabolism were the main interactions among the consistently expressed proteins. Consequently, these upregulated proteins, namely, ferritin, CD209, IL13Rα1, VDAC2, GIMAP7, PSMA1, and two ANXA11s could be considered as potential biomarkers for rainbow trout immune responses. BIOLOGICAL SIGNIFICANCE: This study provides the first identification of immune markers through an analysis of the differential expression of both genes and their corresponding protein products in the spleen of rainbow trout after infection by A. salmonicida, shedding light on the molecular mechanisms triggered in rainbow trout against A. salmonicida infection and providing new molecular targets for further immunological research in fish.

Sphingosine 1-phosphate (S1P) promotes mitochondrial biogenesis in Hep G2 cells by activating Peroxisome proliferator-activated receptor γ coactivator 1α (PGC-1α).

Sphingosine 1-phosphate (S1P), a potent bioactive phospholipid, has been reported to regulate a broad spectrum of biological processes. However, little is known regarding S1P's effects on mitochondrial function. In this study, we investigated the S1P's effects on the Peroxisome proliferator-activated receptor γ coactivator 1α (PGC-1α) signaling pathway and mitochondrial biogenesis in Hep G2 cells. Our results indicate that administration of S1P leads to a significant upregulation of mitochondrial DNA replication and transcription, increased mitochondrial mass, and elevated adenosine triphosphate synthesis. In addition, we found that treatment with S1P stimulates expression of PGC-1α, a master regulator of mitochondrial biogenesis, as well as its downstream targets: nuclear respiratory factor 1 (NRF1) and mitochondrial transcription factor A (TFAM). Moreover, our data demonstrate that S1P's effects on PGC-1α and mitochondrial biogenesis are mediated by the protein kinase A/cAMP response element-binding protein (PKA/CREB) pathway. Importantly, we also revealed that S1P's effects on mitochondrial biogenesis are dependent on its type 2 receptor (S1P2), though not on either its type 1 (S1P1) or type 3 (S1P3) receptors. Based on these observations, we concluded that S1P activates the PKA/CREB pathway through S1P2, which then promotes expression of PGC-1α/NRF1/TFAM and subsequent mitochondrial biogenesis in Hep G2 cells.

Complete mitochondrial genomes of two relatively closed species from Gymnocypris (Cypriniformes: Cyprinidae): genome characterization and phylogenetic considerations.

The complete mitochondrial genome for Gymnocypris eckloni and Gymnocypris przewalskii ganzihonensis is 16,686 and 16,682 bp in size, respectively. Both of the mitochondrial genomes have identical genomic organization and structure including 13 protein-coding genes (PCGs), 22 tRNA genes, 2 rRNA genes and 1 control region as found in other vertebrates, with the exception of 86 bp non-coding nucleotides between tRNA(Thr) and tRNA(Pro). The phylogenetic analyses based on the concatenated nucleotide sequence of 12 PCGs on the heavy strand supported a relatively closed relationship among G. eckloni, G. p. przewalskii and G. p. ganzihonensis. The sequence divergence between G. eckloni and G. przewalskii, as well as within G. przewalskii, is relatively low compared to the average sequence divergence among other cyprinid fishes, suggesting that they are the most closely related species resulted from a relatively recent speciation event. The estimated molecular divergence time is coincided with a relatively recent speciation event occurring during the separation of the Yellow River and Qinghai Lake.