Molecular and phenotypic characteristics of Bardet-Biedl syndrome in Chinese patients.

BACKGROUND: Bardet-Biedl syndrome (BBS) is a type of non-motile ciliopathy. To date, 26 genes have been reported to be associated with BBS. However, BBS is genetically heterogeneous, with significant clinical overlap with other ciliopathies, which complicates diagnosis. Disability and mortality rates are high in BBS patients; therefore, it is urgent to improve our understanding of BBS. Thus, our study aimed to describe the genotypic and phenotypic spectra of BBS in China and to elucidate genotype-phenotype correlations. METHODS: Twenty Chinese patients diagnosed with BBS were enrolled in this study. We compared the phenotypes of Chinese BBS patients in this study with those from other countries to analyze the phenotypic differences across patients worldwide. In addition, genotype-phenotype correlations were described for our cohort. We also summarized all previously reported cases of BBS in Chinese patients (71 patients) and identified common and specific genetic variants in the Chinese population. RESULTS: Twenty-eight variants, of which 10 are novel, in 5 different BBS-associated genes were identified in 20 Chinese BBS patients. By comparing the phenotypes of BBSome-coding genes (BBS2,7,9) with those of chaperonin-coding genes (BBS10,12), we found that patients with mutations in BBS10 and 12 had an earlier age of onset (1.10 Vs. 2.20, p < 0.01) and diagnosis (4.64 Vs. 13.17, p < 0.01), whereas patients with mutations in BBS2, 7, and 9 had a higher body mass index (28.35 Vs. 24.21, p < 0.05) and more vision problems (p < 0.05). Furthermore, in 91 Chinese BBS patients, mutations were predominant in BBS2 (28.89%) and BBS7 (15.56%), and the most frequent variants were in BBS2: c.534 + 1G > T (10/182 alleles) and BBS7: c.1002delT (7/182 alleles), marking a difference from the genotypic spectra of BBS reported abroad. CONCLUSIONS: We recruited 20 Chinese patients with BBS for genetic and phenotypic analyses, and identified common clinical manifestations, pathogenic genes, and variants. We also described the phenotypic differences across patients worldwide and among different BBS-associated genes. This study involved the largest cohort of Chinese patients with BBS, and provides new insights into the distinctive clinical features of specific pathogenic variants.

Microalgae as fishmeal alternatives in aquaculture: current status, existing problems, and possible solutions.

Fishmeal is an indispensable ingredient for most aquatic animals. However, the finite supply and escalating price of fishmeal seriously limit its use in aquaculture. Thus the development of new, sustainable protein ingredients has been a research focus. Microalgae are potential fishmeal alternatives owing to their high protein content and balanced amino acid profile. Studies suggest that suitable replacement of fishmeal with microalgae is beneficial for fish growth performance, but excessive replacement would induce poor growth and feed utilization. Therefore, this paper aims to review research on the maximum substitutional level of fishmeal by microalgae and propose the main issues and possible solutions for fishmeal replacement by microalgae. The maximum replacement level is affected by microalgal species, fish feeding habits, quality of fishmeal and microalgal meals, and supplemental levels of fishmeal in the control group. Microalgae could generally replace 100%, 95%, 95%, 64.1%, 25.6%, and 18.6% fishmeal protein in diets of carp, shrimp, catfish, tilapia, marine fish, and salmon and trout, respectively. The main issues with fishmeal replacement using microalgae include low production and high production cost, poor digestibility, and anti-nutritional factors. Possible solutions to these problems are recommended in this paper. Overall, microalgae are promising fishmeal alternatives in aquaculture.

Waterborne copper exposure decreases fish growth and survival by promoting gills and liver impairments in largemouth bass (Micropterus salmoides).

The study aimed to investigate the effect of Cu exposure (0, 51.3, 164, 513, 1,640, and 5,130 µg/L) on fish growth performance, histology, oxidative stress, inflammation, and apoptosis in largemouth bass (Micropterus salmoides) juveniles. 270 fish (2.69 ± 0.02 g) were randomly divided into 6 groups of tanks for 4 weeks with each group comprising three replicate tanks. The results showed that fish exposed to 1,640 and 5,130 µg/L Cu exhibited a significant reduction in fish growth and survival rate (P < 0.05). Compared to the control, the fish at and above 513 µg/L Cu demonstrated histopathological damages in the gills and liver, such as shorter primary and secondary lamellae, smaller hepatocyte nuclei, and an increase in the number of necrotic cells in the liver. Compared to the control, fish at and above 1,640 µg/L Cu had a significantly higher malondialdehyde content and lower activity levels of total superoxide dismutase, glutathione peroxidase, and catalase in the gills and liver (P < 0.05). Furthermore, high concentrations of Cu (1,640 and 5,130 µg/L) significantly increased hepatic inflammation by upregulating interleukin-1ß and tumor necrosis factor α expression and hepatic apoptosis by increasing cysteinyl aspartate specific protease 3 (caspase-3) and caspase-9 expression (P < 0.05). Pearson correlation analysis showed that fish growth and survival positively correlated with histological and antioxidant defense parameters, and negatively correlated with oxidative stress parameters, hepatic inflammation, and hepatic apoptosis. Taken together, these results suggest that high levels of waterborne Cu can induce growth retardation and mortality by damaging the liver and gill health.

Molecular and phenotypic spectrum of cardio-facio-cutaneous syndrome in Chinese patients.

BACKGROUND: Cardio-facio-cutaneous (CFC) syndrome is a RASopathy subtype that presents with unique craniofacial dysmorphology, congenital heart disease, dermatologic abnormalities, growth retardation, and intellectual disability. This study describes the phenotypic spectrum of CFC in China and its association with CFC syndrome gene variants. RESULTS: Twenty Chinese CFC patients, aged 0.6-9.5 years old, were included in this study and their clinical phenotypic spectrum was compared with that of 186 patients with CFC from non-Chinese ethnicities. All 20 Chinese patients with CFC carried de novo heterozygous BRAF, MAP2K1, and MAP2K2 variants. Two novel variants were detected and consistently predicted to be deleterious using bioinformatic tools. The clinical features of CFC in the Chinese patients included hypertrophic cardiomyopathy (2/20, 10%), pulmonary valve stenosis (2/20, 10%), curly or sparse hair (7/20, 35%), epilepsy (1/20, 5%), and hypotonia (10/20, 50%); these features were less frequently observed in Chinese patients than non-Chinese patients (p < 0.05). In contrast, feeding difficulties (19/20, 95%) were more frequently observed in the Chinese patients. Absent eyebrows and severe short stature were more common in patients with BRAF variants than in those with MAP2K1/2 variants. Facial recognition software was used to recognize most CFC patients using artificial intelligence. CONCLUSION: This study identified novel and common variants in our cohort of 20 Chinese patients with CFC. We uncovered differences in clinical features between Chinese and non-Chinese patients and detected genotype-phenotype correlations among the BRAF and MAP2K1/2 variant subgroups. This is the largest cohort of Chinese CFC patients to our knowledge, providing new insights into a subtype of RASopathy.

A novel heterozygous variant of FOXJ1 in a Chinese female with primary ciliary dyskinesia and hydrocephalus: A case report and literature review.

BACKGROUND: Primary ciliary dyskinesia (PCD) is a type of ciliary dyskinesia that is usually caused by autosomal recessive inheritance and can manifest as recurrent respiratory infections, bronchiectasis, infertility, laterality defects, and chronic otolaryngological disease. Although ependymal cilia, which affect the flow of cerebrospinal fluid in the central nervous system, have much in common with respiratory cilia in terms of structure and function, hydrocephalus is rarely associated with PCD. Recently, variants of Forkhead box J1 (FOXJ1) have been found to cause PCD combined with hydrocephalus in a de novo, autosomal dominant inheritance pattern. METHODS: We performed DNA extraction, whole-exome sequencing (WES) analysis, and mutation analysis of FOXJ1 and analyzed the patient's clinical and genetic data. RESULTS: The patient was a 4-year-old female exhibiting normal growth and development. At 3 years and 2 months of age, the patient experienced hand shaking and weakness in the lower limbs. Cardiac ultrasonography showed a right-sided heart, and cranial magnetic resonance imaging showed obstructive hydrocephalus. The nasal nitric oxide level was 54 nL/min. WES indicated a de novo, heterozygous variant of FOXJ1, c.734-735 ins20. This variant was novel, not included in the Human Gene Mutation and Genome Aggregation Database, and likely pathogenic according to the American College of Medical Genetics and Genomics, causing earlier termination of amino acid translation. The patient underwent a neuroendoscopic third ventriculostomy after the diagnosis of obstructive hydrocephalus. Six months after the operation, the patient's motor deficits had improved. CONCLUSION: This is the first report of a de novo, autosomal dominant pattern of FOXJ1 causing PCD combined with hydrocephalus in China. The patient's clinical symptoms were similar to those previously reported. WES confirmed that a novel variant of FOXJ1 was the cause of the PCD combined with hydrocephalus, expanding the spectrum of the genotypes associated with this condition. Physicians should be aware of the correlation of hydrocephalus and PCD and test for FOXJ1 variants.

Biological functions of circRNAs and their advance on skeletal muscle development in bovine.

The development of skeletal muscle in animals is a complex biological process, which are strictly and precisely regulated by many genes and non-coding RNAs. Circular RNA (circRNA) was found as a novel class of functional non-coding RNA with ring structure in recent years, which appears in the process of transcription and is formed by covalent binding of single-stranded RNA molecules. With the development of sequencing and bioinformatics analysis technology, the functions and regulation mechanisms of circRNAs have attracted great attention due to its high stability characteristics. The role of circRNAs in skeletal muscle development have been gradually revealed, where circRNAs were involved in various biological processes, such as proliferation, differentiation, and apoptosis of skeletal muscle cells. In this review, we summarized the current studies advance of circRNAs involved in skeletal muscle development in bovine, and hope to gain a deeper understanding of the functional roles of the circRNAs in muscle growth. Our results will provide some theoretical supports and great helps for the genetic breeding of this species, and aiming at improving bovine growth and development and preventing muscle diseases.

Effects of dietary phosphorus deficiency on the growth performance, hepatic lipid metabolism, and antioxidant capacity of Yellow River Carp Cyprinus carpio haematopterus.

OBJECTIVE: The study aimed to evaluate the effects of phosphorus (P) deficiency in diets on growth performance, hepatic lipid metabolism, and antioxidant capacity in Yellow River Carp Cyprinus carpio haematopterus. METHODS: In this study, 72 healthy experimental fish (initial weight = 12.0 ± 0.1 g [mean ± SE]) were randomly selected and distributed to two groups, with three replicates in each group. The groups were fed either a P-sufficient diet or a P-deficient diet for 8 weeks. RESULT: The P-deficient feed significantly decreased the specific growth rate, feed efficiency, and condition factor of Yellow River Carp. Fish that were fed the P-deficient feed demonstrated higher contents of triglyceride, total cholesterol (T-CHO), and low-density lipoprotein cholesterol in the plasma and a higher T-CHO content in the liver compared to the P-sufficient diet group. In addition, the P-deficient diet significantly reduced the catalase activity level, decreased the glutathione content, and increased the malondialdehyde content in the liver and in the plasma. Furthermore, P deficiency in the diet significantly downregulated the messenger RNA expression of nuclear erythroid 2-related factor 2 and peroxisome proliferator-activated receptor α, whereas it upregulated the messenger RNA expression of tumor necrosis factor α and fatty acid synthase in the liver. CONCLUSION: Dietary P deficiency reduced fish growth performance, induced fat deposition and oxidative stress, and impaired liver health.

Current status of industrialized aquaculture in China: a review.

Industrialized aquaculture is an essential trend for aquaculture development in China, owing to its considerable advantages in lower water consumption, higher productivity, and sustainability. However, information on its current status has been scarce up to now. This paper reviewed the current status and has identified existing problems as well as proposing possible solutions for the development of industrialized aquaculture in China. This field is still at an early stage of development and is mainly distributed in coastal regions. Major constraints on industrialized aquaculture include high capital and operational costs, the uncompetitive market price of aquatic products, uneven distribution of production and farming areas, a lack of suitably experienced managers and operators for recirculating aquaculture systems, and the coronavirus disease 2019 (COVID-19) pandemic. Possible solutions to these problems include technological innovations in systems optimization, the use of renewable energy sources and biofloc technology, the pollution-free certification of industrial aquaculture products, increased numbers of professionals in water quality control and waste management, and the financial assistance to companies and farmers along the aquaculture industrial chain.

Guar gum improves growth performance, intestinal microbiota homeostasis, and hepatic lipid metabolism in juvenile largemouth bass (Micropterus salmoides) fed high-fat diets.

The study aimed to investigate the effects of guar gum on the growth performance, gut microbiota composition, and hepatic lipid metabolism of largemouth bass (Micropterus salmoides) fed high-fat diets. Experimental fish were fed a normal-fat diet (Control), high-fat diet (HF), or HF diets supplemented with 0.3 %, 1 %, and 3 % guar gum (GG0.3, GG1, and GG3, respectively) for eight weeks. The results showed that HF significantly decreased fish growth performance, increased hepatic lipid accumulation, upregulated the expression of sterol regulatory element binding proteins 1 (SREBP1), and downregulated the expression of liver X receptor alpha (LXRα), cytochrome P450 7A1 (CYP7A1), and CYP8B1, compared to Control. However, these problems of high-fat diets were significantly alleviated by GG 0.3. The intestinal microbial communities of the GG0.3 and Control were similar but distinctly different from that of the HF group. Compared to HF, GG0.3 significantly increased the relative abundances of Firmicutes and Lactococcus and decreased the relative abundance of Tenericutes, Mesomycoplasma, and Phenylobacterium. In addition, the GG0.3 and GG1 treatments significantly enhanced the bile salt hydrolase (BSH) activity in the digesta compared to HF. In conclusion, 0.3 % guar gum supplementation can improve growth performance, intestinal health, and hepatic lipid metabolism in fish fed high-fat diets.

Addition of Chlorella sorokiniana meal in the diet of juvenile rainbow trout (Oncorhynchus mykiss): Influence on fish growth, gut histology, oxidative stress, immune response, and disease resistance against Aeromonas salmonicida.

This study aimed to assess the effects of dietary addition with Chlorella sorokiniana on fish growth, gut histology, antioxidant capacity, immune response, and disease resistance in rainbow trout. Three diets with similar proximate composition and different Chlorella meal levels were formulated. The control diet, 5% Chlorella diet, and 10% Chlorella diet contained 0%, 5% Chlorella meal, and 10% Chlorella meal, respectively. Each diet was assigned to triplicate tanks containing 30 fish (165.3 ± 0.6 g) in each tank. Fish were fed experimental diets for ninety days. The results showed that the addition of 5% Chlorella in the diet significantly increased feed intake by 19.3% and weight gain rate by 17.3% (P < 0.05) without affecting feed efficiency and gut histology. Diets containing Chlorella meal significantly decreased malonaldehyde contents in the plasma after the lipopolysaccharide (LPS) challenge (P < 0.05). Dietary supplementation with Chlorella meal significantly increased lysozyme (LZM) activity levels (in the head kidney) and immunoglobulin M (IgM) (in the head kidney) and complement component 3 (C3) (in the spleen) contents before the LPS challenge, and simultaneously increased LZM activity levels (in the plasma) and C3 contents (in the plasma and head kidney) after the LPS challenge (P < 0.05). Furthermore, dietary administration of Chlorella meal significantly increased the survival rate of fish infected with Aeromonas salmonicida (P < 0.05). In conclusion, C. sorokiniana can be used to improve fish growth, antioxidant capacity, and immunity.

Identification and functional analysis of miRNAs in skeletal muscle of juvenile and adult largemouth bass, Micropterus salmoides.

MicroRNAs (miRNAs) are considered key regulators to post-transcriptionally regulate gene expression affecting multiple biological activities. However, the developmental process of fish skeletal muscles is regulated by complicated molecular mechanism that has not been completely well-described. In this study, two small RNAs libraries from skeletal muscle of juvenile as well as adult largemouth bass (LMB) were obtained and sequenced using deep sequencing to investigate the development-related miRNAs. We identified an overall number of 486 already recognized miRNAs in addition to 43 novel miRNAs. Comparison of two different skeletal muscle development stages led to the identification of 220 differently expressed miRNAs between juvenile and adult LMB containing 116 up-regulated as well as 104 down-regulated miRNAs. Of them, confirmation of some differently expressed miRNAs was performed via a stem-loop qRT-PCR, which exhibited differently expressed level in juvenile and adult LMB. Furthermore, GO and KEGG enrichment analyses of targets of differently-expressed miRNAs were carried out. Additionally, the analysis of miRNAs-targets interaction network showed that miR-181b-5p_R-1, miR-725 and miR-103 as the nodal miRNAs has over 20 target genes. Moreover, miR-103 could bind the 3'-UTR of actr8, which was validated via dual-luciferase reporter assay. It has been reasonably hypothesized that miR-103 may play a crucial role, which regulate skeletal muscle development of LMB. The present study provides the first identification of miRNA expression profiles at two different skeletal muscle development stages in LMB. Results may be valuable in interpreting the regulatory role miRNAs plays in the growth and developmental process of skeletal muscle and its possible use in LMB breeding.

Toxicity of amine-functionalized single-carbon nanotube (NH2 f-SWCNT) to Channel Catfish (Ietalurus Punetaus): Organ pathologies, oxidative stress, inflammation, and apoptosis.

The rapid development of carbon nanotubes (CNTs) in the field of fish disease control and prevention raises concerns about the toxicity and safe use in fish. This study was performed to assess the effect on histological changes, oxidative stress related markers in response to various concentrations of amine-functionalized single carbon nanotubes (NH2 f-SWCNT) (1, 10 and 100 mg kg-1 fish) in Channel Catfish (Ietalurus Punetaus) for up to 10 days. Moreover, pro-inflammatory cytokine genes and apoptotic genes were analyzed to obtain a better understanding of molecular mechanism of NH2 f-SWCNT induced toxicity. As a result, intraperitoneal (i.p.) administration of NH2 f-SWCNT caused dose-dependent and time-dependent injuries in the sampled tissues. In comparison with the control groups, decrease of catalase (CAT) activity and superoxide dismutase (SOD), and increase of malondialdehyde (MDA) and lactin dehydrogenase (LDH) were observed in all treatments. Real-time PCR assay showed inflammatory response with dose-dependent increase of tumor necrosis factor alpha (TNFα) and transient increase of interleukin 1ß (IL-1ß) in the liver. NH2 f-SWCNT administration induced increase of p38 as well as caspase-3 in all treatments compared to the control groups, indicating the involvement of p38-MAPK cascade and caspase-3 cascade in liver cell apoptosis. Overall, we conclude that NH2 f-SWCNT exert effects by direct injury and indirectly oxidative stress, resulting in inflammation and apoptosis, which provides data for understanding of the biological mechanisms underlying the toxicity of CNTs in fish.

Dietary sodium butyrate supplementation attenuates intestinal inflammatory response and improves gut microbiota composition in largemouth bass (Micropterus salmoides) fed with a high soybean meal diet.

The study aimed to investigate the effects of dietary sodium butyrate (NaBT) supplementation on the gut health of largemouth bass (Micropterus salmoides) fed with a high soybean meal diet. Three isonitrogenous and isolipidic diets were formulated: a high fishmeal group (Control); a high soybean meal group (SBM), in which the 30% fishmeal protein in the Control diet was replaced by soy protein; and an NaBT group, in which 0.2% NaBT was added to the SBM diet. Each diet was fed to triplicate tanks (20 fish in each tank). After 8 weeks of feeding trial, the distal intestine and intestinal digesta of the fish in each treatment were sampled. The results showed that fishmeal replacement and NaBT supplementation did not affect fish growth performance. Dietary 0.2% NaBT supplementation improved intestinal morphology, increasing the villus width and villus height and reducing the width of lamina propria. The distal intestine of fish in the control and NaBT groups demonstrated lower activities of total superoxide dismutase (T-SOD) and glutathione peroxidase (GPx) and a lower malondialdehyde (MDA) content, compared with the fish in the SBM group. Moreover, the addition of 0.2% NaBT in the feed significantly decreased the expression of tumor necrosis factor α (TNF-α) and interleukin 1ß (IL-1ß) compared to the SBM diet. PCoA and UPGMA analyses based on weighted UniFrac distances demonstrated that intestinal microbial communities in the NaBT group were closer to those in the control group than to those in the SBM group. In addition, dietary 0.2% NaBT supplementation significantly increased the abundance of Firmicutes and Bacteroidetes and decreased the abundance of Tenericutes at the phylum level. Furthermore, the abundance of Bacteroides, Lachnospiraceae_unclassified, and Lachnospiraceae_uncultured was significantly increased, while that of Mycoplasma was significantly decreased in fish intestine at NaBT group at the genus level. In conclusion, dietary NaBT supplementation had beneficial roles in protecting the gut health of largemouth bass from the impairments caused by soybean meal.

Emerging landscape of circHIPK3 and its role in cancer and other diseases (Review).

Circular RNAs (circRNAs) are a special class of recently re­discovered RNAs, which are covalently closed ring RNA molecules. circRNAs have been reported to possess multiple functions and are considered crucial regulators of several processes, and are therefore gaining increasing attention. In recent years, increasing evidence has shown that circRNAs are implicated in several crucial biological processes via regulation of gene expression, and their dysregulation is also associated with the development of numerous diseases, particularly acting as oncogenic or tumor­suppressor molecules in cancer. Furthermore, circRNAs are involved in cell proliferation, differentiation, apoptosis, invasion and metastasis. In the present review, the biogenesis and functions of circRNAs are described, with a focus on the most recent research advances and the emerging roles of circular homeodomain­interacting protein kinase 3 (circHIPK3) in human diseases. The present review may provide novel avenues for research on the roles of circHIPK3 as a clinical diagnostic and prognostic biomarker, as well as highlighting promising therapeutic targets for certain diseases and cancer.

[Study on the spectra of NTO and its rubidium salt in aqueous solution].

In this paper, the Raman and difference FTIR spectra of NTO crystal, NTO solutions at different pH and saturated aqueous solution of Rb(NTO) x H2O have been studied. The determined structure of crystal suggests that the metal ion with oxygen and nitrogen atoms of [chemical structure see text] C-N, [chemical structure see text] C-NO2, [chemical structure see text] C=O and H2O formed bonds. The highest characterization peaks of Raman and FTIR spectra show that [chemical structure see text] C-NO2, [chemical structure see text] C=O and [chemical structure see text] C-N of NTO- formed week coordination bonds. The vibration spectra of saturation solution basically accord with those of the crystal. The bonds of saturation solution of the coordination compound have not changed. Therefore, the different vibration spectra of the saturation water solution of Rb (NTO) x H2O can be explicated by the formation of the bonds in crystal. The coordination compound formation mechanism of metal ion with NTO in alkali aqueous solution is discussed.

[Raman spectroscopic analysis of dissolution and phase transformation of chloropinnoite in the boric acid aqueous solution].

Raman spectroscopy of dissolution and transformation of chloropinnoite in 4.5% (w.t.%) boric acid aqueous solution at 30 degrees C has been recorded. The Raman spectra of kinetics process have been obtained. The phase transformation product is kurnakovite (2MgO x 3B2O3 x 15H2O). The main polyborate anions and their interaction in aqueous solution have been proposed according to the Raman spectrum. Some assignments were tentatively given and the relations between the existing forms of polyborate anions and the crystallizing solid phases have been gained. A mechanisms of dissolution and crystallization reactions and the formation condition of kurnakovite in Qinghai-Tibet plateau were proposed and discussed.

[FTIR spectral analysis of saturated solution during dissolution and transformation of chloropinnoite in magnesium chloride solution].

FTIR differential spectroscopy was used to identify the polyborate ions in saturated aqueous solutions during dissolution and transformation of 2MgO.2B2O3.MgCl2.14H2O in 0.5%, 12%, and 18% MgCl2 solutions at 30 degrees C, respectively. The recorded IR-spectroscopic frequencies of the borate aqueous solution subtracted that of water as a blank value and their assignments were given. The bands at about 515, 630 and 550 cm-1 are characteristics of B(OH)4-/B2O(OH)6(2-), B3O3(OH)4-/B3O3(OH)5(2-), and B4O5(OH)4(2-), respectively. The existing forms of polyborate anions in saturated aqueous solutions and their interaction have been discussed. The relations between the existing forms of polyborate anions and the crystallizing solid phases have been obtained.