Role of alternative splicing in fish immunity.

Alternative splicing serves as a pivotal source of complexity in the transcriptome and proteome, selectively connecting various coding elements to generate a diverse array of mRNAs. This process encodes multiple proteins with either similar or distinct functions, contributing significantly to the intricacies of cellular processes. The role of alternative splicing in mammalian immunity has been well studied. Remarkably, the immune system of fish shares substantial similarities with that of humans, and alternative splicing also emerges as a key player in the immune processes of fish. In this review, we offer an overview of alternative splicing and its associated functions in the immune processes of fish, and summarize the research progress on alternative splicing in the fish immunity. Furthermore, we review the impact of alternative splicing on the fish immune system's response to external stimuli. Finally, we present our perspectives on future directions in this field. Our aim is to provide valuable insights for the future investigations into the role of alternative splicing in immunity.

The evolution and immunomodulatory role of Zc3h12 proteins in zebrafish (Danio rerio).

Zc3h12 family is an important RNA-binding protein family regulating mRNA of inflammatory cytokines in mammals. However, there are few studies on their post-transcriptional level regulation of inflammatory cytokines in fish. Here, we investigated the evolution of zebrafish Zc3h12 family and explored their immunomodulatory role. Phylogenetic and syntenic analysis indicated the number of zc3h12 family members had increased ranging from a single member in invertebrates to a single copy of four members in mammals. As the most evolutionarily diverse group of vertebrates, the number of zc3h12 family members was more complex and diverse in the teleost, each member experienced different fates and followed different rules in multiple rounds of whole-genome duplication events. Thereinto, zebrafish contained three zc3h12 genes, among which zc3h12aa and zc3h12ab were duplicated from the same gene. Zebrafish Zc3h12 family could recognize the 3'-UTR regions of inflammatory cytokines through binding to the specific RNA secondary structure and negatively regulate their expression. Deletion of either Zc3h12 domains or mutation of the key amino acid in RNAase domain attenuated their modulatory effect, suggesting both domain and RNAase activity are important to the immunomodulatory role. These results elucidated the evolution of Zc3h12 family and uncovered Zc3h12-mediated post-transcriptional regulation of cytokines in zebrafish.

Application of Preoperative Ultrasonography in the Diagnosis of Cervical Lymph Node Metastasis in Thyroid Papillary Carcinoma.

Background: The clinical value and application of preoperative ultrasound contrast in the diagnosis of cervical lymph node metastasis in thyroid papillary carcinoma is investigated. Methods: In total, 126 cases of thyroid papillary carcinoma were selected, the sensitivity and accuracy of color ultrasound and ultrasound contrast were analyzed by comparing preoperative gray-scale ultrasound, color ultrasound, and ultrasound contrast. Results: The accuracies of preoperative color ultrasound and ultrasound contrast in detecting lymph node metastasis were 74 and 82%, respectively, and their sensitivities were 80 and 94%, respectively. Lymph node metastasis was significantly more severe when the tumor diameter was >4 cm. The lymphatic metastatic rate of the patients with multifocal papillary carcinoma was 96.4%, whereas the lymphatic metastatic rate of the patients with thyroid gland lesions was 87.7%. The central foci of cervical lymph node metastasis included the following pathological subtypes: diffuse sclerosis type (89.3%, 25/28), high-cell type (72.2%, 8/11), and papillary type (40.0%, 4/10). Conclusion: Ultrasound contrast is more sensitive than color ultrasound in the diagnosis of cervical lymph node metastasis. Primary lesions ≥4 cm, lesion involvement, outer membrane, and high-risk pathologic subtypes and lesions were considered as the criteria for ultrasound contrast application.

BRAF-activated WT1 contributes to cancer growth and regulates autophagy and apoptosis in papillary thyroid carcinoma.

BACKGROUND: Papillary thyroid carcinoma (PTC) is one of most prevalent malignant endocrine neoplasms, and it is associated with a high frequency of BRAF gene mutations, which lead to lymphatic metastasis and distant metastasis that promote tumor progression. The molecular mechanism of PTC and the role of BRAF mutation in PTC progression and development need to be further elucidated. METHODS: In this study, a comprehensive bioinformatics analysis was performed to identify the differentially expressed genes and signaling pathways in thyroid cancer patients carrying mutant BRAF. Then, we confirmed the prognostic role of WT1 in thyroid cancer patients. Immunohistochemistry was performed to measure the expression profile of WT1 in PTC tissue. Lentivirus shWT1 was transfected into BRAFV600E (mutant) PTC cells to stably inhibit WT1 expression. CCK-8, EdU, immunofluorescence, colony formation, cell migration, cell wound healing, apoptosis and autophagy assays were performed to assess the biological functions of WT1 in BRAFV600E PTC cells. RNA sequencing, immunohistochemistry and immunoblotting were performed to explore the molecular mechanism of WT1 in BRAFV600E PTC cells. RESULTS: The results confirmed that "epithelial cell proliferation", "apoptosis" and "selective autophagy" were closely associated with this BRAF mutant in these thyroid cancer patients. Knocking down BRAF-activated WT1 effectively inhibited the proliferation and migration of BRAFV600E PTC cells. Silencing WT1 significantly inhibited autophagy and promoted the apoptosis of BRAFV600E PTC cells. Mechanistic investigations showed that silencing WT1 expression remarkably suppressed the AKT/mTOR and ERK/P65 signaling pathways in BRAFV600E PTC cells. CONCLUSION: All these results indicate that WT1 is a promising prognostic biomarker and facilitates PTC progression and development of cells carrying the BRAFV600E mutation.

A six-gene-based signature for breast cancer radiotherapy sensitivity estimation.

Breast cancer (BRCA) represents the most common malignancy among women worldwide with high mortality. Radiotherapy is a prevalent therapeutic for BRCA that with heterogeneous effectiveness among patients. Here, we proposed to develop a gene expression-based signature for BRCA radiotherapy sensitivity estimation. Gene expression profiles of BRCA samples from the Cancer Genome Atlas (TCGA) and International Cancer Genome Consortium (ICGC) were obtained and used as training and independent testing dataset, respectively. Differential expression genes (DEGs) in BRCA samples compared with their paracancerous samples in the training set were identified by using the edgeR Bioconductor package. Univariate Cox regression analysis and LASSO Cox regression method were applied to screen optimal genes for constructing a radiotherapy sensitivity estimation signature. Nomogram combining independent prognostic factors was used to predict 1-, 3-, and 5-year OS of radiation-treated BRCA patients. Relative proportions of tumor infiltrating immune cells (TIICs) calculated by CIBERSORT and mRNA levels of key immune checkpoint receptors was adopted to explore the relation between the signature and tumor immune response. As a result, 603 DEGs were obtained in BRCA tumor samples, six of which were retained and used to construct the radiotherapy sensitivity prediction model. The signature was proved to be robust in both training and testing sets. In addition, the signature was closely related to the immune microenvironment of BRCA in the context of TIICs and immune checkpoint receptors' mRNA levels. In conclusion, the present study obtained a radiotherapy sensitivity estimation signature for BRCA, which should shed new light in clinical and experimental research.

Feasibility of using colloidal gold immunochromatography for point-of-care identification of parathyroid glands during thyroidectomy.

AIM: To determine the feasibility of using colloidal gold immunochromatography for rapid identification of parathyroid glands during thyroidectomy. MATERIAL AND METHODS: 127 patients undergoing thyroidectomy were randomly divided into PTH-ICGT group (64 cases) and conventional naked eye group (63 cases). The rate of identification of parathyroid glands and the incidence of hypoparathyroidism were compared between the two groups. RESULTS: PTH-CGI assay results showed that PTH concentration in the parathyroid tissue was (955.3 ±â€¯16.1) ng/L; skeletal muscle tissue [(14.5 ±â€¯1.5) ng/L], thyroid tissue [(15.0 ±â€¯1.3) ng/L], adipose tissue [(15.3 ±â€¯1.2) ng/L], lymph node tissue [(14.0 ±â€¯1.2) ng/L];PTH levels in parathyroid tissues were compared with PTH levels in skeletal muscle, thyroid, fat, and lymph node tissues, respectively. The differences were statistically significant(t values were 23.62, 33.42, 39.34, 30.77, P < 0.0001, respectively); Among the 127 patients undergoing total thyroidectomy, the rate of detection of parathyroid glands was 92.7% in the conventional naked eye group and 96.4% in the PTH-ICGT group. There was no significant difference in the detection rate of parathyroid gland between the two groups (χ2 = 0.7067, P = 0.40). The incidence of temporary hypoparathyroidism after surgery in both groups was 11.3% and 5.7%, respectively (χ2 = 1.093, P > 0.05). The incidence of postoperative permanent hypoparathyroidism in both groups was 3.8% and 0, respectively (Fisher's exact test, P = 0.495). CONCLUSION: PTH-CGI has a high efficiency in identifying parathyroid glands, which may increase the rate of clinical parathyroid detection and reduce the incidence of postoperative hypoparathyroidism.

[Regulatory effects of RhoGTPase on transition of liver sinusoidal capillarization: experiment with mice of schistosomal hepatic fibrosis].

OBJECTIVE: To investigate the regulatory effects of RhoGTPase on the transition of liver sinusoidal endothelial cells and the potential mechanism thereof on the sinusoidal capillarization in schistosomal hepatic fibrosis. METHODS: Eight-eight mice underwent abdominal infection of schistosomal cercaria so as to establish liver fibrosis models. 13 weeks later the mice were divided into 5 groups: Group A (normal control group, n = 10), Group B (group of schistosomiasis, n = 24), Group C (anti-schistosoma control group, treated with biltricide), Group D (group of schistosomiasis + hydroxyfasudil, treated with hydroxyfasudil since the week 14, n = 18), and Group E (biltricide + hydroxyfasudil, treated with biltricide since week 13 and added with hydroxyfasudil since week 14, n = 18). The mice in Group A and 6 mice of Group B were killed in week 13, and 6 mice of Groups B, C, D, and E were killed in weeks 16, 19, and 21 each. The livers were taken out to undergo electron microcopy. Immunohistochemistry was used to detect the expression of p-moesin, connective tissue growth factor (CTGF), RhoA, collagen IV (Col IV), and laminin (LN) protein expressions were assessed by Western blotting, and RT-PCR was used to detect the mRNA expression of CTGF, RhoA, and ROCK II. RESULTS: Compared with Group A, the mRNA levels of RhoA, ROCK II, and CTGF were significantly increased (all P < 0.05) and the protein expression levels of p-moesin, CTGF, RhoA, Col IV, and LN were significantly increased (all P < 0.05) in Group B. After intervention with biltricide and/or hydroxyfasudil, the CTGF mRNA expression was significantly decreased (P < 0.05) in Group E in week 16 and the protein expression levels of CTGF, Col IV, and LN were decreased (all P < 0.05) compared with other groups, and the expression of p-moesin of Group E was significantly lower than that of Group D (P < 0.05). Electron microcopy showed that the liver sinusoids of the mice in Group E was significantly better compared with the other groups, and there was no significant difference between Groups B and D. CONCLUSION: An upregulation of RhoGTPase that contributes to increased CTGF expression and phosphorylation of moesin may induce a transition of liver sinusoidal endothelial cells in schistosomiasis.