Sensoproteomic Characterization of Lactobacillus Johnsonii-Fermented Pea Protein-Based Beverage: A Promising Strategy for Enhancing Umami and Kokumi Sensations while Mitigating Bitterness.

This study investigated the mechanism underlying the flavor improvement observed during fermentation of a pea protein-based beverage using Lactobacillus johnsonii NCC533. A combination of sensomics and sensoproteomics approach revealed that the fermentation process enriched or generated well-known basic taste ingredients, such as amino acids, nucleotides, organic acids, and dipeptides, besides six new taste-active peptide sequences that enhance kokumi and umami notes. The six new umami and kokumi enhancing peptides, with human recognition thresholds ranging from 0.046 to 0.555 mM, are produced through the degradation of Pisum sativum's storage protein. Our findings suggest that compounds derived from fermentation enhance umami and kokumi sensations and reduce bitterness, thus improving the overall flavor perception of pea proteins. In addition, the analysis of intraspecific variations in the proteolytic activity of L. johnsonii and the genome-peptidome correlation analysis performed in this study point at cell-wall-bound proteinases such as PrtP and PrtM as the key genes necessary to initiate the flavor improving proteolytic cascade. This study provides valuable insights into the molecular mechanisms underlying the flavor improvement of pea protein during fermentation and identifies potential future research directions. The results highlight the importance of combining fermentation and senso(proteo)mics techniques in developing tastier and more palatable plant-based protein products.

Screening of a Microbial Culture Collection: Empowering Selection of Starters for Enhanced Sensory Attributes of Pea-Protein-Based Beverages.

Pea-protein-based ingredients are gaining attention in the food industry due to their nutritional benefits and versatility, but their bitter, astringent, green, and beany off-flavors pose challenges. This study applied fermentation using microbial cultures to enhance the sensory qualities of pea-protein-based beverages. Using UHPLC-TOF-MS analyses along with sensory profile comparisons, microbial species such as Limosilactobacillus fermentum, Lactococcus lactis, Lactobacillus johnsonii, Lacticaseibacillus rhamnosus, and Bifidobacterium longum were preselected from an entire culture collection and found to be effective in improving the overall flavor impression by reducing bitter off-notes and enhancing aroma profiles. Notably, L. johnsonii NCC533 and L. fermentum NCC660 exhibited controlled proteolytic activities after 48 h of fermentation, enriching the matrix with taste-active amino acids, nucleotides, and peptides and improving umami and salty flavors while mitigating bitterness. This study has extended traditional volatile analyses, including nonvolatile metabolomic, proteomic, and sensory analyses and offering a detailed view of fermentation-induced biotransformations in pea-protein-based food. The results highlight the importance of combining comprehensive screening approaches and sensoproteomic techniques in developing tastier and more palatable plant-based protein products.

Influence of different abutment materials on the color of implant-supported restoration: A laboratory study.

PURPOSE: To investigate the effect of different abutments and crowns on the color of implant-supported restorations. METHODS: Zirconia and lithium disilicate (e.max) disks with A2 shade were fabricated to represent two crowns. The implant abutments were untreated titanium, opaqued titanium, anodized titanium, A2 shade zirconia and white zirconia. 4.0 mm-thickness zirconia and e.max specimens were used as references respectively. The crowns were placed on tested abutments with a drop of clear glycerin between them and the color was measured using a digital spectrophotometer. CIELab values were recorded to evaluate color differences (ΔE) between tested specimens and the references. RESULTS: Titanium abutments presented higher color differences than zirconia. The ΔE values with untreated titanium were higher than those with opaqued titanium. No differences were found between untreated titanium and anodized titanium for zirconia crowns. The ΔE values of zirconia crowns showed no significant differences between shade A2 zirconia and white zirconia abutments; e.max crowns showed a significant difference. The zirconia crown ΔE values were lower than those of e.max for all titanium and A2 zirconia abutments. Lithium disilicate crowns and zirconia abutments may be more suitable for implant-supported restorations. Opaqued titanium abutment may improve color in esthetic regions when a ceramic abutment cannot be used. CLINICAL SIGNIFICANCE: Lithium disilicate crowns and zirconia abutments may be an effective method to achieve excellent color matching in esthetic regions with implant-supported restorations.

Subtrochanteric fracture after core decompression for osteonecrosis of the femoral head: a case report and literature review.

BACKGROUND: Osteonecrosis of the femoral head (ONFH) is a common clinical disease. Improper treatment can lead to femoral head collapse and hip joint dysfunction. Core decompression is particularly important for early ONFH. However, subtrochanteric fractures after core decompression cause some clinical problems. CASE PRESENTATION: This article describes a 34-year-old male patient with early ONFH. After core decompression, he suffered a subtrochanteric fracture of the femur while bearing weight on the affected limb when going up stairs. He was subsequently treated with open reduction and intramedullary nail fixation. CONCLUSION: When core decompression is used to treat ONFH, the location or size of the drill hole, whether a tantalum rod or bone is inserted, and partial weight-bearing of the affected limb may directly affect whether a fracture occurs after surgery. It is hoped that this case report can provide a reference for clinical orthopedic surgeons in the treatment of early ONFH.

Biomarker identification and risk prediction model development for differentiated thyroid carcinoma lung metastasis based on primary lesion proteomics.

PURPOSE: The rising global high incidence of differentiated thyroid carcinoma (DTC) has led to a significant increase in patients presenting with lung metastasis of DTC (LMDTC). This population poses a significant challenge in clinical practice, necessitating the urgent development of effective risk stratification methods and predictive tools for lung metastasis. EXPERIMENTAL DESIGN: Through proteomic analysis of large samples of primary lesion and dual validation employing parallel reaction monitoring and immunohistochemistry, we identified eight hub proteins as potential biomarkers. By expanding the sample size and conducting statistical analysis on clinical features and hub protein expression, we constructed three risk prediction models. RESULTS: This study identified eight hub proteins-SUCLG1/2, DLAT, IDH3B, ACSF2, ACO2, CYCS and VDAC2- as potential biomarkers for predicting DTC lung metastasis risk. We developed and internally validated three risk prediction models incorporating both clinical characteristics and hub protein expression. Our findings demonstrated that the combined prediction model exhibited optimal predictive performance, with the highest discrimination (AUC: 0.986) and calibration (Brier score: 0.043). Application of the combined prediction model within a specific risk threshold (0-0.97) yielded maximal clinical benefit. Finally, we constructed a nomogram based on the combined prediction model. CONCLUSIONS: As a large sample size study in lung metastatic DTC research, the identification of biomarkers through primary lesion proteomics and the development of risk prediction models integrating clinical features and hub protein biomarkers offer valuable insights for predicting DTC lung metastasis and establishing personalised treatment strategies.

Self-powered Pt/a-Ga2O3/ITO vertical Schottky junction solar-blind photodetector with excellent detection performance.

Self-powered solar-blind photodetectors (PDs) are promising for military and civilian applications owing to convenient operation, easy preparation, and weak-light sensitivity. In the present study, the solar-blind deep-ultraviolet (DUV) photodetector based on amorphous Ga2O3 (a-Ga2O3) and with a simple vertical stack structure is proposed by applying the low-cost magnetron sputtering technology. By tuning the thickness of the amorphous Ga2O3 layer, the device exhibits excellent detection performance. Under 3 V reverse bias, the photodetector achieves a high responsivity of 671A/W, a high detectivity of 2.21 × 1015 Jones, and a fast response time of 27/11 ms. More extraordinary, with the help of the built-in electric field at the interface, the device achieves an excellent performance in detection when self-powered, with an ultrahigh responsivity of 3.69 A/W and a fast response time of 2.6/6.6 ms under 254 nm light illumination. These results demonstrate its superior performance to most of the self-powered Schottky junction UV photodetectors reported to date. Finally, the Pt/a-Ga2O3/ITO Schottky junction photodiode detector is verified as a good performer in imaging, indicating its applicability in such fields as artificial intelligence, machine vision, and solar-blind imaging.

SRSF1 induces glioma progression and has a potential diagnostic application in grading primary glioma.

Glioma is the most common intracranial tumor of the central nervous system in adults; however, the diagnosis of glioma, and its grading and histological subtyping, is challenging for pathologists. The present study assessed serine and arginine rich splicing factor 1 (SRSF1) expression in 224 glioma cases in the Chinese Glioma Genome Atlas (CGGA) database, and verified its expression by immunohistochemical analysis of specimens from 70 clinical patients. In addition, the prognostic potential of SRSF1 concerning the survival status of patients was evaluated. In vitro, the biological role of SRSF1 was assessed using MTT, colony formation, wound healing and Transwell assays. The results revealed that SRSF1 expression was significantly associated with the grading and the histopathological subtype of glioma. As determined using a receiver operating characteristic curve analysis, the specificity of SRSF1 for glioblastoma (GBM) and World Health Organization (WHO) grade 3 astrocytoma was 40 and 48%, respectively, whereas the sensitivity was 100 and 85%. By contrast, pilocytic astrocytoma tumors exhibited negative immunoexpression of SRSF1. Additionally, Kaplan-Meier survival analysis indicated that high SRSF1 expression predicted a worse prognosis for patients with glioma in both the CGGA and clinical cohorts. In vitro, the results demonstrated that SRSF1 promoted the proliferation, invasion and migration of U87MG and U251 cells. These data suggested that immunohistochemical analysis of SRSF1 expression is highly sensitive and specific in the diagnosis of GBM and WHO grade 3 astrocytoma, and may have an important role in glioma grading. Furthermore, the lack of SRSF1 is a potential diagnostic biomarker for pilocytic astrocytoma. However, neither in oligodendroglioma and astrocytoma, nor in GBM was an association detected between SRSF1 expression and IDH1 mutations or 1p/19q co-deletion. These findings indicated that SRSF1 may serve as a prognostic factor in glioma cases and could have an active role in promoting glioma progression.

Surgical site infection following open lobectomy in patients with lung cancer: A prospective study.

OBJECTIVE: To assess the incidence and risk factors of surgical site infection after open pulmonary lobectomy and to quantify their clinical and economic burden. METHODS: A prospective nested case-control study was performed on patients with lung cancer who underwent open lobectomy in the lung cancer center of West China Hospital from January 2017 to December 2019. Demographic, clinical data and medical costs were recorded. Logistic regression was used to evaluate risk factors associated with surgical site infection. A Mann-Whitney U test was carried out to evaluate the differences in medical costs. RESULTS: A total of 1395 patients were eligible, and the surgical site infection incidence was 13.47% (188/1395). Of the 188 instances of surgical site infection, 171 (90.96%) were classified as organ/space infection, 8 (4.25%) as superficial incisional infection and 9 (4.79%) as deep incisional infection. The patients with surgical site infection had significantly higher mortality (3.19% vs. 0.41%, p < 0.001), higher median medical cost (90774.95 yuan vs. 63079.38 yuan, p < 0.001), and longer postoperative length of stay (15 days vs. 9 days, p < 0.001). Multivariate logistic regression analysis indicated that age (odds ratio (OR) = 1.560, p = 0.007), respiratory failure (OR = 5.984, p = 0.0012), American Society of Anesthesiologists score (OR = 1.584, p = 0.005), operating time (OR = 1.950, p < 0.001), and operation team (OR = 1.864, p < 0.001) were independent risk factors for surgical site infection. CONCLUSIONS: The high incidence of surgical site infection indicates that postoperative infections remain a significant clinical burden in patients who underwent open lobectomy. Identifying risk factors timely through prospective surveillance may assist clinical decisions against surgical site infection.

Flavour by design: food-grade lactic acid bacteria improve the volatile aroma spectrum of oat milk, sunflower seed milk, pea milk, and faba milk towards improved flavour and sensory perception.

BACKGROUND: The global market of plant-based milk alternatives is continually growing. Flavour and taste have a key impact on consumers' selection of plant-based beverages. Unfortunately, natural plant milks have only limited acceptance. Their typically bean-like and grassy notes are perceived as "off-flavours" by consumers, while preferred fruity, buttery, and cheesy notes are missing. In this regard, fermentation of plant milk by lactic acid bacteria (LAB) appears to be an appealing option to improve aroma and taste. RESULTS: In this work, we systematically studied LAB fermentation of plant milk. For this purpose, we evaluated 15 food-approved LAB strains to ferment 4 different plant milks: oat milk (representing cereal-based milk), sunflower seed milk (representing seed-based milk), and pea and faba milk (representing legume-based milk). Using GC‒MS analysis, flavour changes during anaerobic fermentations were studied in detail. These revealed species-related and plant milk-related differences and highlighted several well-performing strains delivered a range of beneficial flavour changes. A developed data model estimated the impact of individual flavour compounds using sensory scores and predicted the overall flavour note of fermented and nonfermented samples. Selected sensory perception tests validated the model and allowed us to bridge compositional changes in the flavour profile with consumer response. CONCLUSION: Specific strain-milk combinations provided quite different flavour notes. This opens further developments towards plant-based products with improved flavour, including cheesy and buttery notes, as well as other innovative products in the future. S. thermophilus emerged as a well-performing strain that delivered preferred buttery notes in all tested plant milks. The GC‒MS-based data model was found to be helpful in predicting sensory perception, and its further refinement and application promise enhanced potential to upgrade fermentation approaches to flavour-by-design strategies.

Accuracy of immediate dental implant placement with task-autonomous robotic system and navigation system: An in vitro study.

OBJECTIVES: The aim of this study was to compare the accuracy of dental implant placement in a single tooth gap, including the postextraction site and healed site, using a task-autonomous robotic system and a dynamic navigation system. MATERIALS AND METHODS: Forty partially edentulous models requiring both immediate and conventional implant placement were randomly divided into a robotic system group and a navigation system group. The coronal, apical, and angular deviations of the implants were measured and assessed between the groups. RESULTS: The deviations in immediate implant placement were compared between the robotic system and dynamic navigation system groups, showing a mean (±SD) coronal deviation of 0.86 ± 0.36 versus 0.70 ± 0.21 mm (p = .101), a mean apical deviation of 0.77 ± 0.34 versus 0.95 ± 0.38 mm (p = .127), and a mean angular deviation of 1.94 ± 0.66° versus 3.44 ± 1.38° (p < .001). At the healed site, significantly smaller coronal deviation (0.46 ± 0.29 vs. 0.70 ± 0.30 mm, p = .005), apical deviation (0.56 ± 0.30 vs. 0.85 ± 0.25 mm, p < .001), and angular deviation (1.36 ± 0.54 vs. 1.80 ± 0.70 mm, p = .034) were found in the robotic system group than in the dynamic navigation group. CONCLUSIONS: The position in both immediate and conventional implant placement was more precise with the task-autonomous robotic system than with the dynamic navigation system. Its performance in actual clinical applications should be confirmed in further trials.

Multiple Metastatic Extra-gastrointestinal Stromal Tumors with Plasmoid Differentiation: A Case Report and Review of Literature.

Extra-gastrointestinal stromal tumors (EGISTs) are rare mesenchymal tumors that arise from the abdominal, pelvic or retroperitoneal region, unrelated to the gastrointestinal tract. However, cases with a plasmoid morphology are extremely rare. we hererin report a 49-year-old man with abdominal pain who underwent magnetic resonance imaging that revealed an irregular tumor (103×71 mm) in size, in the space between stomach and pancreas, diagnosed as an EGISIT, we also reviewed the clinicopathological characteristics and immunohistochemical characteristics, molecular genetic features and differential diagnoses previously reported in the literature.

Effects of different grafting materials on volumetric changes in the Schneiderian membrane following lateral maxillary sinus floor elevation: a preliminary study.

OBJECTIVES: To investigate the potential influence of different grafting materials on maxillary sinus membrane dimensions and ostium patency following lateral sinus floor elevation (SFE) as assessed using cone-beam computed tomography (CBCT). MATERIALS AND METHODS: A total of 40 sinuses in 40 patients were included. Twenty sinuses were referred for SFE with deproteinized bovine bone mineral (DBBM), and the remaining 20 sinuses were grafted with calcium phosphate (CP). CBCT was performed prior to and 3 to 4 days after surgery. The dimensions of the Schneiderian membrane volume and ostium patency were evaluated, and potential relationships between volumetric changes and any associated factors were analyzed. RESULTS: The median increase in membrane-whole cavity volume ratios was 43.97% in the DBBM group and 67.58% in the CP group, demonstrating no statistically significant difference (p = 0.17). The rates of increased obstruction after SFE were 11.1% for the DBBM group versus 44.4% for the CP group (p = 0.03). The graft volume was found to be positively correlated with the postoperative membrane-whole cavity volume ratio (r = 0.79; p < 0.01) and the membrane-whole cavity volume ratio increase (r = 0.71; p < 0.01). CONCLUSIONS: The two grafting materials seem to have a similar effect on transient volumetric changes in the sinus mucosa. However, the choice of grafting material should still be made with caution since sinuses grafted using DBBM exhibited less swelling and less ostium obstruction.

Fabricating a surgical template for guiding sinus floor elevation in the maxillary sinus septa with simultaneous implant placement: A dental technique.

The presence of sinus septa, common anatomic structures of the maxillary sinus, may increase the incidence of surgical complications during sinus floor elevation. This article introduces a digital protocol for achieving safe and precise sinus floor elevation with an individualized surgical template that combines implant placement and the lateral sinus window technique. This technique facilitates precise preplanning and preparation of the lateral osteotomy window and the implant site and reduces surgical complications, shortens surgical duration, and improves patient-related outcomes.

Endoscopic management of postoperative fistulas using purse-string suture combined with cyanoacrylate.

A 65-year-old man complained of choking and hoarseness for fifteen days, and was diagnosed with thyroid carcinoma infiltrating esophagus and trachea. Therefore, the patient underwent thyroidectomy, partial esophagectomy, and partial tracheal resection, and histopathology confirmed primary squamous cell carcinoma of the thyroid. Unfortunately, on the tenth postoperative day, an esophagogastroduodenoscopy showed a large fistula (25 mm*20 mm) in esophageal introitus, and diagnosed with tracheoesophageal fistula due to sustained choking. The patient failed to response to conservative treatment within 14 days. Consequently, endoscopic management was performed that the fistula was partly closed by purse-string suture using endoloop and hemostatic clips, then 1 ml of cyanoacrylate (Compon, China) was injected into the fistulous tract through a catheter. Interestingly, the patient's symptom was relieved after the procedure. And, esophagogastroduodenoscopy revealed healing of the fistula 14 days later.

[Study on the effect of alginate impression spray disinfection process on model accuracy].

PURPOSE: To investigate the effect of a new cleaning and disinfection process of oral impressions on the accuracy of alginate dental impression. METHODS: Sixteen young volunteers were selected to make alginate oral impression (2 times for each upper and lower jaw) and oral scandata acquisition (1 time for each upper and lower jaw) to obtain STL data. The 2 pairs of alginate impressions of each subject were numbered group 1, group 2, inwhich group 1 was not sterilized and the oral impression film in group 2 was treated with the method and steps of new oral impression cleaning and disinfection. All the oral impressions of the subjects were perfused with plaster models in the same standard way and demoulded. The obtained plaster models were examined and scanned after appropriate trimming and grinding. The plaster model data obtained from each group of each patient after warehouse scanning were compared with the orally scanned model data for alignment analysis, and root mean square(RMS) was used as a parameter to evaluate the deviation between the test model and the standard model. The chromatograms of the deviations were saved for visualization and analysis to observe the differences in model accuracy from several dimensions. Statistical analysis was performed with SPSS 23.0 software package. RESULTS: The differences in RMS of model accuracy of maxillary dental series of alginate impressions were statistically significant, in which the RMS of the group disinfecting the maxillary oral impression film with a new oral impression cleaning and disinfection process was significantly higher than that of the nonsterilized group (P=0.006), while the difference between model accuracy RMS of the mandibular disinfection group and non-disinfection group was not statistically significant(P=0.874). CONCLUSIONS: The new technical specifications for cleaning and disinfection of dental impressions recommended by the Chinese Stomatological Association have certain effects on the accuracy of dental impressions made of alginate material,and the details in its process should be further improved.

Fish Community Structure and Biomass Particle-Size Spectrum in the Upper Reaches of the Jinsha River (China).

To understand the characteristics of the fish community structure and biomass particle-size spectrum in the upper reaches of the Jinsha River, fish and environmental surveys were conducted in 21 segments of the upper reaches of the Jinsha River in September 2019 and June 2020. A total of 4062 fish belonging to 2 orders, 5 families, 18 genera, and 28 species were collected. Among them, Cyprinidae fish were the most abundant (14 species), accounting for 50.00%. The Shannon index and Pielou evenness index values varied from 0.402-1.770 and 0.254-0.680, respectively. The dominant species of fish were Triplophysa stenura, Schizothorax wangchiachii, and Schizopygopsis malacanthus. Redundancy analysis (RDA) was used to analyse the relationship between the fish community and environmental factors. Velocity, altitude, and dissolved oxygen were the main influencing factors of fish community structure differences in the upper reaches of the Jinsha River. The abundance/biomass curves showed that the fish communities in the upper reaches of the Jinsha River were moderately or severely disturbed. The standardized biomass particle-size spectrum of fish showed that the degree of disturbance of fish in tributaries was much lower than that in the main stream. Compared with the historical data, the fish community structure in the Jinsha River has changed significantly, with the number of exotic species increasing, and the individual fish showing miniaturization and younger ages. It is suggested that habitat conservation strategies be adopted in the upper tributaries of the Jinsha River to provide a reference for the restoration of fishery resources and the conservation of fish diversity in the Yangtze River.

Expression Patterns and Gonadotropin Regulation of the TGF-ß II Receptor (Bmpr2) during Ovarian Development in the Ricefield Eel Monopterus albus.

Bmpr2 plays a central role in the regulation of reproductive development in mammals, but its role during ovarian development in fish is still unclear. To ascertain the function of bmpr2 in ovarian development in the ricefield eel, we isolated and characterized the bmpr2 cDNA sequence; the localization of Bmpr2 protein was determined by immunohistochemical staining; and the expression patterns of bmpr2 in ovarian tissue incubated with FSH and hCG in vitro were analyzed. The full-length bmpr2 cDNA was 3311 bp, with 1061 amino acids encoded. Compared to other tissues, bmpr2 was abundantly expressed in the ovary and highly expressed in the early yolk accumulation (EV) stages of the ovary. In addition, a positive signal for Bmpr2 was detected in the cytoplasm of oocytes in primary growth (PG) and EV stages. In vitro, the expression level of gdf9, the ligand of bmpr2, in the 10 ng/mL FSH treatment group was significantly higher after incubation for 4 h than after incubation for different durations. However, bmpr2 expression in the 10 ng/mL FSH treatment group at 2 h, 4 h and 10 h was significantly lower. Importantly, the expression level of bmpr2 and gdf9 in the 100 IU/mL hCG group had similar changes that were significantly decreased at 4 h and 10 h. In summary, Bmpr2 might play a pivotal role in ovarian growth in the ricefield eel, and these results provide a better understanding of the function of bmpr2 in ovarian development and the basic data for further exploration of the regulatory mechanism of gdf9 in oocyte development.

PPP1R14D promotes the proliferation, migration and invasion of lung adenocarcinoma via the PKCα/BRAF/MEK/ERK signaling pathway.

Protein phosphatase 1 (PP1) inhibitors play a role in tumor progression through different mechanisms. Protein phosphatase 1 regulatory subunit 14D (PPP1R14D) is an inhibitor of PP1. However, the role of PPP1R14D in tumors and its mechanism of action are largely unknown. The purpose of the present study was to investigate the expression, function and mechanism of PPP1R14D in lung adenocarcinoma (LUAD). In the present study, GEPIA database analysis and immunohistochemistry demonstrated that PPP1R14D was highly expressed in LUAD tissues and that the expression of PPP1R14D in LUAD was negatively correlated with the age of patients and positively correlated with the 8th American Joint Committee on Cancer staging among patients. In addition, Kaplan­Meier Plotter database analysis showed that PPP1R14D expression was associated with lower survival rates in patients with LUAD. PPP1R14D knockdown significantly inhibited LUAD cell proliferation, migration and invasion and induced LUAD cell arrest at the G1 phase of the cell cycle. Mechanistic analyses revealed that PPP1R14D knockdown may inhibit cell proliferation, migration and invasion by inactivating PKCα/BRAF/MEK/ERK pathway signaling and its downstream key proteins c­Myc/Cyclin E1­CDK2 and MMP2/MMP9/Vimentin. Moreover, knockdown of PPP1R14D suppressed tumor growth in vivo. All these results showed that PPP1R14D plays an important role in LUAD tumorigenesis and may serve as a potential prognostic factor and therapeutic target in LUAD.

Genetic Variation in Schizothorax kozlovi Nikolsky in the Upper Reaches of the Chinese Yangtze River Based on Genotyping for Simplified Genome Sequencing.

Schizothorax kozlovi Nikolsky is a unique cold-water fish in the upper reaches of the Yangtze River in China and has high economic value. In our study, genetic diversity and population structure analyses were performed on seven wild populations (originating from the Jinsha River, Yalong River, and Wujiang River) in the upper reaches of the Yangtze River by genotyping by sequencing (GBS). The results indicated that a total of 303,970 single-nucleotide polymorphisms (SNPs) were identified from the seven wild populations. Lower genetic diversity was exhibited among the intrapopulations of the three tributaries, and the Wujiang River population had significant genetic differentiation when compared to the Jinsha River and Yalong River populations. Furthermore, the selected SNPs were enriched in cellular processes, environmental adaptation, signal transduction, and related metabolic processes between the Wujiang population and the other two populations. The above results indicate that the populations of S. kozlovi have different degrees of tolerance and selection pressure in response to temperature and altitude. The Wujiang intrapopulation has greater genetic diversity and differentiation than the Jinsha River and Yalong River populations, which demonstrates that the Jinsha and Yalong populations require more attention and resources for their protection. The results of this study will increase our understanding of the diversity of S. kozlovi in the upper reaches of the Yangtze River and provide a basis for the conservation and utilization of wild resources.

Circular RNA expression profiles and CircSnd1-miR-135b/c-foxl2 axis analysis in gonadal differentiation of protogynous hermaphroditic ricefield eel Monopterus albus.

BACKGROUND: The expression and biological functions of circular RNAs (circRNAs) in reproductive organs have been extensively reported. However, it is still unclear whether circRNAs are involved in sex change. To this end, RNA sequencing (RNA-seq) was performed in gonads at 5 sexual stages (ovary, early intersexual stage gonad, middle intersexual stage gonad, late intersexual stage gonad, and testis) of ricefield eel, and the expression profiles and potential functions of circRNAs were studied. RESULTS: Seven hundred twenty-one circRNAs were identified, and the expression levels of 10 circRNAs were verified by quantitative real-time PCR (qRT-PCR) and found to be in accordance with the RNA-seq data, suggesting that the RNA-seq data were reliable. Then, the sequence length, category, sequence composition and the relationship between the parent genes of the circRNAs were explored. A total of 147 circRNAs were differentially expressed in the sex change process, and GO and KEGG analyses revealed that some differentially expressed (such as novel_circ_0000659, novel_circ_0004005 and novel_circ_0005865) circRNAs were closely involved in sex change. Furthermore, expression pattern analysis demonstrated that both circSnd1 and foxl2 were downregulated in the process of sex change, which was contrary to mal-miR-135b. Finally, dual-luciferase reporter assay and RNA immunoprecipitation showed that circSnd1 and foxl2 can combine with mal-miR-135b and mal-miR-135c. These data revealed that circSnd1 regulates foxl2 expression in the sex change of ricefield eel by acting as a sponge of mal-miR-135b/c. CONCLUSION: Our results are the first to demonstrate that circRNAs have potential effects on sex change in ricefield eel; and circSnd1 could regulate foxl2 expression in the sex change of ricefield eel by acting as a sponge of mal-miR-135b/c. These data will be useful for enhancing our understanding of sequential hermaphroditism and sex change in ricefield eel or other teleosts.