Functional analysis of the ube3a response in Japanese flounder (Paralichthys olivaceus) to CSBV infection.

Ube3a is a member of the E3 ubiquitin ligase HECTc family, and its role has been established in neurodevelopmental disorders. However, studies on its role in Japanese flounder are scarce. Thus, in this study, the ube3a of Japanese flounder was cloned, and its role in conferring resistance against Chinook salmon bafnivirus (CSBV) was analyzed. Japanese flounder ube3a encoded a protein containing 834 amino acids. Interestingly, its homology with the Atlantic halibut was determined to be 94%. In addition, there were differential expressions of ube3a in different tissues of Japanese flounder, with the highest expression level observed in the fin, followed by the gills and skin (P ≤ 0.05). Subcellular localization analysis revealed that Ube3a is a cytoplasmic protein. We established an in vitro CSBV infection model using Japanese flounder gill cell line (FG). After ube3a overexpression, the viral load was significantly lower than that of the control group (P ≤ 0.05). Contrastingly, after incubation of FG cells with an E3 ubiquitin ligase inhibitor, the viral load was significantly higher than in the control group (P ≤ 0.01). Then, the expression levels of nf-κb, traf3, and tnf-α after incubation with an E3 ubiquitin ligase inhibitor were examined. The results demonstrated that ube3a may exerted a significant antiviral effect in Japanese flounder via the ubiquitination pathway.

CRISPR-Cas9 identifies growth-related subtypes of glioblastoma with therapeutical significance through cell line knockdown.

BACKGROUND: Glioblastoma (GBM) is a type of highly malignant brain tumor that is known for its significant intratumoral heterogeneity, meaning that there can be a high degree of variability within the tumor tissue. Despite the identification of several subtypes of GBM in recent years, there remains to explore a classification based on genes related to proliferation and growth. METHODS: The growth-related genes of GBM were identified by CRISPR-Cas9 and univariate Cox regression analysis. The expression of these genes in the Cancer Genome Atlas cohort (TCGA) was used to construct growth-related genes subtypes (GGSs) via consensus clustering. Validation of this subtyping was performed using the nearest template prediction (NTP) algorithm in two independent Gene Expression Omnibus (GEO) cohorts and the ZZ cohort. Additionally, copy number variations, biological functions, and potential drugs were analyzed for each of the different subtypes separately. RESULTS: Our research established multicenter-validated GGSs. GGS1 exhibits the poorest prognosis, with the highest frequency of chr 7 gain & chr 10 loss, and the lowest frequency of chr 19 & 20 co-gain. Additionally, GGS1 displays the highest expression of EGFR. Furthermore, it is significantly enriched in metabolic, stemness, proliferation, and signaling pathways. Besides we showed that Foretinib may be a potential therapeutic agent for GGS1, the worst prognostic subtype, through data screening and in vitro experiments. GGS2 has a moderate prognosis, with a slightly higher proportion of chr 7 gain & chr 10 loss, and the highest proportion of chr 19 & 20 co-gain. The prognosis of GGS3 is the best, with the least chr 7 gain & 10 loss and EGFR expression. CONCLUSIONS: These results enhance our understanding of the heterogeneity of GBM and offer insights for stratified management and precise treatment of GBM patients.

Characterization and function of Japanese flounder (Paralichthys olivaceus) slc2a6 in response to lymphocystis disease virus infection.

Slc2a6 is a member of the slc2 family (solute carrier 2 family) and previous reports have indicated its involvement in the inflammatory response. Slc2a6 is regulated by the NF-ĸB signaling pathway. This study investigated the differential expression of slc2a6 in the early embryonic development of Japanese flounder, revealing that the early gastrula stage had the highest level of slc2a6 expression. Moreover, slc2a6 expression was increased in vitro after stimulation by lymphocystis disease virus (LCDV), and in vivo experiments also showed significantly elevated levels in the spleen and muscle tissues following LCDV stimulation. Subcellular localization revealed that Slc2a6 was expressed in both the nucleus and cytoplasm of cells. The pcDNA3.1-slc2a6 overexpression plasmid was successfully constructed; the si-slc2a6 interfering strand was screened and samples were collected. The expression of NF-ĸB signaling pathway-related genes il-1ß, il-6, nf-ĸb, and tnf-α was evaluated in overexpressed, silenced, and LCDV-stimulated samples. The results showed that slc2a6 is involved in viral regulation in Japanese flounder by regulating innate immune responses.

Molecular identification and function characterization of four alternative splice variants of trim25 in Japanese flounder (Paralichthys olivaceus).

Trim25 is a member of Tripartite Motif (TRIM) family. Previous studies report that trim25 modulates antiviral activity by activating RIG-I. In this study we explored the four alternative splicing (AS) variants X1-X4 of Japanese flounder trim25. The sequences of the AS variants were highly conserved. Expression levels of trim25 X1-X4 were increased after 12 h of poly I:C treatment in vitro. In vivo expression of X2-X4 in liver, kidney (except X2) and blood was significantly up-regulated in early stages of poly I:C treatment. Subcellular localization analysis showed that Trim25 X1-X4 were distributed in different cellular organelles. The recombinant vector pcDNA3.1-Trim25 X1-X4 were successfully overexpressed in Flounder cells and the samples were collected. Expression patterns of RIG-I pathway genes dhx58, traf6, traf2, nfkbia and il-8 were explored in vitro and in vivo after poly I:C treatment, as well as overexpressed samples. The findings of this study imply that AS variants of trim25 confer antiviral activity in Japanese flounder by modulating innate immune response.

Application of the Modified Basic Life Support Training Model in Improving Community Residents' Rescue Willingness in Nantong City in China.

Objective: This study explores the application and effect of the modified basic life support training in improving the first-aid level and rescue willingness of community residents in China. Methods: A total of 94 residents of a community in Nantong city were selected as the subjects by cluster sampling to receive the modified basic life support (BLS) training. The BLS knowledge, attitudes, and behaviors of all recruited subjects were evaluated by a questionnaire before and after training. A skill operation assessment was used to evaluate the effectiveness of the modified BLS training. Results: There were statistically significant differences in the BLS rescue willingness, theory, and skill scores before and after the training (P < 0.01). A total of 93.62% of the residents considered the modified BLS training model easier to learn and acceptable than the traditional model, and 92.55% of them thought the training content and teaching arrangement were reasonable. Conclusion: The modified BLS training model could improve the community residents' rescue willingness and skill mastery rates, enhance their first-aid skills and awareness, reduce the risk of disease transmission to a certain extent, and improve the success rate of prehospital first aid to ensure the safety of rescuers and patients.

The impact of policy on availability and price of low-price medicines in public healthcare institutions: A retrospective survey in Nanjing, China.

In an effort to promote rational drug pricing and relieve the pressure of drug shortages, the Chinese government implemented a low-price medicine (LPM) policy in July 2014, and abolished price regulations for most medications in June 2015. This study examines trends in the availability and pricing of LPMs since policy implementation. Data on price and availability of 752 LPMs during 2013-2017 were obtained from the Jiangsu Institute of Medicine Information. Availability was defined as the proportion of facilities in which a medicine was in inventory during each survey period. A price index was constructed based on purchasing prices in 40 public healthcare facilities, using a standard method developed by the International Labour Organization. Mean availability fluctuated slightly but held at low levels (<15%). Levels were conspicuously lower in primary hospitals than in secondary and tertiary hospitals. Our logistic regression model showed that the essential medicine designation was the main factor facilitating availability. The overall price index remained static before implementation of the policy, while there was a marked upward trend after implementation of the policy. Further efforts are needed to improve the pharmaceutical supply system, and simultaneously curb unreasonable inflation in medicine costs.

Molecular characterization and functional analysis of Japanese flounder (Paralichthys olivaceus) thbs2 in response to lymphocystis disease virus.

In mammals, a matricellular protein, thrombospondin 2 (Thbs2) has been reported to play important roles in modulating cell-matrix interactions, vascular integrity and thrombosis formation. However, the role of gene, thbs2 has not yet been studied in teleost. In the present study, this novel fish gene from Japanese flounder was cloned and its function in resistant to lymphocystis disease virus was elucidated. The Japanese flounder thbs2 encoded a 1176-amino acid protein with 91% identity to medaka. Amino acid sequence indicated that Japanese flounder Thbs2 contained 10 typical conserved domains. The thbs2 was expressed in all stages of embryo development, and in hatched larva stage, its expression was significantly higher than that in other stages (P < 0.05). The relative expression level of thbs2 was significantly higher in the head kidney, liver, blood, gill, and heart of the lymphocystis disease virus resistant fish than in sensitive fish (P < 0.05); and in muscle, this difference was at highly significant (P < 0.01). Additionally, the distribution of Thbs2 in tissue was evaluated by immunohistochemical staining. Subcellular localization analysis showed that Thbs2 was distributed throughout the cytoplasm of the cells. Taken together, our results provide new basic data for thbs2 function, especially its role in anti-lymphocystis disease virus immune response.

Analysis of the current situation and trend of home-based individualized nursing for residents in a certain area in China.

OBJECTIVE: The aim of this study is to examine the perception, willingness to engage, and demand of community residents regarding the 'internet + nursing service' in a designated pilot area, aiming to offer insights for the widespread adoption of the 'internet + nursing service' throughout China. METHODS: A survey pertaining to the 'internet + nursing service' was conducted from March to April 2022. The study specifically targeted residents within two sub-districts of a city in the Jiangsu province. The sampling technique employed in this study was stratified random sampling. RESULTS: Out of a total of 400 community residents selected from two sub-districts in this region, 378 provided valid responses, resulting in an effective rate of 94.5%. Within the study cohort, 80 participants (21.16%) demonstrated familiarity with the concept of 'internet + nursing service.' Additionally, 231 participants (61.11%) conveyed their willingness to adopt such services. Regarding service preferences, the primary demands were for health guidance, vital sign monitoring, and basic care. Challenges in implementing the service were attributed to concerns related to medical risks, personal safety for both nurses and patients, and potential breaches of privacy. CONCLUSIONS: Residents in the pilot area exhibited a moderate awareness of the 'internet + nursing service,' with a relatively high willingness to embrace the program. There is a need for further refinement of pertinent laws, widespread dissemination of policies, and enhancements in the quality of nursing services. These measures aim to ensure that a greater number of community residents can avail themselves of improved home-based nursing services.

Evaluation of Female Recipient Infertility and Donor Spermatogonial Purification for Germ Cell Transplantation in Paralichthys olivaceus.

Since the advent of germ cell transplantation (GCT), it has been widely used in shortening the fish breeding cycle, sex-controlled breeding and the protection of rare and endangered fish. In this study, the effectiveness of female sterile recipient preparation and donor stem cell isolation and purification were comprehensively evaluated for spermatogonial stem cell transplantation (SSCT) in Paralichthys olivaceus. The best way to prepare sterile recipients was found to be giving three-year-old fish four intraovarian injections of busulfan (20 mg/kg body weight) combined with exposure to a high temperature (28 °C) after the spawning season compared with the two other ways, which induced apoptosis of most of the endogenous germ cells, resulting in shrinkage of the spawning plate and enlargement of the ovarian lumen. Further analysis showed that both the gonadosomatic index and germ-cell-specific vasa expression were significantly lower than those of the natural-temperature group before treatment (p < 0.05). A high percentage (>60.00%) of spermatogonial stem cells (SSCs) were obtained after isolation and purification and were transplanted into the prepared recipients. After three weeks of SSCT, the numbers of PKH26-labeled SSCs were increased in the ovaries of the recipients. These findings provide a basis for the establishment of an ideal SSCT technique using P. olivaceus females as the recipients, ultimately contributing to the efficient conservation of male germplasm resources and effective breeding.

Synchronously Mature Intersex Japanese Flounder (Paralichthys olivaceus): A Rare Case.

Japanese flounder is usually gonochoristic, with gonads that are either testes or ovaries. Here, we report an unusual case of hermaphroditism in Japanese flounder captured from the Bohai Sea. In the intersex flounder, the membrane of the upper ovary was closely connected to the abdominal muscles and internal organs, and the eggs filled the entire abdomen. The lower ovary was small and closely connected to the testes. The testes contained few fully mature sperm. Both eggs and sperm were capable of fertilization. The levels of several reproduction-related hormones (17ß-estradiol, 11-ketotestosterone, 17α, 20ß-dihydroxyprogesterone, luteinizing hormone, follicle-stimulating hormone, and testosterone) in the intersex flounder were intermediate, between those in females and males. The results showed that the heterozygosity of the intersex flounder was 0.632, and there were 28 single-nucleotide polymorphisms in the cyp21a gene. Compared with that of wild flounder, the activity of 21-hydroxylase was reduced by approximately 20.0%, and expressions of cyp19a, amh, and dmrt1 differed. We present the first report of its kind, detailing the anatomy, hormonal endocrinology, molecular biology, and physiology of the intersex Japanese flounder.

Diagnostic Significance of Combined Calcitoninogen, Platelet, and D-Dimer Assay in Severe Heatstroke: with Clinical Data Analysis of 70 Patients with Severe Heatstroke.

The significance of calcitoninogen detection among inpatients was discussed by analyzing the clinical characteristics of severe heatstroke (HS). HS patients who were admitted to the Second Hospital of Nantong University, Jiangsu Province, China, between July 1, 2015, and October 30, 2020, were reviewed. Patients' clinical characteristics and laboratory data were recorded, and they were divided into three groups, that is, a control group (heat cramps and heat exhaustion), an exertional HS (EHS) group, and a classical HS (CHS) group to compare the differences among them. Receiver operating characteristic (ROC) curves were plotted to evaluate patients' clinical utility. (1) The body temperatures in the EHS and CHS groups were significantly higher than in the control group (all p < 0.05). (2) The D-dimer (DD), procalcitonin (PCT), and Acute Physiology and Chronic Health Evaluation (APACHE) II score of the EHS group were significantly higher compared with the control and CHS groups (all p < 0.05); the platelets (PLT), C-reactive protein (CRP), blood sodium (Na), and intravenous glucose (GLU) of the EHS group were lower than in the control and CHS groups (all p < 0.05). (3) The ROC curve analysis showed the performance results for DD (area under the curve [AUC] 0.670, 95% confidence interval [CI] 0.547-0.777), PCT (AUC 0.705, 95% CI 0.584-0.808), and PLT (AUC 0.791, 95% CI 0.677-0.879). The sensitivity was 40.48%, 100%, and 73.81%, and the specificity was 96.43%, 32.14%, and 78.57%, respectively. Using three combined analyses, an elevated AUC of 0.838, 95% CI 0.731-0.916, with a sensitivity of 71.43% and a specificity of 85.71%, respectively, was revealed. Patients in the EHS group had higher DD, PCT, and APACHE II values, whereas PLT, CRP, Na, and GLU were reduced. The apparent decrease in the PLT, as well as the increase in PCT and DD values, could be considered as early sensitivity indicators of severe HS. A combined test of these three indicators presented significant diagnostic value for detecting severe cases of HS.

Establishment and Characterization of a Spermatogonial Stem Cell Line from Tiger Puffer Fish (Takifugu rubripes).

Tiger puffer fish (Takifugu rubripes) has become the main fish species cultured in China since the last century because of its high economic value. Male and female tiger puffer fish need 2 and 3 years each to reach sexual maturity, which limits the development of breeding research for this species. In recent years, in vitro culture of fish spermatogonial stem cells (SSCs) have shown potential in aquaculture. In the present study, we established a spermatogenic stem cell line from T. rubripes (TrSSCs). TrSSCs were characterized by polygonal morphology, predominantly retained 44 chromosomes, and grew rapidly at 26 °C and in L-15. TrSSCs were still able to grow stably after more than one year of in vitro culture. TrSSCs showed positive alkaline phosphatase staining. TrSSCs expressed germ cell-associated genes, including dnd, ddx4, piwil, gfra1b, sox2, myca, nanog, ly75, and dazl, as determined by semiquantitative assays, and almost all cells were found to express the germ cell genes ddx4 and gfra1b in a fluorescence in situ hybridization assay. In vitro, induction experiments demonstrated the TrSSCs possessed the ability to differentiate into other types of cells. Our research has enriched the fish spermatogonial stem cell resource bank, which will provide an efficient research model for sex determination and sex control breeding in fish, establishing a foundation for subsequent breeding research.

The application value of the Modified Early Warning Score combined with age and injury site scores in the evaluation of injuries in emergency trauma patients.

Objective: To explore the application value of the Modified Early Warning Score (MEWS) combined with age and injury site scores in predicting the criticality of emergency trauma patients. Methods: The traditional MEWS was modified by combining it with age and injury site scores to form a new MEWS combined scoring standard. The clinical data were collected from a total of 372 trauma patients from the emergency department of the Nantong First People's Hospital between June and December 2019. A retrospective analysis was conducted, and the patients were scored using the MEWS combined with age and injury site scores. The patients were grouped according to their prognoses and clinical outcomes. A statistical analysis was conducted based on the ranges of the combined scores, and the results of the combined scores of the different groups were compared. Results: Among the 372 patients, the average score was 3.68 ± 1.25 points in the survival group, 8.33 ± 2.24 points in the death within 24 h group, and 8.38 ± 1.51 points in the death within 30 days of hospitalization group, and the differences were statistically significant (p < 0.05). The average score was 2.74 ± 0.69 points in the outpatient treatment group, 4.19 ± 0.72 points in the emergency stay group, 5.40 ± 0.70 points in the specialist inpatient group, 8.71 ± 2.31 points in the ICU group, and 7.82 ± 1.66 points in the specialist unplanned transfer to ICU group, with the differences between the groups being statistically significant (p < 0.05). The average length of hospital stay for patients with a joint score within the range of 6-8 points was 10.86 ± 2.47 days, with a direct ICU admission rate of 22.00% and an unplanned ICU admission rate of 16.00%. Patients with a joint score >8 points had an average length of hospital stay of 27.05 ± 4.85 days, with a direct ICU admission rate of 66.67% and an unplanned ICU admission rate of 33.33%. Conclusion: Age and injury site are important high-risk indicators for trauma assessment, and using them in combination with the MEWS could improve the assessment of emergency patients with trauma, increasing the accuracy of pre-screening triage and reducing rescue time. Therefore, this joint scoring method might be worthy of clinical promotion and application.

Evaluating the Price, Availability, and Affordability of Essential Medicines in Primary Healthcare Institutions: A Mixed Longitudinal and Cross-Sectional Study in Jiangsu, China.

Objective: This study aimed to evaluate the price, availability, and affordability of essential medicines in primary healthcare institutions in Jiangsu Province. Methods: A mixed longitudinal and cross-sectional survey was conducted in primary healthcare institutions in Jiangsu based on the adjusted World Health Organization and Health Action International methodology. 45 essential medicines were collected from 30 primary healthcare institutions in Nanjing from 2016 to 2020. We also collected information on these medicines in 70 primary healthcare institutions across seven cities of Jiangsu in 2021. The availability, price, and affordability were compared with matched sets. Differences of availability between years and cities were further compared using Wilcoxon rank-sum test. Results: In Nanjing, the variation was significant of availability during the study period. The MPR was generally decreasing between 2016 and 2020, with the median price ratio (MPR) for lowest-priced generics (LPGs) ranging from 1.20 to 2.53 and originator brands (OBs) substantially above international levels. The median availability of generic medicines increased in 2018 and subsequently stabilized at around 55%, and the availability of originator medicines was low. There were no significant regional differences in prices across the sampled cities in Jiangsu, and the median MPR for LPGs was acceptable (1.23), while the median MPR for OBs was 8.54. The mean availability was different across regions (p < 0.001), being higher in Nanjing (54.67%) and Nantong (56.22%), and lower in northern Jiangsu (about 35%). For LPGs, there was little difference in the proportion of medicines with low availability and high affordability (50.00% for urban residents and 40.48% for rural residents). For OBs, there were more than half of rural residents had low availability and low affordability of medicines (58.82%). Conclusions: In terms of yearly changes, the prices of essential medicines have considerably decreased, and the availability of LPGs has slightly increased. However, the availability of medicines was found to be poor and there were regional differences in the availability and affordability of medicines among metropolitan and rural areas. Policy interventions targeting external factors associated with health resource allocation are essential and possible strategies include effective and efficient government investment mechanisms on primary healthcare.

A New Cell Line Derived from the Spleen of the Japanese Flounder (Paralichthys olivaceus) and Its Application in Viral Study.

A new cell line Japanese flounder spleen (JFSP) derived from the spleen of Japanese flounder (Paralichthys olivaceus) was established and characterized in this study. The JFSP cells grew rapidly at 29 °C, and the optimum fetal bovine serum concentration in the L-15 medium was 15%. Cells were subcultured for more than 80 passages. The JFSP cells have a diploid chromosome number of 2n = 68, which differs from the chromosome number of normal diploid Japanese flounder. The established cells were susceptible to Bohle virus (BIV), Viral hemorrhagic septicemia virus (VHSV), Hirame rhabdovirus (HIRRV), Infectious hematopoietic necrosis virus (IHNV), and Lymphocystis disease virus (LCDV), as evidenced by varying degrees of cytopathic effects (CPE). Replication of the virus in JFSP cells was confirmed by qRT-PCR and transmission electron microscopy. In addition, the expression of four immune-related genes, TRAF3, IL-1ß, TNF-α, and TLR2, was differentially altered following viral infection. The results indicated that the cells underwent an antiviral immune response. JFSP cell line is an ideal tool in vitro for virology. The use of fish cell lines to study the immune genes and immune mechanism of fish and to clarify the immune mechanism of fish has important theoretical significance and practical application value for the fundamental prevention and treatment of fish diseases.

The Isolation and Full-Length Transcriptome Sequencing of a Novel Nidovirus and Response of Its Infection in Japanese Flounder (Paralichthys olivaceus).

A novel nidovirus, CSBV Bces-Po19, was isolated from the marine fish, Japanese flounder (Paralichthys olivaceus). The viral genome was 26,597 nucleotides long and shared 98.62% nucleotide identity with CSBV WHQSR4345. PacBio Sequel and Illumina sequencing were used to perform full-length transcriptome sequencing on CSBV Bces-Po19-sensitive (S) and -resistant (R) Japanese flounder. The results of negative staining revealed bacilliform and spherical virions. There were in total 1444 different genes between CSBV Bces-Po19 S and R groups, with 935 being up-regulated and 513 being down-regulated. Metabolism-, immune-, and RNA-related pathways were significantly enriched. Furthermore, CSBV Bces-Po19 infection induced alternative splicing (AS) events in Japanese flounder; the S group had a higher numbers of AS events (12,352) than the R group (11,452). The number of long non-coding RNA (lncRNA) in the S group, on the other hand, was significantly lower than in the R group. In addition to providing valuable information that sheds more light on CSBV Bces-Po19 infection, these research findings provide further clues for CSBV Bces-Po19 prevention and treatment.

Effect and prognosis of emergency nursing path in patients with acute stroke.

OBJECTIVE: To evaluate the effect and prognosis of the emergency nursing path in patients with acute stroke. METHODS: In this prospective study, 108 patients with acute stroke were randomly divided into an observation group (n=54, implementing the emergency nursing path) and a control group (n=54, implementing emergency routine care). We compared the rescue time-related indicators, neurological function, cognitive function and prognosis of the two groups. RESULTS: The time from admission to diagnosis and the time from diagnosis to specialist treatment in the observation group were shorter than those in the control group (all P<0.05). After two weeks of intervention, the score of National Institute of Health Stroke Scale in the two groups of patients were lower than before the intervention, and the observation group was even lower (all P<0.05). After one month of discharge, the score of Activity of Daily Living Scale decreased while the scores of Montreal Cognitive Assessment, Mini-mental State Examination and Glasgow Outcome Scale increased in both groups, with more significant changes in the observation group (all P<0.05). The disability rate in the observation group was lower than that in the control group (P<0.05). CONCLUSION: The implementation of emergency nursing could significantly shorten the time from admission to specialist treatment for patients with acute stroke. It could also promote the recovery of neurological and cognitive functions, which was conducive to the prognosis of patients.

Analysis of the Status Quo and Influencing Factors of Community Residents' Awareness of Basic Life Support and Willingness to Attempt Rescue.

OBJECTIVE: This study aimed to investigate community residents' awareness of basic life support (BLS) and their willingness to attempt rescue. METHODS: From October to December 2020, in the communities of Nantong City, a stratified three-stage random sampling method was adopted to select residents from 12 neighborhood committees over the age of 18 with whom to conduct a cross-sectional questionnaire survey. A self-designed questionnaire was adopted, the contents of which included the general situation of the respondent, knowledge, attitude, and behavior in relation to BLS; the Cronbach's α coefficient of the questionnaire was 0.719. RESULTS: A total of 3000 questionnaires were distributed, of which 2812 were valid, with a valid response rate of 93.73%. Of the 2812 respondents, 41.18% had seen an automatic external defibrillator (AED), 48.83% had experience of' cardiopulmonary resuscitation (CPR), and 25.07% of the respondents had experience of' AEDs. When an accident occurred, 50.50% of residents were willing to attempt rescue, 70.80% were willing to attempt rescue under professional guidance, and 71.23% were willing to attempt rescue after learning BLS techniques. Of the residents who were unwilling to attempt rescue, 32.75% were worried about their lack of ability, 27.91% were concerned about legal issues, 14.01% feared infectious diseases, and 10.35% were unwilling to perform mouth-to-mouth artificial respiration. Age, occupation, education level, and whether they had participated in first aid training were the influencing factors. CONCLUSION: Residents in Nantong have less knowledge of BLS, and their knowledge of CPR is better than that of AEDs. Residents have a strong willingness to learn BLS. Measures need to be taken to improve their understanding of BLS and their application skills. Residents have high levels of willingness to attempt rescue, but a certain percentage of residents have concerns. Interventions can be made to target the different reasons.

The Effect of Infertility on the Liver Structure, Endocrinology, and Gene Network in Japanese Flounder.

The liver can synthesize vitellogenin, the precursor of vitellin, which is needed for oocyte development and maturation. Here, we investigated the effects of infertility on liver structure, hormone regulation, and gene and protein networks in Japanese flounder (Paralichthys olivaceus). Results showed that the liver of infertile fish had fewer vacuoles and significantly lower serum vitellogenin (VTG) level than in liver of fertile fish. Whole transcriptomics analysis between infertile and fertile groups identified 2076 significantly differentially expressed (DE) mRNAs, 431 DE lncRNAs, 265 DE circRNAs, and 53 DE miRNAs. Proteomics analysis identified 838 DE proteins. Integrated analysis of whole transcriptomics and proteomics revealed 60 significantly DE genes and proteins associated with metabolism, immunity, signal transduction, and steroid biosynthesis. Moreover, non-coding RNA (miRNAs, circRNA, and lncRNA) transcripts involved in metabolism, immunity, and signal transduction in infertile liver were identified. In conclusion, this study shows that gonadal infertility is associated with not only changes in histological structure and hormone secretion but also changes in metabolism, immunity, and signal transduction networks in the liver. These results provide valuable information concerning the mechanism underlying infertility in fish.

Toxicity of nickel and cobalt in Japanese flounder.

Nickel and cobalt are essential elements that become toxic at high concentrations. Little is known about nickel and cobalt toxicity in aquatic animals. This study aimed to investigate acute and chronic toxicity of nickel and cobalt in Japanese flounder (Paralichthys olivaceous), with emphasis on oxidative stress reactions, histopathological changes, and differences in gene expression. The lethal concentration for 50% mortality (LC50) in 3 and 8 cm Japanese flounder exposed to nickel for 96 h was found to be 86.2 ± 0.018 and 151.3 ± 0.039 mg/L; for cobalt exposure, LC50 was 47.5 ± 0.015 and 180.4 ± 0.034 mg/L, respectively. Chronic nickel and cobalt exposure caused different degrees of oxidative enzyme activity changes in gill, liver, and muscle tissues. Erythrocyte deformations were detected after acute or chronic exposure to nickel and cobalt. the nickel and cobalt exposure also caused pathological changes such as spherical swelling over other gill patches, rod-like proliferations in the gill patch epithelial cell layer, and disorder in hepatocyte arrangement, cell swelling, and cytoplasm loosening. RNA-Seq indicated that there were 184 upregulated and 185 downregulated genes in the liver of Japanese flounder exposed to 15 mg/L nickel for 28 d. For cobalt, 920 upregulated and 457 downregulated genes were detected. Among these differentially expressed genes, 162 were shared by both nickel and cobalt exposure. In both nickel and cobalt, pathways including fatty acid elongation, steroid biosynthesis, unsaturated fatty acid biosynthesis, fatty acid metabolism, PPAR signaling, and ferroptosis were significantly enriched. Taken together, these results aided our understanding of the toxicity of nickel and cobalt in aquatic animals.