Environmentally relevant concentrations of Mn2+ disrupts the endocrine regulation of growth in juvenile Yunlong groupers (Epinephelus moara♀×Epinephelus lanceolatus♂).

Even though manganese is a bioelement essential for metabolism, excessive manganese levels in water can be detrimental to fish development and growth. Therefore, the aim of this study was to evaluate the effects of Mn2+ (0, 0.5,1, 2, and 4 mg·L-1) exposure for 30 d on the growth performance, growth hormone/insulin-like growth factor (GH/IGF) axis, hypothalamic-pituitary-thyroid (HPT) axis, and monoaminergic neurotransmitters of Epinephelus moara♀×Epinephelus lanceolatus♂(Yunlong grouper). Compared with the control and low Mn2+concentration groups of (0.5 and 1 mg·L-1), the high concentration of Mn2+ (4 mg·L-1) significantly reduced body weight (BW), body length (BL), weight gain rate (WGR), and specific growth rate (SGR), increased the feed coefficient rate (FCR) and mortality of Yunlong groupers (P < 0.05). Further, the levels of GH and IGF, along with the expression of ghra and ghrb were significantly reduced after exposure to 2 and 4 mg·L-1 Mn2+for 30 d, whereas the expression of sst5 was significantly up-regulated after exposure to 2 and 4 mg·L-1 Mn2+for 20 and 30 days. Moreover, Mn2+exposure increased thyroid hormone (T3) and thyroid stimulating hormone (TSH) contents, accompanied by increased mRNA levels of dio1 and dio2, however, the T4 level was decreased. Finally, dopamine (DA) and serotonin (5-HT) levels significantly decreased after long-term exposure to higher concentrations of Mn2+, and the levels their metabolites changed as well, suggesting that the synthesis and metabolism of DA and 5-HT were affected. Accordingly, changes in the GH/IGF and HPT axes-related parameters may be the cause of growth inhibition in juvenile groupers under Mn2+ exposure, indicating that the relationship between endocrine disorder and growth inhibition should not be ignored.

[Clinical application of targeted sealing with high viscosity bone cement and secondary injection of low viscosity bone cement in vertebroplasty].

OBJECTIVE: To observe the clinical efficacy of targeted sealing with high viscosity bone cement and secondary injection of low viscosity bone cement in the treatment of OVCFs patients with the fracture lines involved vertebral body margin. METHODS: The elderly patients who underwent vertebroplasty for osteoporotic vertebral compression fractures from January 2019 to September 2021 were selected as the screening objects. Through relevant standards and further CT examination, 56 patients with fracture lines involving the anterior wall or upper and lower endplates of the vertebral body were selected for the study. There were 21 males and 35 females, aged from 67 to 89 years old with an average of (76.58±9.68) years. All 56 patients underwent secondary injection of bone cement during operation. Only a small amount of high viscosity cement was targeted to seal the edge of the vertebral body for the first time, and low viscosity cement was injected to the vertebral bodies during second bolus with well-distributed. The operation time, bone cement volume and bone cement leakage were recorded, and the pain relief was evaluated by visual analogue scale (VAS). RESULTS: All patients were followed up for more than 3 months and the surgeries were successfully complete. The operation time was (50.41±10.30) min and the bone cement volume was (3.64±1.29) ml. The preoperative VAS was (7.21±2.41) points, which decreased significantly to (2.81±0.97) points 3 days after operation(P<0.05). Among the 56 patients, 2 cases(3.57%) had bone cement leakage, 1 case leaked to the paravertebral vein, and 1 case slightly bulged to the paravertebral through the crack when plugging the vertebral crack. Both patients had no obvious clinical symptoms. CONCLUSION: In vertebroplasty surgery, targeted sealing of high viscosity bone cement and secondary injection of low viscosity bone cement can reduce intraoperative bone cement leakage and improve the safety of operation.

Diagnostic performance of C-TIRADS combined with SWE for the diagnosis of thyroid nodules.

Objective: To explore the value of the optimal parameters of shear wave elastography (SWE) to enhance the identification of benign and malignant thyroid nodules by C-TIRADS. Methods: The two-dimensional ultrasonography images and SWE images of 515 patients with a total of 586 thyroid nodules were retrospectively analyzed. The nodules were divided into the D ≤10 mm and D >10 mm groups according to size and were graded by C-TIRADS. With the pathological results as the gold standard, the receiver operating characteristic (ROC) curves were drawn, and the area under the curve (AUC) was calculated to compare the diagnostic performances of C-TIRADS, SWE, and the combination of the two on the benign and malignant thyroid nodules. Results: The ROC showed that the AUC of the maximum elastic modulus (0.875) was higher than that of the mean elastic modulus (0.798) and elasticity ratio (0.772), with an optimal cutoff point of 51 kPa, which was the optimal parameter to distinguish the malignant from the benign nodules (P < 0.001). In the D ≤10 mm group, the AUC of TIRADS combined with SWE (0.955) was elevated by 0.172 compared with the application of C-TIRADS alone (0.783), and the difference was statistically significant (P < 0.05). In the D >10 mm group, the AUC of TIRADS combined with SWE (0.904) was elevated by 0.076 compared with the application of C-TIRADS alone (0.828), and the difference was statistically significant (P < 0.05). Among all nodules, the application of C-TIRADS alone had a sensitivity of 88.14%, a specificity of 74.56%, and an accuracy of 85.50% in diagnosing benign and malignant thyroid nodules, while the sensitivity, specificity, and accuracy were 93.22%, 90.35%, and 92.66%, respectively, in combination with SWE. Conclusion: The diagnostic performance of SWE in combination with TIRADS was better than that of SWE or C-TIRADS alone. Here, SWE enhanced the diagnostic performance of C-TIRADS for the benign and malignant thyroid nodules, most significantly for nodules with D ≤10 mm.

Bioaccumulation of manganese and its effects on oxidative stress and immune response in juvenile groupers (Epinephelus moara ♀ × E. lanceolatus ♂).

We evaluated the effects of Mn in juvenile Yunlong groupers (Epinephelus moara ♀ × E. lanceolatus ♂). The groupers were exposed to Mn2+ (0, 0.5, 1, 2, and 4 mg/L) for 30 days after which they were assessed. The results indicate the accumulation of Mn in fish depended on dose and time. Mn2+ accumulation in tissues occurred in the following order: liver > gills > intestine > muscle. The concentrations of SOD and CAT in the fish significantly increased after 10 and 20 days of treatment with 4 mg/L Mn2+ but decreased after 30 days. Similarly, GSH and GPx levels increased after 10 days of exposure to 2 and 4 mg/L Mn2+ but decreased after 20 and 30 days of exposure. Additionally, malondialdehyde levels significantly increased after exposing the fish to 2 and 4 mg/L Mn2+ for 10, 20, and 30 days. In addition, liver HSP70 and HSP90 levels significantly increased at days 20 and 30 in all fish exposed to Mn2+. In addition, when Mn2+ concentration was 1, 2, and 4 mg/L, liver C3 and C4 levels were significantly increased after 10, 20, and 30 days. Conversely, the levels of LZM and IgM significantly decreased. Mn2+ also significantly upregulated the expression of genes associated with immunity (tlr3, tnf-α, il-1ß, and il-6) in the fish, which suggests that it induces immunotoxicity by altering the immune response. Overall, the findings showed that Mn2+ can disrupt grouper health by bioaccumulating in the fish and subsequently inducing oxidative stress and immune responses. These results can help elucidate the mechanism by which manganese induces toxicity in marine fish. Additionally, they provide a new perspective regarding the detrimental effects of heavy metals in fish.

ISOC1 is a novel potential tumor suppressor in hepatocellular carcinoma.

Isochorismatase domain-containing 1 (ISOC1) plays a carcinogenic role in various tumors. However, its expression and role in hepatocellular carcinoma (HCC) have not been elucidated. This is the first study to investigate the involvement of ISOC1 in HCC growth and migration. ISOC1 expression was analyzed using public databases and clinical samples, and clinical specimens were analyzed by real-time quantitative polymerase chain reaction, western blotting, and immunohistochemistry. ISOC1 was also overexpressed in two HCC cell lines (Huh7 and HepG2) to explore how ISOC1 affects HCC cells. Finally, a nude mouse xenograft tumor model was used to investigate the role of ISOC1 in HCC cell tumorigenicity. ISOC1 was downregulated in HCC tissues compared to that in matched paracancerous tissues, and low ISOC1 expression was associated with a poor prognosis. The proliferation and single-cell colony-forming ability of the ISOC1-overexpressing cell lines Huh7 and HepG2 were significantly inhibited. Moreover, ISOC1 overexpression suppressed the migration and invasion abilities of HCC cells in vitro, and ISOC1 upregulation hindered tumor growth in the xenograft tumor model in vivo. Therefore, ISOC1 is a potential HCC suppressor protein.

Alterations in hematological and biochemical parameters, oxidative stress, and immune response in Takifugu rubripes under acute ammonia exposure.

Ammonia is a major pollutant in aquatic environments and poses a considerable threat to the survival of fish. In this study, we investigated the toxic effects of ammonia on the hematological and biochemical parameters, oxidative stress, and immune responses in Takifugu rubripes. Juvenile T. rubripes (average weight 246.17 ± 3.54 g) were exposed to different concentrations of ammonia (0, 5, 50, 100, and 150 mg/L) for 96 h. The results showed that the hematological parameters (hemoglobin, hematocrit, red blood cell, and white blood cell count) were significantly reduced in response to ammonia exposure. Of the plasma components, such as serum total protein, albumin, glucose, glutamic-oxalacetic transaminase, and glutamic-pyruvic transaminase, were significantly altered in response to ammonia exposure. Additionally, the levels of superoxide dismutase (SOD), catalase (CAT), glutathione (GSH), and glutathione peroxidase (GPx) were increased after exposure to low concentration ammonia exposure. However, when fish were exposed to a high concentration of ammonia, these parameters showed the opposite trend, suggesting that an increase in antioxidant enzymes during the early stages of ammonia exposure may contribute to the removal of the induced reactive oxygen species (ROS) and protect the cells from oxidative damage. However, as the ammonia concentration and exposure time increased, the overproduction of ROS accelerated the depletion of antioxidant enzymes. Ammonia exposure significantly increased the expression of heat shock proteins (HSP70 and HSP90). Ammonia poisoning elevated gene expressions of TLR-3, TNF-α, IL-6, IL-12, and IL-1ß in the gills, causing an inflammatory response. Our findings provide new insights into the mechanisms involved in ammonia-induced aquatic toxicology in marine fish, which may aid in their captive management.

Impact of nitrite exposure on plasma biochemical parameters and immune-related responses in Takifugu rubripes.

Nitrite is a major environmental pollutant in aquatic environments that negatively affects aquatic species. In this study, we investigated the impact of nitrite exposure on plasma biochemical parameters and immune responses in Takifugu rubripes. Fish were exposed to various concentrations of nitrite (0, 0.5, 1, 3, and 6 mM) for 96 h. After 0, 12, 24, 48, and 96 h of exposure, fish blood samples were collected to assay the levels of total protein (TP), albumin (Alb), glutamic-oxaloacetic transaminase (GOT), glutamic-pyruvic transaminase (ALT), complement C3 (C3), complement C4 (C4), immunoglobulin (IgM), and lysozyme activity (LZM). The gills were sampled to analyze the mRNA levels of heat shock protein 70 (hsp70), heat shock protein 90 (hsp90), tumor necrosis factor α (tnf-α), B-cell activating factor (baff), interleukin-6 (il-6), and interleukin-12 (il-12). Levels of GOT, ALT, C3, and C4 were significantly enhanced in the high nitrite concentration group (3 and 6 mM), whereas those of TP, Alb, LZM, and IgM decreased significantly with the same treatments. Nitrite significantly upregulated hsp70, hsp90, tnf-α, il-6, il-12, and baff mRNA levels after 96 h of exposure. These results indicated that nitrite exposure altered the blood physiological status and immune system response, resulting in dysfunction and immunotoxicity in T. rubripes. Furthermore, our results reveal the possible mechanism of aquatic-nitrite-induced toxicity in fish.

Effect of acute exposure to nitrite on physiological parameters, oxidative stress, and apoptosis in Takifugu rubripes.

In the present study, we evaluated the effects of nitrite exposure on hematological parameters, oxidative stress, and apoptosis in juvenile Takifugu rubripes. The fish were exposed to nitrite (0, 0.5, 1, 3, and 6 mM) for up to 96 h. In the high nitrite concentration groups (i.e., 3 and 6 mM), the concentrations of methemoglobin (MetHb), cortisol, glucose, heat shock protein (Hsp)-70, Hsp-90, and potassium (K+) were significantly elevated. Whereas, the concentrations of hemoglobin (Hb), triglyceride (TG), total cholesterol (TC), and sodium (Na+) and chloride (Cl-) ions were significantly decreased. Compared with those of the control groups, the concentrations of the antioxidant enzymes, namely, superoxide dismutase (SOD), catalase (CAT), glutathione (GSH), and glutathione peroxidase (GPx), in the gills were considerably elevated at 12 and 24 h after exposure to nitrite (1, 3, and 6 mM), but reduced at 48 and 96 h. The increase in the antioxidant enzymes may contribute to the elimination of reactive oxygen species (ROS) induced by nitrite during early nitrite exposure, when the antioxidant system is not sufficiently effective to eliminate or neutralize excessive ROS. In addition, we found that nitrite exposure could alter the expression patterns of some key apoptosis-related genes (Caspase-3, Caspase-8, Caspase-9, p53, Bax, and Bcl-2). This indicated that the caspase-dependent apoptotic pathway and p53-Bax-Bcl-2 pathway might be involved in apoptosis induced by nitrite exposure. Furthermore, our study provides insights into how acute nitrite exposure affects the physiological responses and potential molecular mechanism of apoptosis in marine fish. The results can help elucidate the mechanisms involved in nitrite-induced aquatic toxicology in marine fish.

Exposure to nitrite alters thyroid hormone levels and morphology in Takifugu rubripes.

Nitrite (NO2-) can act as a toxic nitrogenous compound with the potential to disrupt endocrine systems in fish. The aim of the present study was to investigate the effects of nitrite on the thyroid endocrine system of Takifugu rubripes. Fish were exposed to 0, 0.5, 1, 3, and 6 mM nitrite concentrations. Blood was collected to assay the concentrations of thyroid-stimulating hormone (TSH), thyroxine (T4), triiodothyronine (T3), free thyroxine (FT4), free triiodothyronine (FT3), and 3,3,5'-triiodothyronine (rT3), as well as the activity of iodothyronine deiodinases (Dio1, Dio2, and Dio3,) after 0, 12, 24, 48, and 96 h of exposure to nitrite. The first branchial arch to the third branchial arch of T. rubripes were sampled and fixed, and thyroid morphology was observed. The results showed that exposure to nitrite significantly increased the concentrations of TSH, T3, FT3, and reduced the concentrations of T4, FT4, and rT3. The activity of Dio1 and Dio2 increased significantly, whereas Dio3 activity decreased significantly. Additionally, thyroid follicles degenerated and became blurred and most colloid material disappeared 96 h after exposure to high nitrite concentrations. Based on these results, high nitrite concentration exposure can disturb thyroid hormone homeostasis, alter thyroid follicle morphology, and result in thyroid endocrine toxicity.

Expression of Fra-1 in human hepatocellular carcinoma and its prognostic significance.

This study aimed to explore the clinical significance and prognostic value of Fra-1 in hepatocellular carcinoma patients after curative resection. Fra-1 expression was investigated using a combination of techniques: immunohistochemistry for 66 samples of hepatocellular carcinoma and quantitative real-time polymerase chain reaction and western blotting assays for 19 matched hepatocellular carcinoma specimens. Fra-1 was present in 38 of 66 (57.6%) tumor tissues, with intense staining in the nuclei. There was also positive staining in 14 of 66 (21.2%) adjacent peritumoral tissues, with weak staining in the cytoplasm. Quantitative real-time polymerase chain reaction and western blotting assays confirmed higher expression of Fra-1 messenger RNA and Fra-1 protein in tumor tissues than adjacent non-tumor tissues for 19 hepatocellular carcinoma samples (p < 0.001). Positive expression of Fra-1 was significantly related to vascular invasion and serum alpha-fetoprotein. Kaplan-Meier survival analysis found that overexpressed Fra-1 was correlated with poor overall survival and disease-free survival. Multivariate analysis identified Fra-1 as an independent prognostic factor. Fra-1 may be involved in the progress of hepatocellular carcinoma and could be a promising molecular candidate in the diagnosis and treatment of hepatocellular carcinoma.

Developmental changes in digestive enzyme activity in American shad, Alosa sapidissima, during early ontogeny.

In order to assess the digestive physiological capacity of the American shad Alosa sapidissima and to establish feeding protocols that match larval nutritional requirements, we investigated the ontogenesis of digestive enzymes (trypsin, amylase, lipase, pepsin, alkaline phosphatase, and leucine aminopeptidase) in larvae, from hatching to 45 days after hatching (DAH). We found that all of the target enzymes were present at hatching, except pepsin, which indicated an initial ability to digest nutrients and precocious digestive system development. Trypsin rapidly increased to a maximum at 14 DAH. Amylase sharply increased until 10 DAH and exhibited a second increase at 33 DAH, which coincided with the introduction of microdiet at 30 DAH, thereby suggesting that the increase was associated with the microdiet carbohydrate content. Lipase increased until 14 DAH, decreased until 27 DAH, and then increased until 45 DAH. Pepsin was first detected at 27 DAH and then sharply increased until 45 DAH, which suggested the formation of a functional stomach. Both alkaline phosphatase and leucine aminopeptidase markedly increased until 18 DAH, which indicated intestinal maturation. According to our results, we conclude that American shad larvae possess the functional digestive system before mouth opening, and the significant increases in lipase, amylase, pepsin, and intestinal enzyme activities between 27 and 33 DAH suggest that larvae can be successfully weaned onto microdiets around this age.

Effects of different salinities on growth performance, survival, digestive enzyme activity, immune response, and muscle fatty acid composition in juvenile American shad (Alosa sapidissima).

The effects of salinity on survival, growth, special activity of digestive enzymes, nonspecific immune response, and muscle fatty acid composition were evaluated in the American shad (Alosa sapidissima). Juveniles of 35 days after hatching were reared at 0 (control), 7, 14, 21, and 28 ppt for 60 days. At the end of the experiment, juvenile American shad presented higher survival and specific growth rate (SGR) in salinity group (7, 14, and 21 ppt) than control group (P < 0.05). The special activity of trypsin and chymotrypsin was highest in fish reared at 21 ppt, while the highest lipase special activity was obtained in control group (P < 0.05). The special activity of alkaline phosphatase (ALP), lysozyme (LZM), superoxide dismutase (SOD), and catalase (CAT) showed significant increases in salinity group (14 and 21 ppt) compared to control group (P < 0.05). Lower muscle ash contents were detected in salinity group (14, 21, and 28 ppt) than control group (P < 0.05), while the contents of crude lipid and crude protein were significantly higher than control group (P < 0.05). The level of monounsaturated fatty acids (MUFA) exhibited a decreasing trend, while an increased level of polyunsaturated fatty acids (PUFA) was detected with the increase of salinity. Among the PUFA, the content of n-3 fatty acids in muscle tissue was found to be increasing with the increasing salinity, especially eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA). Results indicate that appropriate increase in salinity was reasonable and beneficial for juvenile American shad culture after a comprehensive consideration, especially salinity range from 14 to 21 ppt.

Compact KGd(WO4)2 picosecond pulse-train synchronously pumped broadband Raman laser.

We demonstrate an efficient approach to realizing an extra-cavity, synchronously pumped, stimulated Raman cascaded process under low repetition frequency (1 kHz) pump conditions. We also construct a compact KGd(WO4)2 (KGW) crystal picosecond Raman laser that has been configured as the developed method. A pulse-train green laser pumped the corresponding 70 mm long KGW crystal Raman cavity. The pulse train contains six pulses, about 800 ps separated, for every millisecond; thus, it can realize synchronous pumping between pump pulse and the pumped Raman cavity. The investigated system produced a collinear Raman laser output that includes six laser lines covering the 532 to 800 nm spectra. This is the first report on an all-solid-state, high-average-power picosecond collinear multi-wavelength (more than three laser components) laser to our knowledge. This method has never been reported on before in the synchronously pumped stimulated Raman scattering (SRS) realm.

Prognostic value of polarized macrophages in patients with hepatocellular carcinoma after curative resection.

As the most predominant tumour-infiltrating immune cells, tumour-associated macrophages (TAMs) are significant for fostering tumour growth, progression and metastasis. CD68-positive TAMs display dissimilarly polarized programmes comprising CD11c-positive pro-inflammatory macrophages (M1) and CD206-positive immunosuppressive macrophages (M2). The aim of this study is to determine the prognostic significance of diametrically polarized TAMs in hepatocellular carcinoma (HCC) and their application to risk stratification of patients according to their specific prognostic values. This study included 80 consecutive patients with HCC, and we evaluated diametrically polarized functional status of macrophages by immunohistochemical staining of CD68, CD11c and CD206. Prognostic values and clinicopathologic features were assessed in these patients. High versus low CD11c-positive TAM density (P = 0.005) and low versus high CD206-positive TAM density (P = 0.002) were associated with better overall survival, whereas CD68-positive TAM density had no prognostic significance (low versus high, P = 0.065). Furthermore, the presence of these positive staining macrophages did not show any prognostic significance for recurrence-free survival (all P > 0.05). Multivariate Cox regression analysis identified CD11c-positive and CD206-positive TAMs as an independent prognostic factor (P < 0.001, P = 0.031, respectively). Intratumoural infiltration of diametrically polarized TAMs, a novel identified independent prognostic factor for survival in patients with HCC, could be combined with the TNM stage and the Barcelona Clinic Liver Cancer stage to improve a risk stratification system.