Hyoscyamine induces developmental toxicity by disrupting metabolism in zebrafish embryo (Danio rerio).

Hyoscyamine is a kind of tropane alkaloids, which exists in several plants of the family Solanaceae. However, the mechanism underlying such hyoscyamine toxic effects during early development remains unclear. In this study, an untargeted metabolomics approach was used to investigate the toxic mechanisms of hyoscyamine in zebrafish embryos. The LC10 and MNLC of hyoscyamine in zebrafish embryos were determined to be 350 and 313 µg/mL, respectively. Moreover, hyoscyamine exposure increased the accumulation of ROS and MDA, and altered the activity of antioxidant enzymes (CAT, SOD, and GSH) in zebrafish embryos. After exposure, the embryos were extracted, derivatized and analyzed by UHPLC-Q-Orbitrap-HRMS for 3551 metabolites to identify 38 significantly changed metabolites based on the VIP, p value, and fold change results. Metabolic pathways associated with those metabolites were identified using MetaboAnalyst 5.0 as follows: pyrimidine metabolism, purine metabolism, histidine metabolism, beta-Alanine metabolism, and glutathione metabolism. These results suggested that hyoscyamine exposure to zebrafish embryos exhibited marked metabolic disturbance. Such significant perturbations of important metabolites within crucial biochemical pathways may have biologically hazardous effects on zebrafish embryos induced by hyoscyamine.

High fried food consumption impacts anxiety and depression due to lipid metabolism disturbance and neuroinflammation.

Western dietary patterns have been unfavorably linked with mental health. However, the long-term effects of habitual fried food consumption on anxiety and depression and underlying mechanisms remain unclear. Our population-based study with 140,728 people revealed that frequent fried food consumption, especially fried potato consumption, is strongly associated with 12% and 7% higher risk of anxiety and depression, respectively. The associations were more pronounced among male and younger consumers. Consistently, long-term exposure to acrylamide, a representative food processing contaminant in fried products, exacerbates scototaxis and thigmotaxis, and further impairs exploration ability and sociality of adult zebrafish, showing anxiety- and depressive-like behaviors. Moreover, treatment with acrylamide significantly down-regulates the gene expression of tjp2a related to the permeability of blood-brain barrier. Multiomics analysis showed that chronic exposure to acrylamide induces cerebral lipid metabolism disturbance and neuroinflammation. PPAR signaling pathway mediates acrylamide-induced lipid metabolism disorder in the brain of zebrafish. Especially, chronic exposure to acrylamide dysregulates sphingolipid and phospholipid metabolism, which plays important roles in the development of anxiety and depression symptoms. In addition, acrylamide promotes lipid peroxidation and oxidation stress, which participate in cerebral neuroinflammation. Acrylamide dramatically increases the markers of lipid peroxidation, including (±)5-HETE, 11(S)-HETE, 5-oxoETE, and up-regulates the expression of proinflammatory lipid mediators such as (±)12-HETE and 14(S)-HDHA, indicating elevated cerebral inflammatory status after chronic exposure to acrylamide. Together, these results both epidemiologically and mechanistically provide strong evidence to unravel the mechanism of acrylamide-triggered anxiety and depression, and highlight the significance of reducing fried food consumption for mental health.

Methyl Parathion Exposure Induces Development Toxicity and Cardiotoxicity in Zebrafish Embryos.

Methyl parathion (MP) has been widely used as an organophosphorus pesticide for food preservation and pest management, resulting in its accumulation in the aquatic environment. However, the early developmental toxicity of MP to non-target species, especially aquatic vertebrates, has not been thoroughly investigated. In this study, zebrafish embryos were treated with 2.5, 5, or 10 mg/L of MP solution until 72 h post-fertilization (hpf). The results showed that MP exposure reduced spontaneous movement, hatching, and survival rates of zebrafish embryos and induced developmental abnormalities such as shortened body length, yolk edema, and spinal curvature. Notably, MP was found to induce cardiac abnormalities, including pericardial edema and decreased heart rate. Exposure to MP resulted in the accumulation of reactive oxygen species (ROS), decreased superoxide dismutase (SOD) activity, increased catalase (CAT) activity, elevated malondialdehyde (MDA) levels, and caused cardiac apoptosis in zebrafish embryos. Moreover, MP affected the transcription of cardiac development-related genes (vmhc, sox9b, nppa, tnnt2, bmp2b, bmp4) and apoptosis-related genes (p53, bax, bcl2). Astaxanthin could rescue MP-induced heart development defects by down-regulating oxidative stress. These findings suggest that MP induces cardiac developmental toxicity and provides additional evidence of MP toxicity to aquatic organisms.

Hydrogen Sulfide Promotes Osteogenesis by Modulating Macrophage Polarization.

Macrophages, a versatile subset of immune cells, are essential for successful bone repair. Hydrogen sulfide (H2S) is a gasotransmitter associated with tissue development and repair. Emerging evidence demonstrates that H2S is involved in bone formation under physiology condition and bone regeneration under pathology condition. However, whether hydrogen sulfide mediates osteogenesis by influencing macrophages is unknown. Here, we aimed to investigate the effects of hydrogen sulfide on macrophage polarization and the subsequent impact on bone regeneration. In the present study, we found that the H2S-donor GYY4137 stimulated M0/M1 macrophages to express high level of CD-206 and IL-10 but decreased the levels of i-NOS and TNF-α in M1 macrophages. Furthermore, coculture of GYY4137-treated M0 macrophages with pro-osteoblastic MC3T3-E1 cells significantly increased the viability of the MC3T3-E1 cells. Importantly, the formation of mineralized particles in MC3T3-E1 cells was significantly promoted following coculture with IL-4-treated and GYY4137-treated M0 macrophages. Collectively, our study demonstrated that hydrogen sulfide increased macrophages M2 polarization and subsequently promoted bone regeneration.

Scopoletin Induced Metabolomic Profile Disturbances in Zebrafish Embryos.

Scopoletin, a typical example of a coumarin compound, exists in several Artemisia species and other plant genera. However, the systemic metabolic effects induced by scopoletin remain unclear. In the present study, we evaluated the metabolic profiles in scopoletin-exposed zebrafish embryos using UHPLC-Q-Obitrap-HRMS combined with multivariate analysis. Compared with the control group, 33 metabolites in scopoletin group were significantly upregulated, while 27 metabolites were significantly downregulated. Importantly, scopoletin exposure affected metabolites mainly involved in phosphonate and phosphinate metabolism, vitamin B6 metabolism, histidine metabolism, sphingolipid metabolism, and folate biosynthesis. These results suggested that scopoletin exposure to zebrafish embryos exhibited marked metabolic disturbance. This study provides a perspective of metabolic impacts and the underlying mechanism associated with scopoletin exposure.

Experimental research into the potential therapeutic effect of GYY4137 on Ovariectomy-induced osteoporosis.

BACKGROUND: Evidence has shown that endogenous H2S plays an important role in the physiological and pathophysiological processes of many organs. The study aimed to explore whether exogenous H2S has a potential therapeutic effect on a rat ovariectomy-induced model of osteoporosis. METHODS: The OVX osteoporosis model was established in female Sprague-Dawley rats by full bilateral ovariectomy. The rats were randomly divided into four groups, with the two experimental groups receiving an intraperitoneal injection of GYY4137 or sodium alendronate. The level of H2S in the plasma was determined and common laboratory indicators to diagnose osteoporosis, such as alkaline phosphatase (ALP) activity and the levels of osteocalcin (OCN), calcitonin, parathyroid hormone and leptin were measured. The bone mineral density (BMD) of the 4th and 5th lumbar vertebrae was measured using dual-energy X-ray absorptiometry. The maximum stress of femoral fracture was obtained through a three-point bending test of the femur. RESULTS: The OVX osteoporosis model was successfully established. GYY4137 was injected to increase the level of H2S in the plasma in one group, designated OVX-GYY during the observation period (p < 0.05). At 12 weeks, the BMD value of the fourth lumbar vertebra in the OVX-GYY group had increased (p < 0.05). The BMD femur value in the OVX-vehicle group had decreased (p < 0.05). Bilateral ovariectomy leads to biochemical disorders related to bone metabolism and hormone levels in rat plasma (all p < 0.05). Ovariectomy also reduced blood calcium, blood phosphate and calcitonin, and increased parathyroid hormone and leptin. The opposite results were obtained for the groups with alendronate sodium or GYY4137 treatment (all p < 0.05). CONCLUSIONS: Through the slow release of H2S, GYY4137 did an excellent job of simulating endogenous neuroendocrine gaseous signaling molecules. Exogenous H2S had a regulatory effect on osteoporosis in ovariectomized rats, showing potential value for the treatment of human postmenopausal osteoporosis.

GYY4137 stimulates osteoblastic cell proliferation and differentiation via an ERK1/2-dependent anti-oxidant mechanism.

OBJECTIVE: Oxidative stress plays a critical role in the development of osteoporosis. Hydrogen sulfide (H2S), produces anti-oxidant effect in various biological systems. The present study found that GYY4137, a slow H2S releasing compound, stimulated both mRNA level and activity of alkaline phosphatase, the marker of osteoblast differentiation. This research aims to explore the mechanism on how GYY4137 stimulates osteoblastic cell proliferation and differentiation via an ERK1/2-dependent anti-oxidant approach. METHODS: The MC3T3-E1 osteoblast-like cell line was cultured in plate. After pretreatment with GYY4137 (100 µM) for 30 min, the cells were washed twice with PBS solution and then incubated in freshly prepared low serum medium containing 400 µM H2O2 for 4 h. Cells viability was evaluated with the MTT. Cell apoptosis was evaluated by the Hoechst 33342. Then, ALP activity, NO and the superoxide dismutase (SOD) activity is determined by assay kit accordingly, ALP mRNA is identified by RT-PCR. ERK1/2 was analyzed by Western blot. The ROS production was measured with a fluorescence reader. All data was analyzed by SPSS 16.0. RESULTS: We found in the present study that GYY4137, a slow H2S releasing compound, stimulated both mRNA level and activity of alkaline phosphatase, the marker of osteoblast differentiation. RT-PCR shows that GYY4137 stimulated the transcriptional levels of Runx2, a key transcription factor associated with osteoblast differentiation. These data suggest that GYY4137 may stimulate osteoblastic cell proliferation and differentiation. Moreover, GYY4137, which alone at 1-1000 µM had no significant effect, protected MC3T3-E1 osteoblastic cells against hydrogen peroxide (H2O2)-induced cell death and apoptosis. This was mediated by its anti-oxidant effect, as GYY4137 reversed the reduced superoxide dismutase activity and the elevated productions of reactive oxygen species and nitric oxide in the osteoblastic cells treated with H2O2. Western blotting analysis showed that the protective effects of GYY4137 were mediated by suppression of ERK1/2. CONCLUSIONS: GYY4137 stimulates osteoblastic cell proliferation and bone differentiation via an ERK1/2-dependent anti-oxidant mechanism. Our findings suggest that GYY4137 may have a potentially therapeutic value for osteoporosis.

[Clinical study of three-dimensional strapping reduction in treatment of patellar fracture].

Objective: To investigate the effectiveness of three-dimensional strapping reduction in treatment of patellar fracture. Methods: Between January 2015 and June 2015, thirty-two patients were randomly allocated to three-dimensional strapping reduction group (trial group) and towel clamp reduction group (control group). There was no significant difference in age, gender, damage side, interval from injury to opreration, fracture pattern, and cause of injury ( P>0.05). The operation time, fluoroscopy time, fracture healing time, postoperative Hospital for Special Surgery (HSS) scores, and complications were collected and analysed. Results: All incisions healed at stage I. All patients of 2 groups were followed up 10-14 months (mean, 12.4 months). The operation time and fluoroscopy time of trial group were both shorter than those of control group ( t=6.212, P=0.000; t=6.585, P=0.000). X-ray films showed that the fractures in both groups healed successfully and there was no significant difference in healing time between groups ( t=1.973, P=0.058). Bone nonunion, infection, and failure fixation were not found in both groups. HSS scores of trial group (91.6±3.8) was higher than that of control group (86.4±5.5) ( t=-3.105, P=0.004). Conclusion: Compared with towel clamp reduction, the three-dimensional strapping reduction in treatment of patellar fracture has the advantages of shorter operation time and fluoroscopy time, better knee function after operation, and satisfactory fracture healing.

Hydrogen sulfide protects MC3T3-E1 osteoblastic cells against H2O2-induced oxidative damage-implications for the treatment of osteoporosis.

Osteoporosis is a bone disease that leads to an increased risk of fracture. Oxidative damage is an important contributor to the morphological and functional changes in the development of osteoporosis. We found in this study that hydrogen sulfide (H2S), a novel endogenous gaseous mediator, protected MC3T3-E1 osteoblastic cells against hydrogen peroxide (H2O2)-induced oxidative injury. NaHS, an H2S donor, increased cell viability and reduced cell apoptosis caused by H2O2. NaHS also stimulated osteoblast proliferation by enhancing both transcription and activity of alkaline phosphatase in MC3T3-E1 osteoblastic cells. Moreover, treatment with NaHS stimulated the transcriptional level of osteocalcin, the main bone matrix protein, and the protein expression of collagen, a major constituent of bone tissue. The above effects were mediated by the antioxidant effect of H2S. NaHS reversed the reduced superoxide dismutase activity, decreased reactive oxygen species production, and suppressed NADPH oxidase activity in H2O2-treated osteoblasts. In addition, NaHS treatment also produced anti-inflammatory effects via inhibition of the production of nitric oxide and TNF-α, suggesting an anti-inflammatory effect of H2S. Cell viability and Western blotting analysis demonstrated that the protective effects of H2S were mediated by p38 and ERK1/2 MAPKs. In conclusion, H2S protects osteoblastic cells against oxidative stress-induced cell injury and suppression of proliferation and differentiation via a MAPK (p38 and ERK1/2)-dependent mechanism. Our findings suggest that H2S may have a potentially therapeutic value for osteoporosis.

[Biocompatibility of polylactic-co-glycolic acid for culturing bFGF gene-transfected bone marrow stromal cells and application of the cell complex for repairing rabbit cartilage defect].

OBJECTIVE: To evaluate the biocompatibility of polylactic-co-glycolic acid (PLGA) for culturing bFGF gene-transfected bone marrow stromal cells (BMSCs) and assess the feasibility of this cell complex for repairing cartilage defect in rabbits using tissue engineering method. METHODS: BMSCs transfected by bFGF gene were cultured on PLGA matrix to assess the biocompatibility of PLGA. The cell complex was then implanted into the cartilage defect in rabbits, and its effect in cartilage defect repair was evaluated by histological observation and immunohistochemical staining. RESULTS: BMSCs transfected by bFGF gene grew normally on PLGA matrix. After implantation, the complex showed good effect for cartilage defect repair in rabbits. CONCLUSION: PLGA has good biocompatibility with the transfected BMSCs, and the cell complex can be used for repairing rabbit cartilage defect and may potentially serve as a substitute of cartilage autograft.

[Construction of eukaryotic expression vectors of basic fibroblast growth factor and transfection in rabbit bone marrow stromal cells].

OBJECTIVE: To investigate approach and possibility of transferring basic fibroblast growth factor (bFGF) gene into rabbit bone marrow stromal cells (BMSCs). METHODS: The eukaryotic expression vectors harboring bFGF cDNA were constructed and transfected into rabbit BMSCs mediated by liposome. The transcription and expression of bFGF gene in the transfected BMSCs were detected by means of morphological observation, immunohistochemistry, enzyme-linked immunosorbent assay (ELISA) and RT-PCR. The changes in the biological characteristics of the transfected MSCs were also observed. RESULTS: Stable overexpression of bFGF protein was detected in the transfected BMSCs, which showed differentiation towards chondrocyte lineage. CONCLUSION: Stable expression of bFGF gene in transfected BMSCs can induce cell differentiation into chondrocyte lineage.

[Biomechanic study on length of plate in treatment of tibial shaft fracture].

OBJECTIVE: To research the biomechanical effect of different length bone plates on treatment of tibial shaft fracture. METHODS: Forty-five tibia specimens from fresh adult corpse (20-40 years old) were donated (30-38 cm in length, 34 cm on average) and were divided into 3 groups randomly (n = 15). Under the following three conditions, the experiment was made separately. Compression stress-strain indexes of whole tibia were determined under the reverse, three spots curving and compression. The vertical elastic strain was 0-1 000 N, the reverse angle was 0-3 degrees, and three bending stress was 0-400 N. Then the center-section squint non-damage bone fracture model was made, fracture was fixed by 6, 10, 14 stainless steel AO LC-DCP, respectively. The compression stress-strain indexes were determined under reverse angle, three spots curving and compression, statistical analysis was done. RESULTS: The vertical direction strain value of 6, 10 and 14 hole steel plate under vertical compressions, was 0.449 +/- 0.241, 0.093 +/- 0.003, 0.139 +/- 0.005, respectively; showing significant difference between 10 and 14 hole steel plates and 6 steel plate (P < 0.01) and no significant difference between 10 and 14 hole steel plate (P > 0.05). The lateral strain value of 6, 10 and 14 hole steel plate was 0.120 0 +/- 0.000 4, 0.127 5 +/- 0.010 0, 0.237 0 +/- 0.000 6 respectively, indicating a significant difference between stell plates of 6 and 10 hole and 14 hole steel plate (P < 0.01) and no difference between 6 and 10 hole steel plate (P > 0.05). The torque of 6, 10 and 14 hole steel plate was (5.066 +/- 2.715) x 10(-3), (5.671 +/- 2.527) x 10(-3) and (4.570 +/- 2.228) x 10(-3) Nm, respectively and three spot curving vertical direction strain value was 0.049 +/- 0.009, 0.124 +/- 0.017, 0.062 +/- 0.009, respectively. There were significant differences between various steel plates (P < 0.01). CONCLUSION: For the fixation of tibial oblique fracture, 14 hole steel plate's stabilities of anti-vertical compression, anti-reverse and anti-curving are better than those of 6 hole steel plate under the condition of the same material quality, thickness, width and screw quantity used.