Risk factors for and outcomes of heatstroke-related intracerebral hemorrhage.

Some patients with heatstroke also experience intracerebral hemorrhage (ICH). However, clinical case reports of heatstroke-induced ICH are rare. The risk factors for cerebral hemorrhage after heatstroke remain unknown. The present study evaluated the clinical characteristics and risk factors of patients with heatstroke-related ICH. In this retrospective observational study, we collected data on all ICHs after heatstroke occurred between 2012 and 2022. The characteristics of patients with heatstroke-induced ICH were described. The risk factors for cerebral hemorrhage after heatstroke were examined using logistic regression analysis. In total, 177 patients were included in this study, and 11 patients with ICH secondary to heatstroke were identified. Variables with P values of <.05 in univariate models, comparing the cerebral hemorrhage and control groups, included heatstroke cause, temperature, heart rate, respiratory rate, vasopressor use, mechanical ventilation use, Acute Physiology and Chronic Health Evaluation II, total bilirubin, creatinine, platelet count, prothrombin time, procalcitonin, creatine kinase, disseminated intravascular coagulation (DIC) occurrence, and DIC score. Multivariate logistic regression showed that heatstroke patients with higher DIC scores (odds ratio, 18.402, 95% confidence interval, 1.384-244.763, P = .027) and higher creatine kinase levels (odds ratio, 1.021, 95% confidence interval, 1.002-1.041, P = .033) were at a higher risk of developing ICH. The death rate was higher in the cerebral hemorrhage group than in the control group (P = .042). Heatstroke-related cerebral hemorrhage may be associated with elevated creatinine levels and DIC severity (International Society on Thrombosis and Hemostasis score) after heatstroke, and heatstroke with cerebral hemorrhage may accelerate death.

Does the Low-Carbon City Pilot Policy Improve the Urban Land Green Use Efficiency?-Investigation Based on Multi-Period Difference-in-Differences Model.

Improving urban land green use efficiency (ULGUE) is an effective way to increase social, economic, and ecological benefits and achieve regional sustainable development goals. This study takes three batches of low-carbon pilot cities construction as a quasi-natural experiment and investigates the impact of low-carbon pilot construction on ULGUE through the multi-period difference-in-differences method and spatial Dubin difference model (SDM-DID). The results show that (1) from 2006 to 2019, ULGUE in China increased. From the aspect of space, ULGUE in China gradually decreased from west to east, showing an obviously high agglomeration phenomenon in Beijing-Tianjin-Hebei and the Pearl River Delta; (2) after the robustness test, parallel trend test, and endogenous test, it is found that the conclusion that the low-carbon pilot construction can effectively improve ULGUE is still relevant and can indirectly improve ULGUE in the local region through fund allocation, talent gathering, and industrialization; and (3) the national ULGUE has significant positive spatial correlation. The results of the SDM-DID model confirm that the low-carbon pilot policy can produce the significant spatial spillover and drive the common advance of ULGUE in neighboring regions. Therefore, the resources and environmental conditions in each city are supposed to be taken into full consideration theoretically. Furthermore, it is necessary to effectively promote the development of ULGUE by strengthening the linkage of green production factors between different cities, so as to make meaningful contributions to promoting China's overall green development.

Probiotic effects of Lacticaseibacillus rhamnosus 1155 and Limosilactobacillus fermentum 2644 on hyperuricemic rats.

Hyperuricemia is the main cause of gout and involved in the occurrence of multiple diseases, such as hypertension, metabolic disorders and chronic kidney disease. Emerging evidence suggests that lactic acid bacteria (LAB) have shown the beneficial effects on the prevention or treatment of hyperuricemia. In this study, the urate-lowering effect of two LAB strains, Lacticaseibacillus rhamnosus 1155 (LR1155) and Limosilactobacillus fermentum 2644 (LF2644) on hyperuricemic rats were investigated. A hyperuricemic rat model was induced by the intragastric treatment of potassium oxonate, combined with a high purine diet. The oral administration of LR1155, LF2644, or a combination of LR1155 and LF2644 for 4 weeks significantly prevented the rise of the serum uric acid (UA) induced by hyperuricemia. LR1155 and LF2644 significantly elevated the fecal UA levels, increased the UA content and up-regulated gene expression of UA transporter, ATP-binding cassette subfamily G-2 (ABCG2), in colon and jejunum tissues, suggesting the accelerated UA excretion from the intestine. Besides, LR1155 significantly inhibited the activity of xanthine oxidase (XOD) in liver and serum, benefited the reduce of UA production. In addition, LF2644 strengthened the gut barrier functions through an up-regulation of the gene expressions for occluding and mucin2, accompanied with the reduced inflammatory indicators of lipopolysaccharide (LPS) and interleukin-1ß (IL-1ß) in hyperuricemic rat. Moreover, using 16s rDNA high-throughput sequencing of feces, LR1155 was shown to improve the hyperuricemia induced gut microbial dysbiosis. The genera Roseburia, Butyricicoccus, Prevotella, Oscillibacter, and Bifidobacterium may associate with the effect of LR1155 on microbiota in hyperuricemic rats. Collectively, the results indicated that LR1155 and LF2644 exhibit urate-lowering effects and could be used alone or in combination as a new adjuvant treatment for hyperuricemia.

Screening of lactic acid bacteria strains with urate-lowering effect from fermented dairy products.

Hyperuricemia is a well-known cause of gout and also a risk factor for various comorbidities. Current agents like xanthine oxidase inhibitors prevent hyperuricemia, but usually induce severe side effects. Alternative strategies, such as novel dietary supplementations, are necessary for the management of hyperuricemia. Lactic acid bacteria (LAB) have been used in human diet for a long time with a good safety record. In this study, 345 LAB strains isolated from traditional fermented dairy products were tested for assimilating abilities of guanosine. Two LAB strains, Lacticaseibacillus rhamnosus 1155 (LR1155) and Limosilactobacillus fermentum 2644 (LF2644), showing great capacities of guanosine transformation and degradation were selected. Compared to LR1155, LF2644 showed a better effect with 100.00% transforming rate and 55.10% degrading rate. In an in vivo test, a hyperuricemic rat model was established and the results showed that administration of LR1155 (p < 0.01) or LF2644 (p < 0.01) prevented the rise of serum uric acid with more than 20% decrease when compared with the hyperuricemia rats. In addition, an increased fecal uric acid level was observed in LF2644 or LR1155 treated rats (LR1155-M p < 0.05, others p < 0.01). This study proved that LR1155 and LF2644 can be promising candidates of dietary supplements for prevention or improvement of hyperuricemia. PRACTICAL APPLICATION: The LAB strains tested in this study could be considered as good potential probiotic candidates for dietary supplements because of their urate-lowering effects, which provide a novel antihyperuricemic strategy with advantages of safety and sustainability.

Chlorine poisoning caused by improper mixing of household disinfectants during the COVID-19 pandemic: Case series.

BACKGROUND: Misuse of disinfectants during the coronavirus disease 2019 pandemic has led to several poisoning incidents. However, there are few clinical case reports on poisoning caused by improper mixing of household disinfectants. AIM: To summarize the clinical characteristics and treatment effects of chlorine poisoning caused by improper mixing of hypochlorite bleach with acidic cleaning agents.METHODSWe retrospectively analyzed baseline and clinical data, clinical symptoms, and treatment methods of seven patients with chlorine poisoning who were admitted to the National Army Poisoning Treatment Center. RESULTS: Among the seven patients, the average poisoning time (exposure to admission) was 57 h (4-240 h). All patients were involved in cleaning bathrooms. Chest computed tomography scans revealed bilateral lung effusions or inflammatory changes in five patients. The partial pressure of oxygen decreased in six patients, and respiratory failure occurred in one. Five patients had different degrees of increase in white blood cell count. Humidified oxygen therapy, non-invasive mechanical ventilation, anti-inflammatory corticosteroids, antioxidants, and antibiotics were administered for treatment. The average length of hospital stay was 7 d (4-9 d). All seven patients recovered and were discharged. CONCLUSION: Improper mixing of household disinfectants may cause damage to the respiratory system due to chlorine poisoning. Corticosteroids may improve lung exudation in severe cases, and symptomatic supportive treatment should be performed early.

Process Optimization for Supercritical Carbon Dioxide Extraction of Origanum vulgare L. Essential Oil Based on the Yield, Carvacrol, and Thymol Contents.

BACKGROUND: Origanum vulgare L. essential oil (OEO) is widely known for its good biological activity, but different extraction methods with significant implications on the yield of OEO and the content of the thymol and carvacrol. As an efficient method for extracting essential oils (EO), the supercritical carbon dioxide extraction (SC-CO2) can improve the yield of EOs while protecting their main active components from loss. OBJECTIVE: In this study, the process optimization of SC-CO2 of OEO was carried out. The effects of extraction pressure, temperature, time, and modifier concentration on the composite score of OEO extraction process were investigated. METHOD: Response surface analysis was performed using a Box-Behnken design with three levels and four independent variables. Steam distillation (SD) and lipophilic solvents (n-hexane) extraction (LSE) were compared with SC-CO2 for OEO yields. OEOs extracted by the three methods were qualitatively and semi-quantitatively analyzed by gas chromatography quadrupole-time-of-flight mass spectrometry and gas chromatography-flame ionization detector. RESULTS: The results showed that extraction pressure was the most significant factor affecting the OEO yield, thymol, and carvacrol content. In the optimal conditions (pressure: 217 bar, temperature: 54°C, time: 2 h, modifier concentration: 14%), the yield of OEO reached up to 1.136%, and the contents of thymol and carvacrol reached 53.172 and 41.785 mg/g, respectively. CONCLUSIONS: SC-CO2 was the best extraction method compared to the other two methods. Under the optimal conditions, yield and the content of main components can be effectively improved. It can provide a theoretical basis for the industrial extraction of OEO. HIGHLIGHTS: Taking the comprehensive score as the index, the interaction between the four independent variables in the supercritical fluid extraction process was evaluated by the response surface method. The effects of extraction parameters on the yield of EOs and the contents of thymol and carvacrol were comprehensively investigated.

Automatic Sleep Stage Classification of Children with Sleep-Disordered Breathing Using the Modularized Network.

PURPOSE: To develop an automatic sleep stage analysis model for children and evaluate the effect of the model on the diagnosis of sleep-disordered breathing (SDB). PATIENTS AND METHODS: Three hundred and forty-four SDB patients aged between 2 to 18 years who completed polysomnography (PSG) to assess the severity of the disease were enrolled in this study. We developed deep neural networks to stage sleep from electroencephalography (EEG), electrooculography (EOG) and electromyogram (EMG). The model performance was estimated by accuracy, precision, recall, F1-score, and Cohen's Kappa coefficient (ĸ). And we compared the difference in calculation of sleep parameters among the technicians, the model ensemble, and the single-channel EEG model. RESULTS: The numbers of raw data divided into training, validation, and testing were 240, 36, and 68, respectively. The best performance appeared in the model ensemble of which the accuracy was 83.36% (ĸ=0.7817) in 5-stages, and the accuracy was 96.76% (ĸ=0.8236) in 2-stages. The single-channel EEG model showed the classification satisfyingly as well. There was no significant difference in TST, SE, SOL, time in W, time in N1+N2, time in N3, and OAHI between technician and the model (P>0.05). On the datasets from sleep-EDF-13 and sleep-EDF-18, the average classification accuracies achieved were 92.76% and 91.94% in 5-stages by using the proposed method, respectively. CONCLUSION: This research established the model for pediatric automatic sleep stage classification with satisfying reliability and generalizability. In addition, it could be applied for calculating quantitative sleep parameters and evaluating the severity of SDB.

TRPV4 Regulates Soman-Induced Status Epilepticus and Secondary Brain Injury via NMDA Receptor and NLRP3 Inflammasome.

Nerve agents are used in civil wars and terrorist attacks, posing a threat to public safety. Acute exposure to nerve agents such as soman (GD) causes serious brain damage, leading to death due to intense seizures induced by acetylcholinesterase inhibition and neuronal injury resulting from increased excitatory amino-acid levels and neuroinflammation. However, data on the anticonvulsant and neuroprotective efficacies of currently-used countermeasures are limited. Here, we evaluated the potential effects of transient receptor vanilloid 4 (TRPV4) in the treatment of soman-induced status epilepticus (SE) and secondary brain injury. We demonstrated that TRPV4 expression was markedly up-regulated in rat hippocampus after soman-induced seizures. Administration of the TRPV4 antagonist GSK2193874 prior to soman exposure significantly decreased the mortality rate in rats and reduced SE intensity. TRPV4-knockout mice also showed lower incidence of seizures and higher survival rates than wild-type mice following soman exposure. Further in vivo and in vitro experiments demonstrated that blocking TRPV4 prevented NMDA receptor-mediated glutamate excitotoxicity. The protein levels of the NLRP3 inflammasome complex and its downstream cytokines IL-1ß and IL-18 increased in soman-exposed rat hippocampus. However, TRPV4 inhibition or deletion markedly reversed the activation of the NLRP3 inflammasome pathway. In conclusion, our study suggests that the blockade of TRPV4 protects against soman exposure and reduces brain injury following SE by decreasing NMDA receptor-mediated excitotoxicity and NLRP3-mediated neuroinflammation. To our knowledge, this is the first study regarding the "dual-switch" function of TRPV4 in the treatment of soman intoxication.

Safety and efficacy of a nerve matrix membrane as a collagen nerve wrapping: a randomized, single-blind, multicenter clinical trial.

A new nerve matrix membrane derived from decellularized porcine nerves has been shown to retain the major extracellular matrix components, and to be effective in preventing adhesion between the nerve anastomosis sites and the surrounding tissues in a rat sciatic nerve transection model, thereby enhancing regeneration of the nerve. The effectiveness of the membrane may be attributed to its various bioactive components. In this prospective, randomized, single-blind, parallel-controlled multicenter clinical trial, we compared the safety and efficacy of the new nerve matrix membrane with a previously approved bovine tendon-derived type I collagen nerve wrapping. A total of 120 patients with peripheral nerve injury were recruited from Beijing Jishuitan Hospital, The First Bethune Hospital of Jilin University, and Yantai Yuhuangding Hospital, China. The patients were randomly assigned to undergo end-to-end and tension-free neurorrhaphy with nerve matrix membrane (n = 60, 52 male, 8 female, mean age 41.34 years, experimental group) or tendon-derived collagen nerve wrapping (n = 60, 42 male, 18 female, mean age 40.17 years, control group). Patients were followed-up at 14 ± 5, 30 ± 7, 90 ± 10 and 180 ± 20 days after the operation. Safety evaluation included analyses of local and systemic reactions, related laboratory tests, and adverse reactions. Efficacy evaluation included a static 2-point discrimination test, a moving 2-point discrimination test, and a Semmes-Weinstein monofilament examination. Sensory nerve function was evaluated with the British Medical Research Council Scale and Semmes-Weinstein monofilament examination. The ratio (percentage) of patients with excellent to good results in sensory nerve recovery 180 ± 20 days after the treatment was used as the primary effectiveness index. The percentages of patients with excellent to good results in the experimental and control groups were 98.00% and 94.44%, respectively, with no significant difference between the two groups. There were no significant differences in the results of routine blood tests, liver and renal function tests, coagulation function tests, or immunoglobulin tests at 14 and 180 days postoperatively between the two groups. These findings suggest that the novel nerve matrix membrane is similar in efficacy to the commercially-available bovine-derived collagen membrane in the repair of peripheral nerve injury, and it may therefore serve as an alternative in the clinical setting. The clinical trial was approved by the Institutional Ethics Committee of Beijing Jishuitan Hospital, China (approval No. 20160902) on October 8, 2016, the Institutional Ethics Committee of the First Bethune Hospital of Jilin University, China (approval No. 160518-088) on December 14, 2016, and the Institutional Ethics Committee of Yantai Yuhuangding Hospital, China (approval No. 2016-10-01) on December 9, 2016. The clinical trial was registered with the Chinese Clinical Trial Registry (registration number: ChiCTR2000033324) on May 28, 2020.

Author Correction: Repair of bone defects with prefabricated vascularized bone grafts and double-labeled bone marrow-derived mesenchymal stem cells in a rat model.

An amendment to this paper has been published and can be accessed via a link at the top of the paper.

Evaluation of the Antibacterial Material Production in the Fermentation of Bacillus amyloliquefaciens-9 from Whitespotted Bamboo Shark (Chiloscyllium plagiosum).

Bacillus amyloliquefaciens-9 (GBacillus-9), which is isolated from the intestinal tract of the white-spotted bamboo shark (Chiloscyllium plagiosum), can secrete potential antibacterial materials, such as ß-1,3-1,4-glucanase and some antimicrobial peptides. However, the low fermentation production has hindered the development of GBacillus-9 as biological additives. In this study, the Plackett-Burman design and response surface methodology were used to optimize the fermentation conditions in a shake flask to obtain a higher yield and antibacterial activity of GBacillus-9. On the basis of the data from medium screening, M9 medium was selected as the basic medium for fermentation. The data from the single-factor experiment showed that sucrose had the highest antibacterial activity among the 10 carbon sources. The Plackett-Burman design identified sucrose, NH4Cl, and MgSO4 as the major variables altering antibacterial activity. The optimal concentrations of these compounds to enhance antibacterial activity were assessed using the central composite design. Data showed that sucrose, NH4Cl, and MgSO4 had the highest antibacterial activities at concentrations of 64.8, 1.84, and 0.08 g L-1, respectively. The data also showed that the optimal fermentation conditions for the antibacterial material production of GBacillus-9 were as follows: Inoculum volume of 5%, initial pH of 7.0, temperature of 36 °C, rotating speed of 180 rpm, and fermentation time of 10 h. The optimal fermentation medium and conditions achieved to improve the yield of antibacterial materials for GBacillus-9 can enhance the process of developing biological additives derived from GBacillus-9.

Involvement of the Nrf2-Keap1 signaling pathway in protection against thallium-induced oxidative stress and mitochondrial dysfunction in primary hippocampal neurons.

Thallium ion (Tl+) and its neurotoxic products are widely known to cause severe neurological complications. However, the exact mechanism of action remains unknown, with limited therapeutic options available. This study aims to examine the toxic effects of Thallium (I) Nitrate (TlNO3) on primary hippocampal neurons of E17-E18 Wistar rat embryos, and the potential neuroprotective role of Nrf2- Keap1 signaling pathway against thallium-induced oxidative stress and mitochondrial dysfunction. TlNO3 induces a significant increase in reactive oxygen species levels and mitochondrial dysfunction in primary hippocampal neurons. Furthermore, the Nrf2-Keap1 signaling pathway played a protective role against TlNO3-induced hippocampal neuronal cytotoxicity. Moreover, mitochondrial fusion protein Mitofusin 2 (Mfn2) levels significantly decreased in hippocampal neurons when exposed to TlNO3, indicating that Mfn2 protein levels are linked to TlNO3-induced neurotoxicity. t-BHQ, a Nrf2 and phase II detoxification enzyme inducer, counteracted the oxidative damage in hippocampal neurons by activating the Nrf2-Keap1 signaling pathway after TlNO3 exposure; the activated Nrf2-Keap1 pathway could then maintain Mfn2 function by regulating Mfn2 protein expression. Thus, Nrf2-Keap1 pathway activation plays a protective role in Tl+-induced brain damage, and specific agonists have been identified to have great potential for treating thallium poisoning.

Clinical characteristics and treatment of thallium poisoning in patients with delayed admission in China.

Thallium is highly toxic and its effects are cumulative. The clinical symptoms of thallium poisoning are non-specific, thereby delaying admission and treatment. This study aimed to summarize the clinical features and treatment experience of patients with delayed admission who experience thallium poisoning.We conducted a retrospective descriptive analysis of patients in our hospital from 2008 to 2018 who had thallium poisoning and experienced a delay in hospital admission. The time from symptom onset to admission was assessed. The patients were divided into 3 groups and descriptive analyses of their clinical characteristics, including basic patient information, symptoms, laboratory test results, examination findings, treatment methods, outcomes, and follow-up information, were conducted.A total of 34 patients with thallium poisoning were included: 8 were admitted to the hospital early or with mild delay, 9 had a moderate delay, and 17 had a severely delayed admission. The time from illness onset to admission was 13 (interquartile range, 7.5-26) days. Some patients with delayed admission had significant symptoms associated with central nervous system damage, and changes in magnetic resonance images and electroencephalograms were also noted. After admission, all patients received Prussian blue treatment, and some patients with relatively high blood concentration received blood purification treatments. Following treatment, the blood and urine thallium concentrations of all patients decreased significantly, and their symptoms were alleviated.Our results show that delayed patient admission in cases of thallium poisoning is associated with greater risk of central nervous system damage. Use of Prussian blue combined with blood purification treatments might improve patients' conditions.

NLRP3 inflammasome activation is involved in trimethyltin-induced neuroinflammation.

Trimethyltin (TMT), a neurotoxic organotin compound, is selectively localized within the limbic system. The mechanisms of TMT-induced hippocampal neurodegeneration include inflammatory responses, oxidative stress, and neuronal death. Increasing evidence shows that the inflammatory response, mediated by activated inflammasomes, is involved in apoptosis and cellular dysfunction during brain injury. This study aimed to assess the role of the nucleotide-binding oligomerization domain-like receptor pyrin-domain-containing protein 3 (NLRP3) inflammasome in TMT-induced central nervous system (CNS) injury. In addition, the mechanisms underlying TMT neurotoxicity are similar to those involved in the pathogenesis of multiple neurodegenerative diseases; hence, a study on TMT cytotoxicity may be informative for the understanding of human CNS diseases. Microglia were significantly activated in the rat hippocampal dentate gyrus after TMT treatment. The mRNA expression of pro-inflammatory cytokines, interleukin-1ß and interleukin-18, was induced both in vitro and in vivo. TMT treatment activated the NLRP3 inflammasome in the microglial cell line BV2. NLRP3 RNA interference significantly protected these cells from TMT-induced neuroinflammation. Our results demonstrate that the NLRP3 inflammasome is a key mediator of neuroinflammation and plays an important role in TMT-induced neuroinflammation.

Successful treatment of a patient with severe thallium poisoning in a coma using Prussian blue and plasma exchange: A case report.

RATIONALE: This is the first reported severe thallium poisoning patient successfully treated with Prussian blue (PB) and plasma exchange (PE). PATIENT CONCERNS: A 42-year-old woman in a coma owing to severe thallium poisoning was admitted to our department after day 44 of poisoning. At admission, blood and urine thallium concentrations were 380.0 and 2580.0 ng/mL, respectively. DIAGNOSIS: The patient was diagnosed with toxic encephalopathy induced by thallium poisoning; in addition, she was also diagnosed with bilateral pneumonia, respiratory failure, moderate anemia, hypoproteinemia, and electrolyte imbalance based on her chest X-ray, blood gas analysis, Hb level, albumin levels, and serum electrolyte results. INTERVENTIONS: The patient was intubated and treated with PB (6600 mg/d, 15 days in total) combined with PE (once daily, 5 days in total) as well as other symptomatic supportive care measures. OUTCOMES: After treatments, her blood and urinary thallium concentrations gradually decreased and on the 13th day after admission, the blood thallium concentration decreased to 0 ng/mL. The oxygenation index gradually improved, meantime, the patient gradually regained consciousness, and on the 50th day of admission, the patient's consciousness reverted to a clear-headed state. The patient recovered mostly after 37 months of follow-up. LESSONS: Through this case, we learned that the gradual reduction in blood and urine thallium concentration and the patient's improved condition is correlated with PB and PE treatment. For patients with severe thallium poisoning, this treatment method might be effective; but the exact curative effect is unconfirmed, requiring further research to verify.

A fragment activity assay reveals the key residues of TBC1D15 GTPase-activating protein (GAP) in Chiloscyllium plagiosum.

BACKGROUND: GTPase-activating proteins (GAPs) with a TBC (Tre-2/Bub2/Cdc16) domain architecture serve as negative regulators of Rab GTPases. The related crystal structure has been studied and reported by other members of our research group in 2017 (Chen et al. in Protein Sci 26(4):834-846, 2017). The protein crystal structure and sequencing data accession numbers in Protein structure database (PDB) are 5TUB (Shark TBC1D15 GAP) and 5TUC (Sus TBC1D15 GAP), respectively. In this paper, we analyzed the Rab-GAP specificity of TBC1D15 in the evolution and influence of key amino acid residue mutations on Rab-GAP activity. RESULTS: Sequence alignment showed that five arginine residues of the TBC1D15-GAP domain are conserved among the species Sus/Mus/Homo but have been replaced by glycine or lysine in Shark. A fragment activity assay was conducted by altering the five residues of Shark TBC1D15-GAP to arginine, and the corresponding arginine in TBC1D15 GAP domains from Sus and Homo species were mutated to resemble Shark TBC1D15-GAP. Our data revealed that the residues of G28, K45, K119, K122 and K221 in the Shark TBC1D15-GAP domain had a key role in determining the specificity for Rab7 and Rab11. Mutation of the five residues significantly altered the Shark TBC1D15-GAP activity. CONCLUSIONS: These results revealed that the substrate specificity of TBC1D15 has had different mechanisms across the evolution of species from lower-cartilaginous fish to higher mammals. Collectively, the data support a different mechanism of Shark TBC1D15-GAP in substrate selection, which provides a new idea for the development of Marine drugs.

Osteogenic efficiency in vivo of scaffolding material of prefabricated vascularization.

Osteogenic efficiency of pre-vascularization and non-vascularization decalcified bone scaffolds in bone defect repair was investigated. Twenty-one Sprague-Dawley (SD) mice were randomly assigned to three groups, and a bone defect area of ~1.5 cm in length in the thigh bone of the right posterior limbs of each mouse was made. Pre-vascularization decalcified bone scaffolds in vitro (group A) and non-treatment decalcified bone scaffolds (group B) were separately implanted. The defect vacancy was considered as the blank control (group C). Sampling was made 4 weeks after the operation for the histological examination, and then the osteogenic efficiency was observed by gross sample, imaging, hematoxylin and eosin staining and Masson's staining. When implanting pre-vascularization decalcified bone scaffolds in vitro, the scaffolding material showed an obvious absorption, and more new bone formations and abundant vascular proliferation were observed. In non-vascularization decalcified bone scaffolds implanting, absorption insufficiency of the scaffolding material was observed, fewer new-born bone formations were shown, and the new vessels were very small and few in number. The pre-vascularization decalcified bone scaffolds had a better osteogenic efficiency.

Clinical analysis of 86 botulism cases caused by cosmetic injection of botulinum toxin (BoNT).

This study was conducted to analyze the clinical characteristics of and treatment strategies for botulism among patients receiving cosmetic injection of botulinum toxin (BoNT).A total of 86 botulism patients caused by cosmetic injection of BoNT were enrolled in our study. All of the patients were diagnosed according to their history of cosmetic BoNT injection, clinical symptoms and signs, and other auxiliary examinations (including those on renal and liver functions, blood index detection, and chest X-ray). All of the patients received comprehensive treatments and botulinum antitoxin serum injection.The main symptoms of botulism patients included headache, dizziness, insomnia, fatigue, blurred vision, eye opening difficulty, slurred speech, dysphagia, bucking, constipation, and anxiety. These clinical symptoms occurred 0∼36 days after BoNT injection, especially from 2nd to 6th day after the operation. Furthermore, the usage dose of BoNT was negatively related to latent period. Finally, patients all discharged from our hospital 1∼20 days after treatments, and their symptoms relieved or disappeared.Botulism is a severe side effect for BoNT injection. Injecting botulinum antitoxin serum may be an effective approach to improve clinical outcomes of botulism cases.

Effects of Bacillus amyloliquefaciens and Yarrowia lipolytica lipase 2 on immunology and growth performance of Hybrid sturgeon.

A 12-weeks feeding trial was performed to investigate the possible effects of supplementation of Hybrid sturgeon diet with Bacillus amyloliquefaciens (GB-9) and Yarrowia lipolytica lipase2 (YLL2) single or combined on immune response and growth performance of Hybrid sturgeon (Acipenser schrenkii ♂and Acipenser baeri ♀). For this aim, Hybrid sturgeons were fed with four experimental diets namely: Diet 1 (0-control), Diet 2 (5.0 g/kg GB-9), Diet 3 (4.0 g/kg YLL2), and Diet 4 (5.0 g/kg GB-9 + 4.0 g/kg YLL2), respectively. After fed with varied diets, growth performance, mucosal immune response, leukocytes immune response and serum immunological response were measured. The results indicated that supplementations of GB-9 + YLL2 resulted in a significant increase in final weight, Docosahexaenoic acid (DHA) and Eicosapentenoic acid (EPA) concentration, compared with that of control (p < 0.05). For innate immunity, the results showed that skin mucus lysozyme activity, leukocytes phagocytosis activity and reactive oxygen species level, and serum alternative complement pathway activity, peroxidase and lysozyme activity were significantly higher in supplemented groups compared to the control (p < 0.05). The highest values were recorded in fish fed both YLL2 and GB-9 with respect to the individual application. The present results suggested that the combination of these supplementation could be considered as potential feed-additives for aquaculture farmed fish.

Isolation and characterization of Bacillus sp. GFP-2, a novel Bacillus strain with antimicrobial activities, from Whitespotted bamboo shark intestine.

The abuse of antibiotics and following rapidly increasing of antibiotic-resistant pathogens is the serious threat to our society. Natural products from microorganism are regarded as the important substitution antimicrobial agents of antibiotics. We isolated a new strain, Bacillus sp. GFP-2, from the Chiloscyllium plagiosum (Whitespotted bamboo shark) intestine, which showed great inhibitory effects on the growth of both Gram-positive and Gram-negative bacteria. Additionally, the growth of salmon was effectively promoted when fed with inactivated strain GFP-2 as the inhibition agent of pathogenic bacteria. The genes encoding antimicrobial peptides like LCI, YFGAP and hGAPDH and gene clusters for secondary metabolites and bacteriocins, such as difficidin, bacillibactin, bacilysin, surfactin, butirosin, macrolactin, bacillaene, fengycin, lanthipeptides and LCI, were predicted in the genome of Bacillus sp. GFP-2, which might be expressed and contribute to the antimicrobial activities of this strain. The gene encoding ß-1,3-1,4-glucanase was successfully cloned from the genome and this protein was detected in the culture supernatant of Bacillus sp. GFP-2 by the antibody produced in rabbit immunized with the recombinant ß-1,3-1,4-glucanase, indicating that this strain could express ß-1,3-1,4-glucanase, which might partially contribute to its antimicrobial activities. This study can enhance a better understanding of the mechanism of antimicrobial activities in genus Bacillus and provide a useful material for the biotechnology study in antimicrobial agent development.