Immunogenicity and protective efficacy of a novel bacterium-like particle-based vaccine displaying canine distemper virus antigens in mice and dogs.

Canine distemper virus (CDV) poses a severe threat to both domesticated and wild animals, including multiple carnivores. With the continued expansion of its host range, there is an urgent need for the development of a safer and more effective vaccine. In this study, we developed subunit vaccines based on a bacterium-like particle (BLP) delivery platform containing BLPs-F and BLPs-H, which display the CDV F and H glycoprotein antigens, respectively, using the antigen-protein anchor fusions produced by a recombinant baculovirus insect cell expression system. The combination of BLPs-F and BLPs-H (CDV-BLPs), formulated with colloidal manganese salt [Mn jelly (MnJ)] adjuvant, triggered robust CDV-specific antibody responses and a substantial increase in the number of interferon gamma (IFN-γ)-secreting CD4+ and CD8+ T cells in mice. Dogs immunized intramuscularly with this vaccine not only produced CDV-specific IgG but also displayed elevated concentrations of IFN-γ and interleukin 6 in their serum, along with an increase of the CD3+CD4+ and CD3+CD8+ T cell subsets. Consequently, this heightened immune response provided effective protection against disease development and reduced viral shedding levels following challenge with a virulent strain. These findings suggest that this BLP-based subunit vaccine has the potential to become a novel canine distemper vaccine. IMPORTANCE: Many sensitive species require a safe and effective distemper vaccine. Non-replicating vaccines are preferred. We constructed subunit particles displaying canine distemper virus (CDV) antigens based on a bacterium-like particle (BLP) delivery platform. The CDV-BLPs formulated with theMn jelly adjuvant induced robust humoral and cell-mediated immune responses to CDV in mice and dogs, thereby providing effective protection against a virulent virus challenge. This work is an important step in developing a CDV subunit vaccine.

Therapeutic effect of vitamin D in Hashimoto's thyroiditis: a prospective, randomized and controlled clinical trial in China.

BACKGROUND: The study aims to explore the clinical effects of Vitamin D (VitD) supplements for Hashimoto's Thyroiditis (HT), which are unclear according to other studies. METHODS: Female patients with newly diagnosed HT from January to June in 2018 were included. This study is registered in the Chinese Clinical Trials Registry with registration number ChiCTR1800014619 (URL: https://www.chictr.org.cn/). Patients were randomly assigned to the treatment group and the control group. The treated group were further randomly assigned to a VitD supplement group or VitD & Levothyroxine (L-T4) supplement group. After 6 months, we recorded and compared various indicators between different groups. RESULTS: A total of 179 patients, aged 12 to 75, were used for statistical analysis. A significant decrease in Thyroid Peroxidase Antibody (TPOAb) level was observed (351.70±183.25 vs. 246.37±157.39, P<0.001) in the VitD-treated group compared to the control group after 6 months. Free Triiodothyronine (FT3) and Free Thyroxine (FT4) level were increased (FT3: 4.30±0.64 vs. 4.84±0.9, P<0.001; FT4: 15.15±1.93 vs. 17.38±2.97, P<0.001), and Thyroid-Stimulating Hormone (THS) level was decreased (3.58±1.78 vs. 2.25±1.22, P<0.001) in the VitD-treated group compared to the control group. CONCLUSION: VitD supplementation can effectively slow progression of hypothyroidism, improve thyroid function, and reduce the anti-thyroid antibody level. This suggests it is useful for HT.

Technical guidelines for risk assessment of heavy metals in traditional Chinese medicines.

BACKGROUND: Heavy metals are considered a global concern because they can deteriorate human health. This guideline aims to scientifically evaluate health risk of heavy metals in TCM and to propose a reference for decision making in developing TCM-related health policies. METHODS: Using a multidisciplinary approach, a steering committee oversaw the development of the guideline. To obtain a reasonable and accurate risk assessment, key exposure assessment parameters for TCM, e.g., exposure frequency (EF), exposure duration (ED), and daily ingestion rate (IR) were obtained from surveys. In addition, transfer rates for heavy metals from Chinese medicinal materials (CMM) to decoctions or preparations were examined. RESULTS: Based on the scientific theory of risk control, the guideline was designed systematically, and principles and procedures for the risk assessment of heavy metals in TCM were identified. The guideline can be utilized to assess the risk of heavy metals in CMM and Chinese patent medicines (CPM). CONCLUSION: This guideline may help standardize the risk assessment of heavy metals in TCM, advance regulatory standards for heavy metals in TCM, and ultimately improve human health through scientific TCM usage in clinic.

Arctigenin mitigates insulin resistance by modulating the IRS2/GLUT4 pathway via TLR4 in type 2 diabetes mellitus mice.

Arctigenin (AR), extracted from Arctium lappa L. (Burdock), is a folk herbal medicine used to treat diabetes. However, its mechanism of action has remained elusive. In this study, type 2 diabetes mellitus (T2DM) mice received AR orally for 10 weeks to evaluate its therapeutic effect based on changes in glucose and lipid metabolism, histological examination of target tissues, and liver immunohistochemistry. Furthermore, HepG2 insulin-resistant cells were established to verify the mechanism of AR against diabetes. The results showed that AR treatment reduced blood glucose and lipid levels, reversing liver as well as pancreas tissue damage in T2DM mice. AR reduced the levels of pro-inflammatory cytokines in the serum of T2DM mice, as well as those in insulin-resistant HepG2 cell supernatants, while increasing interleukin-10 (IL-10) levels. The levels of p-p65, phospho-c-Jun N-terminal kinase (p-JNK), induced nitric oxide synthase (iNOS), and cyclooxygenase-2 (COX-2) were reduced in the liver tissue of T2DM mice, accompanied by an upregulation of glucose transporter 4 (GLUT4) and insulin receptor substrate 2 (IRS-2). In vitro studies further showed that AR downregulated toll-like receptor 4-mediated inflammation, while upregulating insulin pathway-related proteins and ultimately improving glucose uptake in insulin-resistant HepG2 cells. In conclusion, AR protected mice from insulin resistance, and its therapeutic effect was likely associated with inhibition of toll-like receptor 4 inflammatory signaling to reactivate IRS-2/GLUT4.

Preparation of Fe/C-MgCO3 micro-electrolysis fillers and mechanism of phosphorus removal.

Iron-carbon micro-electrolysis is effective for the removal of phosphorus in wastewater; however, meeting the stringent emission standards required for treatment is difficult. To meet these treatment standards, modified micro-electrolytic fillers were prepared from iron dust, powdered activated carbon, clay, and additives using an elevated temperature roasting process under an inert atmosphere. The results show that among several additives, the modified micro-electrolytic (Fe/C-MgCO3) fillers using MgCO3 were the most effective at phosphorus removal. The preparation conditions for the Fe/C-MgCO3 fillers and their effects on phosphorus removal performance were investigated. Under the optimal preparation conditions (calcination temperature: 800 °C, Fe/C = 4:1, clay content 20%, and 5% MgCO3), the filler yielded a high compressive strength of 3.5 MPa, 1 h water absorption rate of 25.7%, and specific surface area and apparent density of 154.2 m2/g and 2689.2 kg/m3, respectively. The iron-carbon micro-electrolysis process removed 97% of phosphorus in the wastewater by using the Fe/C-MgCO3 fillers, which was 14% more than the Fe/C filler. Electrostatic adsorption and surface precipitation were identified as the main phosphorus removal mechanisms, and the surface of the Fe/C-MgCO3 filler was continuously updated. These results demonstrated that Fe/C-MgCO3 is a promising filler for phosphorus removal in water treatment.

Integrative Metabolomics and Proteomics Detected Hepatotoxicity in Mice Associated with Alkaloids from Eupatorium fortunei Turcz.

The traditional Chinese herbal medicine Eupatorium fortunei Turcz. (E. fortunei) has been widely adopted to treat nausea, diabetes, siriasis, and poor appetite. However, E. fortunei contains multiple pyrrolizidine alkaloids (PAs). This study aimed to investigate the hepatotoxicity of total alkaloids in E. fortunei (EFTAs) and identify the toxic mechanisms of EFTAs on hepatocytes. Liquid chromatography with a tandem mass spectrometry assay with reference standards indicated that EFTAs mainly consisted of eight PAs whose content accounted for 92.38% of EFTAs. EFTAs markedly decreased mouse body and liver weights and increased the contents of AST and ALT. The histopathological assays demonstrated that, after exposition to EFTAs, the structures of hepatocytes were damaged and the fibrosis and apoptosis in hepatocytes were accelerated. Moreover, EFTAs increased the serum level of inflammatory cytokines and aggravated circulating oxidative stress. A combination of hepatic proteomics and metabolomics was used to investigate the toxic mechanisms of EFTAs. The study revealed that EFTAs seriously disrupted glycerophospholipid metabolism by upregulating the contents of lysophosphatidylglycerol acyltransferase 1 and phosphatidylinositol and downregulating the contents of choline/ethanolamine kinase beta, choline-ethanolamine phosphotransferase 1, phospholipase D4, 1-acylglycerophosphocholine, phosphatidylcholine, and dihydroxyacetone phosphate in the liver, resulting in detrimental inflammation, fibrosis, and apoptosis. This study revealed that EFTAs induced severe hepatotoxicity by disrupting glycerophospholipid metabolism.

Performance enhancement and population structure of denitrifying phosphorus removal system over redox mediator at low temperature.

The development of denitrifying polyphosphate accumulating organisms (DPAOs) presents a strategy to carbon competition between denitrifying bacteria and phosphorus removing bacteria. However, low temperature inhibits the rate of enzyme-catalyzed and substrate diffusion during denitrifying phosphorus removal (DPR). Therefore, the present study assessed the addition of NQS (100 µmol/L) for enhancing the removal of TP and TN in DPR reactors operated at alternating anaerobic and anoxic phases and different influent phosphate concentrations. The results showed that the removal efficiency of TP and TN in NQS-DPR system at 10 °C were 99.9% and 42.0%, respectively, which were 2.1 and 2.0 times higher than that of DPR system. Adding NQS significantly alleviated the increase of pH under anoxic condition and decreased the ORP value of the reactor, which in turn enhanced the PHAs accumulation process. The determination of functional genes (nirK, narG and phoD) showed that Dechloromonas, Lentimicrobium, and Terrimonas were the dominant functional bacteria in NQS-DPR system at 10 °C with the relative abundance of 3.09%, 2.99% and 2.28%, respectively. This study can provide valuable information for the effects of the addition of the redox mediator on denitrifying phosphorus removal technology.

A strategy for quality control of ginkgo biloba preparations based on UPLC fingerprint analysis and multi-component separation combined with quantitative analysis.

BACKGROUND: Many studies have assessed the fingerprint and quantitative analysis of Ginkgo biloba preparations, but the fingerprint mainly focuses on flavonoid glycosides. However, according to our previous study, the differences among diverse manufacturers mainly involve organic acids. METHODS: A novel reverse-phase liquid chromatography assay using diode array detection was developed for evaluating Ginkgo biloba preparations for quality based on a chromatographic fingerprint allowing the simultaneous assessment of eleven compounds, including four organic acids, six flavonol glycosides and one flavonoid aglycone. And the method was applied to 51 batches of Ginkgo biloba preparations from manufacturers in China. Chemometric approaches were performed for evaluating 51 batches of Ginkgo biloba preparations from various manufacturers. RESULTS: The similarity values among the chromatograms of 51 samples ranged from 0.45 to 1.00, showing that the quality of Ginkgo biloba preparations produced by different manufacturers varied greatly. Data analysis of the 51 batches of GBP samples suggested significant variations of the total contents of all 11 targets, also demonstrating the quality difference of GBP samples. There were significant differences in organic acids in particular. CONCLUSION: Combining the chemical fingerprint and quantitative assessment revealed significant variations in the examined commercial products with regard to organic acids. Thus, this study provided a more comprehensive tool for monitoring the quality consistency of Ginkgo biloba preparations.

Conjugated linoleic acid ameliorates hepatic steatosis by modulating intestinal permeability and gut microbiota in ob/ob mice.

Background: Conjugated linoleic acid (CLA) is an effective supplement for reducing fat mass, but its effect on hepatic steatosis remains controversial. Objective: This study aims to evaluate the effect of CLA on liver fat accumulation, inflammation, gut microbiome, and intestinal barrier integrity. Design: Wild-type (WT) mice and ob/ob (OB) mice were randomly divided into four groups according to the treatment with/without 1% CLA: WT, WT mice treated with CLA (WT-CLA), OB, and OB mice treated with CLA (OB-CLA). Lipid metabolism and hepatic fat accumulation were evaluated by changes in histological and biochemical parameters. Gene expressions related to liver inflammation and intestinal barrier integrity were examined. The effect of CLA on the gut microbiota population was investigated. Results: The body weight, fatty tissue mass, and serum lipid levels of the WT-CLA group and OB-CLA group were separately lower than those of the WT group and OB group, but the livers of the WT-CLA group had more fatty lipids, higher triglyceride properties, and saturated fatty acid (FA) composition than those of the WT group, which was contrary to the effect of CLA on OB mice. Real time quantitative PCR results showed that CLA increased hepatic inflammation and intestinal permeability in the WT mice, while it significantly decreased the mRNA expression of liver TNF-α, IFN-γ, and IL-1ß and markedly ameliorated intestinal tight junction proteins in the OB mice. The gut microbiota testing indicated a higher abundance of beneficial bacteria (e.g., Lachnoclostridium, Roseburia, Dubosiella, Oscillibacter, and Anaerostipes) and a lower abundance of pro-inflammatory bacteria (e.g., Tyzzerella and Alistipes) in the OB-CLA group than those of the OB group. Correlation analysis suggested that gut microbiota correlated with liver inflammation, intestinal permeability, and hepatic FA composition. Conclusion: CLA potentially contributed to ameliorating hepatic steatosis in OB mice via modulating liver inflammation, intestinal permeability, and gut microbiota, which suggests CLA is more suitable for people with obesity or overweight.

Association of Fecal Microbiota with Irritable Bowel Syndrome-Diarrhea and Effect of Traditional Chinese Medicine for Its Management.

Changes in intestinal microbiota have been linked to the development of diarrhea predominant irritable bowel syndrome (IBS-D). In order to better elucidate the relationship between intestinal microbiota changes and IBS-D, we compared fecal microbiota of IBS-D rats and healthy control using pyrosequencing of bacterial 16S rRNA gene targeted. Furthermore, we explored the effects of different traditional Chinese medicine (TCM) on intestinal microbiota of IBS-D in dose-dependent manner. Our results showed that there was no significant difference in fecal microbial community diversity among the healthy control group, IBS-D rats and IBS-D rats treated with traditional Chinese medicine, but the fecal microbial composition at different taxonomic levels have changed among these groups. Interestingly, the weight of IBS-D rats treated with moderate doses (13.4 g/kg) of TCM increased significantly, and the diarrhea-related symptoms improved significantly, which may be related to the enrichment in Deferribacteres, Proteobacteria, Tenericutes, Lachnospiraceae, and Ruminococcaceae and the reduction in Lactobacillus in fecal samples.

[Simultaneous determination of six pyrrolizidine alkaloids in different parts of Emilia sonchifolia by UPLC-MS/MS].

This study aims to develop a UPLC-MS/MS method for simultaneous determination of six pyrrolizidine alkaloids(PAs)--intermedine N-oxide(ImNO), lycopsamine N-oxide(LyNO), seneciphylline(Sp), seneciphylline N-oxide(SpNO), senecionine N-oxide(SnNO), and senkirkine(Sk) in different parts of Emilia sonchifolia. UPLC conditions are as follows: ACQUITY UPLC HSS T3 column(2.1 mm×100 mm, 1.8 µm), mobile phase consisting of 0.05% formic acid and 2.5 mmol·L~(-1) ammonium formate in water(A)-0.05% formic acid and 2.5 mmol·L~(-1) ammonium formate in acetonitrile(B) for gradient elution. MS conditions are as below: electrospray ionization(ESI) in the positive ion mode, multiple reaction monitoring(MRM), and the content of the six PAs was calculated with the external standard method. The results suggested the differences in the six PAs among different parts of E. sonchifolia. Sk was detected in all the four parts, with similar content. SnNO also existed in all the four parts, but the content in roots was significantly higher than that in other parts. Sp and SpNO were found in both roots and flowers, with the content higher in the former than in the later. ImNO and LyNO were only found in leaves, and the content was low. Among the six components detected, ImNO, LyNO, and SpNO were found and determined for the first time, which enriched the toxic components and laid a scientific basis for the quality and safety evaluation of E. sonchifolia.

Influence of body mass index at diagnosis on outcome of thyroid cancer in children and adolescents.

PURPOSE: To systemically examine the relationship between body mass index, the extent of thyroid cancer and metastatic burden at diagnosis, and overall outcomes in children and adolescents with thyroid cancer. METHODS: A retrospective series of children and adolescents with differentiated thyroid cancer who underwent total thyroidectomy and 131I therapy was analyzed. Body mass index (BMI) at the time of surgery was assessed. The Chinese age- and sex-specific BMI percentile criteria for screening overweight and obesity in children and adolescents were used to define the overweight and obesity among patients. The relationship between BMI and clinicopathological features and clinical outcomes of the disease was evaluated by logistic regression modeling, incorporating the eighth edition of the American Joint Committee on Cancer TNM staging system and the 2015 American Thyroid Association guidelines. RESULTS: This study included 181 children and adolescents with differentiated thyroid cancer (82.9% females, median age 17 years). The mean BMI was 21.0 ± 3.9 kg/m2; 77.9% of the patients (N = 141) were normal weight, 13.2% (N = 24) were overweight, and 8.8% (N = 16) were obese. No positive associations were noted between BMI and T, N, or M stage on logistic regression analyses. The absence of an association was also demonstrated on analysis by BMI categories. After a median follow-up of 51 months, 114 children and adolescents (63.0%) had achieved excellent response to therapy. No associations were noted for persistent/recurrent disease among BMI groups. CONCLUSION: Our study highlights the absence of increased disease burden at the time of thyroid cancer diagnosis in pediatric patients with high BMI along with the same prognosis compared with normal-weight pediatric patients.

Therapeutic and Prebiotic Effects of Five Different Native Starches on Dextran Sulfate Sodium-Induced Mice Model of Colonic Colitis.

SCOPE: The availability of studies related to the effects of natural macronutrients on inflammatory bowel disease (IBD) remain relatively limited. This study investigates whether and to what extent the consumption of five different native starches alleviate the clinical symptoms and dysbiosis of gut microbiota associated with colitis. METHODS AND RESULTS: Using dextran sodium sulfate (DSS)-induced mouse model of colitis, the potential effects of native potato starch (PS), pea starch (PEAS), corn starch (CS), Chinese yam starch (CYS), and red sorghum starch (RSS) on the clinical manifestations and dysbiosis of gut microbiota are studied. Compared to CS and RSS, the consumption of PEAS, PS, and CYS significantly diminishes clinical enteritis symptoms, including reduced disease activity index, and the alleviated degree of colonic histological damage. Furthermore, the analysis of gut microbiota reveals the significant prebiotic characteristics of PEAS, PS and CYS, as indicated by the maintenance of gut microbiota hemostasis and the inhibition of typically pathogenic bacteria, including Escherichia coli and Helicobacter hepaticus. CONCLUSION: Starches from potato, pea, and Chinese yam alleviate colitis symptoms in a mouse model, and also show significant prebiotic characteristics. These findings suggest a cost-effective and convenient dietary strategy for the management of IBD.

The protective effect of tanshinone IIa on endothelial cells: a generalist among clinical therapeutics.

INTRODUCTION: Tanshinone IIa (TSA) has been approved to treat cardiovascular diseases by the China State Food and Drug Administration. TSA has exhibited a variety of pharmacological effects, including vasodilator, antioxidant, anti-inflammatory, and anti-tumor properties. Endothelial cells play an important physiological role in vascular homeostasis and control inflammation, coagulation, and thrombosis. Accumulating studies have shown that TSA can improve endothelial function through various pathways. AREAS COVERED: The PubMed database was reviewed for relevant papers published up to 2020. This review summarizes the current clinical and pharmaceutical studies to provide a systemic overview of the pharmacological and therapeutic effects of TSA on endothelial cells. EXPERT OPINION: TSA is a representative monomeric compound extracted from Danshen and it exhibits significant pharmacological and therapeutic properties to improve endothelial cell function, including alleviating oxidative stress, attenuating inflammatory injury, modulating ion channels and so on. TSA represents a spectrum of agents that are extracted from plants and can restore the endothelial function to establish the beneficial and harmless molecular therapeutics. This also suggests the possible detection of endothelial cells for very early diagnosis of diseases. In future, precise therapeutic methods will be developed to repair endothelial cells injury and recover endothelial dysfunction.

The prognostic relevance of p53 and Ki-67 to chemotherapy sensitivity and prognosis in triple-negative breast cancer.

BACKGROUND: This study aimed to explore the prognostic function of p53 and Ki-67 protein expression in chemotherapy sensitivity and prognosis in triple-negative breast cancer (TNBC). METHODS: Patients who were confirmed with TNBC in Wenzhou Geriatric Hospital and Wenzhou Hospital of Traditional Chinese Medicine (including the Oncology Department, Tumor Surgery Department, and Gynecology Department) between January 2006 and February 2018 were included in this study. The expression of p53 and Ki-67 detected by immunohistochemistry, the rate of recurrence, and the objective curative effect evaluation at the end of the first-line rescue treatment were recorded for all patients. RESULTS: The patients were followed up to August 2020, and the median follow-up time was 9 years and 4 months. A total of 285 patients with TNBC were enrolled in the study. The patients ranged in age from 19 to 76 years old, with an average age of 53 years. The overall recurrence rate among the patients was 31.58%. The majority of cases (68.07%) were pathological stage I. The overall positive expression rates of Ki-67 and p53 were 53.33% and 56.84%, respectively. In the TNBC recurrence group, the positive rates of p53 and Ki-67 were 71.11% and 82.22%, respectively, which were significantly higher than those in the non-recurrence group. The positive rates of p53 and Ki-67 in the chemosensitive group were 96.05% and 92.11%, respectively, which were significantly higher than those in the non-chemosensitive group. Among all the TNBC patients, 128 patients had positive expression of both p53 and Ki-67, and 101 patients had negative expression of both p53 and Ki-67. The chemosensitivity rate of TNBC patients with positive expression of both Ki-67 and p53 was 98.53%, and that of TNBC patients with negative expression of both Ki-67 and p53 was 0.00%. The difference was statistically significant. The recurrence rate in TNBC patients with positive expression of both Ki-67 and p53 was 53.13%, and that in patients with negative expression of both Ki-67 and p53 was 6.93%. The difference was statistically significant. CONCLUSIONS: The expression of p53 and Ki-67 had prognostic relevance to chemotherapy sensitivity and prognosis in TNBC patients.

Effects of Dietary Supplementation with κ-Selenocarrageenan on the Selenium Accumulation and Intestinal Microbiota of the Sea Cucumbers Apostichopus japonicus.

A 30-day feeding trial was conducted to investigate the effect of κ-selenocarrageenan on the growth performance, selenium accumulation, antioxidant capacity, and intestinal microbiota of sea cucumbers Apostichopus japonicus, with different sizes (70 g ± 10 g and 100 g ± 10 g). Sea cucumbers of each size were randomly assigned into two groups; a diet without supplemented κ-selenocarrageenan was referred to as a control diet, or supplemented with κ-selenocarrageenan at selenium (Se) levels of 2.0 µg/g. Selenium accumulation in the body wall and intestine was determined on days 0, 10, 20, and 30. The survival rate (SR) was significantly higher in the κ-selenocarrageenan-treated group (Se group) than in the control group. After 30 days of feeding, κ-selenocarrageenan supplementation increased the activities of glutathione peroxidase (GSH-Px) and total antioxidant capacity (T-AOC), and decreased malondialdehyde (MDA) levels in A. japonicus. Furthermore, the intestinal microbiota diversity of sea cucumbers was increased by dietary supplementation with κ-selenocarrageenan and the relative abundances of some probiotics (such as Sulfitobacter and Rhodobacteraceae) were also increased. It is suggested that κ-selenocarrageenan could increase the antioxidant capacity and modulate the intestinal microbiota of sea cucumbers A. japonicus. Further researches will be conducted for its optimal administration concentrations in vivo.

Role of aspartate ammonia-lyase in Pasteurella multocida.

BACKGROUND: Pasteurella multocida is responsible for a highly infectious and contagious disease in birds, leading to heavy economic losses in the chicken industry. However, the pathogenesis of this disease is poorly understood. We recently identified an aspartate ammonia-lyase (aspA) in P. multocida that was significantly upregulated under iron-restricted conditions, the protein of which could effectively protect chicken flocks against P. multocida. However, the functions of this gene remain unclear. In the present study, we constructed aspA mutant strain △aspA::kan and complementary strain C△aspA::kan to investigate the function of aspA in detail. RESULT: Deletion of the aspA gene in P. multocida resulted in a significant reduction in bacterial growth in LB (Luria-Bertani) and MH (Mueller-Hinton) media, which was rescued by supplementation with 20 mM fumarate. The mutant strain △aspA::kan showed significantly growth defects in anaerobic conditions and acid medium, compared with the wild-type strain. Moreover, growth of △aspA::kan was more seriously impaired than that of the wild-type strain under iron-restricted conditions, and this growth recovered after supplementation with iron ions. AspA transcription was negatively regulated by iron conditions, as demonstrated by quantitative reverse transcription-polymerase chain reaction. Although competitive index assay showed the wild-type strain outcompetes the aspA mutant strain and △aspA::kan was significantly more efficient at producing biofilms than the wild-type strain, there was no significant difference in virulence between the mutant and the wild-type strains. CONCLUSION: These results demonstrate that aspA is required for bacterial growth in complex medium, and under anaerobic, acid, and iron-limited conditions.

Research on the mechanism of microwave-toughened starch on glucolipid metabolism in mice.

Potato resistant starch (RS) was prepared by microwave-toughening treatment (MTT). This study investigated the beneficial effects of RS on high-fat diet (HFD)-induced hyperlipidemia in C57BL/6J mice by evaluating changes in the gut microbiota. The mice were fed low-fat diet with corn starch, HFD with corn starch, HFD with potato starch (HFP), or HFD with RS (HFR) for 6 weeks. The results showed that the HFR group had lower body weight and total cholesterol, triglyceride, and low-density lipoprotein cholesterol levels compared with the HFP group. Moreover, the brown adipose tissue levels of uncoupling protein 1 (UCP-1), ß3-adrenoceptor (ß3-AR), peroxisome proliferator-activated receptor-γ (PPAR-γ), and PPAR-γ coactivator-1α (PGC-1α) were increased. Our results showed that RS supplementation increased the Bacteroidetes/Firmicutes ratio and the abundance of short-chain fatty acid-producing Allobaculum, Ruminococcus, and Blautia. Our data suggest that RS prepared by MTT may be used as a prebiotic agent to prevent gut dysbiosis and obesity-related chronic diseases, such as hyperlipidemia, and obesity.

Xiao-Yao-San Formula Improves Cognitive Ability by Protecting the Hippocampal Neurons in Ovariectomized Rats.

Xiao-Yao-San (XYS) decoction is a traditional Chinese medicine formula. This study aimed to investigate the effect of XYS on cognitive abilities and its underlying mechanism in ovariectomized rats. Female Sprague-Dawley rats were ovariectomized and treated with XYS (3 g/kg or 9 g/kg) by gavage, with subcutaneous injection of 17-ß estradiol (E2, 2 µg/kg) as a positive drug control and gavage of 1 ml saline (0.9%) as a placebo control. After 6 weeks of treatment, rats were examined using the Morris water maze test. The estradiol level in the serum and hippocampus was measured by ELISA. Golgi staining was performed to observe neuronal morphology in the hippocampus. Apoptosis of hippocampal cells was observed by TUNEL staining. The protein content of N-methyl-D-aspartate receptor (NMDAR) 2A and 2B in the hippocampal CA1 region was determined by Western blot and immunohistochemistry. Expression of estrogen receptor (ER) and PI3K signaling was detected by Western blot. Compared with the sham group, both learning and memory were impaired in ovariectomized rats. Rats treated with E2 or high-dose XYS showed better learning and memory compared with the saline-treated rats. High-dose XYS significantly reduced escape latency in the spatial acquisition trial; meanwhile, the cross times and duration in the probe quadrant were increased in the spatial probe trial. High-dose XYS promoted the de novo synthesis of E2 content in the hippocampus but had no significant effect on the serum E2 level. Golgi staining indicated that high-dose XYS could increase the branch number and density of dendritic spines in the hippocampal CA1 area. TUNEL staining showed that high-dose XYS alleviated ovariectomy-induced neuronal apoptosis. The expression level of NMDAR2A and NMDAR2B in hippocampal CA1 was upregulated by XYS treatment. The beneficial effect of XYS was through activating ERα-PI3K signaling. In conclusion, high-dose XYS treatment can improve the cognitive abilities of ovariectomized rats by protecting the hippocampal neurons and restoring the hippocampal E2 level.

Branch-like gas in a commercial diver's liver: a case report.

We report the case of a 42-year-old commercial diver who presented with palpitations, arthralgia, tachypnea and vomiting after three hours of repetitive dives to 25-30 meters below sea level (msw). He was diagnosed with severe decompression sickness (Type II DCS) based on his dive history, his abrupt ascent to the surface within minutes, and systemic symptoms with mild hypovolemic shock. Besides remarkable cutis marmorata on the torso, the patient was also found positive for diffuse branch-like pneumatosis in the liver, mesentery and intestines on an abdominal computed tomography (CT). His vitals were relatively stable, with a soft distended abdomen and mild tenderness over the right upper quadrant. He was treated with hyperbaric oxygen (HBO2) treatment in addition to essential crystalloid resuscitation. The abdominal pneumatosis resolved completely after two HBO2 sessions. Post-diving intra-abdominal pneumatosis is a rare complication of DCS. In our case it was difficult for dive doctors to diagnose promptly because an emergency abdominal CT was not a routine for potential DCS cases. We propose that a contrast-enhanced abdominal CT, which usually involves a intravenous injection of imaging agent, should be considered in emergency management of these patients, especially when they present with gastrointestinal symptoms.