The protective effect of Jangkanghwan (Korean traditional food) on lipopolysaccharide-induced disruption of the colonic epithelial barrier

Jangkanghwan (JKH) is a Korean traditional food that is a mixture of food ingredients and traditional Korean medicine ingredients, and it has been observed to produce satisfactory anti-inflammatory, antioxidant, and antibacterial effects. In the current study, JKH was administered by gavage to BALB/C mice with lipopolysaccharide (LPS)-induced colonic epithelial dysfunction, and mouse body weight and food intake were recorded. Indexes such as colonic paracellular permeability, serum inflammatory cytokines, and bacterial translocation were used to comprehensively evaluate the regulatory effect of JKH on mouse colonic epithelial function, and qPCR and Western blot were also used to analyze the expression of tight junction (TJ)-related genes, such as occludin, claudin, zonula occludens (ZOs) proteins, and junction adhesion molecules (JAM) in the colonic epithelial tissue. The experimental results indicated that JKH relieved the edema of the liver, spleen, and mesenteric lymph node tissues, and reduced the loss of appetite and diarrhea caused by LPS injection in mice. It increased the amount of mice food intake from 3.7 g/day in the LPS group to 4.7 g per day; the water content in the feces of mice in the JKH group was 13.86% less than that in the LPS group. JKH reduced the inflammatory response in mice caused by LPS, protected the integrity of the colon, the permeability of fluorescent macromolecules was one-fourth of the LPS group, and enhanced the mRNA and protein expression of TJ-related proteins in colon tissue. Our findings highlight that JKH has benefits in intestinal health and relieving systemic inflammation, relevant aspects of its use as a functional food.

Up-regulation of Arl4a gene expression by broccoli aqueous extract is associated with improved spermatogenesis in mouse testes

Introduction: Broccoli (Brassica oleracea) is well recognized due to its properties as an anti-cancer, antioxidant and scavenging free radicals. However, its benefit in enhancing spermatogenesis is not well understood. Objectives: To investigate the effect of broccoli aqueous extract on sperm factors and also expression of the involving genes (Catsper1, Catsper2, Arl4a, Sox5 and Sox9) in sperm factors in mice. Material and methods: Male mice were divided randomly into six groups: (1) Control, (2) Cadmium (3 mg/kg mouse body weight), (3) Orally treated with 200 broccoli aqueous extract (1 g ml-1), (4) Orally treated with 400 µl of broccoli aqueous extract, (5) Orally treated with 200 broccoli aqueous extract plus cadmium, and (6) Orally treated with 400 µl of broccoli aqueous extract plus cadmium. Sperms factors and also gene expression in Catsper1, Catsper2, Arl4a, Sox5 and Sox9 genes were studied. Results: An obvious improvement in sperm number and slight enhancement in sperm motility was observed in mice treated with broccoli extract with and without cadmium. While sperm viability was reduced by broccoli extract, except for 200 µl of broccoli extract with cadmium that was significantly increased. Interestingly, Arl4a gene expression showed an increase in 400 µl broccoli-treated group. Likewise, the Arl4a mRNA level in mice treated with cadmium along with 200 µl broccoli extract was higher than in cadmium-treated mice. Furthermore, broccoli extract enhanced the mRNA level of Catsper2 and Sox5 genes in mice treated with both 200 and 400 µl broccoli extract along with cadmium than the only cadmium-treated group. Conclusion: Generally, improvement in sperm count in broccoli-treated mice provides insight into the pharmaceutical industry to make new products available to infertile men.

Interaction between Chitosan and Chloroquine against Plasmodium berghei and P. falciparum using In-Vivo and In-Vitro Tests

Background: The use of antimalarial drugs with number of compounds in combination form may potentiate each other's activity. Methods: This study was conducted in the School of Public Health, Tehran University of Medical Sciences, Tehran, Iran in 2018. It was based on two methods including in vivo and in vitro tests with aim of considering interaction between chitosan and chloroquine against Plasmodium berghei and P. falciparum parasites using different ratios of the agents with ED50s and IC50s baselines. Results: Administrating 10 and 20 mg/kg (mouse body weight) of chitosan alone to the P. berghei –infected mice up to 4 successive days resulted in 37% and 45% inhibition of P. berghei respectively, while employing the compound with chloroquine in combination form with ratios of 90/10 and 70/30 (chloroquine/chitosan) had a considerable potentiation including 71.58% and 83.85% inhibition effectiveness against P. berghei. Moreover, 20 mg/L (CCM) concentration of chitosan alone could eliminate 69.55% of P. falciparum in culture medium while in combination with chloroquine in ratios of 90/10 (chloroquine/chitosan) had considerable potentiation including 79.14% inhibition effectiveness. Mean survival time of those mice received combination therapy in ratios of 90/10 and 70/30 (chloroquine/chitosan) was longer than those took up mono therapy of either chloroquine or chitosan based on their ED50s doses. Conclusion: Interaction between chloroquine and chitosan showed considerable potentiation in combination form against either P. berghei or P. falciparum using in vivo and in vitro tests respectively. Meanwhile, interaction between the above mentioned agents resulted in a notable survival time for those P. berghei-infected mice treated with the combination.

Polydatin Attenuates 14.1 MeV Neutron-Induced Injuries via Regulating the Apoptosis and Antioxidative Pathways and Improving the Hematopoiesis of Mice

With more powerful penetrability and ionizing capability, high energetic neutron radiation (HENR) often poses greater threats than photon radiation, especially on such occasions as nuclear bomb exposure, nuclear accidents, aerospace conduction, and neutron-based radiotherapy. Therefore, there emerges an urgent unmet demand in exploring highly efficient radioprotectants against HENR. In the present study, high-throughput 14.1 MeV neutrons were generated by the high-intensity D-T fusion neutron generator (HINEG) and succeeded in establishing the acute radiation syndrome (ARS) mouse model induced by HENR. A series of preclinical studies, including morphopathological assessment, flow cytometry, peripheral complete blood, and bone marrow karyocyte counting, were applied showing much more serious detriments of HENR than the photon radiation. In specific, it was indicated that surviving fraction of polydatin- (PD-) treated mice could appreciably increase to up to 100% when they were exposed to HENR. Moreover, polydatin contributed much in alleviating the HENR-induced mouse body weight loss, spleen and testis indexes decrease, and the microstructure alterations of both the spleen and the bone marrow. Furthermore, we found that the HENR-damaged hematopoiesis was greatly prevented by PD treatment in such aspects as bone marrow hemocytogenesis, splenocytes balancing, or even the peripheral blood cellularity. The additional IHC investigations revealed that PD could exert potent hematopoiesis-promoting effects against HENR via suppressing apoptosis and promoting the antioxidative enzymes such as HO-1.

ACUTE SKIN TOXICITY CHARACTERISTICS OF THE DRUG BASED ON MOXIDECTIN

The acute skin toxicity characteristics of the drug for veterinary use “Inspector Mini” were studied at mice and rats. The active ingredient of the drug is moxidectin which belongs to the group of macrocyclic lactones of the milbemycin class. The studies were carried out in the vivarium of VNIIP – FSC VIEV (Moscow, Russia) on 2 experimental and 1 control groups of white outbred male mice of 19–21 g, 10 animals in each group and male rats of 200–230 g, 6 individuals in each. The mass of animals was indicated during application of the drug. The drug was used once without dilution in the form of the provided solution with single-channel mechanical dispensers with a dosing volume of 10–100 μl for mice and 100–1000 μl for rats. The animals in the experimental group 1 were treated at a dose of 10 400 mg/kg (100 μl per 10 g of mouse body weight or 1000 μl per 100 g of rat body weight), animals in the experimental group 2 – at a dose of 5 200 mg/kg (50 μl per 10 g of mouse body weight or 500 μl per 100 g of rat body weight). The animals in control group were not treat with the drug. As a result of the study, it was found that the LD50 of the drug “Inspector Mini” applied to the skin of mice and rats was more than 10 400 mg/kg per animal weight. During clinical examination of laboratory animals from the experimental groups, no signs of intoxication were observed. During the experiment, there was no significant difference (p ≥ 0.05) in the animals weight from the experimental groups within all periods of weighing compared with the control group of analogues.

Very High Dose Factor VIII Needed to Prevent Bleeding in the Presence of a Low Titer Anti-C1 Domain Antibody

Introduction: Up to 30% of patients with severe hemophilia A will develop inhibitory antibodies to factor VIII (fVIII inhibitors). Patients with congenital hemophilia who develop inhibitors usually have a polyclonal antibody response directed against the A2 and C2 domains of fVIII, but antibodies to all domains have been reported. We recently reported that anti-C1 domain antibodies were present in approximately 50% of plasmas from patients with hemophilia A and inhibitors. M6143 is a low titer (180 BU/mg IgG), tight-binding, anti-C1 monoclonal antibody that inhibits fVIII binding to von Willebrand Factor but not phospholipid. Surprisingly in a murine tail-snip bleeding model M6143 was pathogenic despite having a low inhibitory titer of 2.3 BU/ml (0.5 mg M6143/kg mouse body weight) and treatment with 180 U/kg of B-domain deleted (BDD) fVIII. The presence of M6143 resulted in increased clearance of fVIII/M6143 complexes. Given this data we hypothesized M6143 would remain pathogenic until fVIII was added in molar excess of M6143. Methods: 8-12 week old E16 hemophilia A mice received 100 µl injections of M6143 at a concentration of 0.5 mg/kg (65 nM estimated peak plasma concentration), 0.25 mg/kg (~32.5 nM), 0.125 mg/kg (~16 nM), 0.0625 mg/kg (8 nM), or normal saline, followed by BDD fVIII 15 minutes later at a dose of 180 U/kg (~2.5 nM estimated peak plasma concentration), 360 U/kg (~5 nM), 720 U/kg (~10 nM), or 1440 U/kg (~20 nM). Tails were warmed then transected at 4 mm distally two hours post fVIII injection. Blood loss per mouse body weight (mg/g) following tail snips was measured using a pre-weighed 15 ml conical tube containing normal saline. Results: Hemophilia A mice treated with 180 U/kg in the absence of antibody had minimal blood loss. M6143 at 0.5 mg/kg (2.3 BU/ml) produced significant bleeding at fVIII doses ranging from 180-1440 U/kg. Subsequently the concentration of M6143 was decreased while continuing extremely high fVIII dosing of 1440 U/kg (~20 nM). At all decreased concentrations of M6143, with high dose FVIII, bleeding was corrected (Figure 1). Conclusion: M6143 is a minimally inhibitory antibody whose pathogenicity is only overcome in the presence of very high dose fVIII. The amount of fVIII needed relative to the inhibitor titer is paradoxical to the typical clinical strategy that low titer inhibitors can be overwhelmed with modest increases in the fVIII dosing. Inhibitors with similar characteristics to M6143 may be responsible for the subset of patients with low titer inhibitors who do not respond to treatment with fVIII. Figure 1 Figure 1. Disclosures Meeks: CSL Behring: Membership on an entity's Board of Directors or advisory committees; Biogen: Membership on an entity's Board of Directors or advisory committees; Genentech: Membership on an entity's Board of Directors or advisory committees; Grifols: Membership on an entity's Board of Directors or advisory committees; Bayer Healthcare: Membership on an entity's Board of Directors or advisory committees; Shire: Membership on an entity's Board of Directors or advisory committees.