Mouse Tail Research Articles

Incorporation of mixed-dimensional palygorskite clay into chitosan/polyvinylpyrrolidone nanocomposite films for enhancing hemostatic activity

Clay mineral-based hemostatic materials have attracted much attention in recent years, but it is scarce to report the hemostatic nanocomposite films containing natural mixed-dimensional clay composed of natural one-dimensional and two-dimensional clay minerals. In this study, the high-performance hemostatic nanocomposite films were facilely prepared by incorporating the natural mixed-dimensional palygorskite clay leached by oxalic acid (O-MDPal) into chitosan/polyvinylpyrrolidone (CS/PVP) matrix. By contrast, the obtained nanocomposite films exhibited the higher tensile strength (27.92 MPa), lower water contact angel (75.40°), better degradation, thermal stability and biocompatibility after incorporation of 20 wt% of O-MDPal, suggesting that O-MDPal contributed to enhancing the mechanical performance and water holding capacity of the CS/PVP nanocomposite films. Compared with the medical gauze and CS/PVP matrix groups, the nanocomposite films also indicated excellent hemostatic performance evaluated by blood loss and hemostasis time indexes based on the mouse tail amputation model, which might be ascribed to the enriched hemostatic functional sites, and hydrophilic surface, robust physical barrier role of nanocomposite films. Therefore, the nanocomposite film exhibited a promising practical application in wound healing.

Research on mechanisms of acupoint sensitization in gastric ulcer mice from perspective of dorsal root ganglion neuronal excitability

To investigate the relationship between acupoint sensitization on the body surface and neuronal intrinsic excitability of the medium- and small-size dorsal root ganglion (DRG) neurons from the perspective of ion channel kinetics in mice with gastric ulcer. Male C57BL/6J mice were randomly divided into control (n=32) and model groups (n=34). The gastric ulcer model was established by injection of 60% glacial acetic acid (0.2 mL/100 g) into the gastric wall muscle layer and submucosa near the pylorus in the minor curvature of the stomach. In contrast, the same dose of normal saline was injected in the same way in the control group. Six days after modeling, Evans blue (EB) solution was injected into the mouse's tail vein for observing the number and distribution of the exudation blue spots on the body surface. Histopathological changes of the gastric tissue were observed by H.E. staining. Then, whole-cell membrane currents and intrinsic excitability of medium- and small-size neurons in the spinal T9-T11 DRGs were measured by in vitro electrophysiology combining with biocytin-ABC method. In the control group, EB exudation blue spots were not obvious, while in the model group, the blue spots on the body surface were densely distributed in the area of spinal T9-T11 segments, the epigastric region, and the skin around "Zhongwan" (CV12) and "Huaroumen" (ST24) regions, and near the surgical incision region. Compared with the control group, the model group had a high level of eosinophilic infiltrates in the submucosa of gastric tissues, severe gastric fossa structure damage, gastric fundus gland dilation and other pathological manifestations. The number of exudation blue spots was proportional to the degree of inflammatory reaction in the stomach. In comparison with the control group, the spike discharges of type II of medium-size DRG neurons in T9-T11 segments were decreased, and the current of whole-cell membrane was increased, basic intensity was decreased (P<0.05), discharge frequency and discharge number were increased (P<0.01,P<0.000 1); while the discharges of type I small-size DRG neurons were decreased, those of type II neurons increased, the whole-cell membrane current was decreased, and discharge frequency and discharge number were decreased (P<0.01, P<0.000 1). Both the medium- and small-size DRG neurons from the spinal T9-T11 segments involve in gastric ulcer-induced acupoint sensitization via their different spike discharge activities. And intrinsic excitability of these DRG neurons can not only dynamically encode the plasticity of acupoint sensitization, but also can help us understand the neural mechanism of acupoint sensitization induced by visceral injury.

Acute toxicity, analgesic and anti-inflammatory effects of soursop leaf ethanol extract (Annona muricata l.) in vivo

This study tested the peripheral analgesic effect of leaf extract by injecting 1% acetic acid on the peritoneum of white mice; tested the central analgesic effect by dipping the mice's tail in hot water (52 ± 0.5) °C; tested the acute anti-inflammatory effect by injecting carrageenan into the paws of mice causing edema; tested the chronic an- ti-inflammatory effect by injecting Freund's complete adjuvant (FCA) into the paws of mice causing edema. The control drugs used in the trials were diclofenac and codeine. Result: LD50 of the Annona muricata leaf ethanol extract was 0.705 mg/kg; The study showed that at all 3 doses, 35 mg/kg, 70 mg/kg and 140 mg/kg Annona muricata leaf extract had anti-inflammatory effects on acute and chronic inflammation; central and peripheral analgesic effects on mice.

Formulation and Evaluation of Nanogel used for the Treatment of Psorasis

Context: Although several formulation strategies have been developed for the treatment of psoriasis, there is an unmet need for optimization of its therapy. Objective: The objective was to develop a nanogel composed of Acitretin (ACT) loaded nanostructured lipid carrier (ACT-NLC) and to evaluate its potential in imiquimod-induced psoriasis model to ameliorate symptoms of psoriasis. Materials and methods: The developed control (CNGs) nanogels, acitretin loaded nanogels (ActCNGs) were characterized by UV and FTIR. Results and discussion: The systems were found to be spherical in shape with a size range of 98±10, and 138±8 nm having zeta potential values of +28±3 and +27±3 mV for CNGs and ActCNGs respectively. The in vitro haemolysis assay revealed that all the nanogel systems are blood compatible. The systems exhibited higher swelling and release at acidic pH. The ex vivo skin permeation studies using porcine skin confirmed the higher deposition of the systems at epidermal and dermal layers, which was confirmed further by fluorescent imaging. The in vivo anti-psoriatic activity study using Perry's mouse tail model and skin safety studies confirmed the potential benefit of the system for topical delivery of acitretin in psoriasis.

Evaluation of Fractionated Photofrin-mediated Photodynamic Therapy Using Different Light Fluences with Reactive Oxygen Species Explicit Dosimetry (ROSED).

Photodynamic therapy (PDT) is an established modality for cancer treatment, and reactive oxygen species explicit dosimetry (ROSED), based on direct measurements of in-vivo light fluence (rate), in-vivo photofrin concentration, and tissue oxygenation concentration, has been proved to provide the best dosimetric quantity which can be used to predict non-fractionated PDT outcome. This study performed ROSED for Photofrin-mediated PDT for mice bearing radiation-induced fibrosacorma (RIF) tumor. As demonstrated by our previous study, fractionated PDT with a 2-hour time interval can significantly improve the long-term cure rate (from 15% to 65% at 90 days), and it tends to increase as the light dose for the first light fraction gets larger. This study focused on further improving the long-term cure rate without introducing apparent toxicity using combinations of different first light fraction lengths and total light fluences. Photofrin was injected through the mouse tail vein at a concentration of 5 mg/kg. After 18~24 hours, treatment was delivered with a collimated laser beam of 1 cm diameter at 630 nm. Mice were treated using two fractions of light fluences with a 2-hour dark interval. Different dose metrics were quantified, including light fluence, PDT dose, and [ROS]rx. In addition, the total reacted [ROS]rx and treatment outcomes were evaluated and compared to identify the optimal light fraction length and total light fluence.

Helicobacter pylori outer membrane vesicles induce astrocyte reactivity through nuclear factor-κappa B activation and cause neuronal damage in vivo in a murine model

BackgroundHelicobacter pylori (Hp) infects the stomach of 50% of the world’s population. Importantly, chronic infection by this bacterium correlates with the appearance of several extra-gastric pathologies, including neurodegenerative diseases. In such conditions, brain astrocytes become reactive and neurotoxic. However, it is still unclear whether this highly prevalent bacterium or the nanosized outer membrane vesicles (OMVs) they produce, can reach the brain, thus affecting neurons/astrocytes. Here, we evaluated the effects of Hp OMVs on astrocytes and neurons in vivo and in vitro.MethodsPurified OMVs were characterized by mass spectrometry (MS/MS). Labeled OMVs were administered orally or injected into the mouse tail vein to study OMV-brain distribution. By immunofluorescence of tissue samples, we evaluated: GFAP (astrocytes), βIII tubulin (neurons), and urease (OMVs). The in vitro effect of OMVs in astrocytes was assessed by monitoring NF-κB activation, expression of reactivity markers, cytokines in astrocyte-conditioned medium (ACM), and neuronal cell viability.ResultsUrease and GroEL were prominent proteins in OMVs. Urease (OMVs) was present in the mouse brain and its detection coincided with astrocyte reactivity and neuronal damage. In vitro, OMVs induced astrocyte reactivity by increasing the intermediate filament proteins GFAP and vimentin, the plasma membrane αVβ3 integrin, and the hemichannel connexin 43. OMVs also produced neurotoxic factors and promoted the release of IFNγ in a manner dependent on the activation of the transcription factor NF-κB. Surface antigens on reactive astrocytes, as well as secreted factors in response to OMVs, were shown to inhibit neurite outgrowth and damage neurons.ConclusionsOMVs administered orally or injected into the mouse bloodstream reach the brain, altering astrocyte function and promoting neuronal damage in vivo. The effects of OMVs on astrocytes were confirmed in vitro and shown to be NF-κB-dependent. These findings suggest that Hp could trigger systemic effects by releasing nanosized vesicles that cross epithelial barriers and access the CNS, thus altering brain cells.

A Dual-Modality Imaging Method Based on Polarimetry and Second Harmonic Generation for Characterization and Evaluation of Skin Tissue Structures

The characterization and evaluation of skin tissue structures are crucial for dermatological applications. Recently, Mueller matrix polarimetry and second harmonic generation microscopy have been widely used in skin tissue imaging due to their unique advantages. However, the features of layered skin tissue structures are too complicated to use a single imaging modality for achieving a comprehensive evaluation. In this study, we propose a dual-modality imaging method combining Mueller matrix polarimetry and second harmonic generation microscopy for quantitative characterization of skin tissue structures. It is demonstrated that the dual-modality method can well divide the mouse tail skin tissue specimens' images into three layers of stratum corneum, epidermis, and dermis. Then, to quantitatively analyze the structural features of different skin layers, the gray level co-occurrence matrix is adopted to provide various evaluating parameters after the image segmentations. Finally, to quantitatively measure the structural differences between damaged and normal skin areas, an index named Q-Health is defined based on cosine similarity and the gray-level co-occurrence matrix parameters of imaging results. The experiments confirm the effectiveness of the dual-modality imaging parameters for skin tissue structure discrimination and assessment. It shows the potential of the proposed method for dermatological practices and lays the foundation for further, in-depth evaluation of the health status of human skin.

Thymus transplantation regulates blood pressure and alleviates hypertension-associated heart and kidney damage via transcription factors FoxN1 pathway

Previous studies have found that thymus is involved in the process of hypertension. However, whether thymus transplantation alleviates target organ damage in hypertensive mice remains unknown. The aim of this study was to evaluate the effects of thymus transplantation on blood pressure and target organ changes in mice with hypertension. Mice were randomly divided into normal control group (Con), hypertensive group (HTN) and thymus transplantation group (HTN + Trans). Thymus of neonatal mice was transplanted into the renal capsule of the transplantation group. After transplantation, the mouse tail noninvasive pressure was measured and heart function was evaluated weekly. Then mice were euthanized and organs or tissues were harvested at 4 weeks post-transplantation. The blood pressure of HTN + Trans group was lower than that in the HTN group. The expression of FoxN1, Aire, ATRAP, thymosin β4 and the content of sjTREC in thymus of HTN group was decreased and the number of naïve T cells in HTN group was lower compared with other two groups. The ratio of cTEC/mTEC in HTN group was higher than that in Con group and lower than that in HTN + Trans group. Cardiac pathology showed cardiac hypertrophy and fibrosis in HTN group whereas thymus transplantation improved heart function and structure. Altogether, our findings demonstrated thymus transplantation could improve thymus function of hypertensive mice, which increased the expression of thymus transcription factor FoxN1, affected the proportion of T cell subsets, and increased thymosin β4 thereby reducing blood pressure and reversing the progression of target organ damage.

The Effectiveness of Kenikir Leaf Extract (Cosmos Caudatus Kunth) on Bleeding Time in Mice (Mus Musculus) Tails

Background: Continuous bleeding will inhibit the formation of blood clots and cause the wound healing process to be hampered, thereby endangering the patient's life. Kenikir leaves contain flavonoid compounds, tannins, and saponins which have the potential as natural hemostatic agents in shortening bleeding time. Aim: To analyze the effectiveness of kenikir leaf extract (Cosmos caudatus kunth) on bleeding time in the tails of mice (Mus musculus). Material and Methods: This type of research is a laboratory experiment with a post test only control group design method. The samples used were 12 mice and divided into 3 groups, namely negative control (K-), positive control (K+), and treatment (P). All samples were adapted for 7 days, then fasted for 4 hours, then given each ingredient according to the group, then waited for 60 minutes and cut the mice tails along 0.5 cm from the tail end. Blood is dropped on filter paper every 30 seconds until the bleeding stops, the results of the calculation of bleeding time were analyzed using the Anova and LSD analysis. Results: The average bleeding time in the K-, K+, and P groups was 7.25 minutes, 1.87 minutes and 3.37 minutes. Anova analysis results obtained a significance value (p&lt;0.05), which means that there are differences in all groups. LSD analysis showed that there were significant differences between K- and K+ groups and K- and P groups, whereas in the K+ and P groups no significant differences. Conclusion: Kenikir leaf extract can shorten the bleeding time in mice tails.

Age-dependent mechanical properties of tail tendons in wild-type and mimecan gene-knockout mice – A preliminary study

Mimecan, or osteoglycin, belongs to the family of small leucine-rich proteoglycans. In connective tissues mimecan is implicated in the development and maintenance of normal collagen fibrillar organization. Since collagen fibrils are responsible for tissue reinforcement, the absence of mimecan could lead to abnormal tissue mechanical properties. Here, we carried out a preliminary investigation of possible changes in the mechanical properties of tendons in mice lacking a functional mimecan gene, as a function of age. Tail tendons were dissected from mimecan gene knockout (KO) and wild type (WT) mice at ages 1, 4 and 8 months and mechanical properties evaluated using a microtensile testing equipment. Mimecan gene knockout resulted in changes in tendon elasticity- and fracture-related properties. While tendons of WT mice exhibited enhanced mechanical properties with increasing age, this trend was notably attenuated in mimecan KO tendons, with the exception of fracture strain. When genotype and age were considered as cross factors, the diminution in the mechanical properties of mimecan KO tendons was significant for yield strength, modulus and fracture strength. This effect appeared to affect the mice at 4 month old. These preliminary results suggest that mimecan may have a role in regulating age-dependent mechanical function in mouse tail tendon.

Fatty acid oxidation fuels agonist-induced platelet activation and thrombus formation: Targeting β-oxidation of fatty acids as an effective anti-platelet strategy.

Platelet mitochondria possess remarkable plasticity for oxidation of energy substrates, where metabolic dependency on glucose or fatty acids is higher than glutamine. Since platelets metabolize nearly the entire pool of glucose to lactate rather than fluxing through mitochondrial tricarboxylic acid cycle, we posit that majority of mitochondrial ATP, which is essential for platelet granule secretion and thrombus formation, is sourced from oxidation of fatty acids. We performed a comprehensive analysis of bioenergetics and function of stimulated platelets in the presence of etomoxir, trimetazidine and oxfenicine, three pharmacologically distinct inhibitors of β-oxidation. Each of them significantly impaired oxidative phosphorylation in unstimulated as well as thrombin-stimulated platelets leading to a small but consistent drop in ATP level in activated cells due to a lack of compensation from glycolytic ATP. Trimetazidine and oxfenicine attenuated platelet aggregation, P-selectin externalization and integrin αIIb β3 activation. Both etomoxir and trimetazidine impeded agonist-induced dense granule release and platelet thrombus formation on collagen under arterial shear. The effect of inhibitors on platelet aggregation and dense granule release was dose- and incubation time- dependent with significant inhibition at higher doses and prolonged incubation times. Neither of the inhibitors could protect mice from collagen-epinephrine-induced pulmonary embolism or prolong mouse tail bleeding times. However, mice pre-administered with etomoxir, trimetazidine and oxfenicine were protected from ferric chloride-induced mesenteric thrombosis. In conclusion, β-oxidation of fatty acids sustains ATP level in stimulated platelets and is therefore essential for energy-intensive agonist-induced platelet responses. Thus, fatty acid oxidation may constitute an attractive therapeutic target for novel antiplatelet agents.

Artificial cerebrospinal fluid restores aspirin-inhibited physiological hemostasis through recovery of platelet aggregation function.

Optimal hemostasis provides safety and reliability during neurosurgery which improves surgical outcomes. Previously, artificial cerebrospinal fluid (aCSF) and its component sodium bicarbonate were found to facilitate physiological hemostasis by amplifying platelet aggregation. This study aimed to verify whether aCSF amplifies platelet-dependent hemostasis in the presence of antiplatelet agents. We prepared platelet-rich plasma (PRP) or washed platelets using aspirin (acetylsalicylic acid, (ASA)) or normal saline (NS). We evaluated samples treated with a commercially available aCSF solution or NS for amplification of aggregation, activation of integrin αIIbβ3, phosphatidylserine (PS) exposure, P-selectin (CD62P) expression, and formation of microparticles (MPs). We assessed the effect of aCSF on in vivo hemostasis in the presence of ASA by measuring the tail bleeding time in ASA-or NS-injected C57BL/6N mice. Compared with NS, aCSF amplified ASA-inhibited platelet aggregation by recovering platelet activation including PS exposure, MP release, CD62P expression, and integrin αIIbβ3 activation. When using washed platelets, aCSF almost completely counteracted the inhibition of platelet aggregation by ASA. Prolonged bleeding time from the amputated tail of ASA-injected mice was significantly shortened by the treatment with aCSF compared to NS. Sodium bicarbonate also directly amplified ASA-inhibited platelet aggregation. aCSF and sodium bicarbonate facilitate physiological hemostasis through the recovery of inhibited platelet aggregation even in the presence of ASA. The utilization of aCSF in the operative field may be advantageous for facilitating hemostasis in patients with impaired platelet function and contribute to improving outcomes of neurosurgery.

A snake venom-analog peptide that inhibits SARS-CoV-2 and papain-like protease displays antithrombotic activity in mice arterial thrombosis model, without interfering with bleeding time

Background(p-BthTX-I)2 K, a dimeric analog peptide derived from the C-terminal region of phospholipase A2-like bothropstoxin-I (p-BthTX-I), is resistant to plasma proteolysis and inhibits severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) strains with weak cytotoxic effects. Complications of SARS-CoV-2 infection include vascular problems and increased risk of thrombosis; therefore, studies to identify new drugs for treating SARS-CoV-2 infections that also inhibit thrombosis and minimize the risk of bleeding are required.ObjectivesTo determine whether (p-BthTX-I)2 K affects the hemostatic system.MethodsPlatelet aggregation was induced by collagen, arachidonic acid, and adenosine diphosphate (ADP) in the Chronolog Lumi-aggregometer. The coagulation activity was evaluated by determining activated partial thromboplastin clotting time (aPTT) and prothrombin time (PT) with (p-BthTX-I)2 K (5.0–434.5 µg) or 0.9% NaCl. Arterial thrombosis was induced with a 540 nm laser and 3.5–20 mg kg− 1 Rose Bengal in the carotid artery of male C57BL/6J mice using (p-BthTX-I)2 K. Bleeding time was determined in mouse tails immersed in saline at 37 °C after (p-BthTX-I)2 K (4.0 mg/kg and 8.0 mg/kg) or saline administration.Results(p-BthTX-I)2 K prolonged the aPTT and PT by blocking kallikrein and FXa-like activities. Moreover, (p-BthTX-I)2 K inhibited ADP-, collagen-, and arachidonic acid-induced platelet aggregation in a dose-dependent manner. Further, low concentrations of (p-BthTX-I)2 K extended the time to artery occlusion by the formed thrombus. However, (p-BthTX-I)2 K did not prolong the bleeding time in the mouse model of arterial thrombosis.ConclusionThese results demonstrate the antithrombotic activity of the peptide (p-BthTX-I)2 K possibly by kallikrein inhibition, suggesting its strong biotechnological potential.

Anti-leishmanial, immunomodulatory and anti-oxidative activity of quercetin against cutaneous leishmaniasis caused by Leishmania major

Objective: To evaluate the in vitro and in vivo efficacy of quercetin and its immunomodulatory and anti-oxidative activity against Leishmania major (L. major). Methods: L. major promastigotes and amastigotes were incubated with different concentrations of quercetin to estimate EC50. For in vivo study, the base of tails of mice was infected with L. major. After developing ulcers in the inoculation site, mice were treated with 50 mg/kg quercetin orally for 28 consecutive days. The wound-healing potential of quercetin was evaluated by histopathological analysis of tissue sections stained by hematoxylin and eosin as well as Masson's trichrome. In addition, the levels of tumor necrosis factor-α, interleukin-6, malondialdehyde, and adiponectin, the ferric reducing ability of plasma, as well as superoxide dismutase and glutathione peroxidase activities were measured. Results: The EC50 values of quercetin against L. major promastigotes and intracellular amastigotes were 0.27 and 0.85 μM, respectively. Histopathological analysis showed that fewer inflammatory cells, more fibroblasts, and more collagen deposition were observed in tissue sections of quercetin-treated mice. In addition, treatment with quercetin markedly increased glutathione peroxidase activity, the ferric reducing ability of plasma and adiponectin levels while decreasing malondialdehyde, interleukin-6, and tumor necrosis factor-α levels. Conclusions: Quercetin shows anti-leishmanial activity, immunomodulatory, anti-oxidative, and anti-inflammatory effects. Therefore, it may be further explored as an effective drug in treating leishmaniasis.

Метод взятия крови у мышей из периферической вены хвоста для исследования специфической активности эритропоэтина

One of the main quality indicators showing the effectiveness and safety of recombinant human erythropoietin drugs is their specific activity. The determination of this specific activity is recommended to carry out by the biological method in vivo on polycythemic and normocythemic mice. The main method of blood collection was the method of taking blood from the retroorbital sinus of the eye. The disadvantages of this method are the high risk of complications and the inability to take blood again after a short period of time. The next blood sampling is possible only 10 days after the procedure. This article describes a simple method of taking blood that allows us to select a sufficient amount of blood to study the specific activity of erythropoietin, minimize the traumatism of mouse tail tissues, reduce contamination of the obtained blood with foreign particles, minimize the number of complications, the possibility of repeated blood sampling after a short period of time.

Determination of Catecholamines in a Small Volume (25μL) of Plasma from Conscious Mouse Tail Vein.

The determination of plasma catecholamine levels is commonly used as a measure of the sympathetic nervous system's response to stress and is highly important for diagnosis, therapy, and prognosis of cardiovascular diseases, catecholamine-secreting tumors arising from the chromaffin cells of the sympathoadrenal system, and affective disorders. Diseases in which catecholamines are significantly elevated include pheochromocytoma, Parkinson's disease, Alzheimer's disease, neuroblastoma, ganglioneuroblastoma, von Hippel-Lindau disease, baroreflex failure, chemodectina (nonchromaffin paraganglioma), and multiple endocrine neoplasia. Plasma norepinephrine levels provide a guide to prognosis in patients with stable, chronic, and congestive heart diseases. The method described here for the determination of plasma catecholamines is based on the principle that plasma catecholamines are selectively adsorbed on acid-washed alumina at pH 8.7 and then eluted at a pH between 1.0 and 2.0. Upon injection, catecholamines in elutes were separated by a reversed phase C-18 column. After separation, the catecholamines present within the mobile phase enter the electrochemical detector. Electrochemical detection occurs because electroactive compounds oxidize at a certain potential and thereby liberate electrons that create measurable current. Catecholamines readily form quinones under these conditions, get oxidized, release two electrons, and create current. The electrochemical detector detects this electrical current that linearly correlates to the catecholamine concentration loaded into the ultra-performance liquid chromatography instrument. A 15-min mixing time during the adsorption and desorption steps was found to be optimal. If the washing step was omitted, the catecholamines could not be eluted from the acid-washed alumina. To prevent dilution, the alumina had to be centrifuged and not aspirated to dryness after the washing step. We report here that by changing the range in the electrochemical detector, plasma catecholamines were measured with only 12.5μL of plasma and more reliably with 25μL of plasma. The detection limit was 1ng/mL. This assay method is very useful as blood can be collected from the tail vein in a conscious mouse and the same mouse can be used for time-dependent or age-dependent studies.

A case of rickets in an artificially raised white-tailed eagle (Haliaeetus albicilla) chick at a zoo.

A two-week-old white-tailed eagle presented with an inability to stand and flex its limbs. Despite hatching naturally and owing to lack of parental attention, the bird was raised indoors by zookeepers with no access to sunlight. Palpation and radiographic examination of the bilateral tibiotarsus and femur bone revealed pronounced deformation and curvature, and bilateral decreased bone densities, respectively. The reduced calcium concentration in the blood was treated with calcium gluconate injections and calcium-supplemented feeds. Chopped mouse tails were fed directly, and whole pink-skinned nude mice were fed weekly. The zookeeper also gently massaged the bird and dressed it with a bandage. Sunlight exposure was provided daily. Saliva containing chicken feed was obtained from the mother. The bird could stand properly after four weeks of treatment, and the blood calcium concentration was restored to normal levels.

Acute Toxicity Study of Intravenously Administered Capecitabine Resealed Erythrocytes in Mice

Introduction:- Capecitabine resealed erythrocytes were prepared as a potential targeted drug delivery system for the treatment of hepatocellular carcinoma. This system has been developed to increase the therapeutic efficacy of drug and to reduce its adverse effects. This paper demonstrated the acute toxicity study of the formulation in Swiss albino mice. Methodology:- Dose of Capecitabine was 2000 mg/kg orally with zero toxicity as per the previous studies. The doses of formulation selected were 1/5th, 1/10th, 1/20th of the oral dose calculated on the basis of the % encapsulation efficiency of drug in formulation. The doses were given intravenously in single dose in mice tail vein. All the animals were observed for 14 days period for mortality, behavioral symptoms and locomotor activity. Results:- After sacrification of animals, serum was collected to analyze hematological and biochemical parameters. The isolated liver was observed for histopathological changes in mice. Here, we found that all the three to be selected would be safe for further in- vivo screening of Hepatocellular carcinoma. Conclusion:- So, we concluded that, all the three doses of formulation low (0.90mg/kg), medium (1.79mg/kg) and high (3.58mg/kg) with no signs of mortality, behavioural changes and no necrosis in liver histopathology. This revealed that, all the three selected doses were safe for further in vivo studies.

The Effects of Synthetic SREBP-1 and PPAR-γ Decoy Oligodeoxynucleotide on Acne-like Disease In Vivo and In Vitro via Lipogenic Regulation.

Acne vulgaris has a pathogenesis that involves increased sebum production and perifollicular inflammation. Sterol regulatory element-binding protein-1 (SREBP-1) and peroxisome proliferator activated receptor-γ (PPAR-γ) are transcription factors that regulate numerous genes involved in lipid biosynthesis. To improve a new therapeutic approach, we designed the SREBP/PPAR decoy oligodeoxynucleotide (ODN), a synthetic short DNA containing complementary sequences for the SREBP and PPAR transcription factors. We aim to investigate the beneficial functions and the molecular mechanisms of the synthetic SREBP/PPAR decoy ODN in lipogenic models. C. acnes was intradermally injected with a 1.0 × 107 colony forming unit/20 μL. The synthetic SREBP/PPAR decoy ODN or scrambled decoy ODN (10 μg) was transferred via the mouse tail vein injection. SZ95 cells were transfected with 2 μg of synthetic ODNs. After transfection, the SZ95 cells were cultured in serum-free medium containing 20 ng/μL of insulin-like growth factor-1 (IGF)-1 for 24 h. To investigate the expression of gene and signaling pathways, we performed Western blotting. The distribution of the chimeric decoy ODN was confirmed by EMSA. Lipid levels were assessed by Nile red and Oil Red O staining. The cytokine levels were measured by ELISA kit. This study showed that C. acnes-injected mice and IGF-1-stimulated SZ95 cells exhibited increased expression of SREBP-1 and PPAR-γ compared to the normal controls. In contrast, the administration of the SREBP/PPAR chimeric decoy ODN significantly suppressed the upregulation of lipogenic genes. Furthermore, the SREBP/PPAR decoy ODN decreased the plasma cytokines and cytokine levels of total protein. These results suggested that the SREBP/PPAR decoy ODN exerts its anti-lipogenic effects by regulating lipid metabolism and by inhibiting lipogenesis through the inactivation of the SREBP and PPAR pathways. Therefore, the synthetic SREBP/PPAR ODN demonstrates substantial therapeutic feasibility for the treatment of acne vulgaris.

Integrated metabolomics and molecular docking reveal berberrubine inhibits thrombosis by regulating the vitamin K catalytic cycle in mice

ObjectiveNatural product berberine was reported to inhibit platelet activation and thrombosis by suppressing the class Ⅰ PI3Kβ/Rasa3/Rap1 pathway. This study aims to investigate the effects and mechanisms of berberrubine, a main metabolite of berberine, to inhibit thrombus formation. MethodsCarrageenan-induced mouse tail thrombosis model was used to evaluate the effects of berberrubine hydrochloride (BBB) on thrombus formation in vivo. Non-targeted metabolomics was performed with UPLC-Q-TOF/MS to explore the potential mechanisms of BBB in inhibiting thrombosis. The effects of BBB on bleeding risk and prothrombin time were determined. And molecular docking was used to identify the possible target of BBB. ResultsAfter oral administration, BBB significantly inhibited carrageenan-induced thrombosis in mice without prolonging bleeding time. The results of non-targeted metabolomics showed that oral BBB could regulate ‘Phenylalanine, tyrosine and tryptophan biosynthesis’ and ‘Ubiquinone and other terpenoid-quinone biosynthesis’, which is closely related to the vitamin K catalytic cycle. Molecular docking revealed BBB could combine and interact with vitamin K epoxide reductase (VKOR) and γ-Glutamyl carboxylase (GGCX), which was mutually confirmed with the experimental results that oral BBB could significantly prolong prothrombin time. ConclusionsIntegrated metabolomics and molecular docking reveal BBB inhibited thrombosis by regulating the vitamin K catalytic cycle. Our research is helpful in deeply understanding the antithrombotic material basis of oral berberine, and also could provide scientific evidence for developing new antithrombotic drugs based on BBB in the future.