Caudal Vein Injection Research Articles

Effect of intestinal microbiota transplantation on cerebral ischemia reperfusion injury in aged mice via inhibition of IL-17.

This study investigates the effects and mechanisms of intestinal microbiota transplantation on cerebral ischemia reperfusion injury in aged mice. We constructed a middle cerebral artery occlusion model after fecal microbiota transplantation from young C57 mice to aged C57 mice for 30 consecutive days via enema. The neurological deficit score, cerebral infarction volume, fecal flora composition, and IL-17 levels in the colon, brain, and serum were evaluated in young mice, aged mice, and aged mice that received fecal microbiota transplantation. Moreover, we administered rIL-17A through caudal vein injection to verify its effect on cerebral ischemia reperfusion injury in aged mice. We find that aged mice exhibited larger cerebral infarction volume and more severe neurological deficit than young mice after middle cerebral artery occlusion. Bacteroidetes increased and firmicutes decreased significantly in the feces of aged mice after microbiota transplantation. Furthermore, the transplanted mice showed improved neurological function and reduced infarction volume after middle cerebral artery occlusion compared with the control aged mice. We also find that the neuroprotective effect of the microbiota transplantation was reversed by pre-treatment of rIL-17A. In summary, intestinal microbiota transplantation can alleviate cerebral ischemia reperfusion injury in aged mice by restoring their microbiota environment and inhibiting IL-17 in the gut, serum, and brain tissue.

A Quantitative LC-MS/MS Method for the Simultaneous Determination of the Presence of R-α-lipoic Acid and S-α-lipoic Acid After Protein Precipitation in Rat Plasma and its Application in a Toxicokinetic Study

Background: Lipoic acid is the only known chiral antioxidant that is both lipidsoluble and water-soluble. It is often used as a treatment for peripheral polyneuropathy caused by diabetes, alcohol, and chemicals. However, only a few long-term toxicological studies have been conducted on R-α-lipoic acid, which is a bioactive ingredient in lipoic acid. Objective: In this study, a simple, efficient, sensitive and stable LC-MS/MS method was used to determine RLA in rats, using deu-lipoic acid as an internal standard. Method: The samples to be detected were plasma samples treated with protein precipitation and the simultaneous determination of the presence of R-α-lipoic acid and S-α-lipoic acid was conducted using LC-MS/MS. An isocratic elution program with a mobile phase composed of acetonitrile and 0.1% formic acid water solution (52/48) used for chromatographic separation was set up using a CHIRALPAK® IE C18 (250 mm × 4.6 mm, 5 μm) column with a flow rate of 0.9 mL/min. A negative electrospray ionization source was chosen, and the multiple monitoring (MRM) mode was applied. Results: R-α-lipoic acid and S-α-lipoic acid both were found to be present at a linear range of 5- 5000 ng/mL. The plasma samples were stable under various storage conditions and temperatures. The toxicokinetics study indicated that there were gender differences and that R-α-lipoic acid showed bioaccumulative toxicity after long-term daily administration. In addition, R-α-lipoic acid and S-α-lipoic acid were not converted into each other in the rats. Conclusions: The method established was successfully used for the long-term toxicokinetic study of R-α-lipoic acid administered to rats through caudal vein injection. The toxicokinetics results indicated the presence of gender differences and the toxic accumulation of R-α-lipoic acid. The two enantiomers were not converted into each other in the rats.

Management of Airway Remodeling in a Mouse Model of Allergic Airways Inflammation Using Extracellular Vesicles from Human Bone Marrow-Derived Mesenchymal Stromal Cells

HIGHLIGHTS Extracellular vesicles (EV) from human bone marrow were able to reduce airway inflammation in a mouse model. Treatment applied by Intratracheal injection was more effective than systemic administration by caudal vein injection. IT administration of EVs appears to act therapeutically by restoring the basal BAL cell counts and attenuating early lung remodeling.

Molecularly Engineered Aptamers Targeting Tumor Tissue and Cancer Cells for Efficient in Vivo Recognition and Enrichment

Molecularly Engineered Aptamers Targeting Tumor Tissue and Cancer Cells for Efficient in Vivo Recognition and Enrichment

Protective Effect of Human Umbilical Cord Mesenchymal Stem Cells in Glucocorticoid-induced Osteonecrosis of Femoral Head.

To evaluate the effect of human umbilical cord mesenchymal stem cells (hUC-MSCs) on preventing rats from glucocorticoid-induced osteonecrosis of femoral head (GCONFH) in the early stage in vivo and to investigate the possible mechanism of hUC-MSCs in regulating the balance of osteogenesis and adipogenesis. All rats were randomly divided into 3 groups: control group (C group), model group (M group), and intervention group (I group). The model of GC-ONFH was developed by a sequential administration of lipopolysaccharide and methylprednisolone. The rats in the I group were treated with caudal vein injection of hUC-MSCs. Six weeks later, the blood samples were obtained to measure the activity of alkaline phosphatase (ALP) and the content of triglyceride (TG) in serum, and the femoral heads were harvested and observed by hematoxylin-eosin staining, Micro-CT, Western blot and real-time quantitative polymerase chain reaction. After intervention of hUC-MSCs, the necrosis rate of femoral head decreased from 83% (10/12) to 33% (4/12), the rate of empty bone lacuna was significantly decreased, the activity of ALP increased significantly, the content of TG decreased significantly, the bone density increased obviously, the expression of RUNX2 and Col I increased significantly and the expression of PPARγ decreased significantly. These results revealed that caudal vein injection of hUC-MSCs can effectively reduce the incidence of GC-ONFH in rats by increasing ALP activity and reducing TG content in serum, increasing bone mineral density, promoting the expression of RUNX2 and Col I, and inhibiting the expression of PPARγ.

Pancreatic Cancer Progression Is Regulated by IPO7/p53/LncRNA MALAT1/MiR-129-5p Positive Feedback Loop.

Background: Pancreatic cancer is a malignancy with poor prognosis. Importin 7 (IPO7) is a soluble nuclear transport factor, which has been linked to the pathogenesis of several human diseases. However, its role and underlying mechanism in pancreatic cancer are still obscure.Methods: Immunohistochemical staining and quantitative real-time polymerase chain reaction (qPCR) were performed to determine IPO7 expression in pancreatic cancer tissues and adjacent tissues. Western blot was used to measure IPO7 expression at the protein level in cell lines. Cell Counting Kit-8 (CCK-8), 5-bromo-2′-deoxyuridine (BrdU), flow cytometry, and Transwell assays were employed to explore the biological functions of IPO7. Subcutaneous xenograft transplanted tumor model and caudal vein injection model in mice were also established to validate the oncogenic role of IPO7. Western blot and qPCR were utilized to detect the regulatory function of IPO7 on p53 and metastasis-associated lung adenocarcinoma transcript 1 (MALAT1), respectively. Interaction between MALAT1 and miR-129-5p and interaction between miR-129-5p and IPO7 were verified by bioinformatics prediction, qPCR, dual-luciferase reporter gene experiment, RNA immunoprecipitation (RIP), and pull-down assay.Results: Upregulation of IPO7 in pancreatic cancer tissues was associated with adverse prognosis of the patients with pancreatic cancer. Knocking down IPO7 remarkably suppressed cancer cell proliferation and metastasis, while it promoted apoptosis. Overexpression of IPO7 facilitated the malignant phenotypes of pancreatic cancer cells. Mechanistically, IPO7 could repress the expression of p53 and induce the expression of MALAT1 but reduce miR-129-5p expression. Furthermore, miR-129-5p was identified as a posttranscriptional regulator for IPO7, and its inhibition led to IPO7 overexpression in pancreatic cancer cells.Conclusion: IPO7 is a novel oncogene for pancreatic cancer, and IPO7/p53/MALAT1/miR-129-5p positive feedback loop facilitates the progression of this deadly disease.

Effects of Cells Self-aggregation in the Treatment of Neurogenic Erectile Dysfunction With Traditional Single Cell Suspension of Adipose-derived Stem Cells

ObjectiveTo clarify the effects of cellular self-aggregation of adipose-derived stem cells (ADSCs) on erectile function (EF). MethodsA model of neurogenic erectile dysfunction was performed using bilateral cavernous nerve crush injury in rats. ADSCs suspensions (1 × 106/0.2 ml), were administered via intracavernous injection (ICI) after being allowed to shelve for 0 minute (ICI 0) or 60 minutes (ICI 60) in vitro, as well as cell aggregates isolated from ICI 60 (ICI A). The caudal vein injection group (CVI 60) was used to evaluate whether cell self-aggregation was beneficial to EF when introduced into the peripheral circulation. One day after the transplantation, the distribution of cells was observed. EF and histopathological changes were evaluated after 4 weeks. ResultsApproximately 85% of ADSCs self-aggregated into cell clusters at 60 minutes. The ICI 60 had more significant improvements in EF and more visualized ADSCs retained in the corpus cavernosum (CC) than ICI 0 and CVI 60 (P <.05), but no significant difference between ICI 60 and ICI A. In the CVI 60 group, the cell clusters formed by self-aggregation could hardly reach the CC and were mostly found in lung tissue. Immunofluorescence staining showed increased the content of expressing biomarkers of smooth muscle, nerve within the CC tissue in the ICI groups when compared to the CVI group. ConclusionADSCs self-aggregation before ICI may be an influential factor in the treatment of neurogenic erectile dysfunction. Its potential mechanism may be through improving cell retention in the CC.

Ultra-high-performance liquid chromatography coupled with quadrupole time of flight mass spectrometry method for quantifying polymer poloxamer 124 and its application to pharmacokinetic study.

Poloxamer is a commonly used pharmaceutical excipient. It is a high molecular polymer formed using polypropylene oxide and polyethylene oxide units. Specifically, poloxamer 124 is one of the smaller molecular weight in the poloxamer series; however, its pharmacokinetic behaviors in vivo are still unclear. In this study, a method for quantifying poloxamer 124 in rat plasma through ultra-high-performance liquid chromatography coupled with quadrupole time of flight mass spectrometry was developed. The intravenous dosage of PL124 was 10mg/kg. Plasma was collected at different times. The calibration curve was linear in the range of 0.1-5 μg/mL for the poloxamer 124 (r ≥ 0.9956) with the lower limit of quantitation of 0.1 μg/ml. The relative standard deviation of the intraday and interday precisions was below 8.0%, and the relative error of the accuracy was within ±12.0%. The extraction recovery, matrix effect, and stability were satisfactory in rat plasma. The validated method was successfully applied to a pharmacokinetic study of poloxamer 124 in rats. Results indicated that poloxamer 124 could be rapidly absorbed and eliminated through caudal vein injection. This study is helpful for the further study of poloxamer 124.

CTRP3 ameliorates fructose-induced metabolic associated fatty liver disease via inhibition of xanthine oxidase-associated oxidative stress

ObjectivesThe incidence of metabolic associated fatty liver disease (MAFLD) induced by high fructose consumption is dramatically increasing in the world while lacking specifically therapeutic drugs. The present study aimed to investigate the effect of complement C1q/tumor necrosis factor-related protein-3 (CTRP3) on fructose-induced MAFLD and its potential mechanisms. MethodThe animal models with MAFLD were built with Sprague-Dawley (SD) rats drinking 10 % fructose solution for 12 weeks. Then, specific hepatic CTRP3 overexpression was conducted by a single caudal-vein injection of CTRP3-expressing adenoviruses. Rats were sacrificed two weeks later. ResultsDrinking 10 % fructose solution for 12 weeks successfully built the rats models with MAFLD. Fructose feeding markedly decreased hepatic CTRP3 expression in rats. However, CTRP3 overexpression in liver alleviated hyperuricemia, dyslipidemia, liver function injury, intrahepatic triglyceride (TG) accumulation and histological changes of hepatic steatosis in rats fed with fructose. CTRP3 overexpression also inhibited hepatic XO activity in liver and improved subsequent oxidative stress, accompanied with downregulation of gene expression of sterol-regulatory element binding protein 1c (SERBP-1c) and fatty acid synthase (FAS). ConclusionCTRP3 attenuates MAFLD induced by fructose, which maybe partially attribute to rescued oxidative stress related with xanthine oxidase overactivity.

Establishment of a male Wistar rat model of nanobacteriainduced kidney stones

Purpose: To establish a male Wistar rat model of nanobacteria (NB)-induced kidney stones.&#x0D; Methods: Sixty male Wistar rats were randomly divided into control group (NC group) given caudal vein injection of saline + saline gavage, and NB-induced stone group (NBS group) given caudal vein injection of NB + saline gavage.&#x0D; Results: Compared with NC, serum creatinine, blood uric acid, urea nitrogen and urinary calcium levels in NBS group increased between weeks 3 and 8 (p &lt; 0.05). Kidney index (kidney weight/body weight ratio) in the NBS group was higher than that in NC group from weeks 8-10. At week 8, urine pH and serum phosphorus in NBS group were higher than those in NC group (p &lt; 0.05). Between weeks 6 and 7, serum calcium in NBS group was higher than that in NC group (p &lt; 0.05). Calcium crystals in NBS rats were distributed mostly in the distal and proximal convoluted tubules. However, no such crystals were observed in NC rats. Similarly, no such pathological changes were seen in the renal tissue of NC group. Calculus analysis showed that stone formation was higher in NBS group than in NC group (p &lt; 0.05). There was no significant difference in micro-CT between the two groups (p ˃ 0.05).&#x0D; Conclusion: The successful establishment of the Wistar rat kidney stone model using NB cultured from urine of upper urinary tract stone patient is potentially useful for further etiological studies on kidney stone formation.

LINC00958 promotes endometrial cancer cell proliferation and metastasis by regulating the miR‐145‐3p/TCF4 axis

Long noncoding RNAs (lncRNAs) exert an essential regulatory role in cancer progression. This work focuses on the role of LINC00958 in endometrial cancer (EC). LINC00958 expression in EC tissues was examined by GEPIA database and TCGA-UCEC dataset. LINC00958, miR-145-3p, and TCF4 mRNA expression levels in EC tissues and cells were examined by qRT-PCR. Western blot was employed to determine TCF4, E-cadherin, and N-cadherin protein expression levels. After LINC00958 was overexpressed or silenced, cell proliferation was determined using Cell Counting Kit 8 (CCK-8) and bromodeoxyuridine (BrdU) incorporation experiments. Cell migration and invasion were examined by Transwell experiment. Dual-luciferase reporter gene or RNA immunoprecipitation (RIP) experiments were executed to validate the targeting relationships among LINC00958 and miR-145-3p and TCF4. The effects of LINC00958 on EC cell proliferation and metastasis were investigated in vivo using a nude mouse subcutaneous graft model and a caudal vein injection model. LINC00958 was remarkably upmodulated in EC. Moreover, its overexpression was strongly linked to unfavorable overall survival of the patients. Functional experiments confirmed that in vitro knockdown of LINC00958 suppressed EC cell proliferation and metastasis. LINC00958 was validated to decoy miR-145-3p and repressed its expression, and TCF4 was uncovered to be a target gene of miR-145-3p and negatively modulated by miR-145-3p. Furthermore, the function of LINC00958 was dependent on its regulation of miR-145-3p and TCF4. LINC00958 acts as an oncogenic lncRNA to regulate EC progression by modulating the miR-145-3p/TCF4 axis. Knockdown of LINC00958 impedes tumor growth and metastasis in vitro and in vivo, opening a new avenue for therapeutic intervention.

AFGF Targeted Mediated by Novel Nanoparticles-Microbubble Complex Combined With Ultrasound-Targeted Microbubble Destruction attenuates Doxorubicin-Induced Heart Failure via Anti-Apoptosis and Promoting Cardiac Angiogenesis.

The purpose of this study was to evaluate the protective effect of acidic fibroblast growth factor targeted mediated by novel nanoparticles–cationic lipid microbubbles complex (aFGF–NP + CPMBs) combined with ultrasound targeted microbubble destruction (UTMD)on doxorubicin–induced heart failure (HF)and its mechanism. Heart failure rats induced by intraperitoneal injection with doxorubicin (DOX) to achieve cummulative dose of 15mg/kg for continuous 6 weeks showed left ventricular dysfunction, seriously oxidative stress, cardiomyocyte apoptosis, and decrease of myocardial vascular density. In contrast, aFGF–NP + CPMBs combined with UTMD therapy (3ug/kg, caudal vein injection, twice a week, 6weeks)prominently ameliorated left ventricular dysfunction by increased ejection fraction (EF) and fractional shortening (FS), decreased brain natriuretic peptide (BNP); strengthened the ability of antioxidant stress confirmed by increasing the activity of SOD and reducing the production of MDA; exerted the effect of anti–cardiomyocyte apoptosis and promotion angiogenesis by inhibited Bax expression and increased Bcl–2 expression and platelet endothelial cell adhesion molecule (CD31) expression. Taken together, the research suggested that aFGF targeted mediated by novel nanoparticles–cationic lipid microbubbles complex combined with UTMD should be a promising targeted treatment for heart failure.

Hypoxia Preconditioning Promotes the Proliferation and Migration of Human Urine-Derived Stem Cells in Chronically Injured Liver of Mice by Upregulating CXCR4.

Our previous studies reported that urine-derived stem cells (USCs) possess a strong self-renewal ability and multidirectional differentiation potential and thus are an ideal candidate cell source for hepatocellular transplantation. USC transplantation may repair the pathological changes of chronic liver injury to a certain extent, and hypoxia pretreatment may improve the recovery efficiency of USCs. Therefore, the present study aimed to investigate the possible mechanism of the improved recovery efficiency of hypoxia-pretreated USCs. A chronic liver injury model was established by intraperitoneal injection of carbon tetrachloride into nude mice. USCs were transplanted via caudal vein injection. Hematoxylin and eosin staining and Masson's staining were performed to determine the pathology of the liver. Immunofluorescence and frozen section biopsy were performed to determine differentiation and cell fusion in vivo. Cell coculture was used to detect cell fusion in vitro. The proliferative ability of USCs was evaluated using cell viability and colony formation assays, and the migratory functions of USCs were evaluated using wound healing and transwell assays. The degeneration of hepatocytes and the level of fibrosis in the hypoxia transplantation group were improved compared with the normoxia transplantation group. It was found that exogenous USCs may be differentiated into functional hepatocytes or fused with hepatocytes in vivo. C-X-C motif chemokine (CXC) ligand 12 (CXCL12) expression levels in liver tissue of the chronic liver injury model were upregulated compared with those in the control group. The expression of CXC receptor 4 (CXCR4) in hypoxia-pretreated USCs was also significantly upregulated. The results suggested that USCs fused with different types of liver cells and that hypoxia treatment promoted the fusion rate in vitro by upregulating CXCR4 signaling. Furthermore, hypoxia pretreatment promoted cell proliferation, migration, and cell fusion by inducing CXCR4 signaling, leading to USC-elicited liver tissue recovery following injury in vivo.

Improvement of Vascular Function by Knockdown of Salusin-β in Hypertensive Rats via Nitric Oxide and Reactive Oxygen Species Signaling Pathway.

PurposeSalusin-β, a multifunctional vasoactive peptide, has a potentially important function in the pathological development of hypertension. However, the exact functional role of salusin-β and the underlying mechanism in this process are still not fully understood. The current study aimed to investigate the effects of silencing salusin-β on vascular function and vascular remodeling, as well as its signaling pathways in spontaneously hypertensive rats (SHR) and Wistar-Kyoto rats (WKY).MethodsSilencing salusin-β was performed by caudal vein injection of adenovirus expressing salusin-β short hairpin RNA (shRNA). Acetylcholine (ACh)-induced endothelium-dependent relaxation was used to evaluate vasodilator function, and high K+ solution-induced constriction was used to evaluate vasoconstriction function.ResultsSalusin-β levels in plasma and its protein expression in mesenteric artery (MA), coronary artery (CA), and pulmonary artery (PA) of SHR were higher than those in WKY. The salusin-β level and expression were decreased effectively by salusin-β shRNA. Knockdown of salusin-β decreased arterial blood pressure (ABP) and high K+ solution-induced vascular constrictions, and improved the endothelium-dependent relaxation and vascular remodeling in SHR. The improved effect of silencing salusin-β on ACh-induced relaxation in SHR was almost blocked by the nitric oxide synthase (NOS) inhibitor L-NAME. Compared to WKY, the endothelial NOS (eNOS) activity and level, and nitric oxide (NO) level were decreased, while NAD(P)H oxidase activity and reactive oxygen species (ROS) levels in MA, CA, and PA of SHR were increased, which were all redressed by salusin-β knockdown.ConclusionThese results indicate that knockdown of salusin-β improves endothelium-dependent vascular relaxation and vascular remodeling and decreases ABP and vasoconstriction in SHR, which might be accomplished by increasing eNOS activation and NO release while inhibiting NAD(P)H oxidase derived-ROS generation. Scavenging salusin-β improves vascular function and then prevents the development and progression of vasculopathy of hypertension.

A safe and efficient bioactive citrate-lysine/miRNA33 agonist nanosystem for high fat diet-induced obesity therapy

The increased prevalence of obesity is recognized as a serious public health problem affecting millions of people. Although some anti-obesity drugs are currently licensed for use, these drugs involve diverse side effects such as diarrhea, vomiting, and low efficiency. Here we developed a safe and efficient biomaterials-based anti-obesity nanosystem (PCG-EPL/miR33agonist), which was formed with poly (citric acid)-glycerol-polylysine (PCG-EPL) and miR33 agonist by self-assembly. The PCG-EPL could efficiently load, protect and deliver the miR33 agonist into the adipocytes and decreased the obesity-related IL-1β expression in adipocytes in vitro. The high-fat diet induced obese rat model experiment showed that the PCG-EPL/miR33agonist via caudal vein injection effectively reduced the body weight by enhancing lipid metabolism and decreasing the inflammatory factors expressions (IL-1β, TNF-α and IL-6) in vivo, without suppressing the appetite of rat. Compared with the obese mice, mice in PCG-EPL/miR33 had higher average food intake but lower energy conversion rate. PCG-EPL/miR33agonist significantly inhibited the growth of adipocytes. Noteworthy, this weight loss strategy is not only safe and efficient, but also does not need diet control. Thus, PCG-EPL may serve as an ideal platform for RNA drug delivery in anti-obesity therapy, especially for those who need to lose weight but unable to autonomously control their diet.

Baicalein attenuates cardiac hypertrophy in mice via suppressing oxidative stress and activating autophagy in cardiomyocytes.

Baicalein is a natural flavonoid extracted from the root of Scutellaria baicalensis that exhibits a variety of pharmacological activities. In this study, we investigated the molecular mechanisms underlying the protective effect of baicalein against cardiac hypertrophy in vivo and in vitro. Cardiac hypertrophy was induced in mice by injection of isoproterenol (ISO, 30 mg·kg-1·d-1) for 15 days. The mice received caudal vein injection of baicalein (25 mg/kg) on 3rd, 6th, 9th, 12th, and 15th days. We showed that baicalein administration significantly attenuated ISO-induced cardiac hypertrophy and restored cardiac function. The protective effect of baicalein against cardiac hypertrophy was also observed in neonatal rat cardiomyocytes treated with ISO (10 μM). In cardiomyocytes, ISO treatment markedly increased reactive oxygen species (ROS) and inhibited autophagy, which were greatly alleviated by pretreatment with baicalein (30 μM). We found that baicalein pretreatment increased the expression of catalase and the mitophagy receptor FUN14 domain containing 1 (FUNDC1) to clear ROS and promote autophagy, thus attenuated ISO-induced cardiac hypertrophy. Furthermore, we revealed that baicalein bound to the transcription factor FOXO3a directly, promoting its transcription activity, and transactivated catalase and FUNDC1. In summary, our data provide new evidence for baicalein and FOXO3a in the regulation of ISO-induced cardiac hypertrophy. Baicalein has great potential for the treatment of cardiac hypertrophy.

FITC-Labeled Alendronate as an In Vivo Bone pH Sensor.

pH is a critical indicator of bone physiological function and disease status; however, noninvasive and real-time sensing of bone pH in vivo has been a challenge. Here, we synthesized a bone pH sensor by labeling alendronate with the H+-sensitive dye fluorescein isothiocyanate (Aln-FITC). Aln-FITC showed selective affinity for hydroxyapatite (HAp) rather than other calcium materials. An in vivo biodistribution study showed that Aln-FITC can be rapidly and specifically delivered to rat bones after caudal vein injection, and the fluorescence lasted for at least 12 h. The fluorescence intensity of Aln-FITC binding to HAp linearly decreased when the pH changed from 6 to 12. This finding was further confirmed on bone blocks and perfused bone when the pH changed from 6.8 to 7.4, indicating unique pH-responsive characteristics in the bone microenvironment. Aln-FITC was then preliminarily applied to evaluate the changes in bone pH in a nude mouse acidosis model. Our results demonstrated that Aln-FITC might have the potential for minimally invasive and real-time in vivo bone pH sensing in preclinical studies of bone healing, metabolism, and cancer mechanisms.

Detecting Establishment of Shared Blood Supply in Parabiotic Mice by Caudal Vein Glucose Injection.

Parabiosis is an experimental method for surgically combining two parallel animals along the longitudinal axis of the body. We present a protocol for detecting the successful establishment of blood chimerism in parabionts by a caudal vein injection of glucose. Parabiotic mice were constructed. Glucose was injected into the donor mouse through the tail vein, and the fluctuation of blood glucose level was measured in both mice using a blood glucometer at different time points. Our results showed that after glucose injection, the blood glucose level in donor mice increased sharply after 1 min and decreased slowly thereafter. Meanwhile, the blood glucose level of the recipient mice peaked 15 min after injection. Similar results were obtained with Evans blue, used as a positive control for glucose. The synchronous fluctuation of blood glucose levels indicates that blood flow between the two mice was established successfully.

Human umbilical cord mesenchymal stem cells suppress lung cancer via TLR4/NF-κB signalling pathway

The present study investigated the inhibitory effect of human umbilical cord mesenchymal stem cells (hucMSCs) on the growth of lung cancer in mice and its relationship with the TLR4/NF-κB signalling pathway. A mouse model of Lewis lung cancer was constructed by inoculating mice with Lewis lung cancer cell suspension under the axilla. Twenty mice were randomly and evenly divided into experimental group and control group. The experimental group received a caudal vein injection of 1 × 106 hucMSCs each week for two consecutive weeks, and the control group, an equal amount of saline at the same time. Two weeks after treatment with hucMSCs, the mice were sacrificed and the tumour volume and tumour weight were recorded. Enzyme-linked immunosorbent assay was used to determine the IL-1, IL-6 and TNF-α contents in lung cancer tissues; real-time polymerase chain reaction, to measure the level of mRNA; and Western blotting, to determine the protein expression. The results showed that injection of hucMSCs inhibited the growth of Lewis lung cancer tissues, reduced Lewis lung cancer tissue weight, enhanced thymus index, and suppressed inflammatory responses. hucMSCs inhibited the expression of inflammatory factors IL-1, IL-6 and TNF-α by activating TLR4/NF-κB signalling pathway, and facilitated the expression of Bax. Moreover, huMSCs induced the expression of apoptogenic factor Bax by activating TLR4/NF-κB signalling pathway, and inhibited the expression of antiapoptotic factor bcl-2. The present study demonstrates that hucMSCs reduce inflammatory responses by mediating TLR4/NF-κB signalling pathway, promote apoptosis of tumour cells, and inhibit tumour proliferation in mice with Lewis lung cancer.

Magnesium isoglycyrrhizinate protects against concanavalin A-induced immunological liver injury in a mouse model

Abstract Background: To evaluate the protective effects of magnesium isoglycyrrhizinate on a mouse model of concanavalin A (ConA)-induced immunological liver injury. Materials and Methods: Forty-eight mice were randomly divided into a normal control group, a model group, three dose groups of magnesium isoglycyrrhizinate (12.5, 25, 50 mg/kg) and a dexamethasone group (2.5 mg/kg). Magnesium isoglycyrrhizinate was intraperitoneally injected for 5 consecutive days, and the model of immunological liver injury was established on the fifth day after caudal vein injection of ConA (20 mg/kg). Blood was collected to detect the activities of alanine transaminase (ALT) and aspartate transaminase (AST) as well as the levels of tumor necrosis factor-α (TNF-α) and interferon-γ (IFN-γ). The levels of neopterin (NP) and malondialdehyde (MDA) and the activities of myeloperoxidase (MPO) and superoxide dismutase (SOD) in liver tissues were measured, and histopathological changes were observed. Results: The serum levels of ALT and AST in the model group increased. Hepatic lobules had necrotic foci and inflammatory cell infiltration. The plasma levels of TNF-α and IFN-γ increased. In liver tissues, the levels of NP, MDA and MPO rose, but that of SOD decreased. Magnesium isoglycyrrhizinate significantly attenuated the activities of ALT and AST (P&lt;0.05). Histopathological staining showed that inflammation of the liver was relieved significantly. Magnesium isoglycyrrhizinate also decreased the levels of NP, MDA and MPO in liver tissues (P&lt;0.05), raised that of SOD and reduced the plasma levels of TNF-α and IFN-γ (P&lt;0.05). Conclusion: Magnesium isoglycyrrhizinate protected against ConA-induced immunological liver injury in mice, probably through immune regulation and antioxidation.