ConA Group Research Articles

Doppler Evaluation of Uterine Blood Flow in Patients with Unexplained Recurrent Pregnancy Loss.

This study aimed to analyze uterine artery and spiral artery hemodynamics in patients with unexplained recurrent pregnancy loss (URPL) with varying pregnancy outcomes. 174 pregnant women with URPL and 144 pregnant women without adverse pregnancy histories were enrolled in this retrospective study. Based on pregnancy outcomes, these patients were divided into two groups: normal pregnancy outcomes (URPL-N, n=138) and adverse pregnancy outcomes (URPL-A, n=36). Control group participants were categorized into normal pregnancy outcomes (CON-N, n=129) and adverse pregnancy outcomes (CON-A, n=15). We compared uterine artery and spiral artery hemodynamics during different stages of gestation and the predictive value of these parameters for pregnancy outcomes. URPL-N group had fewer pregnancy losses and lower BMI compared to URPL-A group (P< 0.05). Spiral artery hemodynamics in URPL-N and CON-N groups were lower than those in URPL-A and CON-A groups during the mid-luteal phase, 11-13weeks, 15-17weeks, and 19-21weeks of gestation, respectively. Uterine artery hemodynamics ((Pulsatility index (mPI), resistive index (mRI), and systolic-to-diastolic ratio (mS/D)) in the mid-luteal period were lower in URPL-N group than URPL-A group. Similarly, in CON-N group were lower than CON-A group. The URPL-A and CON-A groups had higher uterine artery and spiral artery hemodynamics when compared to the URPL-N and CON-N groups. Spiral artery hemodynamics exhibited larger areas under the ROC curve compared to uterine artery parameters. Abnormal hemodynamics in these arteries may contribute to URPL and adverse pregnancy outcomes. Spiral artery hemodynamics are more reliable predictors of pregnancy outcomes than uterine artery parameters.

Dietary supplementation with pseudostellaria heterophylla polysaccharide enhanced immunity and changed mRNA expression of spleen in chicks

In recent years, increasing interest has focused on natural components extracted from plants, among which plant polysaccharides as natural immunomodulators that can promote animal immunity. The present study was performed to investigate the effect of feed supplement Pseudostellaria Heterophylla Polysaccharide (PHP) on serum Immunoglobulins, T lymphocyte subpopulations, Cytokines and Lysozyme (LZM) activity in chicks. In addition, the influence of PHP on splenic gene expression was investigated by transcriptome sequencing. Four hundred 7-day-old Gushi cocks were randomly divided into four groups in a completely randomized design. The chicks were fed with a basal diet supplemented with 0 (CON-A), 100 (PHP-L), 200 (PHP-M) and 400 (PHP–H) mg/kg PHP. Blood and spleen samples were collected from 6 randomly selected chicks in each group at 14, 21, 28, and 35 days of age. The results showed that compared to the CON-A group, the PHP-M group exhibited significant increases in the levels of IgA, IgG, IgM, CD3, and LZM in the serum at 14, 21, 28, and 35 days (P < 0.05), and at 28 d, there was a significant quadratic relationship between the levels of dietary PHP and the levels of IgG, IgM, IFN-γ, IL-2, CD3, and LZM. Furthermore, a total of 470 differentially expressed genes (DEGs) were identified in spleen from PHP-M and CON-A at 28 d. These DEGs were significantly enriched in the Phagosome, Intestinal immune network for IgA production and Cytokine-cytokine receptor interaction pathways. The present investigation highlights the ameliorating effect of dietary PHP on immunological variables and spleen of chicks, the study suggests that PHP supplementation can enhance immunity and positively impact spleen mRNA expression in chicks.

Elemental and molecular characterization of degrading blood pools.

Blood is a commonly encountered type of biological evidence and can provide critical information about the crime that occurred. The ability to accurately and precisely determine the time since deposition (TSD) of a bloodstain is highly sought after in the field of forensic science. Current spectral methods for determining TSD are typically developed using small volume bloodstains, we investigate the applicability to larger volume blood pools where drying and degradation mechanics are different. We explored the differences that exist between the surface and bulk of dried segments from fragments collected from 15 mL dried blood pools and identified heterogeneity using RGB colour analysis and hierarchical cluster analysis (HCA). The physical, molecular, and atomic differences between the layers were further investigated using scanning electron microscopy (SEM), X-Ray photoelectron spectroscopy (XPS), attenuated total reflection-Fourier transform infrared (ATR-FTIR) spectroscopy, and Raman spectroscopy. SEM identified different morphology on the surface and the bulk indicative of density-dependant cellular settling. XPS revealed that iron was not present on the surface but rather was present in the bulk where the red blood cells had settled. The oxidation state of the iron was quantified over three weeks in which it transitioned from entirely Fe2+ to primarily Fe3+, as expected for ex vivo degradation of hemoglobin. Further, indications of amide saponification occurring at the blood-air interface were identified in the increased quantity of the C-O moiety relative to CO, and the formation of free amines and OC-ONa groups over time. ATR-FTIR and Raman spectroscopy provided insights into differences in the molecular composition of the layers, suggesting that the surface consists of more nucleic acids, lipids, and glycoproteins than the bulk, which was dominated by proteins (p < 0.001% using principal component analysis (PCA)). Additionally, spectral band trends previously reported to have applicability to the estimation of TSD were observed for the bulk portion of the blood pool as the Hb underwent predictable time dependant changes from oxyHb to metHb. PCA was performed based on all spectral data which demonstrated statistically significant differences between the surface and bulk, as well as proof-of-concept for linear TSD estimation models.

Investigation of the synergetic regulation of O2/Ar preheating treatment and sodium salt addition on semichar combustion characteristics

Using Raman spectroscopy, temperature-programmed desorption (TPD) and laser-induced breakdown spectroscopy (LIBS), the relationship between microstructural characteristics and molecular emissions was determined. It can be concluded that the enhancement of C2 and CN emissions was related to the generation of small aromatic rings and C(O) groups. Compared with the Ar preheating treatment, the presence of additional amount of O2 and sodium salt accelerated the dissociation of the large aromatic ring skeleton, leading to the significant enhancement of small aromatic rings, C(O) and CONa groups. Subsequently, the combustion performance of preheated semichar was investigated by the TG-DTG method. The experimental results demonstrated that pretreatment under oxidizing atmospheres (without and with sodium salt addition) enhanced the comprehensive combustion index (S) and the combustion stability (Rw) of semichars. The corresponding combustion activation energy (E) was weakened. Furthermore, the DFT method was used to analyse the combustion behaviours of semichars. This result demonstrated that preheating treatment (with or without sodium participation) altered the distribution of the Hirshfeld atomic charge (ADCH) of preheated samples, and the combustion essence of semichar was the fracture and recombination of aromatic clusters. For the semichar preheated in the O2/Ar atmosphere, the energy gap of the most optimized pathway was 436.26 kJ/mol. When sodium participated in the oxidizing pretreatment, the energy gap of the most optimized pathway was 413.80 kJ/mol. The positive effect of sodium on the combustion performance of semichar was identified, which was in good agreement with the experimental results.

Huganbuzure Granule Attenuates Concanavalin-A-Induced Immune Liver Injury in Mice via Regulating the Balance of Th1/Th2/Th17/Treg Cells and Inhibiting Apoptosis.

In Uygur medicine, Huganbuzure granule (HBG) is one of the classical prescriptions for liver protection. However, its role in immune liver injury remains unknown. This study evaluates the effect of HBG on concanavalin-A- (ConA-) induced immune liver injury and investigates its protective underlying mechanism. BALB/c mice were randomly divided into five groups (n = 24 mice per group): control, ConA, 1.6 g/kg HBG + ConA, 3.2 g/kg HBG + ConA, and 6 mg/kg prednisolone + ConA. HBG was intragastrically administrated once daily for ten consecutive days, prior to ConA (20 mg/kg) injection. The levels of alanine aminotransferase (ALT), aspartate aminotransferase (AST), total bilirubin (TBIL), superoxide dismutase (SOD), and malondialdehyde (MDA) in mouse serum were measured after ConA injection. Moreover, liver-related mRNA levels were evaluated by qPCR. The detection of liver-related proteins was assessed by immunohistochemistry and western blot analysis. Compared with the ConA group, HBG reduced the mRNA expression of IL-17A and IFN-γ and the protein expression of T-bet and ROR-γt. In addition, HBG increased the mRNA expression of IL-4 and TGF-β and protein expression of GATA3 and Foxp3, indicating that HBG regulated the balance of Th1/Th2 and Th17/Treg. Furthermore, HBG alleviated immune liver injury by reducing oxidative stress, inhibiting apoptosis, and decreasing the expression of p-JNK, p-ERK, p-p38, p-JAK1, p-STAT1, p-STAT3, and IRF1. Our data suggested that HBG attenuated ConA-induced immune liver injury by regulating the immune balance and inhibiting JAK1/STATs/IRF1 signaling, thereby reducing apoptosis induced by JNK activation. The findings indicate that HBG may be a promising drug for immune liver injury.

Magnesium isoglycyrrhizinate ameliorates concanavalin A-induced liver injury via the p38 and JNK MAPK pathway

Context Acute liver failure is a serious disease caused by a variety of factors, and immunological injury is an important pathological process. Comprehensive liver treatment efficacy is poor, and the mortality rate is high. Magnesium isoglycyrrhizinate (MgIG) is a new glycyrrhizin drug extracted from the traditional Chinese medicine licorice. The mechanism by which MgIG regulates ConcanavalinA (ConA)-induced immunological liver injury in mice is not completely clear. Materials and methods Immunological liver injury was induced in mice by ConA injection, and the inflammatory macrophages model was induced by lipopolysaccharide (LPS). MgIG was administered 30 min prior to ConA and LPS treatment. The mice in the different groups were sacrificed 12 h after treatment, and macrophages were measured at 30 min, 1 h, and 2 h after induction. Macrophages, liver, and blood samples were then collected for analysis. Results After drug administration, the MgIG group showed a marked decrease in serum transaminase levels, reduced apoptosis and hepatic inflammatory responses compared to the ConA group. Furthermore, there was a significant reduction in inflammatory cytokine levels in the serum and liver tissue. In vitro, the expression of inflammatory cytokines was distinctly reduced after MgIG administration. In addition, MgIG pretreatment reduced the expression of inflammatory cytokines and regulated the phosphorylation of p38 and JNK proteins in the MAPK pathway. Conclusion These findings demonstrated that MgIG protects against ConA-induced immunological liver injury by markedly alleviating liver inflammation, and this provides guidance for the clinical amelioration of liver inflammation induced by immunological factors.

Role of intestinal barrier in the pathogenesis of autoimmune hepatitis

Objective To observe and analyze the role of intestinal barrier in the pathognesis of autoimmune hepatitis (AIH), to explain the pathogenesis of AIH and to explore the intestinal based new treatment strategies. Methods A total of 14 AIH patients from January to December 2017 at Tianjin Medical University General Hospital (six patients without liver cirrhosis, and eight patients with liver cirrhosis) and 10 healthy controls were enrolled. The serum levels of D-lactic acid (D-Lac) and diamine oxidase (DAO) were detected by enzyme-linked immunosorbent assay. Real time fluorescence quantitative polymerase chain reaction was used to detect the relative expression levels of connexin (zonula occluden-1 (ZO-1), occludin), cytokines (interleukin(IL)-2, interferon(IFN)-γ, IL-4, IL-10) and Toll-like receptor 4 (TLR4) in terminal ileal tissues of each group. The relative expression of secretory immunoglobulin A (sIgA) in the terminal ileum was determined by Western blotting. Thirty BALB/c mice were selected and divided into blank control group, dextran sulfate sodium (DSS) group, concanavalin A (ConA) group, DSS+ ConA group, and DSS+ bacterium+ ConA group, with six mice in each group. The relative expression levels of ZO-1, occludin in mouse colonic tissues, serum alanine aminotransferase (ALT) and aspartate aminotransferase (AST) levels, and inflammatory activity degree of liver tissues (Knodell score) of each group were measured. T-test and one-way analysis of variance were performed for statistical analysis. Results The serum D-Lac and DAO levels of AIH with liver cirrhosis group and AIH without liver cirrhosis group were both higher than those of healthy control group ((1 768.2±147.1) μg/L, (436.2±197.0) μg/L vs. (100.2±10.9) μg/L, and (11.5±2.5) U/L, (5.4±0.9) U/mL vs. (3.5±0.9) U/mL), and the levels of D-Lac and DAO of AIH with liver cirrhosis group were the highest; and the differences were statistically significant (t=5.512, 36.010, 4.088 and 9.443, F=396.958 and 46.640, all P<0.01). The relative expression levels of ZO-1 and occludin in the terminal ileal mucosa of AIH with liver cirrhosis group were lower than those of healthy control group (0.20±0.14 vs. 1.67±0.51, 0.12±0.09 vs. 0.90±0.21), and the relative expression of ZO-1 in AIH without liver cirrhosis group was lower than that in healthy control group (0.99±0.37 vs. 1.67±0.51); and the differences were statistically significant (t=8.641, 7.407 and 2.295, all P<0.05). The relative expression levels of IL-2 and IFN-γ in terminal ileal tissues of AIH with liver cirrhosis group were higher than those of healthy control group (1.11±0.43 vs. 0.24 ±0.16, and 3.50 ± 1.90 vs. 0.32±0.30), however the relative expression of sIgA in terminal ileal tissues was lower than that of healthy control group (0.506±0.024 vs. 1.081±0.102); and the differences were statistically significant (t=4.679, 3.981 and 5.493, all P<0.05). While the relative expression levels of IL-10 in AIH with liver cirrhosis group and AIH without liver cirrhosis group were lower than that in healthy control group (0.30±0.20, 0.42±0.24 vs. 0.84± 0.23), and the relative expression levels of TLR4 in ileum mucosa of the both groups were higher than that of healthy control group (8.74 ±5.13, 6.74 ±3.65 vs. 0.89 ± 0.70); and the differences were statistically significant (t=3.095, 4.816, 3.856 and 3.685, all P<0.05). The relative expression levels of ZO-1 and occludin of DSS+ ConA group were lower than those of ConA group (0.14±0.08 vs. 0.98±0.13, and 0.09±0.02 vs. 0.98±0.16), however serum ALT, AST levels and the Knodell score were all higher than those of ConA group ((5 496.67±618.83) U/L vs. (3 325.00±1 030.06) U/L, (8 825.00±1 165.35) U/L vs. (5 433.33±1 691.14) U/L, and 18.00±2.00 vs. 9.33±3.01); and the differences were statistically significant (t=13.480, 13.520, 4.227, 4.045 and -2.892, all P<0.05). The relative expression levels of ZO-1 and occludin in DSS+ bacterium+ ConA group were higher than those in DSS+ ConA group (0.46±0.08 vs. 0.14±0.08, and 0.53±0.15 vs. 0.09±0.02), while serum ALT and AST levels were lower than those of DSS+ ConA group ((4 343.33±252.16) U/L vs. (5 496.67±618.83) U/L, and (6 123.33±1 086.60) U/L vs. (8 825.00±1 165.35) U/L); and the differences were statistically significant (t=6.928, 7.122, 4.228 and 4.153, all P<0.01). Conclusions AIH patients have increased intestinal permeability and impaired intestinal barrier which is more serious in patients with liver cirrhosis than in patients without cirrhosis. The intestinal barrier injury can aggravate ConA-induced immune-mediated liver injury. While the protection and repair of intestinal barrier can alleviate immune-mediated liver injury induced by ConA. Key words: Hepatitis, autoimmune; Concanavalin A; Intestinal mucosal barrier; Dextran sulfate sodium; Bifidobacterium

Methylprednisolone Decreases Mitochondria-Mediated Apoptosis and Autophagy Dysfunction in Hepatocytes of Experimental Autoimmune Hepatitis Model via the Akt/mTOR Signaling.

Autoimmune hepatitis (AIH) is characterized by massive immune cell-mediated hepatocyte destruction. Glucocorticoids, particularly methylprednisolone (MP), are the most effective treatment for AIH; however, the mechanism underlying the effects of glucocorticoid treatment has not been fully elucidated. The present study explored the effects of MP on damaged hepatocytes in mice with concanavalin A (ConA)−induced experimental autoimmune hepatitis (EAH). C57BL/6 mice were divided into three groups: a normal control group (injected with normal saline), a ConA (20 mg/kg) group, and a ConA + MP (3.12 mg/kg) group. The serum levels of liver enzymes, cytokines, activated T cells, and apoptosis− and autophagy−associated marker proteins were determined 12 h after ConA injection. Human hepatocyte cell line LO2 was used to verify the effects of ConA and MP in vitro. MP treatment significantly decreased inflammatory reactions in the serum and liver tissues and activated the Akt/mTOR signaling pathway to inhibit apoptosis and autophagy in hepatocytes in vivo. Transmission electron microscopy (TEM) revealed fewer autophagosomes in the MP-treated group than in the ConA-treated group. MP treatment obviously suppressed apoptosis and mitochondrial membrane potential (ΔΨm) loss in hepatocytes in vitro. Furthermore, ConA treatment increased the levels of LC3-II, p62/SQSTM1, and Beclin-1, while bafilomycin A1 did not augment the levels of LC3-II. MP treatment decreased the levels of LC3-II, p62/SQSTM1, and Beclin-1 and upregulated the levels of phosphorylated (p)-Akt and p-mTOR. In conclusion, MP ameliorated mitochondria-mediated apoptosis and autophagy dysfunction in ConA-induced hepatocyte injury in vivo and in vitro via the Akt/mTOR signaling pathway.

Retinoic acid modulates IL-4, IL-10 and MCP-1 pathways in immune mediated hepatitis and interrupts CD4+ T cells infiltration

AimsImmune mediated liver injury includes activation of different immune pathways that requires various modalities to control their consequences. The current study involves evaluation of retinoic acid (RA) modulatory effects on immune responses induced in concanavalin A (ConA) model of acute hepatitis. Main methodsMice were divided as follows: Control group; RA group: received 35 mg/kg RA; ConA group: received 15 mg/kg ConA; ConA + RA group: received ConA and RA as described. Liver function biomarkers were measured in addition to malondialdehyde as lipid peroxidation biomarker. Liver tissue sections were scored for necro-inflammation, neutrophils infiltration, CD4+ T cells infiltration and NF-κb positive cells. Effect on hepatic levels of TNF-α, IL-4, IL-10 and MCP-1 was evaluated as well. Key findingsInjection of RA before ConA significantly (p < 0.001) decreased ALT, AST and LDH levels compared to their levels in ConA group. Hepatic infiltration of neutrophils and CD4+ T cells was markedly (p < 0.001) reduced by RA. Hepatic injury, necrosis and expression of NF-κb were significantly decreased by RA when injected before ConA challenge. A significant decrease in the measured cytokines TNF-α and IL-4 was observed in ConA + RA group in addition to a decrease in MCP-1 level. On the other hand, IL-10 was significantly increased in the latter group compared to ConA group. SignificanceRA can protect against ConA-induced hepatitis through: interrupting early inflammatory response as neutrophils, monocytes and CD4+ T cells infiltration, modulating IL-4 level and subsequent production of TNF-α and NF-κb activation, mitigating second inflammatory responses through increasing IL-10 liver production.

The potential antifibrotic impact of apocynin and alpha-lipoic acid in concanavalin A-induced liver fibrosis in rats: Role of NADPH oxidases 1 and 4.

Liver fibrosis results from chronic inflammation that precipitates excessive accumulation of extracellular matrix. Oxidative stress is involved in its pathogenesis. This study aimed to elucidate the potential antifibrotic effect of the NADPH oxidase (NOX) inhibitor, apocynin against concanavalin A (ConA)-induced immunological model of liver fibrosis, and to investigate the ability of the antioxidant, alpha-lipoic acid (α-LA) to potentiate this effect. Rats were treated with apocynin and/or α-LA for six weeks. Hepatotoxicity indices, oxidative stress, insulin, NOXs, inflammatory and liver fibrosis markers were assessed. Treatment of animals with apocynin and α-LA significantly ameliorated the changes in liver functions and histopathological architecture induced by ConA. Liver fibrosis induced by ConA was evident where alpha-smooth muscle actin and transforming growth factor- beta1 were elevated, which was further confirmed by Masson's trichrome stain and increased hydroxyproline. Co-treatment with apocynin and α-LA significantly reduced their expression. Besides, apocynin and α-LA significantly ameliorated oxidative stress injury evoked by ConA, as evidenced by enhancing reduced glutathione content, antioxidant enzymes activities and decreasing lipid peroxides. ConA induced a significant elevation in serum insulin level and inflammatory markers; tumor necrosis factor-alpha, interleukin-6 and nuclear factor kappa b. Furthermore, the mRNA tissue expression of NOXs 1 and 4 was found to be elevated in the ConA group. All these elevations were significantly reduced by apocynin and α-LA co-treatment. These findings indicate that using apocynin and α-LA in combination possess marked antifibrotic effects, and that NOX enzymes are partially involved in the pathogenesis of ConA-induced liver fibrosis.

Effect of G-CSF in vitro Stimulation on Distribution of Peripheral Lymphocyte Subsets in the Healthy Persons

To investigate the effect of granulocyte-colony stimulating factor (G-CSF) in vitro stimulation on the distribution of lymphocyte subset in healthy human. Peripheral blood mononuclear cells (PBMNCs) were collected from 8 healthy volunteers by density gradient centrifugation on Ficoll-PaqueTM. In vitro 200 ng/ml G-CSF or 200 ng/ml G-CSF plus 10 µg/ml ConA directly act on PBMNCs, then the colleted cells were cultivated for 3 days. Lymphocyte subsets were stained with the corresponding fluoresce labeled antibodies and detected by flow cytometry. The levels of T cells in G-CSF group and G-CSF+ConA group were both higher than that in the control group (P<0.001, P<0.05). However, there were not significantly different in B cells and NK cells levels among the 3 groups. Furthermore, analysis of the effect of G-CSF on T cell subsets indicated that the levels of CD4+T cells and CD8+T cells in G-CSF group were both significantly higher than those in control group (P<0.01, P<0.05), Treg cells was not different between G-CSF and control group. Compared with the control group, the level of CD4+T cells, CD8+T cells and Treg cells in G-CSF+ConA group significantly increased (P<0.05, P<0.01, P<0.01). Analysis of G-CSF receptor (G-CSFR) expression showed that G-CSFR expression on T cells in G-CSF+ConA group dramatically increased, as compared with control group (P<0.01). The levels of CD4+T cells and CD8+T cells in healthy human peripheral blood can be increased by G-CSF stimulation. ConA can enhance the level of T cells and induce G-CSFR expression on T cells.

Protective effect of intraperitoneal transplantation of human liver-derived stem cells at different times against concanavalin A-induced acute liver injury in mice

Objective: To investigate the protective effect of intraperitoneal transplantation of human liver-derived stem cells at different times against concanavalin A (ConA)-induced acute liver injury in mice. Methods: A total of 88 male C57BL/6 mice were randomly divided into normal control group (group C), ConA model group (group M), and human liver-derived stem cells (HYX1)+ConA group (group E); according to the interval between phosphate buffer/HYX1 injection and ConA injection, Groups M and E were further divided into 3-hour groups (M1 and E1 groups), 6-hour groups (M2 and E2 groups), 12-hour groups (M3 and E3 groups), 24-hour groups (M4 and E4 groups), and 48-hour groups (M5 and E5 groups). The levels of alanine aminotransferase (ALT), aspartate transaminase (AST), and total bilirubin (TBil) in peripheral blood were measured, liver tissue sections were used to observe pathological changes, and the Ishak score for liver inflammation was determined. The independent samples t-test was used for comparison between groups, and P < 0.05 was considered statistically significant. Results: The levels of ALT, AST, and TBil in group C were (36.25±1.16) U/L, (120.20±5.77) U/L, and (2.20±0.23) μmol/L, respectively; the levels of ALT, AST, and TBil and Ishak score were (8 721.23±837.39) U/L, (8 110.31±290.10) U/L, (8.41±0.10) μmol/L, and (13.32±1.30), respectively, in group M1, (8 334.31±666.50) U/L, (7 560.20±760.34) U/L, (10.40±0.80) μmol/L, and (12.67±0.81), respectively, in group M2, (8 960.75±551.93) U/L, (8 535.62±675.14) U/L, (10.95±1.43) μmol/L, and (14.57±0.65), respectively, in group M3, (8 618.57±886.40) U/L, (11 440.54 ± 1 327.86) U/L, (13.30±1.86) μmol/L, and (13.21±1.06), respectively, in group M4, and (10 170.13±1 112.37) U/L, (11 470.56±1 108.40) U/L, (12.75±1.55) μmol/L, and (15.07±1.58), respectively, in group M5. The levels of ALT, AST, and TBil and Ishak score were (1 016.35±163.47) U/L, (952.30±103.91) U/L, (7.77±0.62) μmol/L, and (3.50±0.21), respectively, in group E1, (42.10±6.20) U/L, (126.72±13.33) U/L, (3.41±0.53) μmol/L, and (2.01±0.40), respectively, in group E2, (44.21±4.30) U/L, (216.71±35.88) U/L, (3.47±0.44) μmol/L, and (2.13±0.25), respectively, in group E3, (2 909.69±212.14) U/L, (2 988.43±333.70) U/L, (7.03±0.93) μmol/L, and (4.70±0.50), respectively, in group E4, and (7 874.26±799.60) U/L, (10 940.54±947.35) U/L, (10.53±1.09) μmol/L, and (8.60±0.83), respectively, in group E5. Groups M1-M5 had significantly higher levels of ALT, AST, and TBil than group C (all P < 0.01), and groups M1-M4 had significantly higher levels of AST and ALT than groups E1-E4 (all P < 0.01), while there were no significant differences in the levels of AST and ALT between groups M5 and E5 (both P > 0.05). The pathological sections of liver tissue showed that compared with group M, group E had significant reductions in the degree of necrosis and Ishak score (both P < 0.05). Conclusion: Intraperitoneal transplantation of human liver-derived stem cells has a protective effect against ConA-induced acute liver injury in mice, and the injection at 6 and 12 hours in advance has the best protective effect.

Effects of long noncoding RNA TUG1 on migration and invasion of osteosarcoma cells

Objective To study the effects of TUG1 on migration and invasion of osteosarcoma cells. Methods The osteosarcoma Saos-2 cells were classified into three groups: TUG1 small interfering RNA (siRNA group), blank control group (Con-A), and the negative control-siRNA group (Con-B). The transfection reagent oligofectamine was mixed with TUG1 siRNA and nonsense siRNA, respectively. The TUG1 knocking down effects were examined by real-time polymerase chain reaction (PCR). The migration of osteosarcoma cells was determined by wound healing test, and the invasion ability was studied by Transwell assay. Results As compared with Con-A and Con-B groups, the expression level of TUG1 mRNA was decreased in siRNA group. After osteosarcoma cells were transfected with TUG1 siRNA, the results of wound healing test showed that the space between scratches of Con-A, Con-B and siRNA groups was (326±11) μm, (335±15) μm, and (588±11) μm respectively (F=22.761, P=0.000). The results of the Transwell showed that the membrane cell number in Con-A, Con-B and siRNA groups was (125.0±2.2), (118.0±1.8), and (56.0±1.8) cells respectively (F=173.000, P=0.000). Conclusion TUG1 might be associated with migration and invasion of osteosarcoma cells. Key words: Osteogenic sarcoma; Long noncoding RNA; TUG1; Migration; Invasion

Effects of upregulatioon of Forkhead box protein M1 on migration, invasion and epithelial-mesenchymal transition of human ovarian cancer cell

Objective To explore the effects and mechanism of over-expression of Forkhead box protein M1 (FoxM1) gene on the migration, and invasion and epithelial-mesenchymal transition (EMT) of human ovarian cancer cells. Methods FoxM1 over-expression of the virus vector was constructed. All HO-8910 cells were divided into three groups, the control group (Con-A), Lentiviral vector(LV) control group (Con-B) and over-expression group FoxM1. After three group HO-8910 cells was transfected with phosphate buffer (PBS), Lentiviral vector (LV), and LV-FoxM1, the protein levels of FoxM1 were determined by Western blotting, respectively. The migration and invasion ability was evaluated by wound healing assay and Transwell assay, respectively. The E-Cadherin and N-Cadherin of protein of cancer cells was examined by Western blotting assay. Results The results from Western blotting assay showed that the levels protein of FoxM1 were greatly increased in HO-8910 cancer cells transfected with LV-FoxM1. The results of the wound healing test showed that the migration distance of Con-A, Con-B, and LV-FoxM1 group was(156.80±2.26), (161.60±1.81) and (278.60±2.62) μm, respectively (P<0.01); The results of the Transwell invasion assay showed that the membrane cell number of Con-A, Con-B, and LV-FoxM1 group was (68.56±1.69) and (67.27±1.36) and (126.38±1.56), respectively (P<0.01); Compared with the Con-A, there were E-cadherin protein expression decreased; however, N-cadherin protein increased in HO-8910 cancer cells transfected with LV-FoxM1. Conclusion FoxM1 overexpression can promote human ovarian cancer cell migration and invasion, and its mechanism may be related to the promotion of epithelial-mesenchymal transition. Key words: Ovarian carcinoma; Forkhead box protein M1; Migration; Invasion; Epithelial-mesenchymal transition

Anti-hepatitis, antioxidant activities and bioactive compounds of Dracocephalum heterophyllum extracts.

Background Dracocephalum heterophyllum was a traditional Tibetan medicine possesses various pharmacological effects involved in anti-inflammatory, antibacterial activities. However, its anti-hepatitis, antioxidant activity and bioactive compounds have not been reported, the objective of this research work was to investigate the pharmacological activity and bioactive compounds of D. heterophyllum extracts.ResultsIn the present study, the anti-hepatics and antioxidant activities of four D. heterophyllum extracts (i.e. petroleum ether extracts, ethyl acetate extracts, n-BuOH extracts, and water extracts) were conducted. The main chemical constituent of petroleum ether and ethyl acetate extracts were also isolated using chromatographic techniques and identified by NMR spectroscopic methods. The anti-hepatitis assay showed that the petroleum ether and ethyl acetate extracts of D. heterophyllum significantly prolonged the mean survival times and reduced the mortality of mouse hepatitis model induced by concanavalin A (ConA). The levels of alanine transaminase, aspartate transaminase in blood serum could be decreased obviously by ethyl acetate extracts compared with ConA group (P < 0.01). The histological analysis demonstrated that the ethyl acetate extracts could inhibit apoptosis and necrosis caused by ConA. In addition, the antioxidant activities of the four extracts of D. heterophyllum were measured by DPPH assay, ABTS assay, anti-lipidperoxidation assay, ferric reducing antioxidant power assay, ferrous metal ions chelating assay and determination of total phenolic contents. The results showed that the ethyl acetate extract had the highest antioxidant activities, followed by petroleum ether extract. Finally, nine mainly compounds were isolated from the Petroleum ether and ethyl acetate extracts, including four triterpenes: oleanolic acid (1), ursolic acid (2), pomolic acid (3), 2α- hydroxyl ursolic acid (4), three flavonoids: apigenin-7-O-rutinoside (5), luteolin (8), diosmetin (9) and two phenolic acids: rosmarinic acid (6), methyl rosmarinate (7).ConclusionThe Ethyl acetate extract of D. heterophyllum had the highest anti-hepatitis and antioxidants activities, followed by petroleum ether extract. The bioactive substances may be triterpenes, flavonoids and phenolic acids, the ethyl acetate extracts of D. heterophyllum may be possible candidates in developing anti-hepatitis medicine.Electronic supplementary materialThe online version of this article (doi:10.1186/s40529-016-0133-y) contains supplementary material, which is available to authorized users.

MRI-based assessment of liver perfusion and hepatocyte injury in the murine model of acute hepatitis.

ObjectiveTo assess alterations in perfusion and liver function in the concanavalin A (ConA)-induced mouse model of acute liver failure (ALF) using two magnetic resonance imaging (MRI)-based methods: dynamic contrast-enhanced MRI (DCE-MRI) with Gd-EOB-DTPA contrast agent and arterial spin labelling (ASL).Materials and methodsBALB/c mice were studied using a 9.4 T MRI system. The IntraGateFLASHTM and FAIR-EPI pulse sequences were used for optimum mouse abdomen imaging.ResultsThe average perfusion values for the liver of the control and ConA group were equal to 245 ± 20 and 200 ± 32 ml/min/100 g (p = 0.008, respectively). DCE-MRI showed that the time to the peak of the image enhancement was 6.14 ± 1.07 min and 9.72 ± 1.69 min in the control and ConA group (p < 0.001, respectively), while the rate of the contrast wash-out in the control and ConA group was 0.037 ± 0.008 and 0.021 ± 0.008 min−1 (p = 0.004, respectively). These results were consistent with hepatocyte injury in the ConA-treated mice as confirmed by histopathological staining.ConclusionsBoth the ASL and DCE-MRI techniques represent a reliable methodology to assess alterations in liver perfusion and hepatocyte integrity in murine hepatitis.

Effects of upregulatioon of trophoblast cell-surface antigens 2 on migration, invasion and epithelial-mesenchymaltransition of human prostate cancer cells

Objective To explore the effects of over-expression of trophoblast cell-surface antigens 2(TROP-2)gene on the migration and invasion of human prostate cancer cells and the action mechanism. Methods The TROP-2 gene was cloned into the eukaryotic expression vector pcDNA3. 1, and TROP-2 over-expression of eukaryotic gene expression plasmid was constructed. All PC-3 cells were divided into three groups: the control group(Con-A), empty vector control group(Con-B)and over-expression group TROP-2(pcDNA3. 1-TROP-2).After PC-3 cells in three groups were transfected with phosphate buffer(PBS), vector pcDNA3. 1, or pcDNA3. 1-TROP -2, the protein levels of TROP-2 were determined by Western blotting, respectively.The migration and invasion ability was evaluated by Transwell.The E-Cadherin and N-Cadherin protein expression in PC-3 cells was examined by Western blotting. Results TROP-2 gene eukaryotic expression vector was successfully constructed. The Western blotting showed that the expression levels of TROP-2 protein were greatly increased in PC-3 cancer cells transfected with pcDNA3. 1-TROP -2.The Transwell migration assay revealed that the number of migrating cells in Con-A,Con-B, and pcDNA3. 1-TROP-2 groups was 132. 6±2. 2, 130. 8±1. 8 and 189. 6±2. 6, respectively (P< 0. 01).The Transwell invasion assay indicated that the number of migrating cells in Con-A, Con-B, and pcDNA3. 1-TROP-2 groups was 118. 16±1. 96, 117. 52±1. 85 and 166. 38±1. 65, respectively(P< 0. 01).As Compared with the Con-A group, the E-cadherin protein expression was decreased,and the N-cadherin protein expression was increased in PC-3 cancer cells transfected with pcDNA3. 1-TROP-2. Conclusion TROP-2 overexpression can promote migration and invasion of human prostate cancer cells probably promoting the epithelial-mesenchymal transition. Key words: Trophoblast cell-surface antigens 2; Prostate carcinoma; Invasion; Epithelialmesenchymal transition

Natural killer T cells play a necessary role in modulating of immune-mediated liver injury by gut microbiota.

Gut microbiota are implicated in many liver diseases. Concanavalin A (ConA)-induced hepatitis is a well-characterized murine model of fulminant immunological hepatic injury. Oral administration of pathogenic bacteria or gentamycin to the mice before ConA injection, liver injury and lymphocyte distribution in liver and intestine were assessed. Our data show that administration of pathogenic bacteria exacerbated the liver damage. There was more downregulation of activation-induced natural killer T (NKT) cells in the liver of pathogenic bacteria-treated ConA groups. Also, there was a negative correlation between the numbers of hepatic NKT cells and liver injury in our experiments. Moreover, intestinal dendritic cells (DCs) were increased in pathogenic bacteria–treated ConA groups. The activation of DCs in Peyer's patches and the liver was similar to the intestine. However, depletion of gut gram-negative bacteria alleviated ConA-induced liver injury, through suppressed hepatic NKT cells activation and DCs homing in liver and intestine. In vitro experiments revealed that DCs promoted NKT cell cytotoxicity against hepatocyte following stimulation with pathogenic bacteria. Our study suggests that increased intestinal pathogenic bacteria facilitate immune-mediated liver injury, which may be due to the activation of NKT cells that mediated by intestinal bacterial antigens activated DCs.

Effects of forkhead box protein M1 down-regulation by RNA interfence on growth,and don and invasion of human ovarian cancer cell

Objective To study the effects of RNA interference forkhead box protein M1 (FoxM1) on growth,and clon formation,and invasion of huaman ovarian cancer cell.Methods Using checks The FoxM1 gene mRNA levels were exmined by real-time quantitative polymerase chain reaction (Real-time PCR) method of eight human ovarian cancer cell lines.FoxM1 small interfering RNA (siRNA)were built and synthesized by using chemical methods.The cells were classified into three groups:blank control group (Con-A),the empty plasmid group (Con-B) and FoxM1 siRNA group (siRNA).Among them,the siRNA group was transfected with FoxM1 siRNA.was used to detect cancer cells FoxM1 mRNA and protein levels in each group were detected by Real-time PCR and Western blotting,respectively.The cell viability of each group was measured by cell counting kit-8 (CCK-8) assay.The clones were evaluated by colony formation assay.The invasion ability was evaluated by Transwell method.Results The mRNA levels of FoxM1 gene is highest in the SKOV-3 and HO-8910PM cell lines.After SKOV-3 cell line was transfected by FoxM1 siRNA,the results of the CCK-8 assay showed that the A Values of Con-A,Con-B and FoxM1 group in 72 h were 2.455 ± 0.033,2.442 ± 0.025 and 1.312 ± 0.028,respectively (P ＜ 0.05) ; the results of colony formation assay showed that clones of Con-A,Con-B and FoxM1 group were 100 ±5,93 ±8 and 51 ±3,respectively (P＜0.05) ; the results of the Transwell showed that the membrane cell number of Con-A,Con-B and FoxM1 group were 93 ± 4,86 ± 3 and 36 ± 1,respectively (P ＜0.05).After HO-8910PM cell line was transfected by FoxM1 siRNA,the results of the CCK-8 assay showed that the A Values of Con-A,Con-B and FoxM1 group in 72 h were 2.691 ±0.039,2.560 ±0.033 and 1.455 ± 0.027,respectively (P ＜ 0.05) ; the results of colony formation assay showed that clones of Con-A,Con-B and FoxM1 group were 146 ± 7,145 ± 8 and 85 ± 2,respectively (P ＜ 0.05) ; the results of the Transwell showed that the membrane cell number of Con-A,Con-B and FoxM1 group were 449 ± 15,445 ± 11 and 151 ± 9,respectively (P ＜ 0.05).Conclusion FoxM1 gene might play an important role in growth,proliferation and invasion of human ovarian cancer,and FoxM1 gene may be an important target in human ovarian cancer. Key words: Ovarian cancer; Forkhead box protein M1; Proliferation; Invasion; RNA interfence

Impacts of pseudogene Cripto-3 down-regulationion by RNA interference on proliferation and invasion of human colon cancer cells

Objective To study the effects of RNA interference pseudogene Cripto-3 (Cr-3) on proliferation and invasion of colon cancer cells.Methods Cr-3 small interfering RNAs (siRNA,S1,S2 and S3) was built and synthesized by using chemical methods.The SW620 cells were classified into three groups:blank control group (con-a),the empty plasmid group (con-b) and Cr-3 siRNA group (siRNA).Among them,the siRNA group was transfected with Cr-3 siRNA.The cell viability of each group was measured by methyl thiazol tetrazolium (MTT) assay.The invasion ability was evaluated by transwell method.All the above were for statistical analysis.Results S1,S2 and S3 could downregulate Cr-3 mRNA with the inhibition rate of 83.5％,61.6％ and 42.8％ respectively.After SW620 cells were transfected by S1,MTT assay revealed that the relative proliferation rate in con-a,con-b and Cr-3 siRNA groups at72 h was (0.927 ±0.037)％,(0.922 ±0.035)％ and (0.663 ±0.018)％ (A) respectively (P＜0.05).Transwell showed that the number of cells penetrating the membrane in con-a,con-b and Cr-3 siR-NA group was 38 ± 4,40 ± 3 and 8 ± 1 respectively (P ＜ 0.05).Conclusion Cr-3 gene might play an important role in proliferation and invasion of human colon cancer cells,and down-regulation of Cr-3 by siRNA could inhibit invasion ability and proliferation of human colon cancer cells. Key words: Colon carcinoma; Invasion ; RNA interference