Objective To study the effect of p38MAPK on the activity and c-myc protein expression in rat acetaldehyde-induced hepatic stellate cell(HSC),and to investigate the alcoholic liver fibrosis related mechanism.Methods The different concentrations of SB203580 as the p38 specific blocker was adopted to conduct the intervention on rat acetaldehyde-induced HSC.The cellular mor-phological change was observed by the microscope.The cell proliferation was detected by MTT,the cell cycle was analyzed by flow cytometry(FCM),and the expression of c-myc protein was examined by the SABC method.Results (1)after acetaldehyde stimula-tion,HSC was increased in size and proliferated rapidly,but with the added SB203580 concentration increase,the cellular prolifera-tion was slowed down,the cells size was diminished and the deformed cells were increased.(2)The proliferation of acetaldehyde-in-duced HSC was inhibited by different doses of SB203580,and the higher concentration has the more significant inhibiting effect.(3) With the SB203580 concentration increase,the cells at the phase G0 and G1 were increased,while the cells at the phase S were de-creased,at the same time the expression positive rate of c-myc protein was decreased.Conclusion Blocking p38MAPK pathway ac-tivity could inhibit the proliferation of acetaldehyde-induced HSC,which may be related to the down-regulation of C-myc protein ex-pression and blocking the DNA synthesis in cells entering from G0/G1 phase to S phase.

Objective To report a peliosis hepatic in child and review literature and discuss.Methods Case history was inquired.Physical,labtoratory,imagement and histopathology of liver biopsy(HE staining) were examed.Results A 4-year old girl appeared dermatitis with erythema and herpes at local skin where was bit by insect before onset.The girl appeared fever,cough,then abdominal pain,hepatomegaly,pleural effusion and ascites.Lab examination revealed slight elevation of aspartate transaminase,?-glutamyltranspeptidase and alkaline phosphatase.The liver B-mode ultrasonography and CT scan revealed hepatomegaly with density heterogeneity of the parenchyma.The liver biopsy revealed many small capsule filled with blood cells.Conclusions Clinical characteristics of the disease are fever,upper abdomen pain,janundice,ascites and hepatomegaly.The diagnosis shall be combined with the pathologic biopsy of liver.

Objective To evaluate the effects of compound glycyrrhizin on the percentage of Th17 cells and expression of interleukin(IL)-17A in peripheral blood of patients with psoriasis vulgaris, and to explore their relationship with therapeutic effects. Methods A total of 30 patients with mild to moderate progressive psoriasis vulgaris were randomly and equally divided into two groups:group 1 treated with compound glycyrrhizin injection, antihistamines and topical drugs, group 2 treated with antihistamines and topical drugs. Twelve healthy human subjects served as the control group. Blood samples were collected from the patients 1 day before start of treatment and after 3 weeks of treatment, and from the control group. Flow cytometry and enzyme-linked immunosorbent assay (ELISA)were performed to determine the percentage of Th17 cells and expression level of IL-17A respectively in the peripheral blood samples. Results Before the treatment, the percentage of Th17 cells and expression level of IL-17A were both significantly higher in the two patient groups than in the control group (all P<0.05). After the treatment, group 1 showed significant decreases in the percentage of Th17 cells and expression level of IL-17A compared with those before the treatment (both P<0.05), while no significant differences were observed in group 2 between pre-and posttreatment IL-17A expression level or Th17 cell percentage (both P>0.05). Furthermore, both Th17 cell percentage and IL-17A expression were significantly different between the two patient groups after the treatment (both P< 0.05). Conclusion Compound glycyrrhizin may treat psoriasis vulgaris by regulating Th17 cell percentage and IL-17A expression in peripheral blood.

Objective To study the alterations of CD19+CD5+CD1dhigh B, Th1 and Th17 cells in non-obese diabetic ( NOD) mice and the correlation between B10 cells and type 1 diabetes in NOD mice. Methods Flow cytometry ( FCM) was used to measure the levels of CD19+CD5+CD1dhigh B, CD19+IL-10+B, CD4+IFN-γ+Th1, CD4+IL-17+Th17 and CD4+CD25+Foxp3+T cells in NOD mice ( 4 weeks old NOD mice:group A, n=10;8 weeks old NOD mice:group B, n=10; NOD mice with diabetes: group C, n=10) and age-matched C57BL/6 mice ( control group, n=20 ) .Hematoxylin-eosin staining of pancreatic tissues was performed for histopathological assessment of the development of insulitis in NOD mice.Results (1) Histopathological analysis showed that mice from A, B and C groups respectively showed no insulitis, insulitis and obvious insulitis with no intact islets.(2) The highest levels of B10 cells in NOD mice were ob-served in group B, followed by those in group C and group A (P<0.01).No significant differences with the levels of B10 cells were found among different tissues of 4 weeks old NOD mice (P>0.05).More B10 cells were detected in pancreatic lymph nodes than in other tissues of 8 weeks old NOD mice, the levels of which were also higher than those in pancreatic lymph nodes of mice form group C ( P<0.01) .The highest levels of B10 cells were detected in peripheral lymph nodes among all tissues samples collected from NOD mice with diabetes (P<0.01).(3) The levels of Th1 and Th17 cells in mice from group C were remarkably in-creased as compared with those in mice from group A and B (P<0.01).(4) The percentages of CD4+CD25+Foxp3+T cells in mice from group C showed no differences with those in mice from A and B groups. No significant difference with Treg cells were observed between NOD mice and age-matched C57BL/6 mice (P>0.05).Conclusion The percentages and distribution of B10 cells in NOD mice changed with age and the development of insulitis.The decrease of B10 cells might participate in the development of type 1 diabe-tes in NOD mice.

Objective Bevacizumab ( BM ) is an angiogenesis inhibitor widely used in cancer therapy, but its off-target effect of proteinuria may lead to discontinuation of treatment.This study was to explore the mechanisms of BM inducing proteinuria in mice. Methods Twenty-four healthy mice were randomly divided into four groups, saline control, low-dose BM, medium-dose BM, and high-dose BM, treated by injection of normal saline and BM at 10, 35, and 60 mg per kg of the body weight, respectively, though the tail vein.At 4 weeks after injection, 24-hour urine was collected to determine the total urine protein and blood obtained from the eyeballs for biochemical analysis.Then all the mice were sacrificed and the kidneys harvested for observation of pathologic changes in the glomeruli as well as for immunohistochemistry, Western blotting, and real-time PCR analysis. Results Compared with normal saline,BM obviously elevated the level of 24-hour urine protein, with statistically significant differences between the control and the medium-and high-dose BM groups (0.23 ±0.02 vs 1.14 ±0.13 and 1.43 ±0.10, P<0.01), but not between the control and the low-dose BM (0.23 ±0.02 vs 0.29 ±0.07, P>0.05).No significant differences were observed among the four groups in the levels of Cr, BUN, AST and ALT (P>0.05).Under the optical microscope, the kidneys showed normal structures in the control group, no signifi-cant pathologic changes in the low-dose BM, and vacuolus-like alteration with atrophic glomerular endothelial cells in the medium-and high-dose BM groups.Immunohistochemical analysis demonstrated the expressions of VEGF and podocin were moderately or strongly positive in the control and low-dose BM groups, by weakly positive or negative in the medium-and high-dose BM groups.Compared with the control group, the expression of the VEGF protein in the renal tissue was significantly decreased in the high-dose BM group (0.76 ±0.09 vs 0.39 ±0.05, P<0.01) but had no remarkable difference from that in the low-dose (0.81 ±0.10) or medium-dose BM (0.64 ±0.08) group (P>0.05), and the expression of the podocin protein was significantly reduced in the medium-dose BM (0.67 ±0.07 vs 0.43 ±0.10, P<0.05) and high-dose BM (0.67 ±0.07 vs 0.19 ±0.04, P<0.01), but not in the low-dose BM group (0.67 ±0.03) (P>0.05).The mRNA expressions of VEGF and podocin were not significantly changed in the low-dose BM group as compared with the control (1.07 ±0.61 and 1.12 ±0.09 vs 1.23 ±0.25 and 1.17 ±0.19, P>0.05) but remarkably de-creased in the medium-dose (0.82 ±0.38 and 0.71 ±0.18) and high-dose BM groups and (0.47 ±0.64 and 0.42 ±0.09) groups (P<0.01). Conclusion Bevacizumab damages glomerular filtration membrane and induce proteinuria partially by down-regulating the protein and mRNA expressions of VEGF and podocin.

Objective To investigate the phenotypes and percentages of B 10 cells in different tis-sues from wild-type mice and to identify their biological functions .Methods The percentages of B10 cells derived from different tissues of mice and their responses to lipopolysaccharide ( LPS) stimulation were ana-lyzed by flow cytometry .Magnetic-activated cell sorting ( MACS ) and fluorescence-activated cell sorting (FACS) were used to purify B10 cells, CD4+CD25-T cells and Treg cells.CD4+CD25-T cells and Treg cells labeled by CFSE were co-cultured with or without B10 cells, and then their proliferation were evaluated after 72 h.Results (1) A subset of CD19+CD5+CD1dhigh regulatory B cells was identified in spleen , pe-ripheral blood and lymph nodes from wild-type mice , of which the highest frequency was detected in spleen (3.95%±0.79%, P<0.05).The isolated B cells from different tissues were stimulated by LPS , PMA, ionomycin and monensin (L+PIM) in vitro to express IL-10.Among them, splenic CD19+IL-10+B cells showed the highest expression of IL-10 (P<0.05).(2) Prolonged LPS stimulation (48 h) to CD5+CD1dhigh B cells enhanced the expressions of IL-10 (P<0.01).(3) CD19+CD5+CD1dhigh B cells inhibited the prolif-eration of CD4+CD25-T cells in vitro in a dose-dependent manner (P<0.01), but increased the secretion of IL-10 by CD4+T cells (P<0.01) and the proliferation of Treg cells in vitro (P<0.01).Conclusion Com-pared with other tissues , the percentage of B10 cell subset in spleen is the highest in wild-type mouse , and B10 cells subset can be activated through Toll-like receptor ( TLR ) signaling pathway .The responses of CD4+CD25-T cells and Treg cells in co-culture with B10 cells are regulated by B 10 cell subset through an increased IL-10 production .B10 cells might be a useful cell population for the treatment of inflammatory au-toimmune diseases.

The purpose of this study is to explore depression metabolic markers in rat hippocampus and to investigate the anti-depressant effect of genipin and its mechanisms using nuclear magnetic resonance (NMR) metabonomics. Chronic unpredictable mild stress (CUMS) procedure was conducted to establish the depressive rat model. At the beginning of the third week, genipin low dose (25 mg x kg(-1)), middle dose (50 mg x kg(-1)), high dose (100 mg x kg(-1)), and venlafaxine (50 mg x kg(-1)) were given to the CUMS rats separately once daily for two weeks except control and model groups. Rat hippocampus was analyzed by 1H NMR based metabonomics after drug administration for 2 weeks. Significant differences in the metabolic profile of rat hippocampus of the CUMS treated group and the control group were observed with metabolic effects of CUMS including decreasing in glycine and N-acetylaspartate, increasing in inositol, glutamate, lactate, glutamine, taurine and alanine. Genipin showed ideal antidepressive effects at a dose of 50 mg x kg(-1) in rats, decrease of inositol, glutamate, lactate, alanine were observed, while glycine and N-acetylaspartate were increased. Important influence has been found on normal nervous system function of these significant changed metabolites, which suggests that the antidepressant effect of genipin may be played by enhancing the activity of neurons in hippocampus, repairing and improving the function of the neuron. The metabonomics approach is an effective tool for the investigation of the anti-depressant effect and pharmacologic mechanisms of genipin.
Alanine ; metabolism ; Animals ; Antidepressive Agents ; isolation & purification ; pharmacology ; Aspartic Acid ; analogs & derivatives ; metabolism ; Behavior, Animal ; drug effects ; Chronic Disease ; Depression ; drug therapy ; metabolism ; physiopathology ; Gardenia ; chemistry ; Glutamic Acid ; metabolism ; Glycine ; metabolism ; Hippocampus ; drug effects ; metabolism ; Inositol ; metabolism ; Iridoids ; isolation & purification ; pharmacology ; Lactic Acid ; metabolism ; Magnetic Resonance Spectroscopy ; Male ; Metabolomics ; Plants, Medicinal ; chemistry ; Rats ; Rats, Sprague-Dawley

Objective To explore the effects of thiamine and riboflavin on H2O2-induced DNA oxidative damage in human umbilical vein endothelial cell line ECV304.Methods ECV304 cells were incubated with 10,100,500,1000 mg/L of thiamine or 20,100,300,500 nmol/L of riboflavin for 24 h,and then oxidative damage of cells were induced by 25 mol/L H2O2 for 30 min.DNA damage was detected with single cell gel electrophoresis(SCGE)assay.ECV304 cells incubated without H2O2,thiamine and riboflavin were served as negative controls,and those incubated with H2O2 and without thiamine and riboflavin were served as positive controls.Results H2O2 induced DNA damage,and the indices of percent of DNA damage cells,percent of tail DNA,tail length and Olive tail moment were increased.The indices of cells pretreated with 10,100,500 mg/L of thiamine or 20,100,300 nmol/L riboflavin were significantly decreased(P0.05).Conclusion Proper supplementation of thiamine and riboflavin may decrease H2O2-induced DNA oxidative damage,while excess thiamine and riboflavin supplementation may be harmful to DNA and enhance the susceptibility to H2O2 potentially.

Objective To investigate the changes of IL‐1β,IL‐2 ,IL‐4 ,IL‐6 ,IL‐10 and INF‐γexpressions in bone marrow of chro‐nic myeloid leukemia(CML)patients .Methods The IL‐1β,IL‐2 ,IL‐4 ,IL‐6 ,IL‐10 and INF‐γexpression levels were detected by u‐sing flow cytometry in 30 cases of CML chronic phase(CML‐CP) ,21 cases of CML accelerated phase(CML‐AP) ,15 cases of CML blastic phase(CML‐BP) ,42 cases of CML remission after treatment and 7 cases of non‐remission .Then the detection results were compared with those in the control group .Results The expression levels of INF‐γ and IL‐2 in each CML groups were lower than those in the control group(P<0 .05) ,while the expression levels of IL‐1β,IL‐4 ,IL‐6 and IL‐10 were higher than those in the con‐trol group(P<0 .05) .With the disease condition progression ,the INF‐γand IL‐2 levels were gradually decreased ,i .e .,CML‐BPCML‐AP> CML‐CP ,the difference among groups had statistical significance (P<0 .05) .After complete remission by treatment ,the INF‐γand IL‐2 levels were risen to some extent ,but which were still lower than those in the control group(P<0 .01) ,the expression levels of IL‐1β,IL‐4 ,IL‐6 and IL‐10 were decreased to some extent ,but likewise were higher than those in CML‐CP(P< 0 .05);the levels of IL‐1β,IL‐4 ,IL‐6 ,IL‐10 ,IL‐2 and INF‐γ had no statistical difference between the non‐remission group and CML‐BP group(P>0 .05) .Conclusion The changes of serum cytokines in bone marrow microenvironment of CMLpatients have certain significance to the occurrence ,development and prognosis of CML ;the de‐tection of IL‐1β,IL‐2 ,IL‐4 ,IL‐6 ,IL‐10 and INF‐γlevels in bone marrow is hopeful to provide new ideas and theoretical basis for im‐mune therapy and prognosis judgment of CML patients .

The optimum condition of shaking-flask p roducing enzyme were the tempe rature 26℃,initial pH 6 4,fermentation period 19 hours,medium volume 15mL m e dium/300mL Flask.soymilk-clotting enzyme was obtained from ammonium sulfate p r ecipitation.The optimum temperature and pH for the soymilk-clotting activity wa s 70℃and 5 8.The enzyme was easy to lose activity in acid or alkaline circumst a nce.About 60% of the original activity remained after 1 hour at 60℃.Ca 2+ ,Fe 2+ , Mg 2+ ,Na +increased the clotting activity,whereas Zn 2+ ,Al 3+ ca use inhibition.