AIM: To find whether lipoxin A_4 (LXA_4) inhibits cell proliferation induced by TNF-? in rat mesangial cells, and to explore the molecular mechanisms of signal pathways of LXA_4 actions. METHODS: Cultured rat mesangial cells were growth-arrested and exposed to TNF-? with or without preincubation with LXA_4. Proliferation of mesangial cells was measured by MTT methods. Activities of STAT_3 were analyzed by electrophoretic mobility shift assay. Expression of cyclin E mRNA was assessed by RT-PCR. Cyclin E proteins were determined by Western blotting analysis. RESULTS: TNF-?-induced proliferation and increased mRNA and protein expression of cyclin E in mesangial cells were inhibited by LXA_4 in a dose-dependant manner. TNF-?-stimulation of the STAT_3-binding activities in mesangial cells was down-regulated by lipoxin A_4. CONCLUSION: Inhibitory effect of LXA_4 on TNF-?-induced mesangial cell proliferation is mediated by Jak_1/STAT_3 signal pathway.

AIM: To examine whether lipoxin A_4 (LXA_4) induces apoptosis of rat renal interstitial fibroblasts and explore the mechanisms. METHODS: Rat renal interstitial fibroblasts (NRK-49F cells) were incubated in RPMI-1640 medium supplemented with 5% fetal calf serum and exposed to LXA_4 at the concentration of 10 nmol/L, 100 nmol/L or 1 ?mol/L for 24 hours. Prior to experiment, some cells were transfected with calpain 10 antisense oligodeoxynucleotide. Apoptosis of cells was recognized by double staining using fluorescent dye acridine orange and ethidium bromide, observed under laser scanning confocal microscopy and counted by flow cytometry following propidium iodide and annexin staining. Activity of caspase-3 was measured by colorimetric assay. The expression of calpain 10 mRNA was determined by RT-PCR. RESULTS: LXA_4 at the concentration of 100 nmol/L or 1 ?mol/L induced 9.83% or 33.82% apoptosis of cells, respectively. Treatment of cells with LXA_4 up-regulated the expression of calpain 10 mRNA and increased the activity of caspase-3. The transfection of the cells with calpain 10 antisense oligodeoxynucleotide inhibited the LXA_4-induced apoptosis, activity of caspase-3 and expression of calpain 10. CONCLUSION: LXA_4 at high concentration induceds apoptosis in rat renal interstitial fibroblasts via up-regulating of calpain 10 mRNA expression.

ObjectiveTo investigate the change of lipoxin A4 (LXA4), leuotriene B4(LTB4) in blood and urine and leukocyte 15-lipoxygenase (15-LO) of the children with acute poststreptococcal glomendonephritis (APSGN) and to evaluate its significance. MethodsBlood and urinary levels of LXA4 and LTB4 were measured with ELISA within 3 days (acute phase), 10 to 14 days (early resolution phase) and 6 to 8 weeks (late resolution phase) respectively after onset of APSGN in 22 patients. In 8 children with APSGN, expression level of leukocyte 15-LO mRNA was examined with RT-PCR. Leukocyte LTB4 synthesis was assessed with ELISA. Chemotactic effect of LTB4, LXA4 and 15-S-hydroxyeicosatetraenoic acid (15-S-HETE) on neutrophils was determined by in vitro chemotaxis assay. Twenty-two healthy children were served as control. ResultsBlood and urinary levels of LXA4 and leukocyte 15-LO mRNA were up-regnlated in acute phase, further increased in early resolution phase, and decreased in late resolution phase of APSGN, which were stir higher than those in the controls (P＜0.01). Blood and urinary levels of LTB4 were increased in acute phase (P＜0.01) and then were decreased in early resolution phase and hte resolution phase of APSGN, which were still higher than those in the controls (P＜0.01). Administration of 15-S-HETE or LXA4 in vitro inhibited LTB4-induced chemotactic effect on neutrophils of the patients,and inhibited the production of leukocyte LTB4. ConclusionsChanges of blood and urinary levels of LXA4 and LTB4 in early resolution phase of APSGN are contrary. 15-S-HETE and LXA4may play a role in anti-inflammation and resolution of APSGN via inhibiting LTB4.

Objective To examine whether lipoxin A4(LXA4) has an antagonistic effect on interleukin (IL)-1?-induced synthesis of IL-6 in glomerular mesangial cells, and to explore its mechanism. Methods Cultured glomerular mesangial cells (GMCs) of rat were treated with IL-1?, with or without preincubation with LXA4. Protein secretion of IL-6 in supernatants was examined analyzed by enzyme-linked immunosorbent assay (ELISA). Expression of IL-6 mRNA was determined by RT-PCR. The expression of Src homology 2 (SH2) containing protein-tyrosine phosphatase 2 (SHP-2) was assessed by immunoblotting. Activities of DNA-binding of nuclear factor-kappa B (NF-?B) were measured by electrophoretic mobility shift assay (EMSA). Results The secretion of protein and expression of mRNA of IL-6 in GMCs stimulated by IL-1? were inhibited by LXA4 in a dose-dependent manner. LXA4 reduced the phosphorylation of SHP-2 and activities of NF-?B. Pretreatmnet of GMCs with NF-?B inhibitor pyrrolidine dithio-carbamate (PDTC) blocked both the secretion of IL-6 protein and activation of NF-?B induced by IL-13- Conclusion LXA4 antagonists IL-1?-induced synthesis of IL-6 in GMCs through the pathway of SHP-2/NF-?B signal transduction.

AIM: To observe the effect of continuous inhaled nitric oxide (NO) on airway resistance in guinea pigs. METHODS: 36 healthy male guinea pigs were divided into control group, isoproterenol group and two inhaled NO (20?10 -6 and 60?10 -6 ) groups. Respiratory resistance (R_E) and dynamic compliance(C_ dyn )were recorded before and after evoked by histamine at different doses. RESULTS: After injections of intravenous histamine at 80,120 and 160 ?g/kg, the R_E of inhaled NO groups were apparently lower than that of control group. Compared with control group, the C_ dyn of inhaled NO groups were significantly higher after administration of histamine at 80, 120 and 160 ?g/kg. After given histamine at more than 80 ?g/kg,the R_E of inhaled NO groups were higher and the C_ dyn lower than those of isoproterenol group. CONCLUSION: Inhalation of 20?10 -6 NO and 60?10 -6 NO can inhibit the increase in airway resistance induced by higher doses (80,120 and 160 ?g/kg )of intravenous histamine, but the effect of intravenous isoproterenol seems stronger.

Myocardial ischemia-reperfusion injury (MI/RI) is an inflammatory cascade process involving the interaction of multiple factors.In recent years,more and more evidence suggests that NOD-like receptor pyrin domain containing 3 (NLRP3) inflammasome,an important component of the innate immune system,is closely associated with the inflammatory damage of MI/RI.Furthermore,blockage of NLRP3 inflammasome or the release of its downstream pro-inflammatory cytokines may provide new therapeutic targets for this disorder.

Objective To investigate the application value of perioperative enhanced recovery after surgery (ERAS) combined with laparoscopic common bile duct exploration (LCBDE) in the treatment of choledocholithiasis.Methods The clinical data of 84 patients with choledocholithiasis who were admitted to the Yijishan Hospital from January 2011 to December 2013 were prospectively analyzed.A single-blind,randomized,controlled study was performed in the 75 patients who were allocated into the control group and the enhanced recovery after surgery group (ERAS group) based on a random number table.All the patients underwent LCBDE,the patients in the control group received conventional perioperative management and the patients in the ERAS group received perioperative management according to enhanced recovery rehabilitation program.All the patients were followed up by outpatient interview till postoperative month 6.The clinical features,liver function and residual stones in the patients were observed.The operation time,postoperative complications,postoperative intestinal function recovery,duration of hospital stay and hospital expenses in the two groups were compared.Measurement data with normal distribution were presented as x ± s.Comparison between groups were evaluated with an independant sample t test.Count data were analyzed using the chi-square test.Results All the 75 eligible patients undergoing successful operation were randomly divided into the control group (35 patients) and the ERAS group (40 patients).The operation time and volume of intraoperative blood loss in the control group and the ERAS group were (185 ±46)minutes and (124 ±28)mL,(178 ±37) minutes and (114 ±32)mL,respectively,with no significant difference (t =0.729,1.431,P ＞ 0.05).There were 12,14 and 10 patients in the control group and 5,6 and 4 patients in the ERAS group with postoperative incision pain,vomit and infection,showing a significant difference (x2=5.054,5.966,4.241,P ＜ 0.05).The level of white blood cell,alanine aminotrausferase and direct bilirubin in the control group and in the ERAS group were (11.4 ± 3.5) × 109/L,(128 ± 33)U/L,(38 ±14) μmol/L and (10.6 ± 3.0) × 109/L,(135 ± 35) U/L,(44 ± 16) μmol/L at postoperative day 1,compared with (7.8 ±2.9) × 109/L,(48 ± 14) U/L,(21 ± 8) μmol/L and (6.9 ±2.1) × 109/L,(43 ± 13) U/L,(20 ±7) μmol/L in the 2 groups at postoperative day 4,respectively,showing no significant difference between the 2 groups (t =1.018,-0.872,-1.767,1.553,1.836,1.044,P ＞ 0.05).The postoperative first flatus day,time of food intake,time of postoperative infusion and duration of hospital stay were (42 ± 13)hour,(45 ±14) hours,(6.8 ±2.3)days and (11.3 ±4.5)days in the control group,and (35± 11)hours,(19 ±7)hours,(4.2 ± 1.8) days and (9.6 ± 2.4) days in the ERAS group,with a significant difference between the 2 groups (t =2.741,10.524,5.485,2.077,P ＜ 0.05).The total hospital expenses was (18 729 ± 3 127) yuan in the control group,which was significantly greater than (16 981 ±2 756) yuan in the ERAS group (t =2.574,P ＜ 0.05).The liver function of all the patients was recovered at the postoperative month 1.Four patients with residual stones in the 2 groups were detected by T-tube cholangiography,and were cured by removal of gallstones by choledochoscopy.There were no complications of the abdominal pain,jaundice and fever in all the patients till the end of follow-up.Conclusion ERAS combined with LCBDE for the treatment of choledocholithiasis is safe and feasible,with the advantages of low morbidity,quick recovery,short duration of hospital stay and less hospital expenses.

Objective To examine whether lipoxin A4(LXA4) induces apoptosis of rat renal interstitial fibroblasts and explore the mechanism concerned.Methods Rat renal interstitial fibroblasts (NRK 49F cells)were incubated in RPMI 1640 medium supplemented with 5%fetal calf serum and exposed to LXA4 at the concentration of 10 nmol/L, 100 nmol/L or 1 ?mol/L for 24 hours. Prior to experiment,some NRK 49F cells were transfected with Smac antisense oligodeoxynucleotide. Apoptosis of NRK 49F cells was recognized by double staining using fluorescent dye acridine orange and ethidium bromide,and observed under laser scanning confocal microscopy and counted by flow cytometry following propidium iodide and annexin staining. Activity of caspase 3 was measured by colorimetric assay. The expression of Smac was determined by Western blotting analysis.Results LXA4 at the concentration of 100 nmol/L or 1 ?mol/L induced apoptosis of 9 83%or 33 82%of NRK 49F cells respectively, and reduced the cells of S and G2～M phase and increased the cells of G0～G1 phase in a dose dependent manner. Treatment of NRK 49F cells with LXA4 up regulated the expression of Smac protein and increased the activity of caspase 3. The transfection with Smac antisense oligodeoxynucleotide inhibited the LXA4 induced apoptosis and expression of Smac in NRK 49F cells. Conclusion LXA4 at high concentration can induce apoptosis of rat renal interstitial fibroblasts via the up regulation of Smac expression.

Objective To investigate the characteristic of MRS in patients suffered diffuse axonal injury (DAI)in acute and suba-cute stage,and the correlation between MRS changes and the severity of disease.Methods We reported MRI and proton magnetic resonance spectroscopy studies of 3 6 head-inj ured patients in acute and subacute stage.Proton magnetic resonance spectra were ac-quired from the white matter and gray matter of bibateral frontal lobe that on conventional MRI appeared normal by using 2D MRSI at 3.0T MRI .30 volunteers as contronl were studied at the same time.Results In patient group,N-acetylaspartate/creatine ratios were (2.14±0.15)and (1.71±0.08)choline/creatine ratios were(1.35±0.13)and (1.03±0.08)for the white matter and the gray matter,respectivily.The brain N-acetylaspartate/creatine ratio was reduced and the choline/creatine ratio was increased in pa-tient group compared with the controls.The increase in the choline/creatine ratio was significant even in the moderate and severe in-j ured groups.Furthermore,there was a significantly correlation between the severity of head inj ury and the N-acetylaspartate/cho-line ratio,and changes in patients with metabolite ratios of the GCS score had a strong correlation.Conclusion We conclude that DAI patients with normal conventional imaging manifestations of the frontal lobe appear metabolite ratio change,suggesting the existence of local cerebral inj ury,and it has a strong correlation between the prognosis and MRS changes.MRS may provide an useful method that can tell us the severity of the brain inj ury in patients with DAI .

In this study, the anti-HBV effects of tea polyphenols (TP) were examined. After cells were exposed to TP for 3, 6, 9 days, amounts of HBsAg, HBeAg and HBV-DNA released into the supernatant of the cultured HepG2 2.2.15 cells were detected. TP, to some extent, inhibited the secretion of HBsAg and strongly suppressed the secretion of HBeAg in a dose-dependent (P<0.01) and time-dependent manner, with 50% maximal inhibitory concentration (IC50) value being 7.34 microg/mL on the 9th day, but the time-dependence was not significant (P=0.051). Expression of HBV-DNA in the supernatant of the cell culture also was significantly decreased in a dose-dependent fashion (P<0.01). The IC50 of TP in inhibiting HBV DNA was 2.54 microg/mL. It concluded that TP possessed potential anti-HBV effects and may be used as a treatment alternative for HBV infection.
Antiviral Agents/*pharmacology ; DNA, Viral/analysis ; Dose-Response Relationship, Drug ; Flavonoids/*pharmacology ; Hep G2 Cells ; Hepatitis B Surface Antigens/analysis ; Hepatitis B e Antigens/analysis ; Hepatitis B virus/*drug effects ; Inhibitory Concentration 50 ; Phenols/*pharmacology ; Tea/*chemistry