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.

Objective An investigation was conducted to assess the effects and mechanism of celecoxib [a selective cyclooxygenase(COX) 2 inhibitor] on a rat colitis model induced by trinitrobenzene sulfonic acid (TNBS). Methods The rats were divided into four groups. Group 1 and group 2 were experimental groups. Group 3 and group 4 were control groups. Colitis was induced by intracolonic administration of TNBS (25 mg/ml) in a vehicle of 50% ethanol (0.25 ml) in rats of experiment groups. Three hours before induction of colitis ,the rats were beginning and continuing to treat orally with celecoxib (1.25 mg/kg, group 1) and distilled water (1 ml/0.3 kg, group 2) twice per day for 7 days , respectively. The rats in group 4 were treated orally with celecoxib (1.25 mg/kg) twice per day for 7 days. Group 3 served as healthy control. All rats that survived until the end of the experiment (7 d) were killed and the severity of damage were assessed. The prostaglandin E2 (PGE2) concentrations of colonic mucosa were tested by radioimmunoassay. Results The colonic damage scores were 11.15?3.3 in group 1 and 8.50?2.82 in group 2. Both were significantly higher than that of group 3 (0.62?0.09)( P

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.

Objective To develop a new methodology of prenatal sonography scoring to assess and dignose clubfoot.Methods Thirty-one fetuses suspected clubfoot(50 foots) and 62 normal fetuses(124 foots) were scanned with both 2D and 3D sonography,and were divided into normal feet group and clubfeet group.Our scoring system was made according to the Pirani-scoring for children,while considering the morphology of mid-foot and hind-foot of infants.The doctors would give final diagnosis for all fetuses after delivery or induced labor,and analyse the correlations among the abnormal signs,sonographic score and the clinical outcomes.Results The curve lat border,the medial crease,the empty heel and the post crease can be observed in the 3D reconstruction images.These four abnormal signs had close correlations with the clinical results.Conclusions The new prenatal clubfeet scoring system by 3D sonography is a promising methodology with clinical values.

OBJECTIVE To study whether immunosuppressant(IS,cyclophosphamide,CPA) have the function of prevention and treatment of congenitalautoimmune sensorineural hearing loss or not.METHODS The female guinea pigs were immunized by crude conspecific inner ear antigens(CIEAgs),and took orally IS or not at the same time during gestation.Then the pregnant guinea pig's and their offspring's hearing function were measured and inner ear histopathologic changes were observed.Some of offspring borne by the female guinea pigs which immunized with CIEAgs and not treated with IS showed hearing loss and were treated by IS,and their hearing functions were measured and inner ear histopathologic changes were observed with same techniques.RESULTS After immunizing with CIEAgs,some of female guinea pigs which immunized with CIEAgs and not treated with IS showed different degrees of hearing loss(the thresholds of acoustic nerve compound active potential and cochlear microphonic potentials elevated) and immune inflammations in inner ear tissues.All females guinea pigs in experimental group which immunized with CIEAgs and treated with IS at same time,and their offspring have no any hearing loss and inner ear histopathologic changes.After IS therapy,the hearing function improved(mainly at the low-frequency region) in some offspring guinea pigs,which borne by the female guinea pigs which immunized with CIEAgs and not treated with IS.CONCLUSION IS could effectively prevent offspring's congenital sensorineural hearing loss induced by their mother's special antibodies against inner ear tissues antigens.IS showed effective result for treatment of congenitalautoimmune sensorineural hearing loss,but the curative effect was limited.

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.

Background: Dampness-heat syndrome is a major syndrome type in patients with a sharp deterioration of chronic renal failure (CRF). Qingshen Decoction, a compound traditional Chinese herbal medicine, could relieve the clinical symptoms of CRF patients, and was considered to have a certain reversal effect on rapid deterioration of renal function. Objective: To observe the changes of serum interleukin-8 (IL-8) and IL-18 levels in CRF patients with a sharp deterioration and dampness-heat syndrome, and to explore the curative efficacy of Qingshen Granule. Design, setting, participants and interventions: Sixty CRF patients with a sharp deterioration and dampness-heat syndrome from Department of Nephrology, the First Affiliated Hospital of Anhui College of Traditional Chinese Medicine, were randomly divided into treatment group (30 cases) and control group (30 cases), with another 20 healthy individuals as normal control. The patients in the treatment and control groups were all treated with Jiedu Xiezhuo II, a compound traditional Chinese herbal medicine, given as retention enema. Qingshen Granule was additionally administered to the patients in the treatment group with 1 dosage each time and 3 times a day. The treatment course was one month. Main outcome measures: The levels of serum IL-8 and IL-18 in the normal individuals and before and after treatment in the treatment and control groups were detected. Results: The total response rates of treatment group in clinical efficacy assessment and assessment of syndrome of traditional Chinese medicine (86.67% and 86.67%) were higher than those of the control group (56.67% and 60%), and there were significant differences between the two groups (P<0.05). The levels of serum IL-8 and IL-18 in CRF patients before treatment were obviously higher than those in the healthy individuals (P<0.01), and there were no differences in the levels of IL-8 and IL-18 before treatment between the treatment group and control group (P>0.05). After one-month treatment, the levels of serum IL-8 and IL-18 were markedly decreased in the two groups (P<0.01), and the levels of serum IL-8 and IL-18 in the treatment group were markedly lower than those in the control group (P<0.05). There were differences in decreased degrees of IL-8 and IL-18 levels between the two groups (P<0.05). Conclusion: CRF patients with a sharp deterioration and dampness-heat syndrome have high serum IL-8 and IL-18 levels. Qingshen Granule can reduce the levels of serum IL-8 and IL-18, and improve the renal function and ameliorate the clinical symptoms.

Objective To observe the effect of low concentration Aβ1-42 monomer/oligomers and CORM-2 in different concentration on livability of SN56 cells. Methods SN56 cells were cultured in the 96-well plate with uniform concentration, and were divided into control group, Aβ1-42 group, Aβ1-42 + CORM-2 50μM group, and Aβ1-42 + CORM-2 100 μM group. Three lines of cells in Aβ1-42 group were cultured in the surroundings of 10nM,100nM and 1 μM Aβ1-42monomer/oligomers, respectively. Aβ1-42 + CORM-2 50μM group and Aβ1-42 + CORM-2 100μM group had the same culture condition as group Aβ1-42 ,except contain 50μM, and 100μM CORM-2, respectively. Control group didnt have any effect factor. Three days later,the livability of different groups was compared with MTT method. Results The livability of group Aβ1-42 with the increasing concentration of Aβ1-42 was (79.73 ±0.94)% ,(67.99 ±0.79)% ,(60.42 ±0.62)% , respectively. The higher the concentration of Aβ1-42 was,the lower the livability of SN56 cell was. The livability of group Aβ1-42 + CORM-2 50μM/100μM was( 75.15±0.096)%,(63.20 ±0.17)%, (55.33 ±0.19)%; (73.20 ±0.27)%, (64.34 ±0.11 )%, (54.17 ±0.12)% , respectively. Both were lower than group Aβ1-42. And different CORM-2 concentration had discrepancy in the ability of decreasing the cell livability. Conclusion Low concentration of Aβ1-42 can reduce the livability of SN56 cells, and higher concentration has more significant effect; CORM-2 in different concentration both can decrease the livability of SN56 cells,and there is a discrepancy in the intensity.

A comparison of the volatile compounds in Rhizomes Curcumae （Ezhu） and Radix Curcumae （Yujin） was undertaken using gas chromatography mass spectrometi-y （GC-MS）. Ultrasonic extraction and GC-MS methods were developed for the simultaneous determination of five sesquiterpenes, namely, α-pinene, β-elemene, curcumol, germacrone and curdione, in Ezhu and Yunjin. Good linearity （r〉0.999） and high inter-day precision were observed over the investigated concentration ranges. The validated method was successfully used for the simultaneous determination of five sesquiterpenes in Ezhu and Yujin. The quantitative method can be effectively used to evaluate and monitor the quality of Chinese curcuma in clinical use.