Objective To investigate the effects of a novel PPARγ agonist DH9 on Wntβ-catenin pathway in human polycystic kidney cystic-lining epithelial cells (WT9-12).Methods WT9-12 cells were treated with different concentrations of DH9 for 72 hours and the proliferation was assessed by MTT.WT9-12 cells were pretreated with SB216763 or GW9662 for two hours and then treated with DH9 for 72 hours.Western blotting was applied to detect the protein expression of β-catenin,phospho-β-catenin,GSK3β,phospho-GSK3β.Results DH9 could effectively inhibit the proliferation of the cells.60 μmol/L DH9 could facilitate β-catenin down-regulation (P＜0.01) and phospho-β-catenin up-regulation (P＜0.01).Inhibition of GSK3β by SB216763 could protect WT9-12 cells against DH9-facilitated β-catenin repression in a dose-dependent manner despite phosphorylating deactivation,but PPARγ inhibitor GW9662 couldn't.Conclusions DH9can effectively block the proliferation of WT9-12 cells.The effect may be mediated by facilitating the down-regulation of β-catenin via GSK3β-dependent mechanism.

Objective To investigate the relationship between blood concentration of sodium valproate and the anti‐epileptic effect and the influencing factors of blood concentration of sodium valproate ,and to provide evidence for clinical individual adminis‐tration .Methods The blood concentrations of 133 cases of patients treated with sodium valproate were determined in the affiliated Yongchuan hospital of Chongqing medical university ,the monitoring results of blood concentration were statistically analyzed ,and the anti‐epileptic effect of 133 cases of patients were observed and analyzed .Results The epilepsy of 80 cases of patients were con‐trolled ,accounted for 60 .15% of the total number;in the 69 cases of patients within the effective blood drug concentration (50-100 mg/L) ,the epilepsy of 51 cases of patients were controlled ,accounting for 38 .35% .In the 44 cases of patients whose blood drug concentration were less than 50mg/L ,the epilepsy of 26 cases of patients were not controlled .The distribution of blood concentra‐tion between men and women were similar .The blood concentrations among each age group were different ,the blood concentrations of 52 .87% patients in the adult group were below or above therapeutic range that were 39 .13% in the minor group .The adverse reactions were increase with the increase of blood concentration .Conclusion There are differences between the blood concentrations of sodium valproate and clinical effect ,the reasonable individual administration should be conducted according to the patient′s blood concentrations of sodium valproate ,the epilepsy control situation of patients and the patients′age .

Objective To observe the impact of heparanase on glomerular endothelium glycocalyx during sepsis and to investigate the prevention of glycocalyx injury.Methods C57/BL6 mice were injected with lipopolysaccharide (LPS) or tumor necrosis factor-α(TNF-o) and sacrificed one hour later.Glomerular endothelium glycocalyx traced with lanthanum was observed by transmission electron microscope(TEM).Western blotting was used to observe heparanse protein expression of renal cortex tissue.Human renal glomerular endothelial cells (HRGECs) were stimulated with TNF-α and active heparanase protien expression was detected by Western blotting.Mice were administrated with heparin sodium or heparinase Ⅲ and renal endothelium glycocalyx was observed by TEM.Urine during twenty-four hours was collected to measure urinary albumin and creatinine.The ratio of albumin to creatinine was calculated and compared among groups.Results The glomerular endothelium glycocalyx of LPS group and TNF-α group was degradated and the one of podocyte was integrated.Renal cortex tissue heparanase protein expression was significantly increased since one hour after LPS injection (P ＜ 0.01).The protein expression of activited heparanase of HRGECs which were stimulated with TNF-α was increased (P ＜ 0.05).Administration of heparin sodium which could inhibit the activity of heparanase could prevent the glycocalyx form degradation.The ratio of urine albumin to creatinine of heparin sodium group was decreased compared with LPS group (P ＜ 0.05) and the ratio of heparinase Ⅲ group was higher than control group(P ＜ 0.01) as a result of degradation of glomerular endothelium glycocalyx.Conclusions During the early stage of sepsis,TNF-α can induce glomerular endothelium heparanase to increase and active,and consequently the glycocalyx is degradated which leads to albuminuria.Inhibition of heparanase can protect glomerular endothelium glycocalyx and prevent albuminuria.

Background and purpose:Both nature killer cells(NK)and cytoxic T lymphocytes in the body tissues of human are the dominant components of cellular immunity,This study was done to explore the degree of infiltration of NK/T in lung squamous cell carcinoma and its relation to patient survival and prognosis.Methods: CCD8 as the markers cytoxic T lymphocyte(CTL)and CD56 as the markers natural killer(NK)were stained immunohistochemically to detect the distribution and infiltration in the lung squamous cell carcinoma specimens. Results:In 39 of 68 lung neoplasm,whose CTL infiltration was zero or mild,the five-year survival rate was 18%, while in 29 with marked CTL infiltration,the five-year survival rate was 42%.In 46 of 68 lung neoplasm,whose NK infiltration was zero or mild,the five-year survival rate was 14%,while in 22 with marked NK infiltration,the five-year survival rate was 45%.In 48 of 68 lung neoplasm,both the NK and T cells were zero or mild,the five-year survival rate was 33%,while in 20 with marked NK and T cell infiltration,the five-year survival rate was 54%.The five-year survival difference among the patients with NK,T infiltration either marked or zero/mild infiltration were significant(x~2=18.62, P=0.00).Conclusions:The degree of NK and T infiltration is positively correlated with the prognosis and survival time of lung squamous cell carcinoma patients.

Objective To explore related factors of amputation in patients with diabetes foot ulcer(DFU) and provide the evidence of reducing amputation rate.Methods A total of 307 inpatients with DFU in this hospital from October 2013 to December 2015 were collected and divided into amputation group(n=28) and non-amputation group(n=279).The clinical data and laboratory data were treated with single variable analysis and multivariate Logistic regression analysis.Results Single variable analysis showed that hospitalization time,hyper-sensitive C-reactive(hs-CRP) and fibrinogen(FIB) level were significantly increased in amputation group(P＜0.05) when compared with non-amputation group,and the hemoglobin (Hb) level,high-density lipoprotein cholesterol(HDL) and serum albumin (ALB) level in amputation group were markedly lower than in non-amputation group(P＜0.05).Amputation rate was significantly different among different Wagner grades (P=0.000).Multivariate Logistic regression analysis showed that hospitalization time,Wagner grade were independent risk factors of amputation in patients with DFU,OR values were 1.024(95％CI:1.009-1.039) and 2.779 (95％CI:1.753-4.404),respectively.While HDL was a protective factor of amputation in patient with DFU (OR=0.161;95％CI:0.036-0.729).Conclusion The higher the rank of Wanger in DFU patients are,the longer hospital stay and the higher amputation risk become.HDL is the protective factor of DFU amputation.

Objective To design an experiment for method-comparison and bias estimation of multi tests on multi instruments.Methods According to the procedure described by the NCCLS approved guideline EP9-A and improving the method of sample collection,we took 8 patient mixing samples each day to analyze all comparison tests on 11 auto-chemistry analyzer within following 5 days.The duplicates were assessed within the same run.The coefficient of correlation and average bias% were calculated,and the system errors at medical decided levels were assessed.Results Taking ALT as an example,the coefficients of correlation were between 0.994-1.000,and the average bias% were between-0.460%-4.927%,SE at 40U/L was-1.510-1.834 and SE at 300 U/L was-3.101-9.188.Conclusion In all tests that joined the comparison among the different instruments,28 tests were acceptable,2 tests were acceptable after modifying the coefficients,and AMY and LIP were not acceptable.

This study aims to develop a method for determination of beta-elemene, curcumol, germacrone and neocurdione in the volatile oil of Curcuma phaeocaulis, and to provide the basis of the quality control method for the volatile oil of C. phaeocaulis and the related preparations. Based on GC-MS, the 4 main compounds were simultaneously determined, with the internal standard n-tridecane. The Agilent 19091S-433 column (0.25 microm x 250 microm x 30 m) was adopted at the temperature of 250 degrees C, the programmed temperature method (60 degrees C for 1 min, 5 degrees C x min x to 110 degrees C for 5 min, 1 degrees C x min(-1) to 140 degrees C, 5 degrees C x min(-1) to 160 degrees C, 10 degrees C x min(-1) to 240 degrees C) was used. Helium gas was used as the carrier gas at a constant flow rat of 1 mL x min(-1), with an injection volume of 1 RL. Mass spectra were taken at 70 eV; the ion-source temperature was 200 degrees C. The relation time and character acteristic ions for each target compound were determined by full scan mode and SIM, and m/z 85.1, 93.1, 121.1, 107.1 and 180.1 were the detection ions of n-tridecane, beta-elemene, curcumol, germacrone and neocurdione. As a result, beta-elemene, curcumol, germacrone and neocurdione were all detected with good separation. They were all in a good linear relationship within each concentration scope. The average recovery rates were in the range of 98.2%-101%. So, the method can be used to control the quality of the volatile of C. phaeocaulis Val. and the preparations related.
Acetic Acid ; chemistry ; Curcuma ; chemistry ; Drugs, Chinese Herbal ; analysis ; isolation & purification ; Gas Chromatography-Mass Spectrometry ; methods ; Oils, Volatile ; chemistry ; isolation & purification ; Plant Oils ; chemistry ; isolation & purification ; Sesquiterpenes ; analysis ; isolation & purification ; Sesquiterpenes, Germacrane ; analysis ; isolation & purification

ObjectiveTo investigate the predisposing factors and pathogenic fungal species of Malassezia folliculitis in different geographical areas and body sites.MethodsTotally,241 patients diagnosed with Malassezia folliculitis were asked to complete a questionnaire.The content of hair follicles was obtained and subjected to fungal smear and culture examination.Fungal species were identified according to morphological,physiological and biochemical features.Results Of the 241 patients with Malassezia folliculitis,204 (84.65％) were positive for smear examination.A total of 259 specimens were collected from these patients,and fungal culture grew 213(82.24％) strains,of which,209 belonged to Malassezia species,4(1.54％) to Candida species.Among the 209 Malassezia strains,186 were activated and subjected to species identification which resulted in 6 species,including M.furfur (111 strains,59.68％ ),M.sloofiae (43 strains,23.12％ ),M.sympodialis (17 strains,9.14％),M.globosa (9 strains,4.84％),M.pachydermatis (4 strains,2.15％),and M.obtuse(2 strains,1.08％ ).Of the pathogenic fungi of Malassezia folliculitis,M.furfur predominated in the chest,back,abdomen,face and neck,M.sloofiae in the upper limbs,shoulders and vertex,M.globosa in the lower limbs.There were obvious differences in the distribution of pathogenic fungal species at different body sites in a same host,and M.furfur with M.sloofiae or M.sympodialis appeared to be the most common pathogens.ConclusionsIn this study,6 Malassezia species are identified in patients with Malassezia folliculitis in Nantong and Nanjing area,M.furfur and M.sloofiae appear to be the dominant pathogens.

In this study, laser scanning confocal microscopy (LSCM) was used to determine the location and relative quantity of flavonoids in the leaves of Apocynum venetum L. from the top, middle and basal parts of the branch. The leaves of the plants of one, two and three years old, separately, were collected in July. ANOVA and LSD test were employed in the statistical analysis. The results indicated that flavonoids located mainly in xylem conduit of vein, collenchyma, epidermic cells and cuticle. The data of flavonoids contents under statistical analysis showed that difference existed in the leaves of different parts and different ages. This study provided the reliable scientific material about the analysis of the ecological and the exploitation of the leaves of Apocynum venetom L.

Objective To investigate the antiproliferative effect of rosiglitazone, a thiazolidinedione (TZD) on autosomal dominant polycystic kidney disease (ADPKD) cystic lining epithelial cells and to explore the underlying molecular mechanism. Methods ADPKD cysticlining immortalized epithelial (WT9-12) cells were stimulated by rosiglitazone with different concentrations. After treatment, MTT method was performed to detect the level of proliferation; flow cytometry was used to determine the cell cycle distribution and the apoptosis rate. Western blotting was used to detect the protein expressions of mTOR, p70S6K, 4E-Bp1, PPARγ PPARγ siRNA was transfected into WT9-12 cells to knock down the expression of PPARγ Results Treatment of WT9-12 cells with rosiglitazone resulted in a dose-dependent and time-dependent strong inhibition of cell proliferation, an accumulation of cells in the G0/G1 phase (rosiglitazone 50 μmol/L 65.43%,rosiglitazone 100 μmol/L 64.02%, control 49.65% ) and 6% apoptosis at high concentration (rosiglitazone 200 μmol/L). Rosiglitazone reduced the phosphorylation of p70S6K in a dosedependent and time-dependent manner. The levels of phosphorylated mTOR and 4E-Bp1, the latter being a downstream substrate of mTOR related mRNA translation initiation, were not changed by rosiglitazone. Cells were pre-incubated with GW9662, a PPARγ antagonist, before the treatment with rosiglitazone, the inhibition of p70S6 kinase phosphorylation by rosiglitazone was partially prevented by GW9662 (P＜0.01). Then PPARγ siRNA was transfected into WT9-12 cells, in contrast to untransfected control or cells transfected with an irrelevant siRNA, rosiglitazone did not cause an obvious inhibition of p70S6 kinase phosphorylation in PPARγ knock-down.Conclusion Rosiglitazone inhibits the proliferation of ADPKD cystic lining epithelial cells, and down-regulates p70S6 kinase phosphorylation through mTOR-independent and PPARγ-dependent signal pathway.