Objective To explore the effect of CYP3A5 gene polymorphisms on the whole blood trough concentration (C0) of tacrolimus (TAC) in patients with idiopathic membranous nephropathy to identify an economical and optimal initial dosage delivering the best curative effect with minimum drug adverse reaction.Methods Sixty patients with idiopathic membranous nephropathy were enrolled in this study.The CYP3A5 genotype was tested by fluorescence in situ hybridization (FISH).According to CYP3A5 genotype, the patients were divided into three groups (AA, AG, and GG).At the same time, the C0 of TAC was measured by enzyme multiplied immunoassay technique (EMIT).C0 of TAC, daily dosage of TAC and the concentration/dose(C0/D) ratio of TAC were detected after taking medicine at 8, 12, 16 and 24 weeks respectively, so as to corroborate the relation between CYP3A5 gene polymorphisms and the dosage of TAC.Results The oral TAC dosage had great variation among individuals.The occurrence of the CYP3A5 genetic polymorphisms (A6986G) designated as G was 53.33％.D and C0 were significantly different at 8, 12, 16 and 24 weeks respectively (all P ＜ 0.05).To reach the same C0, the patients with AA needed 2-3-fold dosage of TAC than GG;and those with AG needed 1-2-fold dosage of TAC than GG.After 24-week treatment, the effective rate of AA group was markedly lower than AG and GG (16.67％ vs 81.25％, 16.67％ vs 87.50％, all P ＜ 0.001).Among CR, PR and NR, there were no significantly difference on C0 or C0/D of TAC (P ＞ 0.05).Conclusions CYP3A5 genotypes are correlated with blood concentration of TAC.CYP3A5 genotyping may be a new approach to predict the optimal initial dosage of tacrolimus in idiopathic membranous nephropathy.

Objective To study the effect of astragaloside on TGF-β1,SMAD2/3,and α-SMA expression in the kidney tissue of diabetic KKAy mice,and evaluate its potential role in renal interstitial fibrosis.Methods 20 type 2 diabetic KKAy mice were randomly divided into model group and astragaloside group,while 10 male C57BL/6J mice were selected as the control.Astragaloside at 40 mg · kg-1 · d-1 was given when the KKAy mice fed with high-fat diet to 14 weeks old.The mice in the control and model group received normal saline at 40 mg · kg-1 · d-1.Blood glucose meter was used to detect the blood glucose value of each mice at 16th,20th and 24th week.The mice were killed at 24 weeks old and the kidney tissue samples were collected.Pathology morphological changes were observed.Results (1) blood glucose value:cmpared with the control group,the blood glucose value of KKAy mice at 14 week increased significantly,and that of model group also increased significantly at 16th,20th and 24th week (P＜0.05);the blood glucose value of astragaloside group decreased compared with control group (P＜0.05).(2) Morphology of kidney:in the control group,the glomerular and tubular had clear structure,there was no renal interstitial fibrosis;in the model group,the renal glomerular mesangial matrix had broaden,mesangial cell had increased,renal tubular epithelial cell cytoplasm showed vacuole degeneration,renal interstitial inflammatory cell had increaised.In astragaloside group,there were few renal tubular epithelial cell cytoplasm,and there was no obvious fibrosis.(3)TGF-β1,SMAD2/3,and α-SMA expression levels of the kidney issuse:compared with control group,mice in model group up-regulated TGF-β1,SMAD2/3 and α-SMA expression (P＜ 0.05).TGF-β1,SMAD2/3,and α-SMA expression levels in astragaloside group were significantly lower than those in the model group (P＜0.05).There was few phosphorylated SMAD2/3 expression in renal tubular and glomerular nuclei,while that of model group increased (P＜0.01),and compared with model group,that of the astragaloside group decreased (P＜0.05).Conclusion Astragaloside can delay the renal fibrosis process in diabetic mice by influencing the TGF-β/SMADS signaling pathway and down-regulating TGF-β1 and α-SMA expression,thus to relieve renal fibrosis in diabetic mice.

0 05); The protein expression of erbB2 in carcinomas of palate and floor of mouth were obviously higher than the expression in lip cancer and gingival carcinoma( P 0 05).Conclusion The amplification and overexpression of c erbB2 and p53 play important roles in the formation and development of OSCC, the overexpression of c erbB2 protein has more important significance in tumorigenesis of carcinomas of palate and floor of mouth; Not only transcriptional regulation but also translational modulation exists on the expression of c erbB2 and p53. [

Objective Idiopathic pulmonary fibrosis ( IPF) is a chronic inflammatory disease with unknown etiology and is lack of effective therapy. The aim of this study is to explore the function of Ca2+ and PI3K/AKT/mTOR pathway in the pathogenesis of IPF, and the impact of 1,25?( OH) 2 D3 on Ca2+ and PI3K/AKT/mTOR pathway in type Ⅱalveolar epithelial cells of rat with IPF. Methods 150 male SD rats were randomly divided into 2 groups: prevention group ( control groupⅠ, model groupⅠ, medication groupⅠ) and treatment group ( control groupⅡ, model groupⅡ, medication groupⅡ) . The tracheal exposure surgery was operated in control groupⅠ/Ⅱ, and then 200μL sterile physiological saline was administered by intraperitoneal injection of each rats 2 days and 14 days after surgery, separately. Bleomycin(BLM)(5 mg/kg) was in?jected into the trachea of model groupⅠ/Ⅱ, and then vitamin D3 solvent(0.1%ethanol and 99.9%glycol propylene, 1μL/g) was ad?ministered by intraperitoneal injection 2 days and 14 days after surger?y, separately. Bleomycin( BLM) ( 5 mg/kg) was injected into the tra?chea of medication groupⅠ/Ⅱ, and then 1,25?( OH) 2 D3( 2μg/kg) was administered by intraperitoneal injection 2 days and 14 days after surgery, separately. IPF model was built by injecting Bleomycin into the trachea of rats, 1,25?(OH)2D3(2μg/kg) was used to prevent and treat IPF by intraperitoneal injection in medication group. The hydroxyproline content of lung tissue in each group was measured, type Ⅱalveolar epithelial cells were separated from lung tissue and labeled with Fluo?3AM, then concentration of Ca2+ was detected by Laser scanning confocal microscope. The mRNA levels of PI3K, AKT and mTOR in the typeⅡalveolar epithelial cells were tested by RT?PCR. Results Compared with control groupⅠ/Ⅱ at each time point, hydroxyproline content of lung tissue, Ca2+ concentration and expression of PI3K, AKT and mTOR in typeⅡalveolar epithelial cells in model groupⅠ/Ⅱand medication groupⅠ/Ⅱwere sig?nificantly raised( P<0.05 or P<0.01) , but these were significantly reduced in medication groupⅠ/Ⅱcompared with model groupⅠ/Ⅱ( P<0.05 or P<0.01) . Correlation analysis showed that there is significant positive correlation between Ca2+ concentration and mRNA expression levels of PI3K, AKT and mTOR in model groupⅠ/Ⅱ(r=0.5988, r=0.6230, r=0.6603,P<0.01)and medication groupⅠ/Ⅱ( r=0.701 2, r=0.632 3,r=0.740 3,P<0.01) . Conclusion The PI3K/AKT/mTOR pathway plays an important role in devel?opment of IPF. 1,25?( OH) 2 D3 is able to reduce Ca2+concentration in typeⅡalveolar epithelial cells and inhibit the PI3K/AKT/mTOR pathway, and then inhibit the development of IPF.

Objective:To explore the function of IL-9 and PU.1 on genesis and development of pulmonary fibrosis,and the effect of active vitamin D3[1,25(OH)2VD3] on the expression levels of this two factors during the pathogenesis of fibrosis.Methods: 90 male SD rats were randomly divided into model group,treatment group,control group (n=30).Bleomycin(5 mg/kg) was injected into the trachea of rats to establish the model of pulmonary fibrosis in the model group and treatment group,while the control group was injected with isopyknic sterile saline.The treatment group,the model group and the control group were injected intraperitoneally with active vitamin D3,solvent of vitamin D3 (propylene glycol) and sterile saline on the 2nd day after surgery respectively.All injections were carried out once every other day.10 rats were euthanized at 14th,21st and 28th day in each group in turn.After obtaining lung tissues from experimental rats,the pathological change of lung was compared by hematoxylin-eosin staining.The difference of collagen fiber and hydroxyproline content were compared by the Masson staining and basic-hydrolysis method respectively.The mRNA and protein expression of IL-9 and PU.1 in lung tissue were detected by Real-time PCR and immunohistochemical technology respectively.The expression of IL-9 in serum was detected by ELISA.Results: Fibrosis appeared in lungs of experimental rats treated with bleomycin after 14 days,and more and more aggravated with time.At three time points,the hydroxyproline content in model group and treatment group were significantly higher than that of control group,and the treatment group was significantly lower than the model group.At three time points,the expression of IL-9 and PU.1 in model group and treatment group were risen gradually,and obviously higher than that in control group.On the 14th and 21st day,the expression of two factors in treatment group was significantly lower than model group;on the 28th day,there was no statistically significant difference between treatment group and model group(P>0.05).In model group and treatment group,the expression of two factors on 21st day was significantly higher than that on 14th day;there was no statistically significant difference between the 28th day and the 21st day.Conclusion: IL-9 and PU.1 may play a profibrotic role at early stage of pulmonary fibrosis induced by bleomycin.The active vitamin D3 may lower the expression level of PU.1,and then reduce the secretion of IL-9,thus may play an inhibiting effect on genesis and development of pulmonary fibrosis in rats.

Objective:To investigate the effects of hydrogen rich-saline (HRS) on intestinal mucosal barrier in rat with intestinal ischemia/reperfusion injury (IIRI).Methods:Twenty-four healthy male Sprague-Dawley rats, aged 8 weeks, were randomly divided into 3 groups (8 in each group) by random number table method: sham group, model group and HRS group.Rats in HRS group were intraperitoneally injected with HRS (10 mL/kg) at 30 min of ischemia, and the same amount of normal saline was intraperitoneally injected in model group.After 45 min of ischemia and 6 h of reperfusion, rats were sacrificed.Serum and ileum were collected for further detection.Tumor necrosis factor alpha (TNF-α), interleukin (IL)- 1β and IL-17A expression levels in serum were detected by conducting enzyme-linked immunosorbent assay (ELISA). The localization expressions of tight junction protein Occludin was detected by immunohistochemical staining (IHC), while the localization expression of tight junction protein zonula occluden-1 (ZO-1) were detected by immunofluorescence staining (IF). The protein expression of Occludin, ZO-1, and Lysozyme were detected by performing Western blot.The mRNA expression of Lysozyme and α-defensin were detected by real-time PCR (qPCR).Results:ELISA results proved that the levels of serum TNF-α and IL-1β in HRS group rats were significantly lower than those in model group [(62.02±29.97) ng/L vs.(113.40±44.58) ng/L, (21.68±0.35) ng/L vs.(28.29±3.49) ng/L], while the level of IL-17A increased [(28.18±5.28) ng/L vs. (15.10±3.60) ng/L] (all P<0.05). IHC staining: compared with model group, the expression of Occludin in HRS group was uniform and continuous, and the staining was darker.IF results: compared with model group, the fluorescence signal intensity of ZO-1 in HRS group rats significantly increased, and the distribution was clear and continuous.Wes-tern blot results: compared with model group, the expression levels of Occludin and ZO-1 proteins in HRS group rats remarkably increased (0.79±0.06 vs. 0.54±0.04, 0.91±0.11 vs. 0.51±0.13), while Lysozyme protein decreased (1.50±0.40 vs. 2.99±0.80) (all P<0.05). qPCR results revealed that the expression level of Lysozyme mRNA in HRS group rats was lower than that in model group (1.64±0.33 vs. 2.20±0.40), while α-defensin mRNA obviously increased (0.82±0.19 vs. 0.47±0.13) (all P<0.01). Conclusions:HRS protects intestinal mucosal barrier by inhibiting the expression of tight junctions and the secretion of antimicrobial peptides in rat suffering from IIRI.

OBJECTIVE: To explore the regulatory role of SOX2-OT in migration of lung squamous cell carcinoma H520 cells and the underlying mechanisms. METHODS: Wound- healing and Transwell migration assays were performed to examine the changes in migration and invasion capacity of lung squamous cell line H520, which expressed higher levels of SOX2-OT than other lung cancer cell lines, following RNA interference-mediated SOX2-OT knockdown. The transcription levels of epithelial-mesenchymal transition (EMT)-related components was detected by qRT-PCR and immunoblotting. Gli1 gain-of-function analysis was performed in H520 cells with SOX2-OT knockdown and the changes in EMT phenotype of the cells were examined. miR-200c mimic and inhibitor were used to analyze the mechanism by which SOX2-OT positively regulates Gli1 and the mediating role of SOX2. RESULTS: SOX2-OT knockdown significantly lowered the invasiveness and migration capacity of H520 cells and caused changes in EMT phenotype of the cells. Overexpression of Gli1, which was positively regulated by SOX2-OT, reversed the inhibitory effect of SOX2-OT knockdown on migration of H520 cells. Transfection of the cells with miR-200c inhibitor effectively reversed SOX2-OT knockdown-induced down-regulation of SOX2. CONCLUSION The SOX2-OT/SOX2 axis positively regulates migration of lung squamous H520 cells via Gli1-mediated EMT.
Humans ; Epithelial-Mesenchymal Transition/genetics* ; Zinc Finger Protein GLI1/metabolism* ; Cell Line, Tumor ; Cell Movement/genetics* ; Lung Neoplasms/genetics* ; MicroRNAs/metabolism* ; Gene Expression Regulation, Neoplastic ; Cell Proliferation/genetics* ; Neoplasm Invasiveness/genetics* ; SOXB1 Transcription Factors/metabolism*