Objective To investigate the role of increased microRNA-21 (miR-21) in the development of renal tubulointerstitial fibrosis secondary to aristolochic acid induced acute kidney injury.Methods C57BL/6J male mice were intraperitoneally injected with aristolochic acid at a dose of 10 mg/kg.Blood samples and kidneys were harvested at day 1,3,7,14,28 after aristolochic acid treatment.To assess the role of miR-21 in aristolochic acid induced acute kidney injury to chronic kidney disease progression,mice were intravenously injected with anti-miR-21 or anti-scramble (10 mg/kg) at 1 h before aristolochic acid dosing,as well as d5 and d10 after aristolochic acid dosing.Results Increased serum creatinine and severe kidney injury were found at d3 after aristolochic acid treatment.Renal tubulointerstitial fibrosis was developed at d14 after aristolochic acid treatment.Protein expression of α-SMA,vimentin and collagen Ⅰ were significantly up-regulated at d7 and peaked at d14 (P ＜ 0.01),while protein abundance of E-Cadherin decreased at d14 and lasted until d28 (P ＜ 0.01).The abundance of miR-21 increased at d7 after aristolochic acid dosing,peaking at d14 and thereafter maintaining at a high level.Anti-miR-21 intervention relieved renal injury with reduced serum creatinine (P ＜ 0.05) and attenuation of renal tubulointerstitial fibrosis.Besides,the protein expression of α-SMA,vimentin,and collagen Ⅰ/Ⅳ was all down-regulated after anti-miR-21 treatment (P ＜ 0.05).PTEN was up-regulated and the ratio of its downstream genes p-AKT/AKT was decreased.(P ＜ 0.05) Conclusions A single high dose of aristolochic acid leads to acute kidney injury and the development of renal tubulointerstitial fibrosis secondary to AKI.Renal tubulointerstitial fibrosis could be partially reversed by inhibiting miR-21 via PTEN/p-AKT pathway.

Objective To investigate the roles of microRNA-382 (miR-382) in the pathogenesis of renal tubulointerstitial fibrosis (TIF).Methods Human kidney epithelial cells (HK2)transfected with miR-382 inhibitor (antagomiR-382) were used to examine the effect of miR-382 abundance on cell polarity,as well as to test the complementary relationship between miR-382 and its predicted target gene heat shock protein 60 (HSPD1),which was further verified by 3'-untranslated region luciferase assay and site-directed mutagenesis.The role of miR-382 played in the development of renal interstitial fibrosis and redox regulation was examined in a mouse unilateral ureteral obstruction (UUO) model.Locked nucleic acid (LAN)-modified anti-miR-382 was intravenous delivered via tail vein 30 min prior to UUO,and repeated the dosage 24 h after the surgery.For clinical verification,renal biopsy specimens from 12 IgA nephropathy (IgAN) patients were collected,6 patients with moderate to severe TIF and 6 patients without TIF.The relative abundance of miR-382 and HSPD1 protein was analyzed by using in situ hybridization and immunohistochemistry.Results HSPD1 was confirmed to be a new,direct target gene of miR-382 by in vitro 3'-untranslated region luciferase assay and sitedirected mutagenesis.The development of epithelial transition in HK2 cells was accompanied with upregulation of miR-382 [(6.54±0.96) vs (1.12±0.26),P ＜ 0.05].Blocking the expression of miR-382 could reversed the progression of epithelial transition partially.In UUO mice the abundance of miR-382 was up-regulated [(6.89 ± 2.47) vs (1.00±0.42),P ＜ 0.01] while HSPD1 and Trx were downregulated compared with the sham group.Down-regulation of miR-382 was associated with significant decrease in TIF,but increase in HSPD1 and thioredoxin protein compared with UUO group [HSPD1:(0.34±0.10) vs (0.14±0.05);Trx:(0.79±0.18) vs (0.36±0.16);all P ＜ 0.05].The expression of miR-382 was up-regulated and HSPD1 was significantly down-regulated in IgAN patients with TIF.Conclusions miR-382 play an important role in renal tubulointerstitial fibrosis in human and mice.HSPD1 is one of the target genes of miR-382.The down-regulation of HSPD1 and the decrease ability of anti-oxidative stress may be the important mechanism of miR-382 involved in renal tubulointerstitial fibrosis.

Objective To determine the serum level of chernokine CCL27 in patients with psoriasis vulgaris,and to analyse its clinical relevance.Methods A total of 61 patients(40 in progressive stage and 21 in stable stage)with psoriasis vulgaris,with an average disease duration of 37.97±14.34 years,were included in this study.Appropriate thempy was given to these patients.Serum samples were collected from the patients before and after therapy,as well as from 45 healthy human controls.ELISA was applied to examine the serum concentration of CCL27.Clinical severity of psoriasis vulgaris was assessed by psoriasis area and severity index(PASI)score.Results Serum level of CCL27 was 670.02±262.15 ng/L in psoriatic patients,compared to 373.10±92.84 ng/L in the controls(t=8.18.P＜0.01).Increased serum level of CCL27 was observed in patients with progressive psoriasis vulgaris compared to those with stable psoriasis (799.94±214.54 ng/L vs 422.57±135.53 ng/L,t=8.39,P＜0.01).After 8 weeks of therapy,a significant decrease was noticed in the serum level of CCL27 in patients who experienced≥70%reduction in PASI score(t=9.95,P＜0.01).but not in those experiencing a PASI reduction of＜70%(t=1.84,P＞0.05).The serum level of CCL27 was positively correlated with PASI score(r=0.58,P＜0.01).Conclusions The serum level of CCL27 is significantly elevated in patients with psoriasis vulgaris,and it is correlated with the disease severity.

200 citrinin mutants were screened with the inhibition zone method from the transformants library of Monascus ruber M-7 by Agrobacterium-mediate DNA transfer, which contains more than 5,000 transformants.Then 53 mutants, whose citrinin contents ranged from 0.04?g/g to 154.57?g/g in the red fermented rice (RFR), were achieved by high performance liquid chromatography (HPLC).Color values of RFR prepared by these mutants were also detected.The results showed that there was a positive correlation between the citrinin content and the color value among the mutants.These results provide materials and research bases for ferrther studying the relationship between the production of citrinin and pigment of Monascus ruber at molecular level.

Objective To observe the role of rosiglitazone in unclipped kidneys of two-kidney- one-clip hypertensive rats and examine its relationship to angiotensinⅡreceptors.Methods Two- kidney-one-clip hypertensive rats were divided randomly into 4 groups as follows:positive control group (CONT),traditional antihypertensive drugs group (TAHD,reserpine 50?g?kg~(-1)?d~(-1), dihydralazine 6.25 mg?kg~(-1)?d~(-1) and hydrochlorothiazide 6.25 mg?kg~(-1)?d~(-1),regular-dose rosiglitazone group (RRGL,rosiglitazone 5 mg?kg~(-1)?d~(-1)),and high-dose rosiglitazone group (HRGL, rosiglitazone 20 mg?kg~(-1)?d~(-1)).Sham operation rats were as negative controls.Each group had 8 rats. Animals were monitored and sacrificed at 10th week.Results Blood systolic pressure in TAHD group and HRGL group was significantly lower than that in CONT group [TAHD(137?27 ) mm Hg and HRGL (143?16) mm Hg vs CONT (191?25 ) mm Hg,P＜0.05],but no significant difference between the former two groups was found.Nor did the blood systolic pressure between RRGL group [(176?18) mm Hg] and CONT group.At 10th week,rats in SHAM group and treated groups had lower urinary urinary protein excretion rate,glomerular injury score and wall-to-lumen ratio of arteriole than those in CONT group [vs CONT urinary protein excretion rate (44.60?17.40) mg/24 h,P＜0.05; vs CONT glomerular injury score 60.85?33.05,P＜0.05;vs CONT wall-to-lumen ratio of arteriole 2.33?1.01,P＜0.01,except TAHD group].Though with the similar level of blood pressure,blood glucose and lipid,HRGL,compared with TAHD group showed lower urinary protein excretion rate [HRGL (16.78?3.50) mg/24 h vs TAHD (27.94?12.79) mg/24 h,P＜0.05],decreased glomerular injury score (HRGL 18.04?7.76 vs TAHD 27.92?6.39,P＜0.05) and wall-to-lumen ratio of arteriole (HRGL 1.75?0.38 vs TAHD 2.16?0.90,P＜0.05) in the cortexes of unclipped right kidneys.The expression of type 1 angiotensinⅡreceptor (AT1R) mRNA was no difference in HRGL group and TAHD group,but the expression of type 2 angiotensinⅡreceptor (AT2R) mRNA was more intensive in HRGL group.Conclusion Rosiglitazone can protect the kidneys from hypertensive injury,especially in high dose.The beneficial effects seem incompletely dependent on the metabolism modulating and reduction of blood pressure,but in relationship to the upregulation of AT2R mRNA.

OBJECTIVETo investigate the effects of serum containing Gumibao (Chinese character: see text) on the proliferation and differentiation of osteoblast induced by dexamethasone.

METHODSOsteoblasts were extracted from skulls in newly born (within 24 hours) SD rats, and digested with collagenase. The first passage of cells were used for experiments. Cells were cultured in the medium containing different concentrations of dexamethasone (0, 10(-8), 10(-7), 10(-6), 10(-5) ,10(-4) mol/L). Alkaline phosphatase staining were carried out after 1 week and numbers of mineralized nodes with alizarin red staining were observed after 3 weeks. Accordingly, following the treatment of 10(-5) mol/L dexamethasone for 1 week, cells were cultured in the medium with serum containing Gumibao (Chinese character: see text). One week after Cumibao (Chinese character: see text) treatment, cells were stained with Alkaline phosphatase and collagen I and PCNA were examined by Western-blot. However, the observation of numbers of mineralized nodes with alizarin red stain required one more week.

RESULTSHigh concentration of dexamethasone could inhibit the expression of PCNA, collagen I, alkaline phosphatase and reduce the number of mineralized nodes of osteoblast, while serum containing Gumibao (Chinese character: see text) could reverse the inhibition.

CONCLUSIONHigh concentration of dexamethasone could inhibit the proliferation and differentiation of osteoblastic cells, while serum containing Gumibao (Chinese character: see text) could reverse the inhibition.

Animals ; Cell Differentiation ; drug effects ; Cell Proliferation ; drug effects ; Cells, Cultured ; Collagen Type I ; analysis ; Dexamethasone ; pharmacology ; Dose-Response Relationship, Drug ; Drugs, Chinese Herbal ; pharmacology ; Female ; Osteoblasts ; cytology ; drug effects ; physiology ; Proliferating Cell Nuclear Antigen ; analysis ; Rats ; Rats, Sprague-Dawley

OBJECTIVETo establish a reliable model for drug screening and therapy by culturing rat femoral head and inducing cartilage degeneration quickly in vitro.

METHODSThe femoral heads from the same SD rats of two-month old were divided into control group and experimental group respectively. They were cultured with DMEM medium plus 10% fetal bovine serum or DMEM medium plus 10% fetal bovine serum plus 50 ng/ml IL-1β for three days. Femoral heads were fixed in 4% paraformaldehyde, decalcified, dehydrated, embedded in paraffin and cut into slices. Specimens were stained with Toluidine blue and Safranine O-Fast Green FCF. The protein expression levels of type II collagen, MMP13, Sox9 and ADAMTS5 were analyzed by immunofluorescence.

RESULTSBoth the Toluidine blue and Safranine O staining were pale in the margin of femoral heads which were stimulated with IL-1β for three days compared to that in control group. The Fast Green FCF staining was positive at the edge of the femoral head in experimental group, which indicated that cartilage became degenerated. The expression levels of both type H collagen and Sox9 were decreased significantly while the expression levels of MMP13 and ADAMTS5 were increased in experimental group.

CONCLUSIONThe model of cartilage degeneration is established by culturing and inducing the degeneration of the femoral heads quickly in vitro.

Animals ; Cartilage Diseases ; genetics ; metabolism ; Collagen Type II ; genetics ; metabolism ; Disease Models, Animal ; Femur Head ; metabolism ; Humans ; In Vitro Techniques ; Interleukin-1beta ; genetics ; metabolism ; Male ; Matrix Metalloproteinase 13 ; genetics ; metabolism ; Rats ; Rats, Sprague-Dawley ; SOX9 Transcription Factor ; genetics ; metabolism

Animals ; Apoptosis ; drug effects ; Capsules ; Cell Proliferation ; drug effects ; Drugs, Chinese Herbal ; therapeutic use ; Hepatocytes ; pathology ; Liver ; pathology ; Liver Cirrhosis, Experimental ; drug therapy ; pathology ; Male ; Phytotherapy ; Rats ; Rats, Wistar

3 mg/L was significantly worse than in those with hs-CRP≤3 mg/L (18.18%,5.45%;P=0.044,log-rank test). Higher hs-CRP concentration was an independent predictor of death or new vascular event(OR 3.609;95% CI 0.869—14.992;P=0.047).Conclusion Higher hs-CRP concentration in acute phase after ischemic stroke is an independent predictor of death or new vascular event in a year.

Objective: To observe the therapeutic effect on hepatic fibrosis of chronic hepatitis B by Chinese medicine Handan Ganle capsule (HDGLC).Methods: A total of 104 patients with chronic hepatitis B has been treated by HDGLC for 6 months, liver fibrosis indexes and the serum biochemical indexes were detected before treatment, during the curative period and by the end of treatment. Hepatic biopsy was performed before or after the treatment.Results: The improvement rate of clinical symptoms was 79.0%-90.6%, and the recovery rate of ALT was 72.6%, serum fibrosis indexes such as HA, type Ⅳ collagen and LN were significantly decreased along with the extending course of disease (P<0.05). The pathohistological score of liver was decreased from 7.82±6.22 before treatment to 5.16±3.75 after treatment (P<0.05) and the score of hepatic fibrosis was decreased from 7.49±5.45 before treatment to 5.16±4.26 after treatment (P<0.05).Conclusion: HDGLC has remarkable therapeutic and reversing effect on chronic hepatitis B induced hepatic fibrosis.