Objective To observe the effect of Chinese Medicines with the function of antioxidation on lupus nephritis(LN)pa- tients.Methods The 25 LN patients consisting of renal function normal group(15 cases)and renal failure group(10 cases)were treated with Huang Qi(Radix Hedysari)and Shu Dihuang(Radix Rehmanniae Praeparata)as main ingredients for 6 months in suc- cession.Changes of symptom,systemic lupus erythematosus disease activity index(SLEDAI),activity index(AI),lipid peroxide (LPO),superoxide dismutase(SOD),glutathione peroxide(GSH-Px),malondialchehyche(MDA),nitric oxide(NO),blood and urine routine examination,24-hour urine protein quantity,liver and kidney function,anti-double-stranded DNA(anti ds-DNA),complement 3(C_3)and complement 4(C_4),after treatment were observed.Results SLEDAI and AI scores were remarkably different from those before treatment(P

Objective To develop new methods to cultivate, retrieve and purify mouse mesenchymal stem cells (mMSCs). Methods Bone marrow was collected from 2-month-old Kunming mice by flushing femurs and tibias with complete medium of DMEM-LG. Cells were plated in a Petri dish. After 24 hours, non-adherent cells were removed by two to three washes with PBS, adherent cells were further cultured in complete medium and retrieved by trypsinisation with 0.25% trypsin for 5 min at 37 ℃. The treated adherent cells were cultivated with 3?dilution for further generations. CD11b-negative cells were retrieved from the collected adherent cells of 3rd generation by using immunomagnetic microbeads, and continued to be cultured in complete medium. After the cultured cells were retrieved, their morphology and their ability of osteoblastic differentiation and adipocytic differentiation were examined. Results Most of mMSCs from 1st generation were of shuttle shape, some of irregular shape. After treatment with magnetic microbeads and several generations, mMSCs were of spindle, star and irregular shape. These cells were of rich cytoplasma, clear nucleolus, and grew in parallel or vortex. The cultured adherent cells from the first and subsequent generations had plenty of CD11b-positive blooding-making cells. After 20-day osteoblastic induction, mMSCs differentiated into bone cells, which showed orange phosphate in extracellular matrix by Alizarin red S staining. mMSCs could differentiate into lipocytes. The size of cells increased along with fat-developing induction period. These cells showed many orange fatty follicles with O Red Oil dyeing. Conclusion Pure mMSCs can not be retrieved by either adhering method or generation cultivation method separately. The combined methods of adhering, immunomagnetic microbeads, and serial subcultivation is effective in vitro in retrieve mMSCs.

AIM To prepare the andrographolide-loaded solid dispersions.METHODS Differential scanning calorimetry (DSC) and X-ray powder diffraction (XRPD) were used for the analysis of solid dispersions previously prepared from the mixture of andrographolide and PVPK30 (1 ∶ 9) in autoclave.In addition to the evaluation index of in vitro dissolution rate,the influencing factors of pressure,temperature and reaction time were taken into consideration for the preparation optimization by Box-Behnken method.RESULTS Andrographolide was totally dispersed in solid dispersions in an amorphous state,manifesting an inhibited crystal diffraction peaks' formulation.Under the optimal conditions of 21.26 MPa for pressure,40.76 ℃ for temperature,and 1.13 h for reaction time,the in vitro dissolution rate was 85.49％.CONCLUSION After andrographolide is made into solid dispersions,its in vitro dissolution rate is obviously increased.

The culture medium and cultural conditions of solid-state fermentation of Streptomyces Menmyco-93-63 were tested in this study. The suitable medium which contains rice, sorghum, millet bran, and rice hull with the proportion of 2:2:3:3 was developed for the spore production of Streptomyces Men-myco-93-63 using single substrate screening, mixture substrate screening and orthogonal experiments, and the sporulation was up to 2.52?109 CFU/g. And then, initial charge, initial ratio of water to solid, inoculating quantity, and culture temperature impact to sporulation of Streptomyces Men-myco-93-63 were tested. The favorite cultural conditions are developed as the following: the initial charge is 15 g in 500 mL Erlenmeyer flask; initial ratio of water to solid is 1.7:1.0 (V/W, rice hull excluding), inoculating quantity is 7 mL, culture temperature is 28℃.

Objective To evaluate values of magnetic resonance imaging(MRI)in eyes filled with silicone oil.Design Prospective cases series.Participants 40 eyes of 40 patients were filled with silicone oil after ocular injury.Methods MRI was performed in the 40 patients,including axial FSE T_1WI,T_2WI,coronal T_2WI with fat saturation,oblique sagittal T_1WI and axial T_2FLAIR.MRI findings,in- cluding morpbous,signal and complications were analyzed.Oculi axes were measured.Main Outcome Measures Morphous,signal, complications and oculi axes of the eyes filled with silicone oil.Results Affected oculi axis was 2.18cm?0.21cm,normal oculi axis was 2.48cm?0.16cm.The silicone oil in eyes demonstrated isointense signal or slightly hyperintense signal on T_1WI and T_2WI,hypointense signal after fat saturation.Hydrops was found in vitreous cavity in 33 patients,including simple hydrops in 17 patients and complicated other abnormality in 16 patients.Choroidal detachment was found in 11 patients,complicating vitreous hydrops in 5 patients and lo- calized bulge of eyeball wall.Retinal detachments were found in 4 patients,of whom 3 patients complicated with vitreous hydrops.Per- fluorocarbon liquid residual in vitreous cavity,foreign body in anterior chamber,localized thickness of the wall of the globe and meagre- mean of silicone oil in vitreous cavity were found respectively in one patient complicating vitreous hydrops.Except for eye changes, fracture of orbital wall and foreign body in orbit were found in one patient.Conclusions MRI can display the changes of eyes filled with silicone oil,and measure oculi axes biologically and accurately offering important clinical application value.(Ophthalmol CHN,2007,16: 312-315)

Background There have been an abundance of literature on the analysis of the mechanical characteristic of the sclera at the entire seleral level in high myopia.However,some recent studies on high myopia are focused on the mechanical changes of the sclera on a cellular level. Objective This experiment was purposed to study how transforming growth factor-β2(TGF-β2)affected sclerotic desmocytes and the mechanical behaviors of scleral fibroblasts in the posterior part of the eyes in guinea pigs with experimental myopia. Methods Induced myopic animal models were established by wearing-10.00 D concave lens for 30 days in lateral eyes of 2-week-old guinea pigs.The fellow eyes were used as control group.Another 5 matched animals served as normal controls.The scleral fibroblasts of each group were purified with the tissue explant method and passaged for 2 generations in vitro.Cultured cells were identified by immunochemistry with vimentin,desmin,keratin and S-100 antibodies.Different concentrations of TGF-β2(0,1,10,100mg/L)were added into serum-free DMEM for 24 hours,and the viscoelastic properties of scleral fibroblasts were measured by micropipette aspiration technique. Results Compared with the fellow,eyes and normal control eyes,the equilibrium modulus and apparent viscosity in model eyes were significantly higher(P＜0.05).After treatment of TGF-β2,the equilibrium modulus and apparent viscosity in the model group and fellow eyes were positively correlated with the concentrations of TGF-β2(r=0.743,r=0.533,r=0.654,r=0.576,P＜0.05).Following the addition of 1 mg/L TGF-β2 and 10 mg/L TGF-β2,the equilibrium modulus and apparent viscosity of scleral fibroblasts were significantly reduced in model eyes compared with fellow eyes(P＜0.05).No significant difference was found in the equilibrium modulus and apparent viscosity of scleral fibroblasts between model eyes and fellow eyes after treatment with 100 mg/L TGF-β2(P＞0.05). Conclusion TGF-β2 car increase the mechanic indexes in a concentration.dependent manner.1 mg/L,10 mg/L TGF-β2 can lower the equilibrium modulus and apparent viscosity of scleral fibroblasts of normal eye and thus cause more changes in the mechanical behavior of scleral fibroblasts.

AIM To investigate the preventive effects of herbal pair,Scutellariae Radix and Coptidis Rhizoma (SC),on Alzheimer's disease (AD),and its mechanism of action.METHODS Dementia mice induced by 8-week s.i.d subcutaneous injection of D-galactose (100 mg/kg),were simultaneously given respective,intragastric administration of SC crude drug at doses of 5,10,20 g/kg,or piracetam support at 0.75 g/kg,and isometrical distilled water was applied to the mice of normal control group.The mice had their learning and memory abilities checked by Morris water maze at intervals of four weeks and eight weeks since the start of the trial,and their blood and brain tissue biochemical indices measured at the end of the test.RESULTS Significantly shortened latent period in place navigation test and the time of enter into the original platform in the space exploration test were observed in the mice treated with 4-week D-galactose and SC (P ＜0.05 或 P ＜0.01).The 8-week intervention demonstrated SC capacity in the significant promotion of T-SOD activity,decreased blood MDA levels (P ＜ 0.01)and the brain AchE levels,and increased brain GSH-Px activity (P ＜ 0.01).CONCLUSION SC increases the concentration of acetylcholine in brain tissue and protects the central nervous tissue under oxidative stress,highlighting its therapeutic effect on AD.

OBJECTIVETo investigate the effect of farnesoid-X-receptor (FXR) antagonist Z-guggulsterone in an in vivo high-fat fed apolipoprotein E knockout (ApoE(-/-)) mice model of myocardial ischemia/reperfusion (I/R).

METHODSMale ApoE(-/-) mice were randomly divided into three groups: standard ApoE(-/-) group (fed with standard mouse diet for 12 weeks before myocardial I/R procedure, n = 18), high-fat ApoE(-/-) group (fed with high-fat mouse diet for 12 weeks before myocardial I/R procedure, n = 22), and high-fat ApoE(-/-) + FXR antagonist group(fed with high-fat mouse diet for 12 weeks and received FXR antagonist Z-Guggulsterone 30 minutes before myocardial I/R procedure, n = 17). The expression of FXR was detected by real-time quantitative-PCR. Myocardial infarct size was determined by Evans blue/TTC double staining methods. Myocardial apoptosis was determined by in situ TUNEL technique. Markers of the mitochondrial-mediated apoptotic pathway (cytochrome c release, caspase-9 activity, and BAX and BCL-2 levels), endoplasmic reticulum stress apoptotic pathway (caspase-12 activity and CHOP level), and death receptor apoptotic pathway (caspase-8 activity, and Fas and FasL levels) were also measured.

RESULTFXR expression (3.7-fold higher, P < 0.01), myocardial infarct size [(62.1 ± 7.0)% vs. (33.8 ± 5.8)%, P < 0.01] and myocardial apoptosis index[ (36.8 ± 5.7)% vs. (17.2 ± 3.8)%, P < 0.01]were all significantly higher in high-fat ApoE(-/-) group than those in standard ApoE(-/-) group. Compared with high-fat ApoE(-/-) group, myocardial infarct size [(24.4 ± 4.7)% vs. (62.1 ± 7.0)%, P < 0.01] and myocardial apoptosis index [(13.8 ± 2.7)% vs. (36.8 ± 5.7)%, P < 0.01] were significantly reduced in high-fat ApoE(-/-) + FXR antagonist group. Moreover, levels of mitochondrial-mediated apoptotic pathway markers (cytochrome c release, caspase-9 activity, and BAX/BCL-2 levels) and endoplasmic reticulum stress apoptotic pathway markers (caspase-12 activity and CHOP level) were significantly lower in high-fat ApoE(-/-) + FXR antagonist group than those in high-fat ApoE(-/-) group (all P < 0.01). Levels of death receptor apoptotic pathway markers (caspase-8 activity, and Fas and FasL levels) were similar between high-fat ApoE(-/-) group and high-fat ApoE(-/-) + FXR antagonist group.

CONCLUSIONFXR antagonist alleviates myocardial reperfusion injury in cholesterol-fed ApoE(-/-) mice via inhibition of the mitochondrial-mediated and endoplasmic-reticulum stress pathway.

Animals ; Apolipoproteins E ; genetics ; Apoptosis ; drug effects ; Caspase 9 ; metabolism ; Cholesterol, Dietary ; administration & dosage ; Cytochromes c ; metabolism ; Disease Models, Animal ; Endoplasmic Reticulum Stress ; Male ; Mice ; Mice, Knockout ; Myocardial Reperfusion Injury ; metabolism ; pathology ; prevention & control ; Pregnenediones ; pharmacology ; Proto-Oncogene Proteins c-bcl-2 ; metabolism ; Receptors, Cytoplasmic and Nuclear ; antagonists & inhibitors ; metabolism ; bcl-2-Associated X Protein ; metabolism

Objective To investigate the regulating effect and mechanism of microRNA-21 (miR-21) on extracellular matrix (ECM) of vascular smooth muscle cells (VSMCs) by vascular remodeling of hypertension. Methods By narrowing the abdominal aorta in rats, the hypertension models were established and divided into 2-week hypertension group and 4-week hypertension group, and sham-operated group was also established as control. VSMCs from the rat aorta were subjected to 0% (static), 5% (normal) and 15%（hypertensive）elongation strain at a constant frequency of 1.25 Hz and duration of 12 hours, respectively. The expressions of Smad 7 and ECM were detected by Western blotting, and the expression of miR-21 was examined by Real-time RT-PCR. Finally, miR-21 siRNA was used to study the role of miR-21 in the mechanical strain-induced expression of ECM, miR-21 and Smad 7. Results Compared with the sham-operated group, ECM and miR-21 in thoracic aorta of 2-week hypertension group were significantly elevated. Collagen I, collagen III and miR-21 in thoracic aorta of 4-week hypertension group were significantly elevated. Compared with the static and 5% strain groups, the protein expression of collagen I in VSMCs did not show significant change, but the protein expression of collagen III was significantly elevated and Smad 7 expression was significantly decreased in 15% strain group. The cyclic strain also enhanced miR-21 expression in VSMCs. miR-21 inhibitor effectively decreased the expression of miR-21 in VSMCs and protein level of collagen III, while enhanced Smad 7 expression under the static and 15% strain. Conclusions The vascular remodeling of hypertension causes the high expressions of ECM and miR-21. The cyclic strain induces the high expression of miR-21, which via Smad 7 results in enhancing the expression of ECM, collagen III especially, in VSMCs under vascular remodeling of hypertension.

Objective To investigate the influence of iodine excess on expression of TRAIl/TRAIL-sR1 in NOD and Balb/c mice and to study the effect of TRAIl/TRAIL-sR1 on the pathogenesis of experimental autoimmune thyroiditis(EAT). Methods Both Balb/c and NOD mice were divided randomly into control and iodine excess group by feeding with water containing no NaI or 0.05% Nal. The mice were sacrificed after 8 weeks. TRAIL and TRAIL-sR1 mRNA levels were detected by RT-PCR. The function, morphology and apoptosis of thyroids were also observed by ELISA and Tunnel stain. Results Treated by HI, enlarged follicles and flattened epithelium by accumulation of colloid were found in thyroids of both NOD and Balb/c mice. But significant lymphoid cell infiltration and local fibrosis were only found in thyroids of NOD HI group. The relative weight of thyroids of NOD mice in HI group[(104.8±14.5)mg/kg]was heavier than that of control group [(71.8±20.4)mg/kg]. The level of TT4 declined in HI group[(30.77±3.59)mmol/L]compared with control group[(36.43±2.66)mmol/L], meanwhile, the level of TSH was higher in HI group[(6.98±0.66)μg/L]than that in control group [(5.55±0.56)μg/L]. The difference being statistically significant(t=7.773,-9.526,-4.458, all P < 0.05). The relative weight of thyroids of Balb/c mice of HI group[(155.8±20.8)mg/kg]also heavier than that of control group [(105.1±22.0) mg/kg]. The level of TT4 droped in HI group [(19.75±3.32) mmoL/L]was higher than that in control group[(23.46±6.21)mmoL/L], the level of TSH in HI group[(4.14±1.71)μg/L]was higher than that in control group[(3.55±1.41)μg/L], the difference being statistically significant(t=7.554,-7.239,3.140, all P< 0.05). A great deal of apoptotie ceils observed in NOD (3.97±0.91) and Balb/c mice (1.05±0.45) by Tunnel stain were greater than control groups (0.21±0.15, 0.10±0.03), the difference being statistically significant in beth of the two species(t=-7.167,-17.772, both P < 0.05). The apoptosis index of thyroid follicular epithelium in NOD was obviously higher than Balb/c(t=-7.625, P<0.05). The level of TRAIL mRNA did not remarkably change in Balb/c between control group(0.000 59±0.000 39) and HI group(0.001 24±0.000 46, t=-1.940, P>0.05), but it increased apparently in NOD mice HI group(0.018 88±0.005 77) than that of control group(0.009 61± 0.00591, t=-2.71, P<0.05). The level of the expression of TRAIL-sR1 mRNA increased in HI groups of NOD (0.000 53±0.000 15) and Balb/c mice(0.000 42±0.000 09) than that in control groups of NOD(0.000 28± 0.000 05) and Balb/c mice (0.000 17±0.000 06) and the differences were statistically significant between the two species(t=3.050,3.990, all P<0.05). The differences of the expression of TRAIL and TRAIL-sR1 mRNA between the two species were significant(t=-3.37,-4.76, all P<0.05). Conclusions Iodine excess induces colloid goiter in beth species of mice and thyroiditis in NOD mice. The increase of TRAIL and TRAIL-sR1 influenced by iodine excess is one of the molecular bases of follicular epithelium apoptosis and inflammation in thyroids. Genetic factor is a key factor in the pathogenesis of thyroiditis.