Background The pathogenesiof age-related maculadegeneration (AMD) iassociated with the senility of human retinal pigmenepithelium (RPE) cells.Seeking drug to arresRPE cell senility iof significance fothe prevention and treatmenof AMD.Research showed thathe lycium barbarum polysaccharide (LBP) can delay senility,buitinfluence on RPE cell aging iunclear.Objective Thistudy wato discusthe protective effecand mechanism of LBP on RPE cell aging.MethodPorcine retinal neural epithelial layewaisolated,and photoreceptooutesegmen(POS) waextracted by density gradiencentrifugation and marked by FITC.The POwathen co-cultured with RPE cellin the medium containing 0.01,0.10 and 1.00 g/L LBP fo24 hours.The areof fluorescence,representing the amounof POphagocytosed by RPE cells,wameasured undethe fluorescenmicroscope to evaluate the influence of LBP on the phagocytifunction of RPE cells.The POS-induced RPE lipofuscin-uptake cell model waestablished by co-culturing human RPE cellwith porcine POfo3 weeks.The RPE-POco-culture cell model waincubated in medium containing 0.01,0.10 o1.00 g/L LBP,and the autofluorescence caused by lipofuscin up-taken into RPE cellwadetected with flow cytometry.cell counting kiwaused to assescell proliferation and viability (value) 24,48 and 72 hourafteculturing.ResultPorcine POpresented athin rodundethe lighmicroscope and appeared abilayedisc-like structureundethe transmission electron microscope,and itFITC-labeled yellow-green autofluorescence waobserved undethe fluorescenmicroscope.No POwaup-taken into the RPE cellin the normal control group,buthe areof POphagocytosed by RPE cellwagradually enlarged with increasing doseof LBP,showing significandifference among the group(F =21.425,P =0.006).Compared with the POcontrol group,the phagocytosed areincreased avariouconcentrationof LBP+POgroup(P＜0.01).Flow cytometry showed thathe autofluorescence value in the POcontrol group wamore highethan thaof the normal control group.Athe LBP dose increased,the autofluorescence value in the RPE celldeclined gradually and iwaneathe normal value in the 1.00 g/L LBP+ POgroup.The rate of proliferation of the lipofuscin RPE cellvaried with the increase of doseof LBP with the maximal value in the normal RPE group and minimal value in the lipofuscin RPE group,and the rate of proliferation of the lipofuscin RPE cellascended with increasing doseof LBP until neathe normal value in the 1.00 g/L LBP + lipofuscin RPE cellgroup (P＞0.05).ConclusionLBP enhance the anti-aging effecof human RPE cellby strengthening the phagocytiability to POand the ability to remove lipofuscin and by heightening the proliferation of human RPE cells.

Abstract?AIM: To investigate the influence of propylene glycol mannite sulfate ( PGMS ) on the expression of tumor necrosis factor -α( TNF-α) and interleukin-1β( IL-1β) , in diabetic retinopathy by a rat model, to study the mechanism of PGMS against diabetic retinopathy, and provide a reliable theoretical and experimental evidence for the PGMS to be applied to clinical prevention and treatment of diabetic retinopathy.?METHODS: Male Wistar rats were randomized into 4 groups, normal control group, diabetic control group and PGMS in group, the PGMS in groups included the doses of 50mg/kg and 100mg/kg. 1% streptozotocin ( STZ) of 60 mg/kg was intraperitoneally injected in rats to establish the diabetic models. The PGMS with the doses of 50mg/kg and 100mg/kg were used to gavage in different groups of models for 12wk.Twelve weeks later, the animals were sacrificed and retinas were isolated. The aqueous humors and serums were taken, expressions of TNF-αand IL-1βprotein in retinas, aqueous humors and serums were detected by enzyme-linked immunosorbent assay ( ELISA) , respectively.The location and the expression of TNF-αand IL-1βprotein in retina tissue was detected by immunohistochemistry.?RESULTS: Twelve weeks after the use of PGMS, the level of blood glucose was not changed.ELISA showed that the expression of TNF-αand IL-1βprotein in serum and retina was significantly increased in diabetic control group than in normal control group(P<0.05), but in the groups which PGMS was given reduced, lower than those in diabetes mellitus( DM) group, especially as the concentration of PGMS increased ( P<0.05 ).But the levels of aqueous humor's TNF-αand IL-1βproteins in PGMS group were not reduced.Immunohistochemistry showed that the TNF -α protein was almost not expressed in normal control group. But the TNF-αprotein was highly expressed in diabetic control group. The expression mainly located in the ganglion cell layer, the inner plexiform layer, outer plexiform layer and pigment epithelium. The TNF-αprotein was weakly expressed at the group of 50mg/kg PGMS, the TNF-αprotein was almost not expressed at the group of 100mg/kg PGMS.When the normal control group was detected, the IL-1βprotein was weakly expressed in the outer plexiform layer.But the IL-1βprotein was also highly expressed in diabetic control group.The expression mainly located in the inner plexiform layer, outer plexiform layer and pigment epithelium. The IL -1βprotein was weakly expressed at the group of 50mg/kg and 100mg/kg PGMS.?CONCLUSION:PGMS can treat the diabetic retinopathy by downregulating the expressions of TNF-αand IL-1βin early diabetic retinopathy.PGMS maybe have a good control effect on early diabetic retinopathy.

Adult ; Female ; Humans ; Lead ; toxicity ; MMPI ; Male ; Middle Aged ; Occupational Exposure ; Personality ; drug effects

Objective To investigate the meaning of expression of apoptosis related molecules NFKBp65 and p53 and GPX1-mRNA in patients with Keshan disease(KSD).Methods Sixteen chronic Keshan Disease patients were enrolled in KSD group according to electrocardiogram,chest X ray film and clinical examinations on 15,September in 2009,and 23 healthy people were included in control group from physical examination taken in The Second Affiliated Hospital of Xi'an Jiaotong University.Fresh blood(5 ml)was collected from antecubital vein of all subjects in the fasting state.Total mRNA and protein of blood sample were isolated using Trizol.GPX Assay Kit was used to detect GPX enzyme activity,and GPX1-mRNA expression was determined by SYBR Real-Time PCR.Meanwhile,expression of apoptosis related molecules NFKBp65 and p53 were determined by Western blot.Results GPX enzyme activity decreased significantly in KSD group[(108.61±14.10)U]compared with control group[(122.78±11.89)U,t=2.874,P<0.05],GPX1-mRNA level of KSD group(0.553±0.299)notably KSD group(0.802±0.057)compared with control group[(1.065±0.355),t=6.829,P<0.01].p53 increased in KSD group(1.604±0.191)compared with control group[(1.137±0.186),t=3.033,P<0.05].Conclusiom Decreased GPX1-mRNA expression may result in lower GPX enzyme activity of patients with KSD.Thus oxidative damage increases and cadioeyte apoptosis is activated by activating apoptosis signal pathway.

To obtain a number of extracellular penicillinase,the gene(penp)encoding penicillinse of Bacillus cereus ATCC10987 was cloned into expression vector in Bacillus subtilis,transformed into Bacillus subtilis DB104 deficient in two proteases.When recombinant bacteria was cultured in LB medium for 24 hours,the result of SDS-PAGE showed that the molecular weight of the protein was 28kDa and the penicillinase activity reached 339U/ml.By screening seven different fermentation media,the result showed that 4# medium is favorable to producing penicillinase by the recombinant cells than the others,with the yield of the enzyme being 1580U/ml.When the recombinant cells was cultured in 7 liter fermentor for 24h,the penicillinase activity reached 1255.8U/ml.

Objective To explore the relationship between dynamic changes in ultra-micro-structural of pulmonary arteries and endogenous hydrogen sulfide in rats with left-right shunt.Methods Rats in shunt group were subjected to an abdominal aorta-inferior vena cava shunt to create an animal model of pulmonary artery structural remodeling. After 1 day, 3 days, 1 week, 4 weeks and 8 weeks of experiment, the ultra-micro-morphologic changes of pulmonary arteries of rats were observed under electronic microscope and H_2S concentration in serum was evaluated by modified sulfide electrode method.Results The changes of ultra-micro-structure of pulmonary arteries were progressively exacerbated, endothelial cells became swollen and large in size on 3 days, smooth muscular cells increased in size as well as the change of endothelial cells in 1 week, and they changed from contractile phenotype to synthetic phenotype in 4 weeks.Conclusions Shunt exhibited changes of ultra-micro-structure of pulmonary arteries are accompanied by the changes of endogenous H_2S. It is suggested that endogenous H_2S might play a protective role in changes of ultra-micro-structure of pulmonary artery.

Objective To observe the sequence of fat deposit and its relationship with insulin resistance in SD rats fed by high fat diet.Methods Normal 8-week-old male SD rats were randomly divided into normal chow (NC,n=40)and high fat diet(HF,n=40)groups.Triglyceride(TG)in serum,liver and muscle were measured;glucose infusion rate(GIR)and the mRNA level of genes related to lipid metabolism in liver and muscle were determined in different stages.GIR was detected by eugiyeemic-hyperinsulinemic clamp for evaluating the insulin sensitivity.Gene expression was determined by real-time PCR.Results(1)As compared with NC group,serum TG was not increased after high fat feeding for4 and 8 weeks,it began to increase after 12 weeks [0.52(0.15-1.00) mmol/L vs O.31(0.09-0.53)retool/L, P0.05)in skeletal muscle.After 8 weeks,the expression of ACC1 in liver in HF group was increased by 20.6%,CPT-1 was decreased by 27.1%(P

OBJECTIVETo explore the in vitro effect on control of graft-versus-host disease (GVHD) and its mechanism in mice by blockade of CD137-CD137L pathway.

METHODSResponder spleen cells from BALB/c donor mice (H-2(d)) were incubated with stimulator spleen cells from C57BL/6 ( H-2(b)) recipient mice, with or without anti-CD137L mAb. Lethally irradiated C57BL/6 mice were transplanted with donor bone marrow cells plus primary MLC spleen T cells. Group A (Allo-BMT control group): allo-BMT mice not receiving any prevention measures for GVHD. Group B (CsA + MTX control group): CsA and MTX given to C57BL/6 mice after transplantation. Group C (experimental group): donor spleen cells from BALB/c mice treated with anti-CD137L mAb. The percentages of CD3+ CD8+ T and CD3+ CD4+ T cells in the three groups were detected by flow cytometry, and the level of cytokines (IFN-gamma, IL-2, IL-10, IL-4) by RT-PCR.

RESULTSThe incidence of GVHD in group C was 70%, while in group A and group B were 100%. The survival rate was higher and the median survival time was longer of group C than that of group A and B (P < 0.01). All mice in group A died of aGVHD within 15 ds, while 30% of mice in group C survived more than 30 ds. Symptoms and histological signs of GVHD in group C were the mildest among the three groups. The percentage of CD3+ CD8+ T cells and the levels of IFN-gamma were significantly lower (P < 0.01), and the levels of IL-10 were significantly higher in group C than those in group A and B (P < 0.01).

CONCLUSIONTreatment of donor T cells with anti-CD137L mAb in vitro may relieve GVHD, thereby improve the survival time and survival rate, which maybe related to increasing Th1 cytokine (IFN-gamma) and decreasing Th2 cytokine (IL-10) as well as reducing CD3+ CD8+ T cells.

4-1BB Ligand ; immunology ; Animals ; Antibodies, Monoclonal ; immunology ; pharmacology ; Bone Marrow Transplantation ; immunology ; Disease Models, Animal ; Female ; Graft vs Host Disease ; immunology ; prevention & control ; Male ; Mice ; Mice, Inbred BALB C ; Mice, Inbred C57BL ; Transplantation, Homologous ; immunology ; Tumor Necrosis Factor Receptor Superfamily, Member 9 ; immunology

The aim of this study was to investigate the mechanism to reverse the drug resistance of leukemia cells in tetrandrine (Tet) alone or in combination with droloxifen (Drol) by using protein chips and to lay the theoretical basis for the clinical applications. Three monoclonal antibodies against P-glycoprotein (P-gp), the multidrug resistance-associated protein (MRP1) and the breast cancer resistance protein (BCRP) were immobilized onto the agarose gel film-coated glass slides. Protein chips were prepared respectively from K562/A02 cells cultured for 12, 24 and 48 hours with Tet alone or in combination with Drol. The results showed that Tet alone or in combination with Drol could decrease only the expression of P-gp in a time-dependent manner, the effect for 48 hours as follows: Tet + Drol 82.620 +/- 3.227; Tet alone 86.440 +/- 2.906; Drol alone 87.230 +/- 2.049; control 93.670 +/- 2.748 (P < 0.05). However, down-regulation of P-gp by K562/A02 cells cultured with Tet alone or in combination with Drol began at 24 hours (Tet + Drol 85.270 +/- 3.095; control 93.670 +/- 2.748, P < 0.05). The results were coincident with that of FCM. It is concluded that Tet and Drol can downregulate the expression of P-gp in the time-dependent way. There is a significant difference between Tet alone and Tet combined with Drol at 24 hours (P < 0.05). The expression of MRP1 and BCRP are not closely correlated with the reversal mechanism of Tet and Drol, and which may be involved in the mechanism of this combination to reverse multidrug resistance in leukemia.
ATP Binding Cassette Transporter, Sub-Family G, Member 2 ; ATP-Binding Cassette Transporters ; biosynthesis ; ATP-Binding Cassette, Sub-Family B, Member 1 ; biosynthesis ; Antineoplastic Agents ; pharmacology ; Benzylisoquinolines ; pharmacology ; Drug Resistance, Multiple ; Drug Resistance, Neoplasm ; drug effects ; Drug Synergism ; Humans ; K562 Cells ; Leukemia ; metabolism ; pathology ; Multidrug Resistance-Associated Proteins ; biosynthesis ; Neoplasm Proteins ; biosynthesis ; Protein Array Analysis ; Tamoxifen ; analogs & derivatives ; pharmacology

Preparative HPLC was used to prepare ferulic acid, senkyunolide I and senkyunolide H from Ligusticum chuanxiong. The separation was conducted on a Shim-Pack Prep-ODS (20.0 mm x 250 mm, 5 microm) column with the mobile phase of methanol-0.2% glacial acetic acid (50:50)at the flow rate of 5 mL x min(-1). The detection wavelength was 278 nm, and the purity of each compound was detected by HPLC analysis. Spectral data analyses including UV, ESI-MS and NMR were used to identify their structures. This method is simple, fast, which is suitable for preparation of standard reference of ferulic acid, senkyunolide I and senkyunolide H from L. chuanxiong and can meet the requirement of new drug research and development.
Benzofurans ; chemistry ; isolation & purification ; Chromatography, High Pressure Liquid ; methods ; Coumaric Acids ; chemistry ; isolation & purification ; Ligusticum ; chemistry