OBJECTIVEA R1210C mutation of complement factor H (CFH) gene has been associated with age-related macular degeneration (AMD) in Caucasian population. This study was to verify above association in Han Chinese population.

METHODSThe mutation was detected by direct sequencing in 258 patients with wet AMD and 426 matched controls.

RESULTSThe R1210C mutation has not been identified in either sample.

CONCLUSIONThe R1210C mutation in CFH gene is not associated with AMD in Han Chinese population.

Aged ; Aged, 80 and over ; Complement Factor H ; genetics ; Female ; Humans ; Macular Degeneration ; genetics ; Male ; Middle Aged ; Mutation

OBJECTIVETo investigate whether resveratrol (RES) plays a protective role in hypothermic preserved isolated rat hearts and whether it is mediated by regulation of silent information regulator protein-1 (Sirt-1) expression.

METHODSThe Langendorff model of isolated rat heart was used. After stored in different Celsior solution at 4 degrees C for 9 h, SD rat hearts were randomly divided into 7 groups: blank control group;9 h group (soley hypothermic preservation for 9 h); RES group (3, 10, 30 micromol/L RES treatment plus hypothermic preservation for 9 h ), niacinamide (NAM) group (40 micromol/L NAM added in Celsior solution plus hypothermic preservation for 9 h), RES + NAM group (30 micromol/L RES and 40 micromol/L NAM were added in Celsior solution plus hypothermic preservation for 9 h). The morphological changes of cardiomyocytes were detected by the HE staining with the light microscope. The mRNA and protein expression levels of Sirt-1 were detected by Real-Time PCR and Western blot respectively.

RESULTS(1) Compared with the blank control group, myocardiocytes were injured remarkably in the 9 h group and the Sirt-1 mRNA and protein expression levels were decreased significantly (P < 0.01); (2) Compared with the 9 h group, rat myocardial injury was alleviated gradually in 3, 10, 30 micromol/L RES group and the Sirt-1 mRNA and protein expression levels were increased in a dose-dependent manner (P < 0.05); (3) The above protective effects of RES were attenuated by Sirt-1 inhibitor NAM.

CONCLUSIONRES can protect myocardiocytes from injury caused by long range hypothermic preservation and this protective effect maybe mediated by upregulation of Sirt-1 expression.

Animals ; Cryopreservation ; Heart ; drug effects ; Male ; Organ Preservation ; Rats ; Rats, Sprague-Dawley ; Sirtuin 1 ; metabolism ; Stilbenes ; pharmacology

OBJECTIVETo investigate the role of the expression of PPAR-gamma gene in the adipogenesis in hemangioma evolution.

METHODSRoutine immunohistochemistry staining of Perilipin A, the marker antigen of adipocytes, was performed to observe the adipogenesis in hemangioma. Immunofluorescence staining of PPAR-gamma, the important transcription factor in promoting adipogenesis, was carried out to observe its location in hemangioma tissue, with the co-staining of alpha-SMA and CD31. And RT-PCR was used to examine the expression of PPAR-gamma gene in hemangioma in different stages.

RESULTSIn the evolution of hemangioma, the number of adipocytes increased continuously. And the tumor was replaced by fibrofatty tissue finally. PPAR-gamma was located in the nuclei of perivascular cell in hemangioma tissue. The expression of PPAR-gamma gene in hemangioma increased in the evolution of hemangioma, but still was lower than that in normal fat tissue from children.

CONCLUSIONThe expression of PPAR-gamma in the perivascular cells suggests that they may contribute to the adipogenesis in hemangioma involution.

Adipogenesis ; Adipose Tissue ; metabolism ; pathology ; Hemangioma ; metabolism ; pathology ; Humans ; PPAR gamma ; metabolism

The purpose of the present study was to investigate the effect of sodium nitrite (SN) on alcohol-induced acute liver injury in mice. Forty male C57bL/6 mice were randomly divided into 4 groups. Acute alcohol-induced liver injury group were injected intraperitoneal (ip) with alcohol (4.5 g/kg); SN preconditioning group were pretreated with SN (16 mg/kg, ip) for 12 h, and received alcohol (4.5 g/kg, ip) injection; Control and SN groups were treated with saline and SN, respectively. After the treatments, liver index (liver/body weight ratio) was determined. Colorimetric technique was performed to measure the serum alanine transaminase (ALT), aspartate transaminase (AST), liver superoxide dismutase (SOD), glutathione peroxidase (GSH-Px), catalase (CAT) activities, as well as malondialdehyde (MDA) content. The pathological index of liver tissue was assayed by HE and TUNEL fluorometric staining. Using Western blot and immunohistochemistry staining, the expression of hypoxia-inducible factor-1α (HIF-1α) protein was detected. The results showed that, compared with acute alcohol-induced liver injury group, pretreatment with low doses of SN decreased liver index and serum levels of ALT and AST, weakened acute alcohol-induced hepatocyte necrosis, improved pathological changes in liver tissue, increased live tissue SOD, GSH-Px and CAT activities, reduced MDA content and apoptosis index of hepatocytes, and up-regulated HIF-1α protein level in liver tissue. These results suggest that the pretreatment of SN can protect hepatocytes against alcohol-induced acute injury, and the protective mechanism involves inhibition of oxidative stress and up-regulation of HIF-1α protein level.
Alanine Transaminase ; metabolism ; Alcohols ; adverse effects ; Animals ; Apoptosis ; Aspartate Aminotransferases ; metabolism ; Chemical and Drug Induced Liver Injury ; drug therapy ; Glutathione Peroxidase ; metabolism ; Hepatocytes ; drug effects ; Hypoxia-Inducible Factor 1, alpha Subunit ; metabolism ; Male ; Malondialdehyde ; metabolism ; Mice ; Mice, Inbred C57BL ; Oxidative Stress ; Protective Agents ; pharmacology ; Sodium Nitrite ; pharmacology ; Superoxide Dismutase ; metabolism ; Up-Regulation

AIM: The effects of ZX-5, as nitric oxide (NO) donor, on ocular blood flow has been investigated using colored microsphere technique in previous study. The relationship between the production of NO by ZX-5 and ocular blood flow has been evaluated. ZX-5 has been shown to have strong positive effect on increasing choroidal blood flow. However,the effect of ZX-5 on retinal function recovery, the effects of its optical isomers, (R, R)-ZX-5 and (S, S)-ZX-5, on choroidal blood flow and retinal function recovery have not been studied and merit investigation.METHODS: Colored microsphere technique was used for in vivo experiments to determine choroidal blood flow of ocular hypertension (40mmHg) in rabbit eyes. Electroretinography was used to measure the b-wave recovery as an indication of retinal function recovery.RESULTS: (R, R)-ZX-5 increased choroidal blood flow at 10g/L, 50μL instillation into eyes at all time points (P＜0.05).(S, S)-ZX-5 was not effective in increasing choroidal blood flow. ZX-5 and (R, R)-ZX-5 showed significant effects in retinal function recovery after ischemia of the retina at all time points (P＜0.05); whereas (S, S)-ZX-5 did not show significant effect on recovery of b-wave after ischemia at most time points except at 120 and 240 minutes.CONCLUSION: ZX-5 and (R, R)-ZX-5 have high potency in increasing the choroidal blood flow and improving the retinal function recovery. It is hoped that they could be used for the prevention/treatment of ocular blood flow related eye diseases.

OBJECTIVE: To explore expression of β-catenin and NF-κB signaling pathway in synovial tissue of rats with different degrees of knee osteoarthritis (KOA). METHODS: Forty-eight SPF male rats weighed (200±20) g were randomly divided into three groups, namely model group (32 rats), sham operation group (8 rats) and control group (8 rats). KOA model rats were established by Hulth method, and 8 rats were killed at 2, 4, 8, 12 weeks respectively after modeling, in order to establish KOA model rats with moderate, early, mild and severe degree. Sham operation group was only cut off capsule of knee joint and suture to exclude interference factor, control group was untreated. Behavior, synovial hyperplasia and cartilage degeneration of rats among each group were observed. Expression of NF-κB and signaling pathway and β-catenin in synovial tissue of rats were detected by real-time PCR. RESULTS: KOA rat model was successfully established, and synovial hyperplasia was observed in KOA model at mild and early degree, and then gradually decreased; while cartilage degeneration in the early moderate and severe KOA model was significantly expressed, and gradually aggravated with time. The results of PCR showed that expression of β-catenin in 4-week group (8.57±0.46) and 8-week group (4.23±0.09) were higher than those in control group (<0.05); expression of TLR-2 in 2-week group (12.04±4.02) and 4-week group (8.54±2.13) were higher than those in control group(<0.05), and TLR-4 in 2-week group(5.04±0.93), 4-week group (3.29±0.58) and 8-week group (1.63±0.12) were higher than those in control group; expression of NF-κB was significantly higher in 2-week group (10.15±2.04), 4-week group (15.97±4.17), 8-week group (7.69±1.48) and 12-week group (6.70±1.58) than that in control group (<0.05), and expression of IL-1β was significantly higher in 4-week group (2.79±0.25) and 8-week group (2.46±0.32) than that of control group (<0.05). CONCLUSIONS On the RNA expression level, both of β-catenin and NF-κB signaling pathways are involved in synovial inflammation in KOA model rats, and they play a regulatory role in expression of IL-1β, degeneration of KOA.
Animals ; Inflammation ; Male ; NF-kappa B ; Osteoarthritis, Knee ; Rats ; Signal Transduction ; beta Catenin

OBJECTIVETo construct and identify the stable expression system of NIH3T3 fibroblast with eukaryotic expression vector of human transforming growth factor beta3 (pcDNA3.1 (-)/TGFbeta3). So as to investigate the proliferation of NIH3T3 fibroblasts transfected with hTGFbeta3 gene stably.

METHODSThe stable transfection of NIH3T3 fibroblasts with recombinant plasmid expressing hTGFbeta3 was established by using LipofectamineTM2000 and G418 selection. The mRNA and protein expression of TGFbeta3 were detected by the RT-PCR and Western blot method, respectively. Microscope and MTT were adopted to examine the proliferation of the stable expression system of fibroblasts with hTGFbeta3.

RESULTSAfter G418 selection, RT-PCR and Western blot analysis, 7 out of 10 cell lines transfected with pcDNA3.1 (-)/TGFbeta3 expressed with very high level of TGFbeta3, as compared with vector control transfectants that showed no expression, and compared with the other cell lines that expressed relatively low level. The stable transfection of NIH3T3 fibroblasts growth slowed down significantly (P < 0.05).

CONCLUSIONThe stable expression system of NIH3T3 fibroblast with hTGFbeta3 were constructed successfully. The TGFbeta3 gene could inhibit the proliferation of NIH3T3 fibroblasts in vitro.

Animals ; Cell Proliferation ; Fibroblasts ; metabolism ; Humans ; Mice ; NIH 3T3 Cells ; Plasmids ; Transfection ; Transforming Growth Factor beta3 ; genetics ; metabolism

AIMTo look for new active constituents from Chinese medicine "Sheng-ma", rhizome of Cimicifuga foetida L.

METHODSThe compounds were separated and purified by chromatography on silica gel and Sephadex LH-20. Their structures were determined by spectral analysis and chemical reaction.

RESULTSEight compounds were obtained and identified as cimicifugic acid (1), esculetin (2), caffeic acid methyl ester (3), 4-O-acetyl-caffeic acid (4), sinapic acid (5), caffeic acid (6), ferulic acid (7), isoferulic acid (8).

CONCLUSIONCompound 1 is a new compound, and compounds 2-7 were isolated from this plant for the first time.

Caffeic Acids ; chemistry ; isolation & purification ; Cimicifuga ; chemistry ; Coumaric Acids ; chemistry ; isolation & purification ; Drugs, Chinese Herbal ; chemistry ; isolation & purification ; Hydroxybenzoates ; chemistry ; isolation & purification ; Molecular Conformation ; Molecular Structure ; Phenylacetates ; chemistry ; isolation & purification ; Plants, Medicinal ; chemistry ; Rhizome ; chemistry ; Umbelliferones ; chemistry ; isolation & purification

AIMTo look for new active constituents from the aerial part of Cimicifuga foetida L.

METHODSVarious column chromatographic techniques were used for the isolation and purification of the principles. The structures were elucidated on the basis of spectral data and chemical evidences.

RESULTSFive 9,19-cycloartane triterpenoid saponins and one sitosterol saponin were obtained and identified as cimifoetiside I [12 beta-hydroxycimigenol-3-O-beta-D-galactoyranoside, (1)], cimifoetiside II [(23R,24R) cimigenol-3-O-beta-D-galactopyranoside, (2)], cimigenol-3-O-beta-D-galactopyranoside (3), 12 beta-hydroxycimigenol-3-O-beta-D-xylopyranoside (4), 12 beta-hydroxycimigenol-3-O-alpha-L-arabinopyranoside (5), daucosterol (6).

CONCLUSIONCompounds 1 and 2 are new and compounds 4 and 5 were isolated from this plant for the first time.

Cimicifuga ; chemistry ; Drugs, Chinese Herbal ; chemistry ; isolation & purification ; Molecular Structure ; Plant Components, Aerial ; chemistry ; Plants, Medicinal ; chemistry ; Sitosterols ; chemistry ; isolation & purification

This study is to report the determination of the effect of sodium nitrite induced oxygen species (ROS) on the epithelial-mesenchymal transition in hepatoma cells in mice bearing H22 and investigation of its role in hypoxia-inducible factor 1alpha (HIF-1alpha) in this process. Mice hepatocarcinoma cell line H22 was inoculated subcutaneously into right axillary of sixty male Kunming mice and then randomly divided into four groups: control group; low-dose sodium nitrite group (10 mg x kg(-1)), medium-dose sodium nitrite group (20 mg x kg(-1)) and high-dose sodium nitrite group (30 mg x kg(-1)). Sodium nitrite group was given (ig) sodium nitrite with 10-30 mg x kg(-1) x d(-1) for 21 days. Compared with control group, there was no obvious difference between the two groups in the volume or weight of xenografts, but in sodium nitrite treatment group, the activity of SOD and CAT decreased and contents of MDA or nitrite increased in tumor tissue of mice bearing H22; epithelial-mesenchymal transition (EMT) of hepatoma cells was induced, the EMT-phenotype tumors displayed a greater degree of local aggressiveness, with dissection through adjacent fascia and skeletal muscle. The increased expression of HIF-la and vimentin and declination of E-cadherin were confirmed by immunohistochemistry and Western blotting. These data indicate sodium nitrite treatment could improve the epithelial-mesenchymal transition of xenografts in mice bearing H22, which might relate to the fact that ROS mediated signal pathway increased the expression of HIF-1alpha.
Animals ; Cadherins ; metabolism ; Carcinoma, Hepatocellular ; metabolism ; pathology ; Catalase ; metabolism ; Cell Line, Tumor ; Dose-Response Relationship, Drug ; Epithelial-Mesenchymal Transition ; drug effects ; Humans ; Hypoxia-Inducible Factor 1, alpha Subunit ; metabolism ; Liver Neoplasms ; metabolism ; pathology ; Male ; Malondialdehyde ; metabolism ; Mice ; Neoplasm Transplantation ; Random Allocation ; Reactive Oxygen Species ; metabolism ; Signal Transduction ; drug effects ; Sodium Nitrite ; administration & dosage ; pharmacology ; Superoxide Dismutase ; metabolism ; Tumor Burden ; drug effects ; Vimentin ; metabolism