1.Inhibition of mevastatin on inflammation and differentiation of orbital preadipocytes in thyroid-associated ophthalmopathy
Chinese Journal of Experimental Ophthalmology 2011;29(10):907-912
Background Inflammation and adipogenesis are two parallel processes with increasing activity in severe thyroid-associated ophthalmopathy(TAO),and mevastatin was proved to have the inhibiting effect on the differentiation of adipose.Objective The aim of this work was to investigate the effects of mevastatin on the expression of cyclooxygenase-2(COX-2)and peroxisome proliferator activated receptor-γ(PPAR-γ)and differentiation of TAO-derived orbital preadipocytes,and explore its modulation effects on lipopolysaccharide(LPS)induced inflammation and the differentiation of TAO-derived orbital preadipocytes in vitro.Methods The retroorbital adipose tissue was obtained from 4 TAO patients during the surgery.The orbital fibroblasts were cultured from orbital adipose tissues using explant culture method.To study the suppressing effect of mevastatin on inflammatory response,cultured cells were divided into 5 groups.The 1000 μg/L LPS orbital fibroblasts were stimulated for 8 hours in group A,and 1000 μg/L LPS combined with 5 μmol/L,10 μmoL/L or 20 μmoL/L mevastatin were used respectively for the substitute in the group B,group C and group D.The orbital fibroblasts in group E were cultured routinely without any intervention as control.To observe the inhibiting effect of mevastatin on the differentiation of adipose,the group A were then subdivided into group A1-A6.After 1000 μg/L LPS was used to treat the cells for 8 hours,the ceils were induced to differentiate into adipocytes.All orbital preadipocytes from A1 to A6 were stimulated to differentiate into mature adipocytes with cocktail differentiation medium for a 10-day duration.During the procedure,group A2,A3 and A4 were interfered with 5,10 or 20 μmol/L mevastatin,and in the group A5 and A6,10 μmol/L mevastatin were added at the fourth day or eighth day.Intracellular fat accumulation in differentiated adipocytes was determined by oil red O staining.The absorption(A492 nm)was measured in the ceils by enzyme-linked immunosorbent assay(ELISA).Expression of COX-2 and PPAR-γ mRNA was detected by reverse transcription polymerase chain reaction(RT-PCR),and the expression of COX-2 and PPAR-γ protein was detected by Westernblot.The level of PGE2 in the supernatant was detected by ELISA.Results The expression of COX-2 protein and mRNA as well as the PGE2 levels in B,C,D group decreased markedly in comparison with those in A group(P<0.05).With the increase of mevastatin concentration,the expression of COX-2 protein and mRNA as well as the PGE2 levels in B,C,D groups decreased successively(F =228.380,101.745,1586.881,P<0.05).The expression of COX-2 protein and mRNA and PGE2 levels in E group were lower significantly than those in A,B and C groups(P<0.05),but no significant differences were found between E group and D group(P>0.05).The A492 value and the expressions of PPAR-γ protein and mRNA in differentiated cells showed the successively decrease in A1-A4 group with the elevation of mevastatin concentration(P<0.05),and the evidently decreased A492 value and the expressions of PPAR-γ protein and mRNA also were seen in A1 and A5 groups compared with A3 group(P < 0.05).Conclusions Mevastatin inhibits LPS-induced COX-2 expression,PPAR-γ expression,PGE2 secretion and differentiation of TAO-derived orbital fibroblasts in vitro in dose-dependent manner.Mevastatin plays these effect more prominently in early stage of adipocytes differentiation.
2.Effect of different pacing sites on ventricular synchrony evaluated by gated blood pool SPECT
Xue, GONG ; Yan-gang, SU ; Wen-zhi, PAN ; Shu-guang, CHEN ; Hong-cheng, SHI ; Xian-hong, SHU ; Jun-bo, GE
Chinese Journal of Nuclear Medicine 2010;30(5):307-311
Objective To compare the effect of right ventricular outflow tract (RVOT) and right ventricular apex (RVA) pacing on ventricular systolic synchrony using gated blood pool SPECT (GBPS).Methods A total of 50 patients implanted with pacemaker due to high degree or complete atria-ventricular block were enrolled in the study. Twenty-three patients were RVOT paced ( Group A, n = 23) and 27 were RVA paced (Group B, n=27). Twenty-four patients with malignancy, normal echocardiographic findings and no history of cardiac diseases were scheduled for pre-chemotherapy evaluation of cardiac structure and function and were enrolled as control group ( Group C, n = 24). All patients underwent GBPS imaging and the values of phase angle (PS), mean phase of each wall, standard deviation (SD) of mean phase of each wall, lateral-septal motion delay of left ventricle ( LV Sep-Lat Delay), septal-right ventricular (RV) delay of LV ( LV Sep-RV Delay) and LV-RV Delay were acquired. The parameters of ventricular systolic synchrony among the three groups were compared using one-way ANOVA. Results The mean phase of LV lateral wall in Groups A and B were significantly higher than that in Group C: Group A (120.50 ±40.58) ms; Group B (103.23±28.34) ms; Group C (84.63 ±22.38) ms (F=7.72, P <0.05). There was no significant difference between Groups A and B ( t = 1.30, P > 0.05 ). The mean phase of RV in Group A was significantly larger than those in Groups B and C: Group A ( 137.05 ± 39.27) ms, Group B ( 100.85 ± 23.79) ms,Group C (59. 13 ±30.52) ms (F=35.55, P<0.05). PS, SD and LV Sep-Lat Delay in Groups A and B were significantly higher than those in Group C: (85.73 ± 12.00)°vs (89.85 ± 15.61 )°vs (58.95 ±9.87)°, (27.68±10.66) ms vs (26.15 ±13.02) ms vs (15.63 ±8.35) ms, (25.06±34.23) ms vs (2. 62 ± 60. 31 ) ms vs ( - 23.66 ± 31.39) ms, F = 41.54,8.55,6.81, all P < 0.01 ), however, there was no significant difference between Groups A and B ( t = 0. 68, 0.68, 1.30, all P > 0.05 ). LV Sep-RV Delay and LV-RV Delay were significantly different among the three groups ( LV Sep-RV Delay: Group A (57.60 ±56.77) ms, Group B (6.36 ±61.88) ms, Group C ( -41.89 ±35.78) ms; LV-RV Delay:Group A (47.36 ±42.59) ms, Group B ( 3.08 ± 38.81 ) ms Group C ( - 26.50 ± 20.99 ) ms, F = 20. 32,25.38, both P < 0.01 ). Conclusion Both RVA and RVOT pacing increase the segmental phases detected by GBPS, causing inter- and intra- ventricular asynchrony compared with patients without pacemakers.
4.Construction,Identification and Expression of Recombinant Eukaryotic Vector pCAG-IRES-SHIP-GFP on Porliferation of Leukemia Cell Line K562
Lin YANG ; Jian-Min LUO ; Xiao-Jun LIU ; Zhi-Yong CHENG ; Shu-Peng WEN ; Xiao-Yang YANG ; Xue-Wen WU ;
China Biotechnology 2006;0(06):-
The aim was to construct and identify the mammalian expression vector of pCAG-IRES-SHIP-GFP and to detect whether it could express in human acute leukemia cell line K562.The cDNA fragment of SHIP obtained by RT-PCR was inserted into pCAG-IRES-GFP.The recombinant plasmid was confirmed by restriction enzyme digesiton,PCR and DNA sequecing.pCAG-IRES-SHIP-GFP was transfected into K562 cells with lipofectamine 2000.The expression of SHIP was determined by GFP fluorescence and Western blot analysis.FQ-PCR was used to quantitate SHIP mRNA.The expression of p-Akt,Akt were determined by Western blot.PI were tested by flow cytometry and MTT to verify whether exogenous SHIP could inhibit proliferation of K562 cells.The results showed that the correct constrution of the recombinant plasmid pCAG-IRES-SHIP-GFP has been shown by restriction enzyme digestion,PCR and DNA sequencing.pCAG-IRES-SHIP-GFP could express SHIP protein in K562 cells.The K562 cells viability after transfected with SHIP gene droped down.Western blot analysis showed that phospha-Akt308 and Akt473 were reduced to 38.7% and 68% respectively.It was concluded that the vector of pCAG-IRES-SHIP-GFP has been successfully constructed and it can be expressed in K562 cells.The expression of exogenous SHIP gene can lead to apoptosis of K562 cells by down-regulating the p-Akt expression.What found here might be one of the mechanisms involved in the pathogenesis of leukemia.
5.Protective and antioxidant properties of wasp (Vespa magnifica) honeycomb extract: a potential inhibitor against acidified ethanol-induced gastric lesions.
Xue-qing XU ; Le YU ; Shu-wen LIU
Journal of Southern Medical University 2011;31(7):1107-1110
OBJECTIVETo examine the protective effects of wasp (Vespa magnifica) honeycomb extract (WCE) against gastric lesions in rats induced by 60% acidified ethanol, and evaluate its capacity to suppress oxidative stress in the gastric tissue.
METHODSWistar rats were subjected to intragastric administration of 60% acidified ethanol to induce gastric lesions following an 8-day oral pretreatment with WCE at 0, 25, 100 and 150 mg/kg or with saline. The levels of 1,1-diphenyl-2-picrylhydrazyl (DPPH) free radical scavenging, myeloperoxidase (MPO) activity and total antioxidant capacity in the gastric tissues were determined.
RESULTSOral administration of 25, 100 and 150 mg/kg WCE prior to 60% acidified ethanol administration significantly inhibited the formation of gastric lesions (with a reduction by 44.2%-87.1%), decreased the mucosal MPO activity (by 16.4%-56.6%) and increased the total antioxidant capacity of the gastric tissue (by 0.5, 1.47 and 1.83 folds, respectively) in a dose-dependent manner. At a high concentration (above 1 mg/ml), WCE also exhibited a stronger DPPH radical scavenging activity than butylated hydroxytoluene (BHT).
CONCLUSIONThe ethanol extract of wasp honeycombs can suppress the formation of acidified ethanol-induced gastric lesions by reducing free radical oxidation and neutrophils infiltration in the gastric tissue in rats.
Animals ; Antioxidants ; pharmacology ; therapeutic use ; Ethanol ; adverse effects ; Female ; Honey ; Male ; Materia Medica ; pharmacology ; therapeutic use ; Neutrophil Infiltration ; drug effects ; Oxidative Stress ; drug effects ; Peroxidase ; metabolism ; Rats ; Rats, Wistar ; Stomach Ulcer ; chemically induced ; prevention & control ; Wasps ; chemistry
6.Protective effect of soyasaponins on acute liver injury induced by D-galactosamine and lipopolysaccharide in mice.
Hui-Xian XU ; Wen-Xi ZHAO ; Ji-Shu QUAN ; Xue-Zhe YIN
China Journal of Chinese Materia Medica 2013;38(13):2187-2190
OBJECTIVETo investigate the protective effect of soyasaponins on acute liver injury induced by D-galactosamine (GalN) and lipopolysaccharide (LPS) in mice.
METHODThe mice were randomly divided into five groups: the normal control, the model group, the silymarin (positive control) group, and soyasaponins high and low-dose groups. They were administered with drugs once every day for 7 days. At the end of the experiment, GalN and LPS were injected intraperitoneally to all of the groups except for the normal group to establish the acute liver injury model. The pathological changes were detected with hematoxylin & eosin (HE) staining, tumor necrosis factor-alpha (TNF-alpha) was detected by ELISA method, and the alanine aminotransferase (ALT), aspartate aminotransferase (AST), catalase (CAT), glutathione peroxidase (GPx), glutathione S-transferase (GST), reduced glutathione (GSH), malondialdehyde (MDA), nitric oxide (NO), and the activation of Caspase-3 and Caspase-8 were detected by the colorimetric method.
RESULTSoyasaponins could reduce the activities of serum ALT and AST, the acute hepatic injury induced by GalN/LPS, serum TNF-alpha level, hepatic NO and MDA contents, and the Caspase-3 and Caspase-8 activations of liver tissues, and increase the hepatic CAT, GPx, GST and GSH levels.
CONCLUSIONSoyasaponins shows the protective effect on acute liver injury induced by GalN and LPS in mice, which may be related to its antioxidative ability and anti-liver apoptosis.
Alanine Transaminase ; blood ; Animals ; Antioxidants ; metabolism ; Apoptosis ; drug effects ; Aspartate Aminotransferases ; blood ; Caspases ; metabolism ; Chemical and Drug Induced Liver Injury ; metabolism ; pathology ; prevention & control ; Galactosamine ; toxicity ; Lipopolysaccharides ; toxicity ; Liver ; pathology ; Male ; Mice ; Saponins ; pharmacology ; Soybeans ; chemistry
7.Effect of the optimized traditional acupuncture prescription for accouchement on cervical ripening in late-stage pregnant rats.
Li-Wen XUE ; Hong-Wen YUAN ; Ying-Ru CHEN ; Fu-Zheng SHU ; Liang-Xiao MA ; Jiang ZHU
Chinese Acupuncture & Moxibustion 2012;32(12):1105-1107
OBJECTIVETo observe the mechanism of the optimized traditional acupuncture prescription for accouchement on cervical ripening based on the molecular biology by observing related indices of cervical ripening in late-stage pregnant rats.
METHODSTwenty initial pregnant Wistar rats were randomly divided into an electroacupuncture (EA) group (n = 10) and a model group (n = 10), and other 10 non-pregnancy female rats with same lot were selected as a blank control group. EA group was treated with the optimized traditional acupuncture prescription for accouchement on the 20th day of pregnant, which performed EA at bilateral "Hegu" (LI 4) for 20 min and then at bilateral "Sanyinjiao" (SP 6) for 5 min with 2 Hz/50 Hz sparse-dense wave, while the other groups without acupuncture intervention. The contents of matrix metalloproteinase 9 (MMP-9) and interleukin 8 (IL-8) in cervix tissue were detected by ELISA method.
RESULTSCompared with the blank control group, the contents of MMP-9 and IL-8 in the model group were increased significantly (both P < 0.01). Compared with the model group, the contents of MMP-9 and IL-8 in the EA group were increased significantly (P < or = 0.05).
CONCLUSIONOptimized traditional acupuncture prescription for accouchement can increase the contents of MMP-9 and IL-8 in cervix tissue of late-stage pregnant rats so as to promote cervical ripening, and the mechanism of EA in promoting cervical ripening is explained from the perspective of molecular biology.
Acupuncture Points ; Acupuncture Therapy ; Animals ; Cervical Ripening ; metabolism ; Cervix Uteri ; enzymology ; Female ; Humans ; Interleukin-8 ; metabolism ; Matrix Metalloproteinase 9 ; metabolism ; Models, Animal ; Pregnancy ; Rats ; Rats, Wistar
8.Efficacy and Safety Evaluation of Bushen Shuji Granule in Treating Ankylosing Spondylitis Patients: a Clinical Study.
Wei-ping KONG ; Qing-wen TAO ; Ying-ze ZHANG ; Shu YANG ; Yuan XU ; Xiao-xia ZHU ; Yue JIN ; Wen-xue YANG ; Xiao-ping YAN
Chinese Journal of Integrated Traditional and Western Medicine 2015;35(6):673-677
OBJECTIVETo evaluate the short-term efficacy and safety of Bushen Shuji Granule (BSG) in treating ankylosing spondylitis (AS) patients.
METHODSA prospective randomized controlled clinical trial was carried out in 62 active stage AS patients with Shen deficiency Du-channel cold syndrome (SDDCS), who were randomly assigned to the BSG group (treated with BSG) and the control group (treated with Celecoxib Capsule). Twelve weeks consisted of one therapeutic course. Therapeutic effects were evaluated by ASAS20 and ASAS40 (set by Assessments in Ankylosing Spondylitis working group) , BASDA150, Chinese medical (CM) syndrome efficacy evaluation standards. BASDAI, the Bath Ankylosing Spondylitis Functional Index (BASFI), the Bath AS Metrology Index (BASMI), scores for spine pain, scores for pain at night, patient global assessment (PGA) , erythrocyte sedimentation rate (ESR) , and C reactive protein (CRP) were observed before and after treatment.
RESULTSAfter three-month treatment by BSG, ASAS20 standard rate was 63. 33% (19/30 cases) in the BSG group and 66.67% (20/30 cases) in the control group with no significant difference between the two groups (χ2 = 0.073, P > 0.05). The efficacy for CM syndromes was 70.00% (21/30 cases) in the BSG group, higher than that in the control group [40.00% (12/30 cases), χ2 = 5.455, P < 0.05]. Scores for CM syndromes, BASDAI, night pain index, spinal pain index, PGA, CRP were improved in the BSG group (P < 0.05, P < 0.01). The incidence of adverse events in the BSG group was lower than that of the control group.
CONCLUSIONBSG based on Shen supplementing, Du-channel strengthening, blood activating, and channels dredging method had good short-term clinical efficacy and safety in treating AS.
Asian Continental Ancestry Group ; Biomedical Research ; Blood Sedimentation ; C-Reactive Protein ; Drugs, Chinese Herbal ; therapeutic use ; Humans ; Pain ; Prospective Studies ; Safety ; Spondylitis, Ankylosing ; drug therapy
9.GLB1 gene mutation and clinical characteristics of a patient with mucopolysaccharidosis type IVB.
Hong-lin LEI ; Jun YE ; Wen-juan QIU ; Hui-wen ZHANG ; Lian-shu HAN ; Xue-fan GU
Chinese Journal of Pediatrics 2012;50(7):549-553
OBJECTIVETo report the results of clinical characteristics, enzyme activity determination and mutation analysis of GLB1 gene in a Chinese patient with mucopolysaccharidosis (MPS) type IVB (Morquio B disease).
METHODA 14-year-old Chinese boy with MPS type IVB was firstly diagnosed by blood leucocytes galactosamine-6-sulfate sulfatase (GALNS) and β-galactosidase (GLB1) determination, who was characterized by short stature, multiplex skeletal abnormalities, difficulty in walking. PCR-sequencing analysis was applied to detect the mutations in GLB1 of the patient.
RESULTThe patient was characterized by dwarfism, pectus carinatum, kyphosis, normal intelligence, and no neurologic damage of spasms, linguistic capacity and so on. The patient had normal GALNS enzyme activity and very low GLB1 enzyme activity [5.03 nmol/(h·mg) vs. normal value 118 - 413 nmol/(h·mg) ] in leukocytes. A compound heterozygous missense mutations c.442C > T(p.R148C)/c.1454A > G(p.Y485C) in GLB1 gene were detected in this patient. The mutation p.Y485C is a novel variant. With the method of gene analysis of new variant, the mutation p.Y485C was considered to be a pathogenic mutation.
CONCLUSIONThe MPS IVB patient showed severe multiple skeletal deformities, normal intelligence, no neurologic damage and very low GLB1 enzyme activity, who carries compound heterozygous mutations p.R148C/p.Y485C. The mutation p.Y485C in GLB1 gene may be a novel pathologic mutation of MPS type IVB.
Adolescent ; Amino Acid Sequence ; Asian Continental Ancestry Group ; genetics ; Chondroitinsulfatases ; genetics ; metabolism ; DNA Mutational Analysis ; Humans ; Joints ; pathology ; Male ; Molecular Sequence Data ; Mucopolysaccharidosis IV ; enzymology ; genetics ; pathology ; Mutation, Missense ; Pedigree ; Polymerase Chain Reaction ; Radiography ; Spine ; diagnostic imaging ; pathology ; beta-Galactosidase ; genetics ; metabolism
10.Treatment of AIDS patients with Chinese medicinal herbs qudu zengning capsule.
Ze-lin LI ; Zhong-min WANG ; Xue-zhou LIU ; Zu-shu ZHANG ; Zhe WANG ; Shi-wen MA ; Chun-hua CHEN ; Xiao-ling XUE ; Rui-xing WEN ; Yan-chao YUE ; Xin-peng ZHU ; Yi ZENG
Chinese Journal of Experimental and Clinical Virology 2004;18(4):305-307
OBJECTIVETo evaluate the therapeutic effect of QuDu ZengNing Capsule on AIDS.
METHODSQuDu ZengNing Capsule is a capsule containing extract from 4 Chinese medicinal herbs. Totally 1,000 AIDS patients were treated, among them 60 patients were clinically observed weekly. Blood routine tests, liver, heart and kidney function, X-ray, CD4, CD8 cells were examined before and after treatment at 1, 3, 6 month. The patients were treated with 4 capsules t.i.d for 6 months.
RESULTSThe symptoms were improved in most of the patients, the CD4 cells increased from 115.0 to 295.2/ul and the viral load (RNA copies/ml) in most patients reduced markedly or maintained at the same level.
CONCLUSIONThese data indicated that QuDu ZengNing Capsule was effective for treatment of AIDS patients.
Acquired Immunodeficiency Syndrome ; drug therapy ; immunology ; Adult ; CD4 Lymphocyte Count ; Capsules ; Drugs, Chinese Herbal ; administration & dosage ; therapeutic use ; Female ; HIV-1 ; Humans ; Male ; Middle Aged ; Phytotherapy ; Viral Load