1.Study on the Mechanism of DNA Conformation Change Induced by Cr(Ⅵ)-GSH Complex
Journal of Environment and Health 1993;0(03):-
Objective To study the interaction between Cr(Ⅵ)-GSH complex and DNA and to understand the mechanism of DNA conformation change induced by Cr(Ⅵ)-GSH complex. Methods Chromium(Ⅵ) reacts on glutathione forms the chromium (Ⅴ) intermediate complex, and the intermediate complex can induce the conformation change of DNA, in the present paper, ultraviolet and fluorescence spectra, Scatchard equation, fluorescence quenching curve, and fluorescence probe method using ethidium bromide (EB) as a fluorescence probe were employed. Results The chromium (Ⅴ) intermediate complex formed rapidly after the reaction between Cr(Ⅵ) and GSH in Hepes buffer solution at pH=7.4, the complex did not insert into the base pairs on the DNA duplex strand and developed electrostatic binding with the phosphate backbone of DNA, but induced a conformation change of DNA leading to a disruption of duplex structure. In the following time, the complex decomposed and the end product was Cr(Ⅲ), Cr(Ⅲ) can cross-link with DNA, the whole process needs about an hour. Conclusion The chromium(Ⅴ) intermediate complex of the conformation change reaction of chromium(Ⅵ) and glutathione may play an important role in inducing the DNA.
2.Protective effects of tea polyphenols on cerebral nerve cell apoptosis induced by D-galactose and beta-amyloid peptide 25-35
Xian QU ; Bing LI ; Wenhao YANG ; Junhua Lü
Chinese Journal of Tissue Engineering Research 2007;11(43):8805-8808
BACKGROUND: Some researches demonstrate that tea polyphenols (TP) has protective effects on neurotoxicity of hippocampal nerve cells induced byβ-amyloid peptide (Aβ), 6-hydroxydopamine (6-OHDA) and oxidative substances. In addition, clinical preliminary examination indicates that TP plays a certain preventive and therapeutic effects on the reduction of recognition function in high-risk population with Alzheimer disease (AD); however, its target and mechanism are still hot topics.OBJECTIVE: To observe the interfering effects of TP on cerebral nerve cell apoptosis induced by D-galactose and Aβ25~35 in mice.DESIGN: Randomized controlled animal study.SETTING: Department of Pharmacology, Pharmacological College of Jinan University.MATERIALS: The experiment was carried out in the Experimental Center of Jinan University from September 2004 to January 2005. A total of 90 healthy Kumning mice, aged 2 months, each gender in half, weighing 26-28 g, were provided by Guangdong Provincial Medical Laboratory Animal Center. Tea polyphenols was provided by Zhejiang Oriental Tea Science and Technology Corporation (batch number: 20040203); D-galactose by Shanghai Number 2 Reagent Plant (batch number: 20030708); Aβ25~35 by Sigma (batch number: 13/01/2004); vitamin E (Vit-E) by Shanghai Xinyi Pharmaceutical Factory (batch number: 20030708).METHODS: Experimental interference: Mice based on body mass were randomly divided into 6 groups: sham operation group (n =17), model group (n =16), vitamin E group (n =16), low-dose (n =13), moderate-dose (n =14) and high-dose (n =14) tea polyphenols groups. In above-mentioned animals, except those in the sham operation group, all were given 120 mg/(kg·d) D-galactose for 12 consecutive weeks, and Aβ25~35 (4 nmol) was slowly injected into the lateral cerebral ventricle. In sham operation group, the same volume of artificial cerebral spinal fluid (CSF) was internally injected into lateral ventricle. Drug treatment began at the first week. Mice in the sham operation group and model group were given distilled water, and the animals in other groups were given the above-mentioned drugs (100 mg/kg Vit-E, 100, 250 and 625 mg/kg TP), respectively. The volume of perfusion was 10 ml/kg, and the treatment lasted for 12 consecutive weeks. Experimental evaluation: After administration, LW-Ⅱ water maze was used to measure learning and memory condition; brain, liver tissues and serum were obtained to measure activity of superoxide dismutase (SOD) and content of malondialdehyde (MDA); Fura-2/AM loading method was used to measure Ca2+ concentration in erythrocytes and neurons; flow cytometer was used to detect cerebral nerve cell apoptosis.MAIN OUTCOME MEASURES: Cl) Learning and memory ability; (2) SOD activity and MDA content in serum, liver and brain tissues; (3) Ca2+ concentration in erythrocytes and neurons;flow cytometer was used to cerebral nerve cell apoptosis.MAIN OUTCOME MEASURES:①Learning and memory ability;②SOD activity and MDA content in serum,liver and brain tissues;③Ca2+ concentration in erythrocytes and neurons;④cerebral nerve cell apoptosis.RESULTS:All 90 mice were involved in the final analysis.①At 12 weeks after administration,time to swim out of the water maze in the moderete-dose and high-dose TP groups and Vit-E group was shorter than that in the model group,and numbers of errors in passing the blind alleys in the water maze was reduced as compared with those in the model group,and there was significant difference(P<0.05-0.01).②SOD activities in the moderate-dose and high-dose TP groups were increased as compared with that in the model group,but MDA content in the high-dose TP group was decreased as compared with that in the model group.There was significant difference(P<0.05-0.01).③Ca2+ concentration in erythrocytes and neurons in the modemte-dose and high-dose TP groups and Vit-E group was lower than that in the model group,and there was significant difference(P<0.05-0.01).④The rates of brain neurons apoptosis in treatment groups with different doses of TP were 12.6%,18.6%,and 24.1% respectively, exhibiting significant difference as compared with the mice in sham operation group(P<0.05-0.01) CONCLUSION:TP can inhibit cerebral nerve cell apoptosis induced by D-galactose and Aβ25~35 and improve learning and memory ability in model mice.The effects may be related to its action of raising general anti-oxidative ability and improvement of intrecellular Ca overload induced by oxidative stress injury.
3.Diagnostic value of pre-operative and intra-operative insulinomas localization
Yongfu ZHAO ; Wenhao HAN ; Yang WU ; Wenlong ZAI
Chinese Journal of Pancreatology 2008;8(3):157-158
Objective To investigate the clinical significance of pre-operative ancl intra-operative insulinomas localization, and improve the diagnostic accuracy. Methods 75 patients with suspected insulinomas who were treated surgically were enrolled; all the patients had pathological evidence of insulinomas. The data of pre-operative ultrasound, CT, MRI and intra-operative ultrasound and surgical palpation were collected and analyzed. Results The sensitivity of localization procedures was as follows: ultrasonography 30.7% ( 23/75 ), CT 40.0% (24/60) , MRI 45.4% ( 22/48 ) , while surgical palpation was 80.4% , intra-operative ultrasound 96.4%. The accuracy of intra-operative procedures was higher than that of pre-operative procedures. Conclusions It was difficult to accurately localize insulinomas before operation, but intra-operative palpation and iutra-operative US was easy to perform and highly accurate. Therefore, too much emphasis should not be placed on pre-operative imaging tests.
4.In vivo animal study on osteal histocompatibility of carbon fiber-reinforced nano-hydroxyapatite/polyamide 66 composites
Ming LU ; Xuesong ZHANG ; Hui XU ; Wenhao HU ; Xiaoqing YANG
Chinese Journal of Tissue Engineering Research 2015;19(16):2523-2528
BACKGROUND:Compared with hydroxyapatite materials and other nano-hydroxyapatite composites, carbon fiber-reinforced nano-hydroxyapatite/polyamide 66 composites have been significantly improved in the mechanical strength, toughness, elastic modulus and other aspects. It can be used for repairing bone defects of loading parts. OBJECTIVE:To investigate the biocompatibility of carbon fiber-reinforced nano-hydroxyapatite/polyamide 66 composites in bone tissues. METHODS:Eight Bama mini pigs were taken to establish models of thoracic vertebral defects and implanted with carbon fiber-reinforced nano-hydroxyapatite/polyamide 66 composites. At 8, 16 and 24 weeks after implantation,the animals were sacrificed, respectively, for bone mineral density detection and hematoxylin-eosin staining. Blood samples for kidney and liver function tests were taken before and 1 and 8 weeks after implantation. RESULTS AND CONCLUSION: Hematoxylin-eosin staining of bone samples showed that the materials could bond with the bone defect interface without rejection, and could induce osteogenesis of chondrocytes. At 8 weeks after surgery, the broken ends of cancelous bone closed and the composite material was wrapped by granulation tissues. At 16 weeks after surgery, granulation tissues were organized and new bone developed directly from fibroblast cels. The new bone tissues were nearly fused with the end of cancelous bone. At 24 weeks after surgery, new bone tissue became mature lamelar bone, and the end of cancelous bone was connected tightly with the composite material. Bone mineral density of the implanted vertebra showed an increase trend at 8, 16 and 24 weeks after implantation. Over time, the bone mass was increased. The liver and kidney function tests showed that there was no significant difference before and after implantation. It is preliminarily believed that the carbon fiber-reinforced nano-hydroxyapatite/polyamide 66 composite has excelent histocompatibility and bioactivity without hepatic toxicity and nephritic toxicity.
5.Surgical treatment of spontaneous intracerebral hemorrhage
Kunpeng WANG ; Weixing WANG ; Jie YANG ; Wenhao SHUN
International Journal of Cerebrovascular Diseases 2014;22(4):300-303
Intracerebral hemorrhage (ICH) is the second most common type of stroke,and its fatality is high.The baseline hematoma volume and hematoma growth are the predictive factors for the poor outcome of the patients.Previous studies have shown that surgical evacuation of hematoma can reduce the hematoma volume and improve the outcome.However,several recent randomized controlled trials of craniotomy hematoma evacuation for ICH and a Meta-analysis have shown that it is no more beneficial than conservative treatment.The previous evidence of minimally invasive evacuation of hematoma for the treatment of ICH has suggested that it has potential benefits for patients with ICH; however,further research is needed to confirm it.
6.Detection of Serum HGF and MMP-9 and Its Clinical Significance in Pat ients with Systemic Lupus Erythematosus
Yang LIU ; Min ZHENG ; Wenhao YIN ; Zhang BAO
Chinese Journal of Dermatology 2003;0(12):-
Objectives To explore the relationship between serum leve ls of hepatocyte growth factor (HGF) and matrix metalloproteinase-9 (MMP-9) and the disease activity of systemic lupus erythematosus (SLE), and study the mechan isms of these two factors in the pathogenesis of SLE. Methods The serum levels of HGF and MMP-9 were measured by ELISA. The growth curve of normal ECV304 cell line was obtained, and the action concentration of recombinant human hepatocyte growth factor (rhHGF) was determined. MMP-9 expression level in cells was detec ted by flow cytometric analysis. Results The serum level of HGF increased sign ificantly in SLE patients as compared with that in healthy controls (P 0.05). The area of ROC curve was 0.707 and the sensitivity was 66 .7% when using the serum level of HGF as diagnostic standard. The area under ROC curve was 0.984 and the sensitivity was 97.2% when using the serum level of MMP-9 as diagnostic standard. The sensitivity was 66.7% (24/36) when two markers (H GF and MMP-9) were examined simultaneously. Additionally, the action concentrati on of rhHGF was 8 ng/mL, and the expression level of MMP-9 was 39.74% in normal ECV304 cells and increased to 40.32% after rhHGF stimulation. Conclusions It i s suggested that HGF and MMP-9 may be involved in the pathogenesis of SLE, and s erum levels of HGF and MMP-9 might be used as markers for monitoring the disease activity, renal damage, disease progression and improvement in SLE. The sensiti vity might be higher when serum level of MMP-9 is used as diagnostic standard, a nd rhHGF can enhance MMP-9 expression in ECV304 cell line.
7.The relationship between MMP-9 serum level and disease activity in patients with systemic lupus erythematosus
Yang LIU ; Min ZHENG ; Wenhao YIN ; Zhang BAO
Chinese Journal of Rheumatology 2001;0(04):-
Objective To investigate the relationship between serum level of MMP-9(matrix metalloproteinase-9) and the disease activity of systemic lupus erythematosus(SLE).Methods Serum level of MMP-9 of 30 controls and 36 SLE patients were measured by Enzyme Linked Immuno Sorbent Assay.Results Significantly decreased serum level of MMP-9 was found in SLE patients as compared to that in healthy controls(108?113) ng/ml vs (352?155) ng/ml (P0.05).Conclusion The present data suggests that MMP-9 may be involved in the pathogenesis of SLE,and serum MMP-9 level may be a marker of disease activity,renal damage,disease progression and amelioration in SLE.
8.The relationship between serum level of HGF and disease activity in the patients with systemic lupus erythematosus
Yang LIU ; Min ZHENG ; Wenhao YIN ; Zhang BAO
Chinese Journal of Laboratory Medicine 2001;0(02):-
Objective To investigate the relationship between serum level of Hepatocyte growth factor (HGF) and the disease activity of systemic lupus erythematosus (SLE). Methods Serum level of HGF in 30 control and 36 SLE patients were measured by Enzyme Linked Immunosorbent Assay. ResultsSignifi cantly increased sera level of HGF were found in SLE patients as compared to that in healthy controls[ (1 433.3?154.0)ng/L vs (1 142.1?78.8)ng/L,P
9.Genomic copy number variations analysis in six neonates with Pierre Robin sequence
Lin YANG ; Jinwen NI ; Guodong ZHAN ; Huijun WANG ; Chao CHEN ; Guoying HUANG ; Wenhao ZHOU
Chinese Journal of Perinatal Medicine 2011;14(11):670-675
Objective To screen for genomic copy number variants(CNVs)in six neonates with Pierre Robin sequence(PRS)by Affymetrix 2.7 M chip to identify possible loci related to PRS.Methods Six neonates with PRS admitted into the Department of Neonatology,Children's Hospital of Fudan University from June 2009 to May 2010 were enrolled in this study.CNVs were detected by Cytogenetic Whole Genome 2.7 M array.Rare CNVs with potential clinical significance that deletion segments' size >50 kb and duplication segments' size >200 kb were selected based on the analysis of Chromosome Analysis Suite(ChAS)software,false positive CNVs and segments of normal population were excluded.The identified CNVs were compared with those in relative published literatures.Results(1)Among 6 PRS patients,two patients had facial deformation,two had congenital heart defects,one had congenital dysplasia of the laryngeal cartilage and one had choroidal space occupying lesion.(2)Seven rare CNVs whose size from 51-11 956 kb were identified in four neonates,including a 739 kb duplication on lp26.23-p36.22,a 6273 kb deletion on lq43-44,a 51 kb and a 55 kb deletions on 14q32.31,a 1022 kb duplication on 14q11.1-11.2,a 11 956 kb duplication on 20p13 and a 105 kb deletion on 4q23.3.(3)Published literatures showed that deletions of 1q43-44 and 14q32.31 might relate to micro/retrognathia and abnormal palate.Region of chromosome 1q43-q44 contained AKT3 and heterogeneous nuclear ribonucleoprotein U(hnRNPU)genes,and the haploinsufficiency of AKT3 and hnRNPU genes might cause developmental human microcephaly and agenesis of the corpus callosum,speech delay and seizures respectively.Region of chromosome 14q32.31 contained some C/D small nucleolar RNA,and the human imprinted 14q32 domain shared common genomic features with the imprinted 15q11-q13 loci.Conclusions This study established a method to discover whole genome CNVs in identifying novel submicroscopic deletions and duplications.Reviewing of published literatures suggested that deletions of chromosome 1q43-q44 and 14q32.31 might cause Pierre Robin sequence.
10.The diagnostic value of antibodies in synovial fluid for rheumatoid arthritis
Ruixiang XU ; Yang LIU ; Wenhao WANG ; Jing ZHAO ; Guizhi ZHANG ; Jing YUAN
Chinese Journal of Rheumatology 2015;19(4):246-248
Objective To study the concentration of anti-cyclic citrullinated peptide antibody (CCP),rheumatoid factor (RF),immunoglobulin (Ig) in synovial fluid and serum of patients with rheumatoid arthritis (RA),and the positive rate of anti-keratin antibodies (AKA) and anti-perinuclear factor (APF) were also measured.The aim was to explore the diagnostic value of these antibodies in synovial fluid for RA.Methods Thirty RA patients were collected,synovial fluid were withdrawn from the inflamed joints and peripheral blood were collected at the same time.Anti-CCP antibody,AKA,APF and RF were measured.The cut-off value of CCP in synovial fluid was determined through operating characteristic curve (ROC curves) analysis.The antibody levels in serum and synovial fluid were compared.Statistical analyses were performed using t test,and Pearson's correlation test.Results The cutoff value of CCP in RA synovial was higher,which was 29.00 U/ml,with the sensitivity of 87.00% and specificity of 94.10%.There were significant differences in the concentration among anti-CCP antibody [(326±313) U/ml,(496±454) U/ml,t=-2.399,P<0.05],IgG [(57±39) mg/L,(173±86) mg/L,t=-7.4792,P<0.05),RF [(53±36) U/ml,(149±104) U/ml,t=-2.402,P<0.05] in synovial and blood serum respectively in RA patients.Conchsion The diagnosis of RA is enhanced if anti-CCP antibodies,RF and IgG in synovial fluid are examined.They are supplementary to RA diagnosis and are helpful for clinical practice,especially for patient with very recent-onset RA.