1.Transcriptional repression of hDaxx enhanced by adenovirus 12 E1B 55-kDa oncoprotein interacting with hDaxx.
Yan-ping WAN ; Yi-mou WU ; Cui-ming ZHU ; Wei-guo YIN ; Heng-ling CAI ; Min-jun YU
Chinese Medical Journal 2004;117(5):753-757
BACKGROUNDDaxx has been identified as a nuclear protein that involves in apoptosis and transcriptional repression. Daxx co-localizes with the promyelocytic leukemia (PML) protein and regulates transcription. Human Daxx (hDaxx) is a protein that functions as a transcriptional regulation through its interaction with some DNA-associated proteins. The aim of this study was to explore the transcriptional regulatory effect of hDaxx interacting with adenovirus (Ad) 12 E1B (Ad12E1B) 55-kDa oncoprotein.
METHODSThe co-localization of hDaxx-Ad12E1B or hDaxx-PML protein in the nucleus was observed under a confocal microscope. Interaction of hDaxx and Ad12E1B was analyzed by yeast two-hybrid assay. Direct binding of hDaxx and Ad12E1B was analyzed using coimmunoprecipitation and Western blot in vivo and in vitro. The activity of a luciferase reporter gene, which was regulated by an hDaxx modulated thymidine kinase (TK) promoter, was detected in an automat luminometer.
RESULTSAd12E1B, which co-localized with hDaxx in the nuclei of G401-CC3 cells, disrupted the co-localization of hDaxx and PML in the PML oncogenic domains (PODs). hDaxx bound directly to Ad12E1B in vivo and in vitro. hDaxx interacted with Ad12E1B along its full length. Ad12E1B enhanced transcriptional repression activity of hDaxx.
CONCLUSIONAd12E1B disrupts the co-localization of hDaxx with PML in PODs and enhances transcriptional repression activity of hDaxx.
Adaptor Proteins, Signal Transducing ; Adenovirus E1B Proteins ; physiology ; Carrier Proteins ; analysis ; genetics ; Cell Line, Tumor ; Humans ; Intracellular Signaling Peptides and Proteins ; Neoplasm Proteins ; analysis ; Nuclear Proteins ; analysis ; genetics ; Promyelocytic Leukemia Protein ; Repressor Proteins ; physiology ; Transcription Factors ; analysis ; Transcription, Genetic ; Tumor Suppressor Proteins
2.Key quality indicators of the China Comprehensive AIDS Response 2008 - 2011 Program.
Di WU ; Heng ZHANG ; Xin-lei YAO ; Yan CUI
Chinese Journal of Preventive Medicine 2012;46(12):1095-1098
OBJECTIVETo analyze the key indicators of China Comprehensive AIDS Response Program (China CARES) and the effects of this program during 2008 and 2011.
METHODSData were obtained from National AIDS Comprehensive Prevention and Control Information System. The general population HIV test rate and spouse of people living with HIV or AIDS (PLWHA) HIV test rate were chosen as the indicators for HIV testing expansion; anti-retroviral therapy (ART) coverage and PLWHA CD4 test rate during the past 6 months as the indicators for treatment expansion; femal sex worker (FSW) intervention coverage and injection drug users (IDU) intervention coverage as the Indicators for behavior intervention expansion. The Key working quality indicators of 309 program sites were calculated and the results were compared.
RESULTSDuring 2008 and 2011, for China CARES, the Median of general population HIV test rate were 1.8%, 2.5%, 3.2% and 5.5%, the Median of spouse of PLWHA HIV test rate were 80.9%, 85.7%, 91.8% and 100.0%, the Median of ART coverage were 60.0%, 66.7%, 76.1% and 92.0%, the Median of PLWHA CD4 test rate during the past 6 months were 27.7%, 45.4%, 58.6% and 75.3%, the Median of FSW intervention coverage were 43.8%, 67.8%, 73.3% and 90.9%, the Median of IDU intervention coverage were 18.2%, 24.0%, 34.0% and 72.4%. The indicators of China CARES increased steadily from 2008 to 2011 in HIV testing expansion, treatment expansion and behavior intervention expansion; Compared to 2008, the percentage of the China CARES which had great progress of the 6 indicators were 98.4% (304/309), 98.3% (286/291), 94.0% (281/299), 93.5% (288/308), 91.8% (279/304) and 90.0% (223/247). in 2008, 3 indicators which were ART coverage, PLWHA CD4 test rate during the past 6 months and FSW intervention coverage were lower than national average (6.9%, 17.9% and 30.9%), the general population HIV test rate, spouse of PLWHA HIV test rate and IDU intervention coverage were higher than national average (3.4%, 64.3% and 22.8%), with the China CARES project going on, in 2011, key indicators which were general population HIV test rate, spouse of PLWHA HIV test rate, ART coverage, PLWHA CD4 test rate during the past 6 months, FSW intervention coverage and IDU intervention coverage were all higher than national average (6.5%, 80.6%, 81.9% and 55.9%).the proportions of China CARES sites whose 6 indicators were higher than national average in 2008 were 32.7% (101/309), 60.6% (149/246), 37.4% (99/265), 61.1% (181/296), 64.0% (174/270) and 45.3% (73/161), and the proportions increase annually to 2011 were 44.0% (136/309), 89.3% (260/291), 76.6% (229/299), 77.9% (240/308), 91.4% (278/304) and 64.8% (160/247).
CONCLUSIONAn obvious achievements have been made in the areas of HIV/AIDS prevention and control among China CARES sites during 2008 and 2011 than the national average.
Acquired Immunodeficiency Syndrome ; drug therapy ; epidemiology ; prevention & control ; Anti-Retroviral Agents ; therapeutic use ; China ; epidemiology ; Humans ; Quality Indicators, Health Care
3.Deep fascia composite autologous red bone marrow transplantation for the treatment of fracture nonunion.
Hui-Min LING ; Heng-Xuan WU ; Cui-Ye HUANG ; Shi-Qian MA
China Journal of Orthopaedics and Traumatology 2009;22(11):859-861
OBJECTIVEAccording to bone regeneration under the membrane and the bone regeneration deep fascia composite autologous red bone marrow transplantation applied in the treatment of fracture nonunion, in order to find a simple and effective clinical treatment of nonunion.
METHODSSince March 2006 to March 2009,17 patients of fracture nonunion were treated by the deep fascia composite autologous bone marrow transplantation,included 10 males and 7 females, aged from 7 to 52 years old (means 32 years). There were 10 cases of tibia, 5 cases of radius, 2 cases of clavicle. Injured to admission time was from 7 to 36 months (means 12 months). Ten cases underwent operation for 1 time,5 cases for twice and 2 cases for 3 times. The position of nonunion were all at bone shaft and the condition of the skin and soft tissue was good. X-ray film showed 11 cases of hyperplasia nonunion, 6 cases of shrinking. The original fixation were removed and the intramedullary nail or plate fixation were re-used, and fracture ends were sutured closed by autogenous deep fascia and implanted with autologous red bone marrow.
RESULTSSeventeen patients were followed-up for from 5 months to 2 years with an average of 1 year. Fracture healing time was from 12 to 20 weeks (means 16 weeks). According to the criteria of fracture healing to assess efficacy, the results were excellent in 14 cases, good in 2 cases and poor in 1 case.
CONCLUSIONDeep fascia composite autologous autologous red bone marrow transplantation for the treatment of fracture nonunion is suitable at the bone shaft and good condition of skin and soft tissue. The method has been observed that the fracture healing time is short.
Adolescent ; Adult ; Bone Marrow Transplantation ; Child ; Fascia ; Female ; Follow-Up Studies ; Fracture Healing ; Fractures, Bone ; diagnostic imaging ; physiopathology ; surgery ; Humans ; Male ; Middle Aged ; Time Factors ; Tomography, X-Ray Computed ; Transplantation, Autologous ; Young Adult
4.Effect of endoplasmic reticulum stress on chondrocytes in a rat model of knee osteoarthritis
Hao WU ; Zhichao MENG ; Yongping CAO ; Liping PAN ; Xingtong ZHOU ; Xin YANG ; Heng LIU ; Rui WANG ; Yunpeng CUI ; Xiang LI ; Zhuoyang LI
Chinese Journal of Tissue Engineering Research 2017;21(16):2502-2508
BACKGROUND: Endoplasmic reticulum (ER) stress has been proved to be related to the occurrence of diabetes, dilated cardiomyopathy and neurodegenerative diseases. Indeed, it is closely associated with osteoarthritis. OBJECTIVE: To explore the effect of ER stress on the chondrocyte viability as well as the occurrence and development of osteoarthritis in rats. METHODS: Rat chondrocytes were isolated and cultured, and the ER stress in the rat chondrocytes was by 10 mg/L tunicamycin. The expression levels of ER stress markers C/EBP-homologous protein and 78 kDa glucose-regulated protein were detected by western blot assay, and the proliferation and apoptosis of chondrocytes were detected by cell counting kit-8 assay and AnnexinV-FITC flow cytometry, respectively. In the in vivo experiment, 15 Sprague-Dawley rats were selected and subjected to anterior cruciate ligament transection and medial meniscectomy to establish an animal model of osteoarthritis. Tunicamycin, tauroursodeoxycholic acid and PBS (blank control group) were respectively injected into the articular cavity, and then the progression of osteoarthritis was assessed by hematoxylin-eosin staining at 4 weeks after treatment. RESULTS AND CONCLUSION: After addition of tunicamycin, the expression levels of C/EBP-homologous protein and 78 kDa glucose-regulated protein were significantly upregulated, the viability of chondrocytes was decreased gradually, while the apoptotic rate was increased significantly. Results from gross observation and hematoxylin-eosin staining suggested that tunicamycin promoted the progression of osteoarthritis and tauroursodeoxycholic acid delayed the deterioration of cartilage in the rats. These findings indicate that ER stress results in the decreased chondrocyte viability and increased apoptosis, which may be an important pathogenesis of osteoarthritis. Additionally, tauroursodeoxycholic acid can effectively alleviate osteoarthritis induced by ER stress.
5.Effect of surface-enhanced laser desorption/ionization time-of-flight mass spectrometry on identifing biomarkers of endometriosis.
Hong ZHANG ; Jie FENG ; Xiao-hong CHANG ; Zhong-xing LI ; Xiao-yi WU ; Heng CUI
Chinese Medical Journal 2009;122(4):373-376
BACKGROUNDEndometriosis is a common gynecological disease. This study aimed to screen proteins that were expressed differently in patients with endometriosis versus normal controls using proteomic techniques, surface-enhanced laser desorption/ionization time-of-flight mass spectrometry (SELDI-TOF-MS).
METHODSProtein chip SELDI-TOF-MS combines the advantages of microarray and mass spectrometry, and can screen latent markers in sera of patients with endometriosis. Serum samples from patients and normal volunteers were analyzed by SELDI-TOF-MS.
RESULTSAfter comparing the serum protein spectra of 36 patients with 24 normal controls, 24 differently expressed potential biomarkers (P < 0.01) were identified. Using Biomarker Pattern software, we established a tree model of the 60 serum protein spectra. When using the three biomarkers to classify the samples, the sensitivity for diagnosing endometriosis was 91.7%, specificity was 95.8%, and coincidence rate was 93.3%. Then we used serum samples from 12 patients and 8 normal controls to validate the tree model and report the sensitivity for diagnosing endometriosis was 91.7%, specificity was 75%, and coincidence rate was 85%.
CONCLUSIONSSELDI-TOF-MS may be a useful tool in high-risk population screening for endometriosis. The identification and application of the biomarkers need to further study.
Biomarkers ; blood ; Endometriosis ; blood ; diagnosis ; Female ; Humans ; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization ; methods
6.Experimental study on preparation of decellularized artery vascular graft matrix and explantation of carotid artery allografts.
Heng-hua FAN ; Bo-xun ZHANG ; Xiang-dang LIANG ; Ai-yuan WANG ; Bin ZHAO ; Xue-mei CUI ; Yi-yun HU ; Di WU ; Hua ZHOU ; Yan WANG
Chinese Journal of Surgery 2005;43(13):870-874
OBJECTIVETo set up a new process to access the preparation of decellularized artery grafts. And to evaluate the feasibility of decellularized artery allografts was evaluated.
METHODSThis study compared the effects of four extraction chemicals [1% t-octyl-phenoxypolyethoxyethanol (Triton X-100), 1% tri (n-butyl) phosphate (TnBP), and 1% sodium dodecyl sulfate (SDS) and trypsin (0.125, 0.25%) on thoracic artery vascular for 24 h (except trypsin for 2 h). At the base of it, a four-step process, including hypotonic, hypertonic solutions and combining with 1% Triton X-100 and 1% SDS detergents, were performed in rabbit thoracic artery vascular. Histological examination, tensile tests and expanding-burst tests were done on the samples. The decellularized carotid artery allografts were transplanted in other rabbits.
RESULTSTreatment with 1% SDS or 1% Triton X-100 for 24 h could remove most cells with retention of near normal structure. A four-step process could remove all cells with the extracellular matrix well conserved. The pulling mechanical properties and burst pressure of decellularized carotid artery were similar to the control. The decellularized carotid artery allografts (diameter of 2 mm) were patent at explanting up to 2 months.
CONCLUSIONSThe acellular artery vascular graft matrix is well prepared with four-step process including detergents, such as TritonX-100, SDS without compromising the graft structure or mechanical properties significantly. The carotid artery allografts (diameter of 2 mm) decellularized by the process are patent at explanting up to 2 months.
Animals ; Aorta, Thoracic ; cytology ; Bioprosthesis ; Blood Vessel Prosthesis ; Blood Vessel Prosthesis Implantation ; Carotid Arteries ; cytology ; transplantation ; Feasibility Studies ; Female ; Male ; Protease Inhibitors ; pharmacology ; Rabbits ; Sodium Dodecyl Sulfate ; pharmacology ; Tissue Engineering ; methods
7.High-performance liquid chromatography-mass spectrometry for determining olmesartan in human plasma.
Shi-Heng CHEN ; Cui-Fang WU ; Ben-Mei CHEN ; Qi PEI ; Hong-Yi TAN ; Li YANG ; Guo-Ping YANG
Journal of Southern Medical University 2008;28(6):1104-1105
OBJECTIVETo establish a simple and rapid HPLC-MS method for determining the contents of olmesartan in human plasma.
METHODSPlasma were precipitated with trifluoroacetic acid, then analyzed on an HyPurity C(18) column (150 mm 2.1 mm, 5 microm). Samples at 40 degrees celsius;. The mobile phase consisted of water-methanol- acetonitrile(14:60:26) with a flow rate of 0.22 ml/min.
RESULTSThe lower limit of qualification was 25 microg/L. The calibration curve was linear over the range of 25-3200 microg/L (r=0.9998), with the intra-day and inter-day RSD less than 15%.
CONCLUSIONThe method is sensitive, rapid and suitable for the study of pharmacokinetics and bioavailability of olmesartan.
Angiotensin II Type 1 Receptor Blockers ; blood ; Calibration ; Chromatography, High Pressure Liquid ; methods ; Humans ; Imidazoles ; blood ; Mass Spectrometry ; methods ; Reproducibility of Results ; Tetrazoles ; blood
8.Preliminary evaluation of clinical effects of below-knee arterial bypass on diabetic lower limb ischemia.
Ying-feng WU ; Yong-quan GU ; Xue-feng LI ; Heng-xi YU ; Li-xing QI ; Lian-rui GUO ; Shi-jun CUI ; Jian-xin LI ; Jian ZHANG ; Zhong-gao WANG
Chinese Journal of Surgery 2010;48(4):257-260
OBJECTIVETo evaluate medium-long term patency of below-knee bypass on patients who suffered from diabetic lower limb ischemia.
METHODSClinical and follow-up data of 51 patients was retrospectively analyzed who underwent 56 below-knee bypass because of diabetes from November 2001 to December 2006. There were 35 male and 16 female with an average age of 68 years. They endured 26 months ischemic time lag in average, and had suffered from diabetes for 11 years. All of the patients were performed bypass to below-knee (posterior tibial, anterior tibial or peroneal) arteries. Kaplan-meier method was applied. The subgroups of different operative methods and different out-flow vessels were compared by Log-rank tests.
RESULTSAn average follow-up time of 23 months was achieved, and lost-follow-up rate was 15%. The total 1- and 5-year primary patent rates were 68% and 54% respectively, secondary patent rate were 70% and 60% respectively, limb salvage rates were 69% and 65%, survival rates were 82% and 60%. One year (3 years) of patent rate(s) of operative method subgroups of femoral-popliteal-infrageniculate bypass with composite grafts, femoral/popliteal-infrageniculate bypass with artificial grafts and femoral/popliteal-infrageniculate bypass with autologous veins were 70% (50%), 33% (33%) and 70% (70%) respectively. One year (4 years) of patent rate(s) of out-flow vessel subgroups of posterior tibial artery, anterior tibial artery and peroneal artery were 65% (60%), 80% (53%) and 77% (66%) respectively. However, both subgroups did not show any statistic differences by log-rank tests.
CONCLUSIONSPartial or whole autologous veins as bypass grafts should be chosen when infrageniculate bypass is considered in diabetic patients. Considerable patent rates are acceptable no matter what kinds of out-flow vessels are chosen.
Aged ; Diabetic Angiopathies ; surgery ; Female ; Femoral Artery ; surgery ; Follow-Up Studies ; Humans ; Ischemia ; etiology ; surgery ; Lower Extremity ; blood supply ; Male ; Middle Aged ; Popliteal Artery ; surgery ; Retrospective Studies ; Saphenous Vein ; transplantation ; Tibial Arteries ; surgery ; Vascular Surgical Procedures ; methods
9.Hydrogen sulfide in cartilage and its inhibitory effect on matrix metalloproteinase 13 expression in chondrocytes induced by interlukin-1β
Liping PAN ; Yongping CAO ; Licheng WEN ; Weibing CHAI ; Junbao DU ; Hongfang JIN ; Jia LIU ; Xin YANG ; Zhichao MENG ; Heng LIU ; Yunpeng CUI ; Rui WANG ; Hao WU ; Xingtong ZHOU ; Xiang LI ; Zhuoyang LI ; Maimaitijuma TALATIBAIKE
Journal of Peking University(Health Sciences) 2016;48(2):194-202
Objective:To investigate whether endogenous hydrogen sulfide (H2 S)was involved in the pathogenesis of osteoarthritis (OA)and its underlying mechanism,to detect H2 S and its synthases ex-pression in knee cartilage in patients diagnosed with different severity of OA,and to explore the transcrip-tion and expression of gene MMP-13 in chondrocytes treated with IL-1βor H2S.Methods:Synovial fluids of the in-patients with different severity of OA hospitalized in Peking University First Hospital were collected for measurement of H2 S content using methylene blue assay.Articular cartilages of the patients who underwent knee arthroplasty were collected for the cell culture of relatively normal chondrocytes.The chondrocytes were cultured to the P3 generation and H2 S molecular probes were used for detection of endogenous H2 S generation in the chondrocytes.Immunocytochemistry was used to detect the localization of H2 S synthases including cystathionine β-synthase (CBS),cystathionine-γ-lyase (CSE),and mercap-topyruvate sulfurtransferase (MPST)in OA chondrocytes.Western blot was used to quantify the protein expressions of CSE,MPST,and CBS in cartilage tissues of the patients who were diagnosed with OA and underwent knee arthroplasty.The relatively normal human chondrocytes were cultured to passage 3 and then divided into 4 groups for different treatments:(1 )the normal control group,no reagent was added;(2)the IL-1βgroup,5 μg/L of IL-1βwas added;(3)the IL-1β+H2S group,200 μmol/L of NaHS was added 30 min before adding 5 μg/L of IL-1β;(4)the H2 S group,200 μmol/L of NaHS was added. The transcription and expression of gene MMP-13 in chondrocytes of each group were determined with Real-time PCR and Western blot,respectively.And the total NF-κB p65 and phosphorylated NF-κB p65 in chondrocytes were detected with Western blot.Results:The content of H2 S in the synovial fluid of degenerative knee was (14.3 ±3.3)μmol/L.Expressions of endogenous H2 S and its synthases including CBS,CSE and MPST were present in the cytoplasm of chondrocytes.CSE protein expression in Grade 3 (defined by outerbridge grading)cartilage tissues was significantly increased as compared with that of Grade 1 cartilage tissues (1.67 ±0.09 vs.1.26 ±0.11,P<0.05).However,no significant difference of CBS or MPST expression among the different groups was observed.The expression of MMP-13 protein in the IL-1βgroup was significantly higher than that in the normal chondrocytes (1 .87 ±0.67 vs.0.22 ± 0.10,P<0.05 ),and that in the IL-1β+H2 S group was significantly decreased than that in the IL-1βgroup (0.55 ±0.11 vs.1.87 ±0.67,P<0.05),and that in the H2S group had no significant difference compared with that in the normal control group.The transcription of MMP-13 protein in the IL-1βgroup was significantly higher than that in the normal chondrocytes (31.40 ±0.31 vs.1.00 ±0.00,P<0.05), and that in the IL-1β+H2 S group was significantly decreased than that in the IL-1βgroup (24.41 ± 1.28 vs.31.40 ±0.31,P<0.05),and that in the H2S group had no significant difference compared with that in the normal control group.The total NF-κB p65 in the IL-1βgroup was significantly higher than that in the normal chondrocytes (2.13 ±0.08 vs.0.73 ±0.08,P<0.05),and that in the IL-1β+H2S group was significantly decreased than that in the IL-1βgroup (1 .24 ±0.13 vs.2.13 ±0.08,P<0.05 ),and that in the H2 S group had no significant difference compared with that in the normal control group.The phosphorylated NF-κB p65 in IL-1βgroup was significantly higher than that in the normal chondrocytes (1.30 ±0.13 vs.0.19 ±0.04,P<0.05),and that in IL-1β+H2S group was significantly decreased than that in the IL-1βgroup (0.92 ±0.26 vs.1.30 ±0.13,P<0.05),and that in the H2S group had no significant difference compared with that in the normal control group.Conclusion:H2 S affected the cartilage degeneration by partly inhibiting the degradation of extracellular matrix.
10.Functional MRI of human brain in musicians and non-musicians.
Heng-wu CUI ; Shi-zheng ZHANG ; Hai-bo DI ; Hai LIU ; Yi-hong ZHU ; Qiao-wei ZHANG ; Xu-chu WENG ; Yi-zhang CHEN
Journal of Zhejiang University. Medical sciences 2005;34(4):326-330
OBJECTIVETo explore the differences in brain activation between musicians and non-musicians by use of functional MRI.
METHODSTwelve right-handed musicians and twelve right-handed non-musicians were recruited in the study. During a listening task, they were scanned on the Sigma 1.5T scanner (GE) while they were passively listening to several segments of music of "the Butterfly Love" and the white noise with same physical energy.
RESULTBoth musicians and non-musicians demonstrated bilateral transverse gyrus weak activated while listening to the white noise. But when listening to music, they showed bilateral temporal areas strongly activated including superior temporal gyrus, transverse gyrus and some middle temporal areas. Moreover, musicians showed relative left dominance (10/12), whereas non-musicians demonstrated right dominance(11/12). Furthermore,besides bilateral temporal areas, more and stronger activated areas were found in musicians such as cuneus, precuneus,medial frontal and left middle occipital gyrus.
CONCLUSIONThere are different neuro-patterns between musicians and non-musicians.
Adult ; Brain ; anatomy & histology ; physiology ; Humans ; Magnetic Resonance Imaging ; Male ; Music ; Temporal Lobe ; physiology