1.Cloning and Analyzing The Promoter of PPAR?
Chengqiang HE ; Congcong LI ; Yanhong WU ; Liguo AN ; Yunlong LI
Progress in Biochemistry and Biophysics 2006;0(04):-
Peroxisome proliferator-activated receptors (PPARs) are a family of nuclear hormone receptors belonging to the steroid receptor superfamily. Three PPAR isoforms, PPAR?, PPAR? (also known as PPAR?) and PPAR? have been found in the mouse. They can activate expression of many genes, including those involved in lipidmetabolism. PPAR? is ubiquitously expressed, but the level of expression differs markedly between different cell types. PPAR? is expressed in skeletal muscle at 10- and 50-fold higher levels compared with PPAR? and PPAR?, respectively. A role for PPAR? in skeletal muscle is to increase the genes expression with relation to oxidative metabolism. In order to determine the molecular mechanisms governing PPAR? gene expression in muscle, a 2 kb 5′ flanking region was cloned and analyzed. The DNA fragment is able to transcribe GFP in COS7 cells. Dual luciferase assay is used to quantify promoter activity. Deletion analysis of the 2 kb PPAR? promoter fragment in COS7 and NIH 3T3 cells shows that the proximal promoter sequence, nt -197 to +120, confers basal transcriptional activity of the mouse PPAR? gene. Computational analysis of putative cis-acting elements located within the ~2.0 kb mouse PPAR? 5′-flanking sequence was performed using the TRANSFAC database and MatInspector software and 4 potential MEF2A binding sites were found. And there is a potential binding site sharing 100% identity with positive element of MEF2A in the proximal promoter (nt -261). Co-transfection experiments of the PPAR? promoter reporter and pMEF2A expression plasmid (pMEF2A) showed that MEF2A significantly enhanced transcription activity of PPAR? promoter in NIH 3T3. Moreover, the enhancive effect depended on the concentration of plasmid pMEF2A transfected into cells. The results suggested that MEF2A may enhance transcription activity of the PPAR promoter in muscle cells.
2.Suppression of RANKL/OPG pathway activation in FLSs in interfacial membrane through up-regulation of HIF-1α induced by Ti particles
Chengqiang WANG ; Shengmao HE ; Xiaobo XIE ; Qi LI
The Journal of Practical Medicine 2017;33(10):1580-1583
Objective To explore the correlation between HIF-1α expression and RANKL/OPG pathway activation of fibroblast-like synoviocytes(FLSs)induced by Ti particles in aseptic loosening interfacial membrane. Method FLSs were extracted from the synovial tissue collected in surgeries and then co-cultured with Ti particles. QRT-PCR and Western blotting were conducted to measure the mRNA and protein expression of RANKL ,OPG and HIF-1αin FLSs at different concentration and time. Results Genes and protein expression levels of RANKL/OPG and HIF-1α were up-regulated with the increase of concentration of Ti particles. Expression of HIF-1α gene and protein increased time-dependently;the mRNA and protein expression of RANKL/OPG increased firstly and then declined alongside the increase of HIF-1αexpression. Conclusions Ti particles induce the up-regulation of HIF-1α expression and activate RANKL/OPG pathway. Up-regulation of HIF-1α may suppress the activation of RANKL/OPG pathway in FLSs of the interfacial membrane induced by Ti particles.
3.Core decompression combined with beta-tricalcium phosphate bioceramics for early non-traumatic osteonecrosis of the femoral head
Shengmao HE ; Lijun LIN ; Chengqiang WANG ; Qi LI
Chinese Journal of Tissue Engineering Research 2017;21(6):883-887
BACKGROUND:There are numerous laboratory reports concerning β-tricalcium phosphate (β-TCP) bioceramics;however, its application in the treatment of osteonecrosis of the femoral head (ONFH) is rarely reported. OBJECTIVE:To analyze the short-term efficacy of core decompression combined with β-TCP bioceramics in the treatment of ARCO Ⅰ/Ⅱ/ⅢA non-traumatic ONFH. METHODS:Twelve patients (16 hips) suffered from ARCO Ⅰ/Ⅱ/ⅢA non-traumatic ONFH were treated by core decompression combined with β-TCP implantation. The Harris hip scores (HHS) and radiological observation were performed before and after treatment. The HHS at the last follow-up or the HHS prior to the radiographs showing deterioration or severe complication occurrence was recorded. RESULTS AND CONCLUSION:Al patients were followed up for 19 months averagely (11-30 months) and no fracture, infection and other complications occurred. The HHS significantly increased from (73.61±3.70) to (84.88±7.11) points after treatment (P<0.001). The postoperative outcome was excellent in five cases with five hips, good in five cases with nine hips, mild in one case with one hip, poor in one case with one hip, and the excel ent and good rate was 87.5%(14/16). The radiographs of only one case of ARCO ⅡC and one case of ARCO ⅢA showed deterioration, and the latter was given the total hip arthroplasty. These results indicate that the core decompression combined with β-TCP implantation achieve short-term efficacy for ARCO Ⅰ/Ⅱ/ⅢA non-traumatic ONFH.
4.Clinical features and risk factors of neonatal clavicle fracture
Chengqiang ZHANG ; Beiqian QIAN ; Xiaotian LI ; Jimei WANG
Chinese Journal of Perinatal Medicine 2017;20(1):27-30
ObjectiveTo summarize the clinical features and risk factors of neonatal clavicle fracture so as to decrease the incidence of this disease.MethodsA total of 85 cases of neonatal clavicle fracture admitted to the Gynecology and Obstetrics Hospital of Fudan University between January 1, 2013 and December 31, 2015 were enrolled in this study. The full-term neonates with natural delivery by the same midwives or doctors on the same day were selected as control group with 1∶1 matched. Clinical features and risk factors of neonatal clavicle fracture based on three factors: mother factors, labor factors and neonatal factors, were analyzed retrospectively. Two independent samplest-test, rank sum test andChi-square test were used for statistical analysis.ResultsThe incidence rate of neonatal clavicle fractures was 0.14% (85/59 991). There were 45 (53%) males and 40 (47%) females; 80 (94%) cases of vaginal delivery, and 5 (6%) cases of cesarean section. According to the relationship between the location of the fracture and the position of the fetus, the fracture occurred in the anterior shoulder in 65 cases (76%) and in the posterior shoulder in 20 cases (24%);and 18 cases (21%) were in the left and 67 cases (79%) in the right. Fifty-five cases (65%) were diagnosed by physical examination, and 30 cases (35%) by X-ray. Two patients (2%) were complicated with brachial plexus injury. Twenty cases (24%) were diagnosed on the day of birth, 32 cases (38%) on the second day after birth, 16 (19%) on the third day, and four (5%) on the 4-6 days. Thirteen (15%) cases were diagnosed in the outpatient department 42 days after birth. All the fractures were cured without special treatment. Compared with the control group, the birth weight of fracture group was significantly higher [(3 646±361) vs (3 447±409) g,t=2.339], and the ratio of the cases complicated with shoulder dystocia was also higher [15% (13/85) vs 2% (2/85),χ2=9.524] (bothP<0.05). The differences in the factors of maternal age, parity, times of pregnancy, gestational age, forceps delivery, time of the second stage of labor and fetal distress between the two groups were not statistically significant.ConclusionsNeonatal clavicle fracture is related to birth weight and shoulder dystocia. Not all fractures can be diagnosed through physical examination on the day of birth, therefore, daily routine physical examinations after birth are necessary. The prognosis of neonatal clavicle fracture is good.
5.Effect of respiratory movement on cone beam computed tomography images.
Chang GUO ; Renming ZHONG ; Guangjun LI ; Chuanxian JI ; Chengqiang LI ; Hong QUAN ; Sen BAI
Journal of Biomedical Engineering 2014;31(2):314-318
To investigate the impact of respiratory movement to determine the target volume on cone beam CT (CBCT) for lung tumor, we used CIRS dynamic thorax phantom (Model-CIRS008) to simulate the sinusoidal motion of lung tumor. With a constant amplitude, the ratio of the time of near-end-expiratory and near-end-inspiratory (E/I) changed when it was scanned with CBCT. We analyzed the contrast changes of target by extracting the CT value of each pixel on the center line of the target movement direction. The targets were contoured with region growing method and compared with the motion volume generated by the tumor trajectory method. The result showed that the contrast of near-end-expiratory increased and the contrast of near-end-inspiratory decreased with increasing E/I. The contoured volume generated by region growing method decreased with increasing E/I. When E/I = 4, the amplitude A = 1 cm, diameter of 1 cm and 3 cm target volumes were reduced by 48.2% and 22.7%. The study showed that CBCT was not suitable to be used to accurately determine the range of lung tumor movement. The internal target volume (ITV) may be underestimated in CBCT images.
Cone-Beam Computed Tomography
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Humans
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Lung Neoplasms
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diagnostic imaging
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Movement
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Phantoms, Imaging
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Respiration
6.The characteristics and clinical application of the ArcCHECK diode array for volumetric-modulated arc therapy verification
Chengqiang LI ; Guangjun LI ; Chuanxian JI ; Jianghong XIAO ; Chang GUO ; Hong QUAN ; Sen BAI
Chinese Journal of Radiation Oncology 2013;(3):253-257
Objective To evaluate the characteristics and clinical suitability of the ArcCHECK diode array for volumetric-modulated arc therapy (VMAT) verification.Methods The intrinsic sensitivity,short and long term reproducibility,dose and dose rate dependence,dose per pulsed dependence,field sizes dependence and directional response of the diodes were measured.The results of the diodes were compared with the measurement results of an ionization chamber and calculated results of treatment planning system.Gamma index was used to analyze the dose difference between the calculation and measurement for random selected 211 verified VMAT plans.Results The ArcCHECK performed well for all tests except directional dependence,which varies from a minimum of-3.9% (seen only when the beam was incident on the diode at 180°) to a maximum of 7.7% (approximately at 255°).Average gamma analysis passing rates with 3 mm/3% for 113 nasopharyngeal cancer,48 cervical cancer and 50 rectal cancer patients VMAT plans were 93.5%,95.7% and 97.5%,respectively,statistical significance has been found between either two of the groups (t=-12.69-4.88,all P< 0.01).Conclusions With proposed calibration method,the ArcCHECK diode array is very suitable for VMAT pretreatment verification,the complexity of VMAT plan is the main factor affecting the passing rate.
7.Efficacy and safety study on infliximab with disease-modifying anti-rheumatic drugs in refractory psoriatic arthritis
Chengqiang ZHANG ; Gailian ZHANG ; Liyun ZHANG ; Xiaofeng LI ; Honglei DAI ; Fang LI ; Hongyan WEN ; Lihui MA
Chinese Journal of Rheumatology 2011;15(11):759-762
ObjectiveTo explore the efficacy and safety of infliximab combined withdiseasemodifying antirheumatic drugs (DMARDs) in the treatment of psoriatic arthritis.MethodsThis was an openlabeled trial.All subjects fulfilled the Moll and Wright criteria for definite PsA and-had poor response to DMARDs.The patients received combined infliximab and DMARDs.Infliximab 3 mg/kg was infused at weeks 0,2,6,14.After week 14,patients received infliximab 3 mg/kg every 8 weeks.The primary end point was the improvement of psoriatic arthritis response criteria(PsARC) response.The secondary end point was the percentage of patients who had 20% improvement of modified American College of Rheumatology (ACR20)response.Parameters for efficacy for psoriatic rash was defined as the proportion of modified 50% and 75%improvement of psoriasis area and severity index scores (PASI).All adverse reactions in the whole observation period were recorded.Chi-square test and repeated measurement data analysis of variance were used for the statistical analysis.ResultsTwenty-one patients completed the 14 weeks treatment.Five patients completed 26-104 weeks treatment,including 2 cases for 104 weeks.At week 14,the percentage of patients achieving PsARC was 84%,the percentage of patients achieving ACR20 was 77%,and the percentage of patients achieving PASI 50 was 76%.At week 14,tender joint counts,swollen joint counts,patient's assessment of pain,patient's global assessment(PGA),physician's global assessment,dermatology life quality index (DLQI),health assessment questionnaire(HAQ) were significantly improved compared with base-line(P<0.05).Five patients received 26-104 weeks follow-up,including 2 cases for 104 weeks,four patients was stable,the rash and joint symptoms of 1 patient recurred at 104 weeks.The most frequently occurred adverse reactions were upper respiratory tract infection and skin as well as appendage damages.The second most common adverse effect was elevation of liver enzymes.ConclusionThe infliximab combined with DMARDs is effective and safe for the treatment of psoriatic arthritis.
8.The regulatory mechanism of PPAR-γ in TH cell differentiation and its relation with transcription factor T-bet and GATA-3
Fang LIU ; Wenjuan WANG ; Chengqiang JIN ; Hong XIAO ; Biying ZHENG ; Qun CHEN ; Guoming LI
Chinese Journal of Microbiology and Immunology 2009;29(1):11-15
Objective To investigate the role of PPAR-γ in the gene expression of T-bet/GATA-3 in Jurkat T cells,and to explore the mechanisms underling this sensitizing effect of the change of TH cell subpopulation group.Methods Jurkat T cells were stimulated with PPAR-γ agonist pioglitazone.TH cell related cytokine IFN-γ and IL-10 was detected by ELISA,and the expression of transcription factors(T-bet and GATA-3)mRNA was detected by RT-PCR.To prove the PPAR-γ-dependent effect.the PPAR-γ-specific antagonist GW9662 was used.Results Stimulated with agonist PPAR-γpioglitazone.the concentration of IFN-γ and IL-10 and the expression of transcription factor T-bet and GATA-3 mRNA were both significantlY decreased in Jurkat T cells obviously,and these actions were dependent on the time and the concentrations of pioglitazone.Added with antagonist GW9662 at the same time,such inhibitory actions of IFN-γ and T-bet expression were recovered.but not IL-10 and GATA-3.Conclusion Pioglitazone can inhibite T cells proliferation and their secretion of cytokines.Pioglitazone can inhibit TH1 cells from secreting cytokines,and it is a PPAR-γ-dependent effect related to T-bet.The inhibition on TH2 is not a PPAR-γ-dependent effect and it is GATA-3 related.
9.Analysis of multi-leaf collimator leaf position deviation in volumetric modulated arc therapy
Chengqiang LI ; Cheng TAO ; Jian ZHU ; Jie LU ; Tonghai LIU ; Yong YIN
Chinese Journal of Radiological Medicine and Protection 2016;36(12):904-908
Objective To evaluate the position deviation of multi-leaf collimator (MLC) in volumetric modulated arc therapy for nasopharyngeal carcinoma and to verify a method of detecting the leaf position accuracy.Methods Thirty-two volumetric modulated arc plans of nasopharyngeal carcinoma were randomly selected.A clockwise arc with a gantry range of 181° to 179° and a counterclockwise arc with a gantry range of 179° to 181° were uscd for each plan.Each arc contained two leaf groups,groups A and B.By using the Argus software,the actual MLC leaf positions were extracted from the Dynalog files in the Varian Trilogy accelerator.The planed MLC leaf positions were compared with the actual positions,and the differences were calculated between them.Results The proportions of the leaf position errors within ± 2,± 1.5,± 1 and ± 0.5 mm were 99.99%,99.90%,99.07% and 93.98%,respectively.The proportions of the leaf position errors within ± 1 mm were 98.08%,100%,98.97% and 99.01% for the first therapeutic arc (ARC1),second therapeutic arc (ARC2),A and B groups,respectively.The leaf position errors range of ARC1 and ARC2 were-2.95 to 2.99 and-0.22 to 0.23 mm (t =2.35,P < 0.01).The leaf position errors range of leaf bank A and leaf bank B were-2.95 to 2.68 and-2.92 to 2.99 mm(P > 0.05).Conclusions During the volumetric modulated arc radiotherapy,the leaf positions deviation are in control.The Dynalog files can be used as an effective way to analysis the leafposition errors.
10.Diagnostic Value of MRI Combined TRUS for Prostate Cancer at Different Prostate-specific Antigen(PSA)Levels
Guanyu SU ; Xiaopeng MAO ; Chengqiang MO ; Baimou LI ; Xu CHEN ; Jintao ZHUANG ; Shaopeng QIU
Journal of Sun Yat-sen University(Medical Sciences) 2017;38(1):133-137
Objective]To explore the diagnostic value of magnetic resonance imaging(MRI)combined transrectal ultrasound (TRUS)to guide prostate cancer detection at different serum prostate-specific antigen(PSA)levels.[Methods]Totally 278 patients who underwent a systematic biopsy were collected in our hospital from November 2014 to June 2016. Preoperative tests of PSA , MRI,TRUS were performed in all the included patients. According to the PSA level of 4~10 ng/mL,10~20 ng/mL,over 20 ng/mL, 278 cases were divided into three group of A ,B ,C. Retrospective analysis was performed within the three groups of diagnostic accuracy.[Results]In Group A,the areas under ROC for MRI+TRUS and TRUS were 0.73 and 0.59,respectively(P = 0.02). In Group B ,the areas under ROC for MRI+TRUS and TRUS were 0.68 and 0.56 ,respectively (P < 0.001). In Group C ,the area under ROC for MRI+TRUS and TRUS were 0.74 and 0.63,respectively(P < 0.001). There is more significant statistical difference in Group B and C.[Conclusion]MRI combined TRUS has higher diagnostic value in cancer detection than TRUS before biopsy between different PSA levels ,which Indicates that MRI combined TRUS is an effective method for the improvement of prostate cancer detection.