2.Establishment and application of screening methods for non-agonist PPARγ ligand.
Yi HUAN ; Jun PENG ; Yue WANG ; Chun-Ming JIA ; Ke WANG ; Ke-Hua WANG ; Zhi-Qiang FENG ; Zhu-Fang SHEN
Acta Pharmaceutica Sinica 2014;49(12):1658-1664
In-vitro assay methods were established to evaluate transactivation and binding activity of compounds on peroxisome proliferator-activated receptor y (PPARγ). Firstly, plasmids were constructed for transactivation assay of PPARγ response element (PPRE) triggered reporter gene expression, and for cell-based binding activity assay of the chimeric receptor, which was fused with PPARγ ligand binding domain (LBD) and yeast transcriptional activator Gal4. Secondly, by using PPARy competitive binding assay based on time resolved-fluorescence resonance energy transfer (TR-FRET), affinities of compounds and drugs to PPARγ were evaluated. In application of these above methods, the PPARγ activating potency and characteristics of different compounds were evaluated, and a novel benzeneselfonamide derivative, ZLJ01, was found to have comparable binding activity and affinity with the well-known PPARy agonist, but lack of PPRE mediated transactivation activity. In preliminary study on in-vitro hypoglycemic activity, ZLJ1 was found to promote insulin-stimulated glucose uptake by liver cells. Therefore, we believe that combining transactivation and binding activity as well as affinity evaluation, the system could be used to screen non-agonist PPARγ ligand as anovel PPARγ modulator
Genes, Reporter
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Hepatocytes
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Hypoglycemic Agents
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chemistry
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Ligands
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PPAR gamma
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agonists
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chemistry
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Plasmids
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Response Elements
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Sulfonamides
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chemistry
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Transcriptional Activation
3.Effect of yigu capsule contained serum on mRNA and protein expression of estrogen receptor in osteoblast rats.
Rong-Hua ZHANG ; Ke-Ping PENG ; Xiao-Feng ZHU
Chinese Journal of Integrated Traditional and Western Medicine 2005;25(4):333-337
OBJECTIVETo explore the effect of Yigu capsule contained serum on mRNA and protein expression of osteoblast estrogen receptor (ER), for investigating the mechanism of its preventing and treating osteoporosis by means of regulating estrogen.
METHODSOsteoblasts separated from newborn SD rats were cultured, and divided into 3 groups after being passaged, i.e. the drug-serum treated group, the blank serum group and the DMEM medium control group. The relative amount of ERalpha and ERbeta expression were determined with RT-PCR, the affinity (expressed by equilibrium dissociation constant, KD) and number of ER (RT) were analysed by radioligand assay.
RESULTSThe relative amount of ERbeta mRNA expression were increased in the drug-serum group, with significant difference as compared with that in the other two groups (P < 0.05), but no significant difference among the three groups in ERa mRNA expression (P > 0.05). KD in the drug-serum group showed insignificant difference as compared with that in the other two groups (P > 0.05), but RT increased in the drug-serum group and the difference in the three groups was significant (P < 0.01).
CONCLUSIONDrug-contained serum of Yigu capsule can up-regulate the expression of osteoblast ERbeta mRNA and increase the amount of ERs. Regulating estrogen is possibly one of the mechanisms of Yigu capsule in preventing and treating osteoporosis.
Animals ; Capsules ; Cell Proliferation ; drug effects ; Cell Separation ; Drugs, Chinese Herbal ; pharmacology ; Female ; Osteoblasts ; cytology ; metabolism ; RNA, Messenger ; biosynthesis ; genetics ; Rats ; Rats, Sprague-Dawley ; Receptors, Estrogen ; biosynthesis ; genetics ; Serum
4.Pharmacokinetics of salvianolic acid A after single intravenous administration in Rhesus monkey.
Jun-ke SONG ; Wen ZHANG ; Wei-ku ZHANG ; Zhang-ying FENG ; Tao XIE ; Guan-Hua DU
Acta Pharmaceutica Sinica 2015;50(9):1142-1147
Salvianolic acid A (Sal A) is one of the most effective compounds isolated from the root of Salvia miltiorrhiza. Up to now, several studies regarding the pharmacokinetic profiles of Sal A have been reported, however there is no such study reported in monkeys, the species which is more similar to human. The aim of this study is to develop a LC-MS method for the determination of Sal A in monkey plasma and apply it to the pharmacokinetic studies of monkeys. After single intravenous administration of Sal A, the plasma concentration-time curves were observed and the main pharmacokinetic parameters were calculated. The plasma concentration at 2 min (C2 (min)) values were (28.343 ± 6.426), (45.679 ± 12.301) and (113.293 ± 24.360) mg x L(-1) for Rhesus monkeys treated with Sal A at 2.5, 5 and 10 mg x kg(-1). The area under the concentration-time curve (AUC(0-∞)) values were (3.316 ± 0.871), (5.754 ± 2.150) and (13.761 ± 2.825) μg x L(-1) x h, respectively. Furthermore, this method was improved and applied to the simultaneous determination of Sal A, Sal B and Sal C, which provided useful information for preclinical studies and clinical trials of Sal A, Sal B and Sal C.
Administration, Intravenous
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Animals
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Caffeic Acids
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pharmacokinetics
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Chromatography, Liquid
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Drugs, Chinese Herbal
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pharmacokinetics
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Lactates
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pharmacokinetics
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Macaca mulatta
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Mass Spectrometry
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Plant Roots
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chemistry
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Salvia miltiorrhiza
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chemistry
5.IL15 DNA adjuvant enhances cellular and humoral immune responses induced by DNA and adenoviral vectors encoding HIV-1 subtype B gp160 gene.
Ke XU ; Shao-Hua XU ; Xia FENG ; Shuang-Qing YU ; Yi ZENG
Chinese Journal of Virology 2014;30(1):62-65
To enhance the immunogenicity of DNA and adenoviral vector vaccines expressing HIV-1 subtype B gp160, human interleukin 15 (hIL15) DNA adjuvant (pVR-hIL15) was constructed. BALB/c mice received DNA prime/protein boost immunization with pVR-HIVgp160/Ad5-HIVgp160 alone or combined with pVR-hIL15. Cellular and humoral immune responses were evaluated by IFN-gamma enzyme-linked immunosorbent spot assay and enzyme-linked immunosorbent assay, respectively. Compared with those immunized with vaccines alone, the mice immunized with vaccines combined with pVR-hIL15 had significantly increased specific cellular response and antibody titer (P < 0.05). It suggests that the IL15 DNA adjuvant can enhance the immune responses induced by prime-boost regimen using DNA and adenoviral vector encoding HIV-1 subtype B gp160.
Adenoviridae
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genetics
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Adjuvants, Immunologic
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Animals
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Antibodies, Viral
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immunology
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Antibody Specificity
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Female
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Genetic Vectors
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genetics
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HIV Envelope Protein gp120
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immunology
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HIV Envelope Protein gp160
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genetics
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immunology
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HIV Envelope Protein gp41
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immunology
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Humans
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Immunity, Cellular
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Immunity, Humoral
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Interleukin-15
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genetics
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Mice
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Mice, Inbred BALB C
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Vaccines, DNA
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genetics
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immunology
6.Clinical analysis of 56 patients with relapsing polychondritis
Xu-Hua SHI ; Jin-Mei SU ; Zhi-Ke CHEN ; Feng-Chun ZHANG ; Fu-Lin TANG ;
Chinese Journal of General Practitioners 2005;0(10):-
Objective To investigate clinical characteristics of relapsing polychondritis(RP)and to improve early recognition for it.Methods Clinical and laboratory data of 56 patients with RP were analyzed retrospectively.Results Ratio of number of male patients to female ones was 1.2.Age at onset was(46?11)years(ranging from 27 to 71)and average interval between onset and diagnosis was(21? 35)months,(8?6),(16?31)and(29?37)months for patients initial onset with auricle,respiratory tract and joints involved,respectively.Site involved included airway in 40 patients(71.4%),auricle in 32 (57.1%),joints in 32(57.1%),eyes in 27(48.2%),nasal chondritis in 25(44.6%)and inner ear in 13(23.2%).At initial stage of the course,17 patients were misdiagnosed as respiratory infection (30.4%),nine as perichondritis(16.1%),six as pulmonary tuberculosis(10.7%),five as rheumatoid arthritis(8.9%).Seven of 40 patients with airway involvement received metallic stents for their tracheobronchial stenosis.Four patients whose condition never improved after regular therapy all had respiratory involvement.Conclusions Patients of RP with initial onset at non-auricle,non-nasal sites tended to be misdiagnosed.Prevalence of airway involvement was not so low with a poor prognosis in patients of RP.
7.Analysis on the causes of unscheduled suspensions of knee and hip arthroplasty
Yang LI ; Zijian LI ; Ke ZHANG ; Hua TIAN ; Yanqing LIU ; Hong CAI ; Feng LI ; Minwei ZHAO
Journal of Peking University(Health Sciences) 2017;49(2):231-235
Objective:To analyze and summarize the causes of unscheduled suspension of knee and hip arthroplasty and to provide the method for optimizing the patient's pre-operative management and improving the efficiency of medical resources as well as the patient's satisfaction.Methods: The data for this report was retrospectively collected from September 2013 to August 2014 in our hospital,from cases of knee and hip arthroplasty that were suspended before the scheduled operation time.Acquisition data from the collected cases including the patients' gender,age and the surgical procedure.At the same time,the suspension reasons were recorded and analyzed.All the decisions of suspension was made by the surgeons and the anesthesiologists according to the abnormal result of preoperative examinations,after communicating with the patients and their families and obtaining their understandings.Results: In the collecting period,our department scheduled 1 146 cases of knee and hip arthroplasty,among which 1 003 were completed,143 suspended (12.5% suspension rate).Among the causes of suspension,the top four common causes were cardiovascular disease (44/143,31%),other infections (20/143,14%),bacteriuria (18/143,13%) and inappropriate surgical indication (16/143,11%).Other causes include surgeon's reason,Blood system abnormalities,high inflammatory index,deep vein thrombosis,other diseases uncontrolled,abnormal liver function and poor diabetes mellitus control,etc.For the rate of suspension,there was no significant difference between the patients with different genders (male: 15.0%,and female: 11.7%,P=0.149),or age (≤50 years: 13.0%;51-65 years: 11.6%;66-80 years 13.3%;>80 years 11.1%;P=0.864).However compared with knee arthroplasty,hip arthroplasty had a higher suspension rate (knee arthroplasry 11.1%,hip arthroplasry 16.1%,P=0.021).Conclusion: It is important to educate and manage the patients before their knee and hip arthroplasty.Through clear diagnosis,detailed medical history analysis careful physical examination,and targeted outpatient examinations and tests for which priority was focused on cardiovascular or other system diseases we could minimize the occurrence of operative suspension post hospitalization,therefore improving the efficiency of the use of medical resources.
8.The comparison of color Doppler ultrasound and computed tomography angiography in the preoperative planning of lower extremity perforators
Shaoqing FENG ; Wenjing XI ; Jue WANG ; Hua LI ; Ke LI ; Yixin ZHANG
Chinese Journal of Microsurgery 2016;39(1):26-32
Objective To compare the application of CDU and CTA in preoperative planning of lower extremity perforator flaps.Methods From February, 2013 to December, 2014, 40 patients who underwent reconstruction with perforator flaps harvested from lower extremities were treated.The position, caliber and route of the perforator vessels were detected by CDU and CTA pre-operatively.The results of the pre-operative navigation with both imaging techniques assisted the selection of the perforators and the flap design and were verified during the operation.Results CDU had a higher accuracy in terms of identification (95.0% vs.90.0%) and location (95.0% vs.82.5%) of the perforators in the lower extremity.There was no significant difference between the two methods in examination and image analysis time.All the flaps survived, besides 1 flap presented with a 2.0 cm × 2.0 cm distal necrosis which healed after regular dressing change.For 35 cases, the donor sites were closed directly while partial skin grafting were performed in 5 case.Conclusion Pre-operative imaging techniques are capable of offering valuable anatomical information of perforator vessels, which can make flap design more convenient, reduce donor site morbidity and improve the surgical results.The authors recommend CDU for pre-operative planning when harvesting perforator flaps from the lower extremity.
9.Monitoring the post-operative lymphedema in prefabricated flap after resurfacing for facial scar
Ke LI ; Shaoqing FENG ; Hua LI ; Peiru MIN ; Wenjing XI ; Ningfei LIU ; Yixin ZHANG
Chinese Journal of Microsurgery 2015;38(5):451-455
Objective Currently, the prefabricated flap is used for reconstructing the post-burn scar in head and face.The aim is to analyze the nature of the post-op edema in the prefabricated flaps, to determine whether it is lymphedema, and to study the prognosis of the edema.Methods From January, 2011 to March, 2015, 18 cases of post-operation edema in superficial temporal fascia prefabricated flaps were studied and each case was followed for 5 months.We used ultrasound and indocyanine green (ICG) fluorescent imaging to study the nature of the edema.Apart from that, we monitored the recovery process of the post-operation edema by a) measuring the horizontal diameter of the flap by a tape, b) measuring the thickness of the flap by the ultrasound, c) monitoring the condition of edema by ICG and d) monitoring the reconstruction and recanalization of the lymphatic system by ICG.Results Pitting edema always occurred in the prefabricated flap from the first day after operation.These patients were diagnosed as mild lymphedema (7 cases), moderate lymphedema (10 cases), and severe lymphedema (1cases) by ultrasound and ICG florescent imaging.The degree of flap lymphedema improved gradually and it improved significantly from 3 days to 6 days post-op eration.The condition of flap lymphedema became stable from the 12 days post-operation.The fluorescence image of ICG showed that the stage of flap lymphedema improved from 3 weeks post-operation and the transportation capability of lymphatic vessels increased most from the 3 days to 6 days post-op eration.We also found lymphatic vessels reconnected from the 6 days post-operation and lymphatic system reconstructed following the direction to lymph nodes.We could discover the collecting lymphatic vessels from 2 months post-operation and the lymphatic system in prefabricated flap matured from 5 months post-operation.Conclusion The post-operation edema in the prefabricated flaps is lymphedema, the incidence rate of which is high.The lymphatic drainage system recovers in 6 days after the operation.It is reconstructed along the lymph nodes.The edema disappears in 2-3 weeks post-operation, the reconstructed the lymph system matures in 5 months.
10.Study on imaging parameters of multi-slice spiral CT long coverage angiography in intracranial and cervical arteries
Bao-Dong JIANG ; Xiao-Yuan FENG ; Cheng LIU ; Ke LI ; Fu-Hua YU ;
Chinese Journal of Radiology 2001;0(09):-
Objective To explore application of the bolus-triggered technique and optimal match between collimation and pitch of MSCTA in intracranial and cervical arteries.Methods(1)The small-dose tests were performed in 19 healthy volunteers and the theoretical threshold was obtained by the time-density curve.(2)forty healthy volunteers were divided randomly into two groups and the scanning parameters were as follows.Group A:collimation 1.0 mm,pitch 1.750. Group B:collimation 2.5 mm,pitch 0.625. Statistical significance was determined with the X~2 test and t test(?=0.05).Results(1)In 19 volunteers,CT value 4 seconds before the peak was 75 HU,the CT value at the beginning segment of the carotid artery and the C_1 segment of internal carotid artery was in accordance with the standard.(2)Image quality ofⅠ,Ⅱgrades artery structures of group B was superior to that of group A.Image quality ofⅢ,Ⅴ grades artery structures of group A was superior to that of group B.Conclusions(1)MSCTA in intracranial and cervical arteries can display systematically the cerebral and carotid arteries.(2)The bolus-triggered technique can improve the image quality of the target vessels.The image quality of the MSCTA of intracranial and cervical arteries is better with the threshold of 75 HU on the basis of 3.5 ml/s injection rate.(3)On the basis of the same other parameters,the optimal scanning parameters are a collimation of 1.0 mm with a pitch of 1.750.