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
;
agonists
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chemistry
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Plasmids
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Response Elements
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Sulfonamides
;
chemistry
;
Transcriptional Activation
3.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
;
Drugs, Chinese Herbal
;
pharmacokinetics
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Lactates
;
pharmacokinetics
;
Macaca mulatta
;
Mass Spectrometry
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Plant Roots
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chemistry
;
Salvia miltiorrhiza
;
chemistry
4.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
5.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.
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.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.
8.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.
9.Expression of interleukin-17 and interleukin-22 in supernatant of peripheral blood mononuclear cell and tear in dry eye patient
Xu-hua, TAN ; Song, SUN ; Ting-ting, ZHU ; Tian-li, REN ; Zhi-feng, WU ; Ke, WANG
Chinese Journal of Experimental Ophthalmology 2013;(7):668-673
Background T helper cell 17 (Th17),a newly discovered subset of CD4+ T cells,have been found to play an important role in dry eye disease in animal model.Further investigation should be done on the immunopathogenesis of Th17 cells in dry eye patients.Objective This study was to analyze the expression status of interleukin-17 (IL-17) and IL-22 in tear and supernatant of peripheral blood mononuclear cells (PBMCs) of dry eye patients and their correlation with clinical symptom and sign.Methods Twenty Sj(o)gren syndrome (SS)patients,twenty non-Sj(o)gren syndrome (NSS) patients were included in Wuxi Second Hospital from 2010 to 2011,and twenty healthy volunteers were recruited at the same period.All of subjects understood the purpose and procedure of research and written informed consent was obtained form each subject initial of this study.Dry eye symptom questionnairs were self-answered and multiple dry eye disease-related clinical tests,including the breakup time of tear film (BUT),Schirmer Ⅰ test (S Ⅰ t) and corneal fluorescein staining were performed.The periphery blood of 3 ml and tear were collected in all the subjects,and IL-17 and IL-22 levels in supernatant of PBMCs and tear were detected by enzyme-linked immunosorbent assay (ELISA).The correlations between levels of IL-17 and IL-22 with BUT,S Ⅰ t,corneal fluorescein staining and dry eye scores were analyzed.Results The dry eye scores reduced,BUT prolonged,S Ⅰ t increased and corneal fluorescein dye decreased from SS group,NSS group to normal control group,with significant differences among the three groups (dry eye scores:H =40.81,P<0.01 ; BUT:H =40.15,P<0.01 ;S Ⅰ t..H=50.07,P<0.01 ;corneal dye scores:H=40.52,P<0.01).The concentration of IL-17 in the supernatant of PBMCs in the SS patients,NSS patients and normal controls were (964.92±124.83)ng/L,(718.85± 115.89)ng/L and (341.95±85.08) ng/L,showing a statistically significant difference among them (F=162.95,P<0.01).The levels of IL-17 in the tear were (440.69±126.09) ng/L,(364.33±126.85) ng/L and (61.16±11.60) ng/L in the SS group,NSS group and normal control group respectively,exhibiting an elevated level in the SS group and NSS group compared with the control group (F=75.27,P<0.01).In addition,the levels of IL-22 in the supernatant of PBMCs in the SS patients,NSS patients and normal controls were (98.77± 11.27) ng/L,(79.65 ± 11.01) ng/L and (32.78±9.34) ng/L,and those in the tear were (22.22 ± 8.96) ng/L,(14.92 ±4.35) ng/L and (10.47 ± 2.67) ng/L,with significant differences among the three groups (F =206.27,P<0.01 ;F =19.87,P<0.01).The significant correlations were found between the IL-17 and IL-22 concentration in the supernatant of PBMCs and tear with corneal fluorescein staining scores and S Ⅰ t.Conclusions The contents of IL-17 and IL-22 in PBMCs and tear upregulate in the SS and NSS patients,indicating that Th17 plays a key role in the immunity mechanism of dry eye.
10.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
;
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
;
immunology
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HIV Envelope Protein gp41
;
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
;
genetics
;
immunology