2.Cloning and eukaryotic expression of human CD38
Xinyu WEN ; Cuiling SHU ; Yan LI ; Zhongtian QI ; Beife SHEN
Academic Journal of Second Military Medical University 2001;0(09):-
Objective:To clone and express the full length cDNA of human CD38. Methods:The full length cDNA of the human CD38 antigen was amplified from total RNA of Daudi cell by RT-PCR, and it was inserted into pGEM-T. The validity on the sequences was confirmed by automatic DNA sequencing. Inserting the valid CD38 gene into pcDNA3.1(+) plasmid to obtain recombinant mammalian expression vector pcDNA3.1(+)/CD38Z; Using lipofectin gene transfer technique system, recombinant expression vector containing CD38 gene was transfected into COS7 cells. The expression of CD38 molecules on the surface of COS7 cells was detected by FACS and immunohistochemical technique. Results:DNA sequencing showed that the cloned full length cDNA sequence was identical with reported. The result of FACS and immunohistochemical technique indicated that CD38 molecules were expressed on the surface of COS7 cells. Conclusion:The full length cDNA of human CD38 is obtained, recombinant mammalian expression vector pcDNA3.1(+)/CD38Z is successfully constructed, and the CD38 molecules is expressed on the surface of COS7 cells,this may facilitate studies on the biochemistry and function of CD38 antigen.
3.EFFECTS OF 1?,25(OH)_2D_3 ON THE EXPRESSION OF RECEPTOR ACTIVATOR OF NF-?B LIGAND AND OSTEOPROTEGERIN IN RAT OSTEOBLASTS IN VITRO
Jianhong GU ; Yan YU ; Shanshan JIANG ; Yan SHEN ; Wen YAN ; Zongping LIU
Acta Nutrimenta Sinica 1956;0(02):-
Objective To investigate the effect of 1?,25-dihy-droxyvitamin D3 (1?,25(OH)2D3) on the expression of receptor activator of NF-?B ligand (RANKL),osteoprotegerin (OPG) and RANKL mRNA,OPG mRNA,of rat osteobalsts (OB). Method The expression of RANKL and OPG was detected by the method of Immunohistochemistry and ELISA. RANKL mRNA and OPG mRNA were determined through FQ-PCR. Results Compared with the control group and the group with 1 nmol/L 1?,25(OH)2D3,10 and 100 nmol/L 1?,25(OH)2D3 can significantly induce the expression of RANKL and RANKL mRNA. 1,10 nmol/L 1?,25(OH)2D3 can stimulate the expression of OPG and OPG mRNA significantly,while 100 nmol/L 1?,25(OH)2D3 can inhibit the expression of OPG mRNA significantly. There were no difference in the rate of RANKL/OPG and RANKL mRNA/OPG mRNA between the control group and the group with 1 nmol/L 1?,25(OH)2D3,however the rate of RANKL/OPG and RANKL mRNA/OPG mRNA in the group with 10,100 nmol/L were higher than the control group and the group with 1 nmol/L 1?,25(OH)2D3. Conclusion Lower dosage of 1?,25(OH)2D3 (1 nmol/L) had no significant effect on bone turnover,but higher dosage of 1?,25(OH)2D3 (10,100 nmol/L) can enhance bone turnover through facilitating the formation and activity of osteoclasts via enhancing RANKL mRNA/OPG mRNA and RANKL/OPG..
4.Design, synthesis and pharmacological investigation of isoindoline derivatives as 5-HT/NE double reuptake inhibitors.
Hui WEN ; Yuan SHI ; Jing-wen DONG ; Yan-shen GUO ; Jian-Jun ZANG ; Guang-zhong YANG
Acta Pharmaceutica Sinica 2015;50(9):1148-1155
A series of isoindoline derivatives were designed, synthesized, and evaluated for their double inhibitory activities. All of them were new compounds, and their structures were confirmed by 1H NMR and HR-MS. Preliminary in vitro pharmacological tests showed that all compounds exhibited 5-HT or NE reuptake inhibition activity. Among the tested compounds, compound I-3 exhibited potent inhibitory activity against 5-HT and NE reuptake in vitro, and exhibited potent antidepressant activity in vivo. These compounds designed can be further optimized for finding more potent 5-HT/NE dual reuptake inhibitors and antidepressant candidates as well.
Antidepressive Agents
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chemical synthesis
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chemistry
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Biological Transport
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Drug Design
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Isoindoles
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chemical synthesis
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chemistry
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Serotonin Uptake Inhibitors
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chemical synthesis
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chemistry
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Structure-Activity Relationship
5.The significance of antibodies to anti-coagulation related factors and anti-phospholipid antibodies in glomerular microthrombosis in lupus nephritis:a prospective study on 124 patients
Hui ZHENG ; Wen AO ; Yan SHEN ; Xiaowei CHEN ; Min DAI ; Xiaodong WANG ; Yucheng YAN ; Chengde YANG
Chinese Journal of Rheumatology 2009;13(5):296-300,前插1
Objective To investigate the prevalence of glomerular microthrombosis in lupus nephritis (LN) and the significance of antibodies to anti-coagulation related factors and anti-phospholipid antibodies in glomerular microthrombosis (GMT).Methods Kidney biopsy specimens and plasma samples were obtained consecutively from 124 patients with LN. Kidney biopsy specimens were examined for the presence of glomerular microthrombi.Plasma samples from 25 LN patients with GMT (LN-GMT group) and 99 LN patients without GMT (LN-non-GMT group) were tested for lupus anticoagnlant (LA) and antibodies to cardiolipin (ACL),β2 glycoprotein I (β2GP I ),plasmin,thrombin,tissue plasminogen activator (t-PA) and Annexin A II.Results The prevalence of GMT in LN patients was about 20.2%.Compared to LN-non-GMT group,LN-GMT group had elevated SLE disease activity indices (SLEDAI),elevated activity and chronicity indices of kidney tissue injury,and elevated serum creatinine,blood urea nitrogen and proteinuria levels,and also had a higher frequency of hypertension (P<0.01).The positive rates of LA,IgG class anti-β2GP I and anti-thrombin antibodies were higher in LN-GMT group than in LN-non-GMT group (P<0.05).The positive rates of IgG class antibodies to ACL,plasmin,t-PA and Annexin A II in LN-GMT group were not statistically different from those in LN-non-GMT group (P>0.05).No difference was found in the positive rate of any IgM class antibody between the two groups (P>0.05).Conclusion This study has shown that GMT occurs approximately in 20.2% of the LN patients.Patients with GMT have more severe kidney tissue injury and more poor renal outcomes than patients without GMT.LA and antibodies to β2GP I and thrombin play a role in glomerular microthrombosis in lupus nephritis.
6.Role of chemokine ligand 2 in spinal eord in a rat model of tibia bone cancer pain
Youmiao XU ; Wen SHEN ; Yan CHEN ; Hongli YUE ; Jiao LIU ; Dongmei YUE ; Yan YUAN ; Dong HANG
Chinese Journal of Anesthesiology 2011;31(9):1052-1055
Objective To investigate the role of chemokine ligand 2 (CCL2) in the spinal cord expression in a rat model of tibia bone cancer pain.Methods Eighty-four female SD rats weighing 160-180 g were randomly divided into 3 groups ( n =28):control group (group C),sham operation group (group S) and tibia bone cancer pain group (group P).Tibia bone cancer pain was induced by intra-tibial inoculation of Walker-256 breast cancer cells.Paw withdral threshold to mechanical stimulation (MWT) was measured with von Frey filaments at 1 d before and at 1,3,7,10,14 and 21 d after inoculation.Six rats in each group were sacrificed after the measurement of MWT at 1 d before inoculation and at 7,14 and 21 d after inoculation.Lumbar 4-6 segments of the spinal cord were removed for determination of the expression of CCL2 by ELISA.The coexpression of CCL2 with Iba-1 (a specific marker of microglia),GFAP(a specific marker of astrocyte) and NeuN (a specific marker of neuron) was determined by double immunofluorescence assay after the measurement of MWT at 14 d after inoculation in group P.Results Compared with groups C and S,MWT was significantly decreased from 7 d to 21 d after inoculation,the expressive of CCI-2 in the spinal cord up-regulated at 7,14 and 21 d after inoculation in group P ( P < 0.05).CCL2 was expressed in the microglia and astrocyte but not in neuron in the spinal cord dorsal horn in a rat model of tibia bone cancer pain.Conclusion Release of CCL2 from microglia and astrocytes in the spinal cord was involved in mechanical hyperalgesia in a rat model of tibia bone cancer pain.
7.Evaluation of trazodone for treatment of sleep disorders and anxiety in methamphetamine addicts after detoxification
Yue LIU ; Yan-Yan HUANG ; Wen-Wen SHEN ; Jian-Bing ZHANG ; Wen-Hua ZHOU
The Chinese Journal of Clinical Pharmacology 2015;(5):336-338
Objective To observe the effect of trazodone on sleep disorders and anxiety in methamphetamine addicts after detoxification. Methods One hundred and six cases were randomly divided into control group(n=53)and treatment group(n=53).Patients in treatment group were given trazodone, while the controls were given placebo, when the detection of methamphetamine in urine were all negative.All of them were toxicity-free and were accessed by pittsburgh sleep quality index ( PSQI) and hamilton anxiety scale ( HAMA) before treatment and at the end of the first week, the second week and the fourth week.The effect of trazodone on sleep quality and anxiety in methamphetamine addicts were observed.The treatment emergent symptom scale ( TESS ) was used to make safety evaluation.Results Compared with the control group, the data PSQI and HAMA decreased significantly in the treatment group at the end of the second week and the fourth week (P<0.05).Conclusion Trazodone is effective in improving sleep disorders and reducinganxiety of methamphetamine addict after detoxification.It cansignificantly prevent relapse.
8.Effects of phosphatidylinositol-3 kinase-protein kinase B signaling pathway on iodine-intake ability of lactating rats
Da WEN ; Liang XUE ; Lixiang LIU ; Dandan WANG ; Yingnan YAN ; Hongmei SHEN
Chinese Journal of Endemiology 2016;35(6):395-400
Objective To observe the variation of phosphatidylinositol-3 kinase (PI3K),protein kinase B (AKT),sodium iodide symporter (NIS) mRNA and protein expression in rat mammary tissues and serum insulin growth factor Ⅰ (IGF-1) under different iodine nutrition levels,and to study the role of PI3K-AKT signaling pathway in the process of mammary gland intaking iodine during lactation period.Methods Totally 130 Wistar rats (100 female rats,30 male rats) were randomly divided into five groups with 20 female rats in each group:①control group (NI):was feed with normal diet and iodine content 50 μg/L in deionized water;②low iodine group 1 (LI1 group):was feed with low iodine diet and deionized water;③low iodine group 2 (LI2):was feed with low iodine diet and iodine content 5 μg/L in deionized water;④high iodine group 1 (HI1 group):was feed with normal diet and iodine content 3 000 μg/L in deionized water;⑤high iodine group 2 (HI2):was feed with normal diet and iodine content 10 000 μg/L in deionized water.After feeding for 3 months,females were mated with male rats,then male rats were taken out and every female rat was feed individually.Urinary iodine level of rats in lactation period 10 days after giving birth was tested.Blood and mammary tissue samples of rats in lactation period were taken after killing them.Enzyme linked immunosorbent assay (ELISA) was used to detect serum IGF-1 level,real-time fluorescence quantification PCR to detect the mRNA expression of mammary gland PI3K,AKT and NIS,Western blotting to detect mammary gland PI3K,total AKT,phosphorylation AKT (p-AKT) and NIS protein expression.Results The medians urinary iodine of lactation period rats in LI1 and LI2 (3.16,6.36 μg/L) were significantly lower than that in NI group (162.59 μg/L),and were significantly higher in HI1 and HI2 (2 356.27,11 507.29 μg/L) than that in NI group.The differences were statistically significant (all P < 0.01).Compared with control group [(8.84 ± 2.12) μg/L],the content of serum IGF-1 increased significantly in lactation period rats in LI1 and LI2 groups [(13.30 ± 2.37) and (10.90 ± 1.92) μg/L,all P< 0.01].The real-time fluorescence quantification PCR detection results indicated that the differences were statistically significant by comparing NIS,AKT,PI3K mRNA expression of the mammary tissues of lactation period rats in the five groups (F=14.916,36.477,14.994,all P< 0.01).Among them,NIS mRNA expression quantities in LI1 and LI2 groups (0.75 ± 0.40,0.89 ± 0.51) were significantly higher than that in NI group (0.53 ± 0.31),and significantly lower in HI2 group (0.30 ± 0.24) than that in NI group (P < 0.05 or < 0.01).AKT mRNA expression quantities in LI1 and LI2 groups (0.90 ± 0.19,0.64 ± 0.22) were significantly higher than that in NI group (0.43 ± 0.22),and significantly lower in HI2 group (0.29 ± 0.15) than that in NI group (P < 0.05 or < 0.01).PI3K mRNA expression quantity in LI1 group (0.85 ± 0.42) was significantly higher than that in NI group (0.50 ± 0.24),and significantly lower in HI2 group (0.28 ± 0.10) than that in NI group (all P < 0.01).Western blot detection results indicated that the differences were statistically significant by comparing mammary gland NIS protein expression of lactation period rats in the five groups (F=4.510,P< 0.01).Among them,LI1 group (1.67 ± 0.97) was significantly higher than NI group (0.87 ± 0.43,P < 0.05).The differences were statistically significant by comparing the p-AKT protein expression among groups (F =3.528,P < 0.05).Among them,HI2 group (1.10 ± 0.30) was significantly higher than NI group (0.75 ± 0.23,P <0.05).The differences were not statistically significant by comparing total AKT and PI3K protein expression among groups (F =0.558,1.319,all P > 0.05).Conclusion The inhibitory effect of PI3K-AKT signaling pathways on NIS in the mammary gland was weaker than the effect of iodine intake.But the expression of functional p-AKT was gradually increased with the increment of iodine intake,which had been presented inhibit effect on NIS expression in lactating mammary gland.
9.Solitary fibrous tumor of pleura associated with episodic hypoglycemia: report of a case.
Zong-kai ZOU ; Wen-qiao WU ; Hai-yan SU ; Hong-wu SHEN
Chinese Journal of Pathology 2007;36(1):67-67
Antigens, CD34
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metabolism
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Female
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Humans
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Hypoglycemia
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etiology
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Immunohistochemistry
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Middle Aged
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Pleura
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metabolism
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pathology
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surgery
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Solitary Fibrous Tumor, Pleural
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complications
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metabolism
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surgery
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Treatment Outcome
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Vimentin
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metabolism
10.Evaluation of SYTO13 as fluorescent dye for high resolution melting based single nucleotide polymorphism genotyping
Qian WANG ; Wen YAN ; Minghui SHEN ; Kejun MA ; Li′na GAO ; Chongge YOU ;
Chinese Journal of Laboratory Medicine 2017;40(2):88-94
Objective To evaluate the utility of fluorescent dye SYTO 13 for high -resolution melting ( HRM) detection in single nucleotide polymorphism ( SNP) genotyping and its clinical application . Methods This is a performance verification study .36 genotype defined samples were divided into three groups:SNP rs3125734 C>T (class Ⅰ SNP) ,rs255758 A>C (class ⅡSNP) and rs688C>T.These samples were used to evaluate SYTO 13′s SNP genotyping capability of class ⅠSNP, classⅡSNP, and two PCR products of different lengths (52 and 107 bp) covering the same SNP of rs688C>T.The commercial HRM dye of LCGreen Plus was used as the control .The genotyping capability is indicated by the Tm difference(ΔTm) between wild type and homozygous mutant genotypes .The Tm differences between wild genotype and homozygous mutant genotype were compared using the Independent Samples t test.Paired t test was used to evaluate genotyping capability of the two dyes .The clinical applicability is evaluated by synchronously performing PCR amplification and HRM analysis on thirty -five randomly selected DNA samples with known genotypes of the three SNPs .Results The SNPs of class Ⅰ and class Ⅱ can be genotyped directly and clearly with SYTO13 (ΔTmclas Ⅰ =0.36 ±0.05,tclas Ⅰ =14.827,Pclas Ⅰ =0.000;ΔTm clas Ⅱ =0.42 ±0.110,tclasⅡ =9.539,Pclas Ⅱ =0.000).The classⅠSNP genotyping results was better using SYTO13 (ΔTmSYTO13 =0.39 ±0.027), while the SNP genotyping for small amplicon did not discriminated clearly in this study .Long amplicons of class ⅠandⅡSNPs can be identified directly except for several samples which can be genotyped accurately after having performed reexamination .Conclusion SYTO13 can apply for HRM analysis of genotyping classⅠand ⅡSNPs with long amplicon and for clinical routine detection.