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.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.
6.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.
7.Gabapentin reduces mechanical allodynia in a rat model of tibial bone cancer pain
Liping CHEN ; Wen SHEN ; Dongmei YUE ; Xueming HU ; Jiao LIU ; Yan YUAN ; Zhengliang MA
Chinese Journal of Behavioral Medicine and Brain Science 2012;21(8):693-695
ObjectiveTo explore the effects of gabapentin on mechanical allodynia in rats with tibial bone cancer pain (BCP).MethodsForty-two female SD rats were randomized into 7 groups ( n=6):naive group (group N ),sham operation + NS control group (group SN),sham operation + GBP200mg/( kg · d) group (group SG200),BCP + NS control group (group BN),BCP + GBP50mg/( kg · d) group ( group BGS0),BCP +GBP100mg/(kg · d) group (group BG100),and BCP + GBP200mg/(kg · d) group (group BG200).The rats in group N,SN and BN received 5 ml normal saline and the rats in group SG200,BG50,BG100 and BG200 received 200,50,100 and 200 mg/( kg · d) dose of GBP via feeding from day 7 to 13 after operation,respectively.Mechanical withdrawal threshold(MWT) of the right paw and behavioral assays for ambulatory pain were measured just before operation and on days 1,3,5,7,8,10,12 and 14 after operation.ResultsMWT( (3.78 ± 0.38)g) in rats with BCP decreased and behavioral assays for ambulatory pain (0.76 ± 0.44) increased on day 7 after operation,as compared with those in group N ( ( 14.50 ± 1.38 ) g,(0.00 ± 0.00 ) ) and group sham ( ( 10.21 ± 0.88 ) g,( 0.00 ±0.00) ) (P < 0.05 ).There was no apparent praxiological difference between group SN and group SG200 in a week of continuous application of gabapentin(P> 0.05 ).Compared with those in group BN,there was no change on MWT in group BG50 (P > 0.05 ),and however,behavioral assays for ambulatory pain decreased (P < 0.05 ).MWT in group BG100( (5.35 ±0.85)g) and BG200( (5.71 ±0.72) g) increased in day 10 after operation,as compared with those in group BN ( ( 2.61 ± 0.40) g) and group BG50 ( ( 3.28 ± 1.15 ) g) (P < 0.05 ),and the difference was still statistically significant until day 14 (P < 0.05 ).Behavioral assays for ambulatory pain in group BG100 and BG200 decreased from day 8 after operation,as compared with those in group BN and group BG50 (P < 0.05 ).ConclusionGabapentin,in medium to large dosage,can inhibit pain reaction of rats with bone cancer pain.Nevertheless,with the development of cancer,the effect of gabapentin decreases.
8.Diagnostic value of DCE-MRI in adenoid cystic carcinoma and pleomorphic adenoma of head and neck
Xiaodong JI ; Shuo YAN ; Shuang XIA ; Huazheng DONG ; Yu GUO ; Wen SHEN
Journal of Practical Radiology 2015;(5):735-739
Objective To evaluate the diagnostic value of dynamic contrast-enhanced magnetic resonance imaging (DCE-MRI)for the differential diagnosis of the adenoid cystic carcinoma and pleomorphic adenoma of the head and neck.Methods Thirty-one pa-tients with histopathologically proved adenoid cystic carcinoma and pleomorphic adenoma were examined with DCE-MRI,in which 10 cases were benign and 21 cases were malignant.The data of the conventional 3.0T DCE-MRI were improved by the 3D fast spoiled gradient-echo sequence (FSPGR)method.Z-test was performed on the quantitative parameters for benign and malignant le-sions,including volume transfer constant (Ktrans ),rate constant (Kep )and extravascular extracellular space fraction (Ve ).The re-ceiver operating characteristic (ROC)curves were plotted to investigate the diagnosis.Results The mean Ktranss of the adenoid cystic carcinoma group and pleomorphic adenoma group were (0.266 ± 0.103 )min-1 and (0.1 55 ± 0.080)min-1 respectively,and the difference between them were statistically significant (Z =-2.699,P < 0.05).The difference between the keps of the aforementioned two groups was of statistical significance (Z =-2.263,P < 0.05),while that of the ves is without statistical significance (Z =-1.132, P >0.05).The areas under the ROC curves of the Ktrans and kep were 0.813 and 0.763 respectively.Choosing the optimal diagnostic cut-off points corresponding to the maximum Youden indexes 0.173 min-1 and 0.818 min-1 ,the sensitivities of Ktrans and kep for identifying the adenoid cystic carcinoma and pleomorphic adenoma were 90.9% and 81.8%,and the specificities were 77.8% and 66.7%.Ktrans was of the highest sensitivity and specificity for the identification of the adenoid cystic carcinoma and pleomorphic adenoma.Conclu-sion The dynamic contrast-enhanced parameter Ktrans plays a certain role in the differential diagnosis of the adenoid cystic carcinoma and pleomorphic adenoma of the head and neck.
9.Reasons of non-planned decannulation of peripheraly inserted central catheters and their prevention
Jinxin SHEN ; Hongli SU ; Yan LIU ; Xiangyu WEN ; Yuhua LIU ; Jing WANG
Chinese Journal of Clinical Nutrition 2009;17(5):306-307
Objective To analyze the reasons of non-planned decannulation (NPD) of peripheraly inserted central catheter (PICC) and explore the applicable prevention strategies.Methods We retrospectively analyzed the clinical data of 432 patients who underwent PICC cannulations maintenance between July 2005 and June 2008.The reasons of 26 cases of NPD were analyzed.Results The main reasons of NPD included cannulation occlusion,cannulation ectopia or translocation,cannulation exfoliation,cannulation-related infections,mechanical phlebitis,and thrombus formation.The complications usually were caused by the poor cannulation material,incorrect fixation,top of canuulation not reaching superior vena cava,lack of management measures for outpatients,accidents,and others.Effective measures for preventing NPD included:use silica gel cannulation;ensure the top of canuulation reach the superior vena cava;and improve the communications between outpatients and nurses.Conclusion NPD of PICC may be caused by various reasons and effective prevention measures should be taken.
10.Effects of intrathecal methotrexate on activation of microglia in spinal cord in a rat model of tibial cancer pain
Wen SHEN ; Dongmei YUE ; Jiao LIU ; Liping CHEN ; Youmiao XU ; Yan CHEN ; Xueming HU
Chinese Journal of Anesthesiology 2012;(11):1311-1313
Objective To evaluate the effects of intrathecal methotrexate on the activation of microglia in spinal cord in a rat model of tibial cancer pain (TCP).Methods Thirty female Sprague-Dawley rats,aged 5-7 weeks,weighing 150-180 g,were randomly divided into 3 groups (n =10 each):sham operation + artificial cerebrospinal fluid (group SA),TCP + artificial cerebrospinal fluid (group CA),and TCP + methotrexate (group CM).TCP was induced by injecting Walker-256 cancer cells into the medullary cavity of tibia.Artificial cerebrospinal fluid or methotrexate 100μg (15μl) was injected intrathecally over 10 min on 7th day after TCP.Mechanical pain threshold (MPT) was measured before TCP,at 1,3,5 and 7 days after TCP and 2,4,8 and 24 h after administration (T0-8).The rats were sacrificed after measurement of the pain threshold at T8 and the spinal cord was isolated for detection of the activation of microglia (by immunofluorescence) and content of tumor necrosis factor-α (TNF-α) and IL-1β (by ELISA).Results Compared with group SA,MPT was significantly decreased,and the number of activated microglia cells in the spinal cord was increased,and the contents of TNF-α and IL-1β were increased in groups CA and CM (P < 0.05).Compared with group CA,MPT was significantly increased,and the number of activated microglia cells in the spinal cord was decreased,and the contents of TNF-α and IL-1β were decreased in group CM (P < 0.05).Conclusion The mechanism by which intrathecal methotrexate reduces TCP in rats is related to inhibition of the activation of microglia and reduction of the secretion of proinflammatory cytokines TNF-α and IL-1β in the spinal cord.