1.The killing effect of NY-ESO-1-sensitzed dendritic cells-induced cellular toxic lymphocyte on human retinoblastoma cell
Chinese Journal of Experimental Ophthalmology 2012;30(7):586-591
Background Cell immunologic therapy for retinoblastoma(RB)is becoming a hot research topic.Cancer-testis antigen is a human immunogenic protein and is used to treat some tumors.However,its effect on RB has not been investigated.Objective The present study was to discuss the antigen specific anti-tumor effect of cytotoxic T lymphocytes(CTL)induced by the cancer-testis antigen,NY-ESO-1-sensitized dendritic cells(DCs),on human RB.Methods PCR was performed to amplify target gene fragments from the NY-ESO-1 plasmids,and then the target gene fragments were digested with the restriction enzymes SalI and EcoRI.Harvested fragments were inserted into the pDC316 plasmid to construct the recombinant plasmid pDC316/NY-ESO-1.The expression of NY-ESO-1 protein in human RB cells strain,HXO-RB44,was detected by immunofluorescence and Western blot.Monocytes were isolated from 60 ml of peripheral blood from a healthy donor using Ficoll density-gradient centrifugation with a cell density of 1 × 107/ml.DCs isolated from blood were stimulated with recombinant human granulocyte-macrophage colony stimulating factor(rhGM-CSF)and recombinant human interleukin-4(rhIL-4).The recombinant plasmid pDC316/NY-ESO-1 was transfected into DCs and the DCs were co-cultured with T lymphocytes.The resultant CTL were used as effector cells.The growth of the CTL was detected by MTT assay.The CTL were then added into the growth medium used for culturing HXO-RB44 cells and the vitality of the HXO-RB44 cells was assayed by MTT assay.Results The sequence of the cloned DNA fragment of the recombinant plasmid pDC316/NY-ESO-1 was conforms with the sequence of the NY-ESO-1 gene.The expression of the NY-ESO-1 protein in HXO-RB44 cells was tested by immunofluorescence and Western blot.DCs were successfully induced with rhIL-4 and rhGM-CSF from PBMC.The recombinant expression plasmid pDC316/NY-ESO-1 was successfully transferred into DCs.These DCs had high expression of surface molecules such as HLA-DR(42.1%),CD80(54.2%),CD83(39.7%)and CD86 (94.8%).The CTL that was induced by DCs-sensitized with NY-ESO-1 specifically killed HXO-RB44 cells.CTL induced by the sensitized DCs had a stronger cytotoxic effect against HXO-RB44 cells compared with un-sensitized DCs and CTL un-induced with DCs,as shown by MTT asssay(P<0.05).The anti-tumor activity was highest when the ratio of effector to target was 75∶1(P<0.05).Conclusions DCs transfected by the recombinant plasmid pDC316/NY-ESO-1 can induce the proliferation of allogenic CTLs,which showed a specific anti-tumor effect against HXO-RB44 cells.These results present a new type of immunotherapy for the treatment of RB.
2.Effects of tetrandrine and fructose-1,6-diphosphate on the elevated intrasynaptosomal [Ca~2+]_i induced by excitatory amino acids
Zhi DONG ; Qixin ZHOU ; Chunsheng XUE
Chinese Journal of Pathophysiology 1999;0(09):-
AIM: To study the effects of tetrandrine(Tet) and fructose-1,6-diphosphate(FDP) on the elevated intrasynaptosomal [Ca 2+ ] i induced by excitatory amino acids (EAA). METHODS: A rapid method for preparing synaptosomes was used, and intrasynaptosomal free calcium ([Ca 2+ ] i) was measured by using the fluorescent indicator quin-2. RESULTS: L-glutamate (Glu, 100 ?mol/L), aspartate (Asp, 100 ?mol?L -1 ), N-methy1-D-aspartate (100 ?mol/L) and Glu (50 ?mol/L) plus Asp (50 ?mol/L) all elevated intrasynaptosomal [Ca 2+ ] i in a dose-dependent manner. Pretreatment with Tet (10,30,60 ?mol/L) , FDP (15, 30, 75, 150 ?mol/L), MK-801 (10, 20 ?mol/L) and Tet (15, 30 ?mol/L) plus FDP(15, 30 ?mol/L) all attenuated the increase in intrasynaptosomal [Ca 2+ ] i induced by EAAs mentioned as above in a dose-dependent manner, and the effect of Tet plus FDP was most significant. CONCLUSION: Both Tet and FDP inhibited a rise in intrasynaptosomal [Ca 2+ ] i induced by EAAs, which may be one of mechanisms that Tet and FDP pretect cerebral tissues against ischemia injury.
3.Detective Significance of Cerebral Electrical Admittance Plethysmogram in Newborns with Asphyxia
ke-ying, ZHOU ; jin-zhi, SONG ; jing-zhi, LI ; hui, LUO ; xue-chang, XIAO ; zhi-guang, LI ; ben-qing, WU
Journal of Applied Clinical Pediatrics 2006;0(20):-
Objective To study hemodynamics of cerebral blood flow in newborns with asphyxia.Method Bilateral cerebral electrical admittance plethysmogram(BCEAP) was used to explore characteristics of cerebral blood flow in 20 healthy newborns and 20 newborns with asphyxia respectively from first to fourth day after birth.Results Both the ratio of Hs to b-S(Hs/ b-S) and index of admittance differential loop(ADL) Ⅰ+Ⅱ decreased significantly in newborns with asphyxia compared to normal control from first to fourth day after birth(P
4.Study on the correlation between spiral CT features and expression of phosphatase and tensin homlogy deleted on chormosome ten,basic fibroblast grouth factor in gastric carcinoma
Jian-Bo GAO ; Hui LI ; Xue-Jun CHEN ; Xue-Hua YANG ; Hua GUO ; Zhi-Gang ZHOU ; Song-Wei YUE ;
Chinese Journal of Radiology 2001;0(08):-
Objective To investigate the spiral CT features of gastric carcinoma in the invasion and metastasis and its correlation with the expression of phosphatase and tensin horology deleted on chormosome ten(PTEN)and basic fibroblast grouth factor(bFGF).Methods Spiral CT plain scan and triphasic enhanced scans were performed in 83 patients.The postoperative specimens were embedded with paraffin to obtain 5?m thickness tissues and stained with HE and immunohistochemistry.Spiral CT findings were compared with the expression of phosphatase and tensin horology deleted on chormosome ten(PTEN)and basic fibroblast growth factor(bFGF).Results(1)The accuracy of spiral CT in T and N staging of gastric carcinoma was 94.0%(78/83)and 89.2%(74/83),respectively.(2)The expression of PTEN was 47.0%(39/83)in gastric carcinoma.The expression of PTEN in T_(3.4)(40.8%,29/71)and N_(1+2) (38.3%,23/60)gastric carcinoma was significantly lower than that of T_2(10/12)and N_0(16/23)gastric carcinoma,respectively(X~2=7.439,P=0.006;X~2=6.511,P=0.011).(3)The expression of bFGF was 63.9%(53/83)in gastric carcinoma.The expression of bFGF in T(3.4)(70.4%,50/71)and N_(1+2) (71.7%,43/60)gastric carcinoma was significantly higher than that of T_2(3/12)and N_0(10/23)gastric carcinoma,respectively(X~2=7.314,P=0.007;X~2=5.724,P=0.017).(4)Both PTEN-positive expression and bFGF-positive expression were detected in 16 specimens.The expression of PTEN(41.0%, 16/39)was negatively correlated with that of bFGF(30.2%,16/53)(r=-0.447,P=0.000). Conclusion Spiral CT triphasic enhanced scans combined with biologic characteristics can improve diagnostic accuracy of gastric carcinoma in the invasion,metastasis and prognosis.
5.Myricetin attenuates renal fibrosis by activating Nrf2/HO-1 pathway to inhibit oxidative stress
Dong-xue LI ; Zhou HUANG ; Han-yu WANG ; Zhi-hao ZHANG ; Ning-hua TAN ; Xue-yang DENG
Acta Pharmaceutica Sinica 2024;59(2):359-367
This paper investigates the effect of myricetin (MYR) on renal fibrosis induced by unilateral ureteral obstruction (UUO) and common bile duct ligation (CBDL) in mice and its mechanism. The animal experiment has been approved by the Ethics Committee of China Pharmaceutical University (NO: 2022-10-020). Thirty-five ICR mice were divided into control, UUO, UUO+MYR, CBDL and CBDL+MYR groups. H&E and Masson staining were used to detect pathological changes in kidney tissues. Western blot (WB) was used to detect the expression of fibrosis-related proteins in renal tissue, and total superoxide dismutase (SOD) activity detection kit (WST-8) was used to detect the changes of total SOD in renal tissue of CBDL mice.
6.An exploration in the action targets for antidepressant bioactive components of Xiaoyaosan based on network pharmacology.
Yao GAO ; Li GAO ; Xiao-xia GAO ; Yu-zhi ZHOU ; Xue-mei QIN ; Jun-sheng TIAN
Acta Pharmaceutica Sinica 2015;50(12):1589-1595
The present study aims to predict the action targets of antidepressant active ingredients of Xiaoyaosan to understand the "multi-components, multi-targets and multi-pathways" mechanism. Using network pharmacology, the reported antidepressant active ingredients in Xiaoyaosan (saikosaponin A, saikosaponin C, saikosaponin D, ferulic acid, Z-ligustilide, atractylenolide I, atractylenolide II, atractylenolide III, paeoniflorin, albiflorin, liquiritin, glycyrrhizic acid and pachymic acid), were used to predict the targets of main active ingredients of Xiaoyaosan according to reversed pharmacophore matching method. The prediction was made via screening of the antidepressive drug targets approved by FDA in the DrugBank database and annotating the information of targets with the aid of MAS 3.0 biological molecular function software. The Cytoscape software was used to construct the Xiaoyaosan ingredients-targets-pathways network. The network analysis indicates that the active ingredients in Xiaoyaosan involve 25 targets in the energy metabolism-immune-signal transmutation relevant biological processes. The antidepressant effect of Xiaoyaosan reflects the features of traditional Chinese medicine in multi-components, multi-targets and multi-pathways. This research provides a scientific basis for elucidation of the antidepressant pharmacological mechanism of Xiaoyaosan.
Antidepressive Agents
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pharmacology
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Benzoates
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Bridged-Ring Compounds
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Coumaric Acids
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Drug Evaluation, Preclinical
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Drugs, Chinese Herbal
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pharmacology
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Flavanones
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Glucosides
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Glycyrrhizic Acid
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Lactones
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Medicine, Chinese Traditional
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Monoterpenes
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Sesquiterpenes
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Software
7.Advance in the study of compounds inducing the expression of heme oxygenase-1.
Xue-Lei LUO ; Xiao-Xia ZHOU ; Zhi LIU
Acta Pharmaceutica Sinica 2008;43(6):553-558
Heme oxygenase-1 (HO-1) is a cellular stress protein, and its expression plays an important regulatory role in a lot of physiological and pathological processes. Although the expression of HO-1 in most tissues of body is low, a number of clinical and pharmacological experiments have proved that many compounds can induce HO-1 expression. The increase of HO-1 expression is the result of regulating different signaling pathways and transcription factors, and this induction of HO-1 is suggested to be partially therapeutic efficacy of these compounds. This article summarizes some kinds of compounds in this field of research at home and abroad over the last 10 years, and provides a brief analysis of the mechanism.
Animals
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Antineoplastic Agents
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pharmacology
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Antioxidants
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pharmacology
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Coumarins
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pharmacology
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Drugs, Chinese Herbal
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pharmacology
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Enzyme Induction
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drug effects
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Gene Expression Regulation, Enzymologic
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Heme Oxygenase-1
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genetics
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metabolism
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Humans
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Lovastatin
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pharmacology
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Nitric Oxide
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pharmacology
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Peptide Hormones
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pharmacology
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Probucol
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pharmacology
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Signal Transduction
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Transcription Factors
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metabolism
10.Study on the association of serum matrix metalloproteinase-9 and white blood cell count to type 2 diabetes mellitus with macroangiopathy and the interventing treatment
Jia LI ; Lv-Zhi JIANG ; Xing-Jia ZHOU ; Xue-Ning ZHANG ; Zheng-Fang LI ;
Chinese Journal of Endocrinology and Metabolism 2001;0(05):-
Objective To evaluate the relationship between serum matrix metalloproteinase-9(MMP-9), white blood cell(WBC)count and type 2 diabetes mellitus with macroangiopathy and to investigate the mechanism of the protective effects of rusiglitazone(RSG)on blood vessel.Methods Serum MMP-9 was determined by ELISA in 30 normal controls and 80 type 2 diabetic patients(including 40 cases with macroangiopathy and 40 without maeroangiopathy).WBC count and other clinical parameters were also determined.32 type 2 diabetic patients received RSG(4rag qd)for 12 weeks.All parameters were determined after 4 weeks(17 patients)and after 12 weeks(all patients)to observe the changes in MMP-9,WBC and other parameters.Results In the diabetic patients,serum MMP-9 and WBC were markedly higher as compared with normal controls;and MMP-9 and WBC in patients with macroangiopathy[579(440-949)?g/L,(7.51?1.47)?10~9/L]were higher than those [324(275-423)?g/L,(6.22?0.79)?10~9/L]in the cases without macroangiopathy(P