1.Effects of resveratrol on proliferation and differentiation of murine 3T3-L1 preadipocytes and the underlying mechanisms
Sifan CHEN ; Xincai XIAO ; Yanshuang SUN ; Lin ZHENG ; Xiang FENG
Chinese Pharmacological Bulletin 2010;26(1):108-111
Aim To explore the effect and mechanism of resveratrol (Res) on proliferation and differentiation of murine 3T3-L1 preadipocytes.Methods 3T3-L1 preadipocytes were cultured and treated with resveratrol in different dosages.Cell proliferation was analyzed by WST-1 method. Oil red O staining method and spectrophotography were applied to analyze the degree of differentiation. Real-time PCR was applied to detect the mRNA expression of adiponectin and leptin. Western blot was applied to detect the expression of silent information regulator 1 (Sirt1),peroxisome proliferator activated receptor γ (PPARγ) and CCTTA enhancer binding proteinα (C/EBPα).Results Res inhibited proliferation of murine 3T3-L1 preadipocytes in a time-dose dependent manner.The expression levels of adiponectin and leptin mRNA were decreased, and Res also inhibited 3T3-L1 preadipocytes to differentiate into mature adipocytes. Res increased the expression levels of Sirt1 and decreased the expression levels of PPARγ and C/EBPα.Conclusions Resveratrol can inhibit the proliferation and differentiation of 3T3-L1 preadipocytes.The underlying mechanisms may include enhancing expression of Sirt1 and inhibiting expression of PPARγ,C/EBPα which are related to cell differentiation.
2.Neural network injury and motor functional parameters in subacute stroke patients using diffusion tensor imaging
Xiang XIAO ; Le LI ; Yanchun LV ; Qiang LIN ; Dongfeng HUANG
Chinese Journal of Tissue Engineering Research 2014;(15):2421-2426
BACKGROUND:The interaction of neural network and motor function in post-stroke brain tissue remains unclear.
OBJECTIVE:To observe neural network impairment fol owing subacute stroke by using diffusion tensor imaging, and to investigate the relationship with neurological defects and motor dysfunction.
METHODS:A total of 19 patients after subactue stroke and 20 healthy adults were examined with diffusion tensor imaging. The fol owing parameters were compared:fractional anisotropy, apparent diffusion coefficient, asymmetry indices of fractional anisotropy and apparent diffusion coefficient. The neurological defect and motor function were evaluated with the corresponding scales. The 10-meter walking speed was measured. The correlation of diffusion tensor imaging parameters with the scale scores and 10-meter walking speed was analyzed.
RESULTS AND CONCLUSION:The stroke group exhibited reduced fractional anisotropy value asymmetry and fractional anisotropy value in bilateral posterior limbs of the internal capsule. Apparent diffusion coefficient value asymmetry and apparent diffusion coefficient value in the posterior limb of the internal capsule were lower than control unaffected side (P<0.05). Apparent diffusion coefficient value and apparent diffusion coefficient value asymmetry in posterior limb of the internal capsule showed a strong negative correlation with Fugl-Meyer assessment scores of the lower extremities (P<0.05). Diffusion tensor imaging parameters is closely linked with motor dysfunction of the lower extremities in subacute stroke patients. Local stroke lesion-caused neurological defect is the leading cause of motor dysfunction of the lower extremities.
3.Research progress in the study of brain microdialysis in glioma.
Lin LIU ; Xiang-Yi ZHANG ; Yao XIAO ; Xing-Guo ZHANG
Acta Pharmaceutica Sinica 2014;49(4):450-456
Glioma is the most common form of brain cancer. Despite recent advances in the treatment of solid tumors, there are few effective treatments for malignant gliomas due to its infiltrative nature. It has important significance to improve the treatment of glioma through in-depth understanding the intracerebral metabolic characteristics and pharmacokinetics of chemotherapeutics. Brain microdialysis (B-MD), an effective method to monitor central nervous system anticancer drug disposition, conditions of drugs through the blood-brain barrier, basic pathophysiologic metabolism, bioactive compounds and the changes of neurotransmitter in brain, provides the unique opportunity to allow the simultaneous determination of unbound concentrations of drugs in several tissues, and directly measure gliomas biochemistry continuously. B-MD has been able to monitor the change of brain drugs, metabolites and neurotransmitters, dynamic analysis of the drug concentration and pharmacological effect after administration, pharmacodynamic interaction between drugs, receptor mechanism of drug transport, as well as feedback information of internal environment. B-MD is expected to provide reference for clinical individual chemotherapy of glioma, but also provide powerful tools for the evaluation of new anticancer drugs in vivo. In this review, a comprehensive overview of B-MD for studies on glioma is elucidated with special emphasis on its application to neurochemistry and pharmacokinetic studies.
Animals
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Antineoplastic Agents
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pharmacokinetics
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Blood-Brain Barrier
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Brain Neoplasms
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metabolism
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Glioma
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metabolism
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Humans
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Magnetic Resonance Spectroscopy
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Metabolomics
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methods
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Microdialysis
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methods
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Neurotransmitter Agents
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pharmacokinetics
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Pharmaceutical Preparations
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metabolism
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Positron-Emission Tomography
4.Effects of tributyltin chloride on superoxide dismutase from Tegillarca granosa
Xiang XIAO ; Yali HAN ; Huixiang YUAN ; Kebo LIN
Chinese Journal of Marine Drugs 1994;0(04):-
Superoxide dismutase(SOD) was purified from Tegillarca granosa and the effects of tributyltin chloride(TBTCl) on the enzyme activity of the SOD were analyzed at different conditios. The results showed that the effects of TBTCl on SOD activity were more ohvious at room temperature than at four degrees centigrade. 58. 1 % of the enzyme activity was reserved after SOD was incubated with 200?g?mL-1 of TBTC1 for 72 hours at room temperature, while 90. 9% of the enzyme activity was reserved at four degrees centigrade. The enzyme activity of SOD was reduced with the increase of TBTC1 concentration and interaction time. TBTCl could reduce heat-resisting, acid-resisting and alkali-resisting abilities of SOD. With TBTCl concentration increase,SOD isozyme bands in the graph of polyacrylamide gel electrophoresis were getting weaker.
5.Experimental study on microcoagulation of dorsal root entry zone of cervical cord with bipolar forceps
Jian-Ping XIANG ; Xiao-Lin LIU ; Jun HU ;
Chinese Journal of Microsurgery 2006;0(06):-
Objective To investigate the safety and obtain parameters of microeoagulation of dorsal root entry zone (DREZ) of cervical cord with bipolar foreps on animal model,and provide histological base for clinical application of treatment of brachial plexus avulsiol pain using microcoagulation of dorsal root entry zone.Methods On the base of swine's weight and spinal cord size in similar to human being,it was chosen to be experimental animal.The right DREZs of cervical cord were microcoagulated with bipolar forceps.The swines were fed in normal way.Their activities were observed.The mass change of the cervical cord segment were observed after 3 weeks and the cervical cord segment was fixed with 10% fromalin,paraffin sliced,HE dying.Coagulating space,depth and width were measured under microsope.The coagulating parameter were adjusted according to measuring outcome in order to achieving a most avaliable parameter.Results All post-op swine survived.When the microcogulation were made with bipolar forceps adopted following parame- ters:The distance of between the polar was 2.0 mm;The diameter of polar was 0.3 mm.The inserting depth 2 mm,the coagulated power 18 watt,the coagulated time was 2 second,then the width of lesions of DREZ in cross section was 1.15 mm and the depth of lesions was 3.10 mm,which was consistent with the area of hu- man DREZ of cervical cord.Conclusion The experiment on swine suggested,microcoagulation of DREZ by bipolar forceps is safe and no mortal complications when the testified parameters are adoped,and can achieve the area of DREZ of cervical cord in human.
6.Progress in monoclonal antibody humanization based on complementarity-determining region grafting
xiao-lin, ZHU ; feng, CHENG ; guan-xiang, QIAN
Journal of Shanghai Jiaotong University(Medical Science) 2006;0(11):-
Humanized monoclonal antibodies(mAbs) are increasingly widely used in targeted therapy for cancer and some other major diseases.Complementarity-determining region(CDR) grafting makes quantities of humanized mAbs available.Herein,we provide an overview on the strategy and progress of CDR grafting.
7.Construction of NF-?B/p65-targeting RNAi combinant adenovirus vector and its suppressive effect on the expression of p65 in ECV304 cells
Gang CHEN ; Yu-Fang QIAO ; Xu LIN ; Jin YAO ; Miao LIN ; Ting-Ting YOU ; Xiao-Yan SHEN ; Xiang-Qing ZHU ; Lun-Pan MOU ; Li-Xiang LIN
Chinese Journal of Endocrinology and Metabolism 1986;0(04):-
Objective To construct RNAi combinant adenoviral expressive vectors specific to p65 subunit of NF-?B and to observe their gene silencing effect on p65 subunit.Methods Three pairs of complementary. single-strand DNA oligos targeting three various sites of p65 mRNA were designed and synthesized.Annealling was used to generate double-strand oligos(ds-oligos),and then the ds-oligos were cloned into pENTR~TM/u6 to generate the entry clone named pENTR.Recombination reaction in vitro with the pENTR and pAd/BLOCK-iT~TM- DEST was used to creat the adenovirus plasmid which contains the RNAi cassette.Then,the adenovirus plasmids digested with PacI were transfected into HEK293A cells to product adenovirus,and latter infected the HEK293A cells to amplify the adenoviral stock.Plaque forming assay was used to titer the adenoviral stock.The p65 gene silencing effect induced by the RNAi adenovirus was detected by Western blot and immunocytochemistry assay in ECV304 cells.Results The RNAi adenovirus specific to p65 subunit of NF-?B were produced with titer of 3.0 x 10~9pfu/ml to 2.5?10~10pfu/ml.The expression of p65 protein in ECV304 cells could be down-regulated efficiently by the RNAi adenovirus 48-72 h after infection,which would last for more than 6 days after infection.Conclusion RNAi adenovirus is an important tool inhibiting the expression of target gene efficiently.
8.Relationship between MTA1 expression and invasive and metastatic ability of cervical cancer cell
Xiaoyan HAN ; Haili QIAN ; Junjun YANG ; Xueyan ZHANG ; Ming FU ; Xiao LIANG ; Chen LIN ; Yang XIANG
Chinese Journal of Obstetrics and Gynecology 2011;46(9):678-683
ObjectiveTo investigate the relationship between metastasis-associated gene 1 ( MTA1 )expression and invasive and metastatic ability of cervical cancer cell. MethodsThree kinds of plasmids pcDNA3( control group), pcDNA3-MTA1 ( MTA1 group) and pSilencer3. 1-MTA1-siRNA ( MTA1-siRNAgroup) were transfected into human cervical cancer cell line CaSki cells. Reverse transcription (RT)-PCR and western blot were used to detected MTA1 mRNA and protein expressions. The effects of MTA1 expression on CaSki cell growth and proliferation, cell migration, adhesion and invasion, and cell cycles were tested by methyl thiazolyl tetrazolium (MTT), clone formation experiment, wound-healing assay, transwell assay, adhesion assay and flow cytometry, respectively. In animal experiment, three groups of cells were inoculated to BALB/c nude mouse subcutaneously to observe tumor formation ability. ResultsCompared with control group, MTA1 mRNA and protein were significantly overexpressed in MTA1 group, while MTA1-siRNA group showed lower MTA1 expression. Compared with control group, MTA1 group showed significantly accelerated cell growth; while MTA1-siRNA group showed decreased cell growth since the second day (P<0. 05). Clone formation number in control, MTA1 and MTA1-siRNA group were 133 ±6, 169 ± 10 and 57 ±5,respectively. MTA1 group showed accelerated cell formation, while MTA1-siRNA group showed the reverse effect compared with that in control group(P < 0. 05 ). At 24, 48 and 72 hours after wounding, the healing ability of MTA1-siRNA group significantly lagged behind that in the control group, while MTA1 group showed accelerated cell healing ability. The adhesion rate of control, MTA1 and MTA1-siRNA group were (69. 3 ± 3. 6) %, ( 80. 4 ± 5. 6 ) % and ( 39. 2 ± 7.4 ) % separately at 90 minutes after cell seeding. In contrast with control group, MTA1 group promoted the adhesion of CaSki cell to matrigel matrix, while MTA1-siRNA group inhibited the adhesion process (P <0. 05 ). In the migration assay, the number of cells migrated to the bottom side of the membrane in control,MTA1 and MTA1-siRNA group were 153 ± 17,247 ± 38 and 82 ± 10, respectively. The number of cells in the invasion assay were 231 ± 19,354 ± 36 and 76 ± 7, respectively. Compared with the control group, MTA1 group significantly increased the migration and invasion ability, while MTA 1-siRNA group showed lower cell migration and invasion ability (P < 0. 05 ). In cell cycle experiment, no significant differences of cell proportions including G1, S and G2 stage were found among three groups (P > 0.05).In animal experiment, compared with control group,MTA1 group showed accelersted tumor formation and growth,whilethe MTA1-siRNA group showed the reverse effect ( P < 0. 05 ). ConclusionsMTA1 may play its roles to promote cervical cancer cell invasion, migration, adhesion, as well as cell growth and colony formation, while RNA interference against MTA1 may decrease the malignant phenotypes. This study shows that it will be an effective beginning to explore metastasis mechanisms and cancer gene therapy strategy targeting MTA1 in cervical cancer.
9.Acupuncture and moxibustion versus western medicine for benign prostatic hyperplasia:a systematic review
Yuanwu CHEN ; Yuanhao DU ; Jun XIONG ; Pan SUN ; Xiang GAO ; Xiaomiao LIN ; Li XIAO
China Journal of Traditional Chinese Medicine and Pharmacy 2005;0(06):-
Objective:To evaluate the quality of prostate hyperplasia related literature in acupuncture and moxibustion,and to compare the curative effect on prostate hyperplasia between acupuncture and moxibustion and western medicine.Methods:Retrieving Pubmed,Cochrane Library,CBM database,CNKI database Etc.to collect the literature of prostate hyperplasia of clinical randomized or quasi-randomized control trials of comparative study between western medicine and acupuncture treatment.The data was extracted independently by two valuers from literatures fitting the selection criteria.Cochrane evaluation manual 4.2.6 was used to evaluate quality,and RevMan 4.2.8 was used in statistical analysis.Results:A total of six randomized or quasi-randomized controlled trials (total 546 examples) were adopted.6 study adopted the total effective rate of evaluation indexes,Meta-analysis showed that there was a significant difference between acupuncture treatment group and western medicine group [merger RR (fixed effects model)=1.26,95%CI(1.15,1.37),Z=5.13,P
10.Construction of a subtracted cDNA library of differentially expressed genes in human gastric carcinoma induced by diallyl disulfide
Chen HUANG ; Xiaoqiu LIANG ; Hui LIN ; Yousheng HUANG ; Shulin XIANG ; Xiaolan XIAO ; Qi SU
Chinese Pharmacological Bulletin 2003;0(11):-
Aim To construct a subtracted cDNA library o f differentially expressed genes in human gastric carcinoma induced by diallyl dis ulfide(DADS). Methods Differentially expressed cDNA species induc ed by DADS in MGC 803 human gastric carcinoma cell line was determined by using suppression subtractive hybridization (SSH). Then these cDNA species were direct ly inserted into T/A cloning vector to set up the subtractive library. Amplification of th e library was carried out with transformation of E.coli by high voltage electrop erforation. One hundred positive bacteria clones were randomly picked and identi fied using PCR method. Results The amplified library contained more than 1,000 positive bacteria clones. Random analysis of 100 clones with PCR m ethod showed that all clones contained 100~600 bp inserts.Conclusions A subtracted cDNA library of differentially expressed genes in MGC 803 hum an gastric carcinoma cell line induced by DADS is constructed successfully with SSH and T/A cloning techniques. The library is efficient and lays solid foundati on for screening and cloning new and specific tumor correlative genes of human g astric carcinoma, and provides a new idea for further exploring the mechanism of DADS effects on carcinoma cells.