1.Detection of IL-2 and IL-6 in human breast tumor Bcap-37 cells treated by hematoporphyrin nonomethyl ether mediated photodynamic therapy
Lihua LIU ; Minghui HUANG ; Yanchun QIAN ; Hongbo ZHANG
Cancer Research and Clinic 2012;24(8):530-532
Objective To observe IL-2 and IL-6 changes in the breast tumor Bcap-37 cells reated by hematoporphyrin nonomethyl ether mediated photodynamic therapy (HMME-PDT).Methods Cells in logarithmic growth phase were collected among breast cancer cells cultured in conventional methods.According to blank control group or the experimental group (laser irradiation group,photosensitive agent group and HMME-PDT group),PDT in addition to HMME and HMME-PDT were conducted.The changes of IL-2,IL-6 were detected by radioimmunoassay.Results After HMME-PDT,IL-2 was increased as time passed.After 12,24 and 48 h,compared with IL-2 level in the control group,in laser irradiation group or photosensitive agent group,the levels of IL-2 in HMME-PDT group was significantly differences (P < 0.05).But IL-6 levels decreased.The most obvious changes of IL-6 levels happened at 12h and 24h.There was significant differences between IL-6 in HMME-PDT group with the control group,laser irradiation group or photosensitive agent group (P < 0.05).Conclusion HMME-PDT maybe have destruction effect by altering IL-2,IL-6 activity on breast tumor cells,which provides objective indicators for clinical patients to regulate immune function and auxiliary diagnosis.
2.Improvement effect of Sophora Flavones combined with captopril on diabetic cardiomyopathy of rats
Ying CHANG ; Qian LU ; Yan WANG ; Yanchun WANG
Journal of Jilin University(Medicine Edition) 2016;42(4):721-724
Objective:To study the improvement effect of Sophora Flavones combined with captopril on the rat diabetic cardiomyopathy,and to clarify its mechanisms of improving the myocardial fibrosis.Methods:Fifteen rats were selected randomly from 100 male Wistar rats as normal group.The other rats were fed with high fat and high sugar food and intraperitoneally injected with small dose of streptozotocin (30 mg· kg-1 )all at once to establish type 2 diabetes mellitus cardiomgopathy models.Then 72 rat models with type 2 diabetic cardiomyopathy were set up in which 60 rats were selected according to the blood glucose levels and divided into model group, captopril group,Sophora Flavones group,and Sophora Flavones combined with captopril group (combination group) (n=15).8 weeks after intragastric administration,the weights,the cardiac indexes,the activities of serum creatine kinase-MB (CK-MB), lactate dehydrogenase (LDH ), and the contents of nitric oxide (NO), nitric oxide synthase (iNOS),collagen Ⅰ (Col Ⅰ)and collagen Ⅲ (Col Ⅲ)of the rats in various groups were detected. Results:Compared with normal group,the weight of the rats in model group was decreased and the cardiac index was increased (P <0.01),the activities of serum LDH and CK-MB were increased (P < 0.01),the contents of iNOS and NO in the myocardium tissue of the rats in model group were decreased (P <0.01),and the contents of Col Ⅰ and col Ⅲ were increased (P <0.05 or P <0.01).Compared with model group,the weights and the cardiac indexes of the rats in medication groups were decreased (P <0.05),the activities of serum LDH and CK-MB were decreased (P <0.05 or P <0.01),the contents of iNOS and NO in the myocardium tissue were increased (P <0.05),and the contents of Col Ⅰ and Col Ⅲ were decreased (P <0.05 or P <0.01).The effect of combination group was better than single medication groups (P < 0.05).Conclusion:Combination of Sophora Flavones and captopril has a improvement effect on the rat type 2 diabetic cardiomyopathy,and its mechanism may be related to reducing the myocardial injury to improve diabetic cardiomyopathy.
3.Inhibitory effect of Rhodiola total flavonoids on proliferation of cardiac fibroblasts in rats
Ying AN ; Qian LU ; Congfei XU ; Yanchun WANG ; Liqin HAN
Journal of Jilin University(Medicine Edition) 2015;(6):1190-1194
Objective To investigate the inhibitory effect of Rhodiola total flavonoids on transforming growth factor (TGF)-β1-induced proliferation of cardiac fibroblasts (CFB) in the rats, and to explore its mechanisms of improving myocardial fibrosis.Methods 5 μg·L-1 TGF-β1 was used to induce the proliferation of CFB to build the cell models of myocardial fibrosis.The cultured CFB were divided into control group,model group,and low, medium,high doses (25,50 and 100 mg· L-1 )of Rhodiola total flavonoids groups.The cells were treated for 48 h,the vitalities of cells were detected by MTT,the levels of collagen proteinⅠ (ColⅠ)and collagen proteinⅢ(Col Ⅲ)in supernatant were measured by ELISA,and the activities of total superoxide dismutase (T-SOD)and glutathione peroxidase (GSH-Px)were detected,the levels of malondialdehyde (MDA)and glutathione (GSH) were also measured.Results The MTT results indicated that compared with control group,the cell vitality in model group was significantly increased (P < 0.01 ).compared with model group,the cell vitalities of CFB in Rhodiola total flavonoids groups were significantly decreased (P < 0.05 ). Compared with control group, the T-SOD activity in model group was decreased, while the MDA level was significantly increased (P <0.05);compared with model group,the T-SOD activities in 50 and 100 mg· L-1 Rhodiola total flavonoids groups were increased (P <0.01);the MDA levels in 25 and 50 mg· L-1 groups were decreased significantly (P < 0.05). Compared with control group,the GSH-Px activity and GSH level in model group were significantly decreased (P <0.01).Compared with model group,the GSH-Px activities and GSH levels in Rhodiola total flavonoids groups were increased (P <0.05).Compared with control group,the ColⅠand Col Ⅲ levels in model group were significantly increased (P <0.01);compared with model group,the ColⅠand Col Ⅲ levels in Rhodiola total flavonoids groups were significantly decreased (P <0.05).Conclusion Rhodiola total flavonoids can inhibit the proliferation of rat CFB.The development of myocardial fibrosis may be inhibited by Rhodiola total flavonoids through anti-oxidative stress pathway.
4.Effects of lipopolysaccharide binding protein on activation of p38 signaling pathway induced by LPS in macrophages
Yifeng HOU ; Yanchun ZHOU ; Baoling MAO ; Guishen QIAN
Chinese Journal of Pathophysiology 1999;0(09):-
AIM: To investigate the regulatory effects of lipopolysaccharide binding protein(LBP)on activation of p38 signaling pathway induced by lipopolysaccharide(LPS)in alveolar macrophages. METHODS: The LBP from actue phase rat serum was purified by ammonium sulphate precipitation, Bio-Rex70 resin and the MonoQ column. Rat alveolar macrophages were exposed to LPS (0 01 mg/L or 1 mg/L) the various concentrations of LBP(0 mg/L, 0 01 mg/L, 0 1 mg/L,1 mg/L and 10 mg/L) Western blotting were used to detect phospho-p38 in alveolar macrophages RESULTS: SDS-PAGE analysis indicated that the purified preparation of rat LBP showed homogeneity and the molecular weight was 60 kD.The binding of lipopolysaccharide to mononuclear cells were enhanced by purified rat LBP. Stimulation of rat alveolar macrophages with LPS at concentration of 0.01 mg/L was LBP dependent. LBP at concentrations up to 1 mg/L was able to increase the activation of p38. However , when LBP concentrations were further increased to 10 mg/L, the phosphorylation levers of p38 were lower as compared with that in the presence of 1 mg/L. Stimulation of rat alveolar macrophages with LPS at concentrations of 1 mg/L was LBP-independent. CONCLUSION: The activation of p38 induced by LPS at lower concentration(0.01 mg/L ) was LBP-dependent, meanwhile, LPS at higher concentration (1 mg/L ) was LBP-independent.
5.Effects of electroacupuncture on proteasomes of substantia nigra in rats with Parkinson's disease.
Qian TU ; Yan LIANG ; Jun MA ; Shuju WANG ; Feng SHEN ; Yanchun WANG
Chinese Acupuncture & Moxibustion 2015;35(4):355-360
OBJECTIVETo explore the effects and action mechanism of electroacupuncture (EA) on Parkinson's disease (PD).
METHODSForty-eight healthy male SD rats were randomly divided into a normal group, a sham operation group, a model group and an EA group, 12 rats in each one. Rats in the model group and EA group were treated with subcutaneous injection of rotenone (1mg/kg, dissolved in dimethyl sulfoxide and 0. 9 % normal saline) on neck and back for 40 days to establish rat model. Rats in the sham operation group were treated with injection of identical dose of dimethyl sulfoxide and 0. 9 %o normal saline at identical location which did not contain rotenone. After model establishment, rats in the EA group were treated with EA at "Fengfu" (GV 16) and "Taichong" (LR 3) with continuous wave (2 Hz, 1 mA), which was given 20 min per time, once a day for consecutive 28 days. Rats in the remaining groups were treated with fixation and immobilization without any other intervention. The rats behavioristics changes were observed and scored; immunohisto-chemistry was adopted to test the expression of tyrosine hydroxylase (TH); fluorescence spectrometry was used to detect the activities of 20 S β1, β2, β5; western blot method was applied to measure the expression of 20S proteasome and its a subunit.
RESULTSCompared with the normal group and sham operation group, there was significant change of behavioristics in the model group, and TH positive neuron counting was obviously reduced; after treatment, the behavioristics score in the EA group was lower than that in the model group (P<0. 05), and TH positive neuron counting was significantly increased (P<0. 05). Compared with the normal group and sham operation group, the activities of 20 S β1, β2, β5 in model group were significantly reduced (all P<0. 01), and those in the EA group were higher than those in the model group (P<0. 01). Compared with the normal group and sham operation group, the expression of 20S proteasome and its a subunit was reduced in the model group, and that in the EA group was higher than that in the model group (P<0. 05).
CONCLUSIONEA could improve the loss of dopaminergic neurons induced by rotenone to prevent and treat PD, which is likely to be related with protecting the activity and expression of proteasomes in substantia nigra.
Animals ; Disease Models, Animal ; Electroacupuncture ; Humans ; Male ; Parkinson Disease ; enzymology ; therapy ; Proteasome Endopeptidase Complex ; metabolism ; Rats ; Rats, Sprague-Dawley ; Substantia Nigra ; enzymology
7.From Earthworm Research to See How to Make Progress in Dilemma of Traditional Chinese Medicine Resources and Quality
Qian JI ; Hanming ZHANG ; Weirong CHEN ; Yanchun WANG ; Xiang CHEN ; Lei ZHANG
World Science and Technology-Modernization of Traditional Chinese Medicine 2014;(11):2308-2313
Resource of traditional Chinese medicine (TCM) is one of the greatest treasures of China. It made magnif-icent contribution in Chinese lives and breeds. Quality is the foundation of safety, effective and quality control of TCM. With the social development, problem with raising need and falling resource and quality had became more and more serious. It slowed down the speed of TCM modernization and internationalization. This study used Shanghai earthworm, the authentic TCM of Shanghai, as an example. Focus on resource and quality problems, such as species confusion, resource shortage, rough machining, low quality and no standards, this study tried to work out the reason-able and feasible solution. This effort funded a way out of a stalemate of TCM resources and quality. It is important to TCM modernization and internationalization.
8.Effect and mechanism of fluvastatin on the expression of fibronectin in human peritoneal mesothelial cells induced by high-glucose peritoneal dialysate
Yanchun LIU ; Jia LIU ; Yaguang XU ; Xiufen ZHAO ; Jun QIAN ; Bin SUN ; Changying XING
Chinese Journal of Nephrology 2012;(11):888-893
Objective To explore the effect and mechanism of fluvastatin on the expression of fibronectin(FN) in human peritoneal mesothelial cells (HPMCs) induced by high-glucose peritoneal dialysate (HGPDS).Methods Cultured HPMCs were randomly divided into control,HGPDS,HGPDS plus GSK650394 10-5 mol/L (the competitive inhibitor of SGK1),different concentrations of fluvastatin,fluvastatin 10-6 mol/L and GSK650394 10-5 mol/L alone.The morphology change of HPMC was observed by light microscopy.The cellular viability was detected by MTT colorimetry.The mRNA and protein expressions of serum and glucocorticoid-inducible kinase 1 (SGK1) and FN were detected by RT-PCR,Western blotting or ELISA.Results After incubation with HGPDS,the cell morphology changed from typical cobblestone-like appearance to fibroblast-like appearance,and the cell viability was inhibited significantly (P<0.05).Fluvastatin 10-6mol/L and GSK650394 could improved the cell morphology and the cell viability injured by HGPDS (P<0.05).Compared with the normal control group,the mRNA and protein expressions of SGK1 and FN increased significantly in HPMC treated with HGPDS(P<0.05).GSK650394 significantly decreased the high expression of SGK1 and FN (P<0.05),also the fluvastatin had same effects as GSK650394 in dose-dependent manner (P<0.05).Conclusions High-glucose peritoneal dialysate can increase FN expression in human peritoneal mesothelial cells,which can be attenuated by fluvastatin.The protective role of fluvastatin in HPMC may be partially achieved through the signal pathway of SGK1.
9.Drug-resistant genes and virulence factors of carbepenem-resistant Klebsiella peumoniae
Qian YU ; Zhidong HU ; Bin TIAN ; Yanchun LI ; Jing LI ; Hairu XU
Chinese Journal of Clinical Infectious Diseases 2016;(1):52-58
Objective To investigate the resistance mechanism and virulence genes of clinical strains of carbapenem-resistant Klebsiella pneumonia ( CRKP ) .Methods Twenty clinical CRKP strains were collected from Tianjin Medical University General Hospital during May 2014 and May 2015.Vitek-2 Compact system was used for identification of the strains and antibiotic susceptibility test .Modified Hodge Test and EDTA double disk phenotypic test were performed for screening of drug -resistant phenotypes .Drug-resistant genes , capsular serotypes and associated virulence genes were amplified by PCR , and positive products were sequenced and analyzed by DNA sequencing .Results Resistance to beta-lactam antibiotics including cephalosporins and carbapenems was observed in 80.0%and above strains , but more than 70.0%strains were sensitive to tigecycline , amikacin and levofloxacin .KPC gene and NDM gene were found in 7 strains (35.0%) and 8 strains (40.0%), respectively.SHV, the most common extended-spectrumβ-lactamases ( ESBLs ) gene, was found in 16 strains ( 80.0%). DHA plasmid-mediated AmpCβ-lactamase was found in 2 strains (10.0%).Deletions of porin-coding genes OmpK35 and OmpK36 were found in 8 stains ( 40.0%) and 13 strains ( 65.0%), respectively.Carbapenem-resistant genes in combination with ESBLs genes and/or variation of porin was found in 14 strains (70.0%), and ESBLs genes in combination with variation of porin was found in 4 strains (30.0%).Three strains were of capsular serotype K1 and 1 was of K57, and all of them carried KPC genes .Virulence gene rmpA was found in 8 strains and all carried carbapenemases , among which 5 strains with KPC, 2 strains with NDM, 1 strain with both KPC and NDM .Six strains were aerobactin gene positive , among which 4 strains carried KPC genes . FimH-1 was positive in all strains .Conclusions KPC and NDM genes mainly account for resistance in CPKP, and ESBLs with OmpK gene deletion may also be an important cause .Strains with capsular serotypes K1 and K57 carrying KPC genes are common .
10.EGFR siRNA inhibits activation of astrocytes derived from rats through blockade of STAT3 phosphorylation
Hong QIAN ; Lijing LIU ; Yanchun LI ; Ming XIE ; Heng WU ; Shuangxi LIU ; Bin WU
Chinese Pharmacological Bulletin 2016;(2):216-222
Aim To observe the expression of epider-mal growth factor receptor ( EGFR) in cerebral tissues around hematomas after intracerebral hemorrhage, and explore the effects of EGFR on activation of astrocytes derived from rats and the involved mechanisms. Meth-ods The specimens of cerebral tissues around hemo-tomas after intracerebral hemorrhage undergoing hemo-tomas removal operation were collected and then divid-ed into 4 groups according to the time of intracerebral hemorrhage: <1 d, 1 ~5 d, 6 ~10 d and >10 d groups. Each group included 20 cases. At the same time, 20 dropped brain tissues distant to hemorrhage in the operative process were collected as control group. Immunohistostaining and Western blot were used to measure the expression of EGFR. After isolation and culturing, the astrocytes of rat cortex were treated with culture solution ( control group) , CNTF that was used to activate astrocytes, scramble siRNA + CNTF and EGFR siRNA +CNTF for 24h, respectively. The ex-pression of glial fibrillary acidic protein ( GFAP) mR-NA was detected through fluorescence real-time quanti-tative PCR. In addition, the protein levels of GFAP, signal transducers and activators of transcription 3 ( STAT3 ) and phosphorylated STAT3 ( p-STAT3 ) were examined using Western blot. Results With the ex-tension of intracerebral hemorrhage time, positive sig-nal index and protein expression levels of EGFR gradu-ally elevated, reached the peak on 6 ~10d, and then decreased after 10 d. There was statistical difference ( P<0. 01 ) . The expression levels of GFAP mRNA and protein as well as p-STAT3 were significantly in-creased in cells treated with CNTF alone as compared to control group ( P <0. 01 ) , whereas these effects were almost completely reversed by EGFR siRNA transfection ( P <0. 01 ) . Additionally, there was no statistical difference in STAT3 protein levels among groups ( P >0. 05 ) . Conclusions EGFR expression is upregulated in the cerebral tissues around hemotomas after intracerebral hemorrhage. Gene silence of EGFR contributes to suppressing the activation of astrocytes derived from rats, which may be involved in the block-ade of STAT3 phosphorylation.