1.Effect of rosuvastatin and memantine on endothelial progenitor cells in vascular dementia rats
Chenchen SONG ; Nan ZHANG ; Yan CHENG
Chinese Journal of Geriatric Heart Brain and Vessel Diseases 2015;(3):299-302
Objective To observe the effect of rosuvastatin and memantine on EPC and learning or memory ability in VaD rats .Methods Forty adult male SD rats were randomly divided into sham operation group ,VaD model group ,memantine treatment group ,rosuvastatin treatment group , and memantine+rosuvastatin treatment group (combined treatment group) (8 in each group) .A VaD model was established by permanent ligation of bilateral common carotid arteries .The ani‐mals in rosuvastatin treatment group ,memantine treatment group and combined treatment group received gastric rosuvastatin 10 mg/(kg · d) ,memantine 10 mg/(kg · d) and memantine 10 mg/(kg · d)+rosuvastatin 10 mg/(kg · d) for 4 weeks while those in model group received gastric normal saline .Five weeks after operation ,the learning or memory ability was tested by Morris water maze test ,the escape latency and percentage of target quadrant were calculated ,the circulat‐ing EPC were detected by flow cytometry ,and the MVD in hippocampaus was assyed with vWF immunostaining .Results Five weeks after operation ,the learning or memory ability was signifi‐cantly lower in model group than in sham operation group (P< 0 .01) whereas the learning or memory ability ,the percentage of circulating EPC and the MVD in hippocampus were significantly higher in 3 treatment groups than in model group (P<0 .05) .Conclusion Rosuvastatin and me‐mantine can effectively improve the learning or memory ability of VaD rats by mobilizing their cir‐culating EPC and increasing the MVD in their hippocampus .However ,the effect of memantine or rosuvastatin does not differ greatly w hen they are used alone .
2.Delayed hypoxic preconditioning decreases the level of cPKC? protein expression in cerebral cortex of mice
Song HAN ; Chenchen NIU ; Junfa LI
Basic & Clinical Medicine 2006;0(04):-
Objective To explore the role of conventional protein kinase C(cPKCs) in delayed hypoxic preconditioning.Methods The biochemistry techniques of SDS-PAGE,Western bolt and Gel Doc imagine were applied to analyze the effect of repetitive hypoxic exposure(H5,H6) on the level of cPKC?,? membrane translocation and protein expression in murine brain.Results We found that cPKC? protein expression was significantly decreased(P
3.Sustained hypoxia increases membrane translocation of conventional protein kinase C isoforms in SH-SY5Y neuroblastoma cells
Pengyu ZU ; Junfa LI ; Song HAN ; Yanming QU ; Hua LI ; Chenchen NIU ; Qunyuan XU
Chinese Journal of Tissue Engineering Research 2005;9(21):242-245
BACKGROUND: The protein kinase C (PKC) family consists of 3 groups of PKCs, namely the conventional PKC (cPKC), atypical PKC and novel PKC.Accumulating studies conducted in recent years have suggested that PKCs may play important roles in the development of cerebral ischemic/hypoxic preconditioning ( I / HPC ).OBJECTIVE: To observe membrane translocation of hypoxia-activated cPKC isoforms(α, βⅠ, βⅡ and γ) at cellular levels in a cell hypoxia model.DESIGN: Randomized block design.SETTING: Department of Neurobiology, College of Basic Medical Sciences,Capital University of Medical Sciences.MATERIALS: The experiment was completed at the Neurobiological Cell Culture Laboratory of Capital University of Medical Sciences in May 2004.Human neuroblastoma cells with the properties of neurons were maintained and passaged in this laboratory.METHODS: The activation of cPKC isoforms under hypoxic condition and changes of cPKCα, βⅠ, βⅡ and γ membrane translocation(an indicator of PKCs activation) in SH-SY5Y neuroblastoma cells in response to hypoxia (1% 02, 5% CO2 and 94% N2) for 0 to 24 hours were observed using SDS-PAGE, Western blotting and immunocytochemistry.MAIN OUTCOME MEASURES: Effect of sustained hypoxia on cPKC membrane translocation in human neuroblastoma cells was observed with SDS-PAGE, cPKC Western blotting, and immunocytochemistry.RESULTS: cPKCα, βⅠ and βⅡ membrane translocation were increased significantly ( P < 0.05 ) in a time-dependent manner in response to hypoxic exposure, and the increase of cPKCβⅠ was more evident( P < 0. 001 ) after 4hours of hypoxic exposure, whereas no cPKCγ was detected in SH-SY5Y neuroblastoma cells either under normoxic or hypoxic condition. The results suggested that all cPKC isoforms, epically cPKCβⅠ, could be activated by sustained hypoxia, and the absence of cPKCγ in SH-SY5Y neuroblastoma cells may be relevant to the loss of specific biological features of the cultured cells.CONCLUSION: Sustained hypoxia activates the isoforms of cPKCα, βⅠ and βⅡ in human neuroblastoma cells and induces their membrane translocation.cPKCγ isoform may not exist in human neuroblastoma cells, or the cells has lost certain biological characteristics.
4.Cloning, expression and purification of novel gene NBEAL1 and its relationship with pathological grades of glioma
Chenchen BAO ; Hao YANG ; Na LI ; Bin LIU ; Hua SONG ; Ping SHENG ; Guohan HU ; Daxiang CUI
Chinese Journal of Cancer Biotherapy 2010;17(1):77-81
Objective: To construct the expression plasmid of a novel gene human NBEAL1 (neurobeachin like 1), and to study its relationship with the pathological grades of glioma. Methods: Total RNA of human glioma cell line U251 was extracted. NBEAL1 expression plasmid pGEX-KG/NBEAL1 was constructed and transferred into E. coli BL21. Recombinant NBEAL1 protein was induced by IPTG and further purified by GST affinity chromatographic column. The purity of recombinant NBEAL1 protein was examined by Western blotting analysis. A NBEAL1 protein specific monoclonal antibody was prepared and was used to study the relationship of NBEAL1 expression with pathological grades of glioma. Results: The NBEAL1 gene fragment was successfully cloned into pGEX-KG expression plasmid and verified by DNA sequencing. The recombinant NBEAL1 protein was expressed in inclusion bodies, with a yield of more than 30% of total bacterial proteins; the purity of purified NBEAL1 protein was above 95%. Western blotting analysis confirmed that the purified protein containing GST tag and NBEAL protein. NBEAL1 protein was lowly expressed in normal brain tissues and highly expressed in low grade glioma tissues; and the expression of NBEAL1 decreased with the increase of glioma malignancy. Conclusion: The NBEAL1 protein has been successfully cloned, expressed and purified. NBEAL1 protein expression in glioma tissues is negatively associated with the pathological grades of glioma.
5.Inhibitory effects of BRCAA1 gene silencing on gastric cancer MGC-803 cells and its possible mechanism
Bin LIU ; Daxiang CUI ; Tong DU ; Zhiming LI ; Hua SONG ; Hao YANG ; Chenchen BAO ; Hui GAN
Chinese Journal of Cancer Biotherapy 2006;0(06):-
Objective:To investigate the inhibitory effect of breast cancer-associated antigen 1(BRCAA1)gene silencing on gastric cancer MGC-803 cells and the related mechanism.Methods:Plasmid shRNA-BRCAA1 and shRNA-N were constructed and transfected with FuGene HD into gastric cancer cell line MGC-803.The transfection efficiency was examined using fluorescent microscope 24 h later.The total RNAs was extracted 48 h 'after transfection and the expression of BRCAA1 and GAPDH gene were analyzed by real-time PCR.The cell proliferation was assessed by MTT assay 24 h,48 h,and 72 h after transfection.The cell apoptosis was determined by Annexin V-PE/TAAD.The expression of Rb,Bax, Bcl-2 and BRCAA1 proteins was analyzed by Western blotting 48 h after transfection.Results:We found that the transfection efficiency of shRNA-BRCAA1 was(81.2?2.6)%24 h after transfection.Forty-eight hours after transfection with shRNA-BRCAA1 the expression of BRCAAI mRNA decreased by 61.4%;the inhibition rate of MGC-803 cells growth was 45.0%.The cell apoptosis rate of shRNA-BRCAA1 transfection group was significantly higher than those of untransfected group and mock plasmid transfected group([14.4?1.6]%vs[5.4?2.0]%,[4.4?2.5]%,P
6. Systematic evaluation of patient safety culture assessment in nursing home
Chao LYU ; Chenchen ZHAO ; Jie SONG ; Guiyu QU
Chinese Journal of Practical Nursing 2018;34(21):1670-1674
Objective:
To focus on the patient′s safety culture management and related research of nursing home in China.
Methods:
China National Knowledge Infrastructure (CNKI), China Biology Medicine disc (CBM), China Scientific Journal Database by VIP (VIP), Wanfang, PubMed, EBSCO and SpringerLink databases were searched by computer to find out all the literature about patient safety culture evaluation in nursing home. Two investigators independently screened, scrambled and cross-checked data according to inclusion and exclusion criteria.
Results:
Finally 11 articles complied with the inclusion criteria, and conducted a descriptive study of patient safety culture assessments.
Conclusions
The evaluation of patient safety culture is conducive to the development of patient safety culture in nursing home. The study of patient safety culture in China′s nursing home is still in its infancy and needs to be further deepened.
7.Preparation of human malignant melanoma ganglioside ScFv antibody-conjugated quantum dot nanoprobe and its specific binding with human malignant melanoma cells
Xiaomin ZHANG ; Tangde ZHANG ; Chenchen BAO ; Hua SONG ; Na LI ; Bin LIU ; Rong HE ; Zhiming LI ; Daxiang CUI ; Qiushi REN
Chinese Journal of Cancer Biotherapy 2010;17(1):30-35
Objective: To prepare a nanoprobe, anti-human melanoma ganglioside single chain variable fragment (GD/ScFvMEL) antibody conjugated with CdTe quantum dot, and to observe its ability to specifically bind human malignant melanoma cells. Methods: The GD/ScFvMEL gene was cloned into pET32a (+), and the plasmid was then transformed into E. coli BL21 (DE3) for GD/ScFvMEL protein antibody expression. The expressed GD/ScFvMEL antibody was purified by denaturing method and further refolded by modified dialysis method. The purified GD/ScFvMEL antibody was analyzed by SDS-PAGE. The GD/ScFvMEL-QDs nanoprobe was prepared by conjugating GD/ScFvMEL antibody with CdTe quantum dot, and its specificity was observed by incubating with MGC-803 cells and melanoma A375 cells. Results: The recombinant pET32a-GD/ScFvMEL was constructed and confirmed by PCR, restriction endonuclease analysis and DNA sequencing. The proportion of expressed GD/ScFvMEL antibody in total bacteria proteins was about 40% as detected by SDS-PAGE. The purified- and refolded-GD/ScFvMEL antibody was effectively conjugated with CdTe quantum dot, and the resulting GD/ScFvMEL-QDs nanoprobe was successfully prepared. The GD/ScFvMEL-QDs nanoprobe could specifically bind melanoma A375 cells, but could not bind stomach cancer MGC-803 cells. Conclusion: We have successfully prepared an anti-human melanoma ganglioside single-chain antibody-CdTe quantum dot nanoprobe, which can specifically bind melanoma cells.
8. Clinical study of different adsorbents with dual plasma molecular adsorption system in the treatment of hepatic failure
Guosheng YAN ; Lili LI ; Shaoli JIANG ; Song MENG ; Chenchen WU
Chinese Journal of Hepatology 2019;27(1):51-55
Objective:
To investigate the effects of two different sorbents(Carbon perfusion apparatus and Resin perfusion apparatus)in Double plasma molecular absorb syetem for liver failure treatment.
Methods:
A total of 152 cases with liver failure who were admitted to The Sixth People's Hospital of Zhengzhou, from June 2016 to May 2018 were selected and divided into DPMARS Carbon group (77 cases) and Resin group (75 cases). The two groups were observed in terms of liver function, prothrombin activity(PTA),Plasma albumin ,tumor necrosis factor alpha and interleukin-6 were detected and compared between the two groups before and after treatment.
Results:
①The clinical symptoms improved in different degree in two groups, the recovery rate of Carbon cans Carbon perfusion apparatus group and Resin group separately were89.6% (69/77)、90.7% (68/75)(
9.Effect of tunicamycin combined with cisplatin on proliferation and apoptosis of human nasopharyngeal carcinoma cells in vitro.
Lele SONG ; Linyan MA ; Xudong ZHANG ; Zhiwen JIANG ; Hao LIU ; Chenchen JIANG
Journal of Southern Medical University 2012;32(6):766-771
OBJECTIVETo study the effects of tunicamycin (a glycosylation inhibitor) combined with cisplatin on the proliferation and apoptosis of human nasopharyngeal carcinoma cells and explore the molecular mechanism.
METHODSNasopharyngeal carcinoma CNE-1 and CNE-2 cells cultured in vitro were treated with different concentrations of tunicamycin with or without cisplatin. The inhibition of cell proliferation was examined using MTT assay and colony formation assay, and the cell apoptosis was analyzed using flow cytometry with propidium iodide staining. The expressions of Bax, Bcl-2, and GRP78 in cells treated with 3 µmol/L tunicamycin with or without 6.00 µmol/L cisplatin were measured with Western blotting.
RESULTSTreatment with tunicamycin or cisplatin obviously inhibited the proliferation of CNE-1 and CNE-2 cells. Treatment with 3 µmol/L tunicamycin for 24, 36 and 48 h resulted in a viability of 72.13%, 51.97%, and 37.56% in CNE-1 cells and 85.61%, 56.95%, and 43.66% in CNE-2 cells, respectively, and the combined treatment with 6 µmol/L cisplatin lowered the cell viability to 67.97%, 47.76%, and 34.68% in CNE-1 cells and 56.89%, 37.05%, and 29.30% in CNE-2 cells, respectively. Tunicamycin at 0.3 µmol/L combined with 0.6 µmol/L cisplatin showed an obviously enhanced inhibitory effect on colony formation of CNE-1 and CNE-2 cells. Tunicamycin treatment (3 µmol/L) of CNE-1 and CNE-2 cells for 48 h induced an apoptosis rate of only 8.89% and 8.67%, but when combined with 6 µmol/L cisplatin, the cell apoptosis rate increased to 37.02% and 32.25%, significantly higher than that in cells with cisplatin treatment alone (7.25% and 6.36%, respectively). Compared with tunicamycin and cisplatin alone, the combined treatment significantly increased Bax expression and decreased Bcl-2 expression in the cells; tunicamycin up-regulated the expression of GRP-78 and enhanced the activity of caspase-3.
CONCLUSIONTunicamycin can inhibit the proliferation of CNE-1 and CNE-2 cells and enhance cisplatin-induced cell death, the mechanism of which may involve excessive endoplasmic reticulum stress response and increased activity of caspase-3.
Apoptosis ; drug effects ; Carcinoma ; Caspase 3 ; metabolism ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Cisplatin ; pharmacology ; Endoplasmic Reticulum Stress ; drug effects ; Heat-Shock Proteins ; metabolism ; Humans ; Nasopharyngeal Neoplasms ; metabolism ; pathology ; Proto-Oncogene Proteins c-bcl-2 ; metabolism ; Tunicamycin ; pharmacology ; bcl-2-Associated X Protein ; metabolism
10.Effect of low-molecular-weight heparin combined with paclitaxel on the invasiveness and migration of nasopharyngeal carcinoma cells in vitro.
Pei ZHANG ; Surong ZHAO ; Lele SONG ; Longjian PU ; Zhiwen JIANG ; Hao LIU ; Chenchen JIANG
Journal of Southern Medical University 2012;32(11):1529-1535
OBJECTIVETo investigate the effect of low-molecular-weight heparin (LMWH) combined with paclitaxel (PTX) on the invasiveness and migration of nasopharyngeal carcinoma cells and explore the molecular mechanisms.
METHODSMTT assay was used to detect the growth inhibition induced by LMWH and PTX in CNE1 and CNE2 cells. Wound healing assay and Transwell migration assay were employed to assess the effects of the drugs on the cell migration, and Transwell invasion assay was used to evaluate the cell invasiveness. The cellular expression of matrix metalloproteinase-9 (MMP-9) and tissue inhibitor of matrix metalloproteinase-1 (TIMP-1) were analyzed by Western blotting. ELISA was used to determine the expression of heparanase (HPA) in the culture medium of the cells.
RESULTSMTT assay showed an obvious suppression of CNE1 and CNE2 cell proliferation in response to LMWH and PTX treatments. Treatment with 200 U·ml LMWH combined with 0.1 µmol·L PTX for 24 h resulted in the inhibition rates of migration of 66.70% and 70.53% in CNE1 and CNE2 cells, respectively significantly higher than the rates in cells with PTX treatment alone. The combined treatment with LMWH and PTX for 24 h also caused a significantly higher inhibition rate of cell invasion than LMWH and PTX alone. LMWH enhanced the down-regulation of MMP-9 and HPA induced by PTX.
CONCLUSIONLMWH can enhance the inhibitory effect of PTX on the migration and invasion of nasopharyngeal carcinoma cells, the mechanism of which may involve the down-regulation of MMP-9 and HPA expressions.
Carcinoma ; Cell Line, Tumor ; Cell Movement ; drug effects ; Cell Proliferation ; drug effects ; Glucuronidase ; metabolism ; Heparin Lyase ; metabolism ; Heparin, Low-Molecular-Weight ; administration & dosage ; pharmacology ; Humans ; Matrix Metalloproteinase 9 ; metabolism ; Nasopharyngeal Neoplasms ; pathology ; Paclitaxel ; administration & dosage ; pharmacology ; Tissue Inhibitor of Metalloproteinase-1 ; metabolism