1.Reversal of Multidrug Resistance of Human Colon Cancer Cells by Dihydroartemisin
Pengyu TAO ; Mingjie SHI ; Yongzhuo HUANG ; Huiyuan WANG ; Qin XU
Journal of Guangzhou University of Traditional Chinese Medicine 2016;33(5):698-703
Objective To investigate the multidrug-resistance reversal action and mechanism of dihydroartemisin (DHA) on human colon cancer cell line HCT8/ADR. Methods The cytotoxicity of dihydroartemisin combined with doxorubicin(DOX) was determined by methyl thiazolyl tetrazolium(MTT) assay and cell apoptosis was observed by flow cytometry. Western blot assay was used to measure the autophagy. Results The combined treatment with dihydroartemisin and doxorubicin significantly enhanced the cytotoxicity in HCT8/ADR cells and effectively increased the apoptotic level. Autophagy was also induced by the combined treatment , which maybe played a crucial role in the regulation of doxorubicin-sensitization of HCT8/ADR cells. Conclusion The results indicated that dihydroartemisin can reverse multidrug resistance through increasing the doxorubicin-sensitivity of HCT8/ADR cells.
2.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.
3.Pituitary crisis in a multiple myeloma patient during treatment
Xiaoxiao XU ; Zhongshu MA ; Yong YU ; Lianyu ZHANG ; Peifang LIU ; Pengyu ZHANG ; Shuxia FANG ; Yizhuo ZHANG
Chinese Journal of Clinical Oncology 2013;(18):1130-1133
Multiple myeloma is common in the older population and is treated mainly with chemotherapy. However, chemotherapy-related side effects imitate the clinical manifestations of Sheehan's syndrome, which leads to misdiagnosis and missed diagnosis, particularly for older patients without a clear history of postpartum hemorrhage. Therefore, when older women with malignant myelomas show refractory hyponatremia and gastrointestinal disorders while under chemotherapy, a diagnosis of Sheehan's syndrome should be considered. The early detection of the disorder will guarantee timely individualized treatment.
4.Study on Anti-tumor Activities of Trichosanthin Mediated by Low Molecular Weight Protamine
Jianming LIANG ; Feng ZENG ; Yingzhi CHEN ; Jiao TAN ; Pengyu TAO ; Qin XU ; Yongzhuo HUANG
Journal of Guangzhou University of Traditional Chinese Medicine 2014;(5):780-785,854
Objective To investigate the anti-tumor activities of cell-penetrating peptide ( CPP) - mediated trichosanthin ( TCS) , which is a recombinant protein obtained from Radix Trichosanthis. Methods Cysteine residue was introduced to the C-terminus of TCS by protein recombinant technique, and then with the newly-formed terminal as the modification site, TCS was coupled with CPP. As a target protein, CPP-mediated TCS was isolated and purified by affinity chromatography. The expression of the target protein and its responsiveness to reducing substances were detected by using the sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The cellular uptake rate of CPP-mediated TCS was determined by using cell uptake test, and its anti-tumor activity was measured by using methyl thiazolyl tetrazolium (MTT) assay. Results The TCS-CPP compound had been successfully developed in this study, and showed certain reducing responsiveness. After modified with CPP, TCS had higher cellular uptake rate and stronger anti-tumor effect on HeLa and MCF-7 cells. Conclusion TCS modified by CPP can enhance the anti-tumor activities of TCS.
5.Glutamine regulates the proliferation and survival of small cell lung cancer H446 cells
Pengyu XU ; Jiayin LI ; Yajing MIAO ; Cuicui GAO ; Yao SHEN ; Fang JIN ; Xiaofei QIU
Tianjin Medical Journal 2016;44(12):1423-1427
Objective To investigate the effects of glutamine (Gln) on proliferation and survival of small cell lung cancer H446 cells, and further to explore the potential mechanism. Methods The proliferation of H446 cells was detected at different time points (0, 24, 48, 72 and 96 h) by CCK-8 assay in Gln (+) group and Gln (-) group, and an optimal time was selected. Under the optimal time, Annexin V-FITC/PI staining, CellTiter-Glo? assay kit and flow cytometer were used to detect cell survival, cellular adenosine triphosphate (ATP) and reactive oxygen species (ROS) levels. Gln (-) group was used as the control group, under the condition of Gln deficiency, cellular ATP, cell proliferation and survival were detected after adding oxaloacetic acid (OAA) or dimethyl-α-ketoglutarate (DM-αKG). Gln (-) group was used as the control group, cellular ROS, cell proliferation, colony and survival were detected after treated with ROS scavenger N- acetyl cysteine (NAC). With different concentrations (0, 2, 5, 10 μmol/L) of glutaminase inhibitor BPTES, the optimal concentration was selected through the colony assay. The cellular ATP and ROS levels and cell proliferation were detected under the optimal concentration. H446 cells were treated with bis-2-(5-phenylacetamido-1,2,4-thiadiazol-2-yl) ethyl sulfide (BPTES), ROS inducer hydrogen peroxide (H2O2) or the combination of them, and cell survival ratio was compared between two groups. Results The proliferation levels of H446 cells at 24, 48, which were decreased most significantly in 72 h in Gln (-) group. When 72 h was used as the optimal time, the cell survival ratio and ATP level were decreased, and the ROS level was increased, in Gln (-) group compared with those of Gln (+) group (P<0.05). There was a higher survival ratio in H446 cells in Gln (-)+OAA group and Gln (-)+DM-αKG group than that of Gln (-) group (P<0.05), but there were no significant differences in cell proliferation and ATP levels between Gln (-) group, Gln (-)+OAA group and Gln (-)+DM-αKG group. The ROS level was reduced, the cell proliferation, colony level and survival ratio were increased in Gln (-)+NAC group compared with those of Gln (-) group (P<0.05). Cloning assay showed that 10μmol/L was the optional concentration. Under this concentration, the proliferation and ATP level were decreased in Gln(+)+BPTES group (P<0.05), and cellular ROS level was up-regulated compared with Gln(+) group. The survival ratio was significantly lower in BPTES+H 2O2 group compared with BPTES (+) group or H2O2 (+) group. Conclusion Glutamine deficiency inhibits the proliferation and survival ratio of H446 cells through enhancing ROS level. BPTES and H2O2 show synergistically inhibitory effect on the survival of H446 cells.
6.Analysis of " Internet plus healthcare" related policies in the eastern, central and western regions of China before and after the COVID-19 pandemic
Jingjing XIONG ; Yunyun HUANG ; Pengyu XU ; Xingyue YU ; Zehong ZHANG
Chinese Journal of Hospital Administration 2021;37(11):883-888
Objective:To analyze the commonalities and differences of " Internet plus healthcare" related policies in the eastern, central and western regions of China before and after the COVID-19 pandemic and to provide references for further improving China′s " Internet plus healthcare" policy.Methods:Collecting the policies of " Internet plus healthcare" promulgated by 31 provinces(autonomous regions, municipalities)in China from April 2018 to December 2020 on the websites of various government departments, extracting keywords based on content analysis methods, and calculating degree centralities using UCINET 6 software. A comparison of the policies of the eastern, central and western regions between the pre epidemic stage(April 2018 to December 2019)and the COVID-19 pandemic stage(January 2020 to December 2020)was carried out.Results:In the pre epidemic stage, Internet diagnosis and treatment, " Internet plus medical service price" and quality supervision were the focus of attention in all regions. But the eastern region paid more attention to the policy planning of Internet hospital operation and Internet medical application, the central region focused on the transformation and efficiency improvement of medical service process by Internet technology, and the western region paid more attention to encourage the application of telemedicine to solve the difficulty of seeing a doctor. In the pandemic stage, the " Internet plus service price" was the policy focus in all regions, mostly in medical insurance payment, and the policy difference was not obvious among the eastern, central and western regions.Conclusions:Before and after the COVID-19 pandemic, the policy of " Internet plus healthcare " promulgated by the local governments in eastern, central and western regions had different emphasis and different characteristics. Our suggestion was to improve the policy system, standardize industry supervision, strengthen information security, orderly interconnect medical insurance, and actively promote the exchange of policy experiences among the provinces.
7.Pain-point analysis of outpatient service based on online patient comments
Jingjing XIONG ; Shuo YANG ; Pengyu XU ; Jiayue ZHU
Chinese Journal of Hospital Administration 2021;37(4):317-321
Objective:To explore the pain points of outpatient service in hospital from the patient′s perspective, and to provide new ideas for optimizing management of service quality.Methods:Ten top hospitals in China were selected to collect the poor evaluation texts of outpatient services published by patients on the Dianping.com from January 1, 2017 to December 31, 2019. The user′ pain-point analysis model was introduced, the pain-point index word collection was constructed according to the word frequency, the pain-point index was quantitatively analyzed combined with the sentimental value, and the content of the index words was analyzed through conceptual annotation.Results:The analysis of 1 259 poor evaluation texts showed that patients′ poor evaluation of outpatient service quality mainly focused on four aspects: interpersonal interaction, process factors, human-computer interaction and environmental factors. The poor attitude of medical staff in interpersonal interaction and the long queuing time and short treatment time in process factors were the most prominent.Conclusions:Online patient comment analysis is suitable for the identification and perception of pain points in outpatient service, and is helpful for the hospital to choose the direction and items of service improvement.
8.Influence of sulforaphane preconditioning in myocardial cold ischemia-reperfusion injury through PI3K/Akt signaling pathway in rats
Xue YI ; Pengyu WU ; Shuliang YANG ; Yan HAN ; Xuehui YANG ; Shubo ZHANG ; Tao XU ; Yizhe GAO ; Muhuo GAO ; Zhanqing LI
Journal of Jilin University(Medicine Edition) 2017;43(5):918-922,后插5
Objective:To explore the effect of sulforaphane (SFN) preconditioning on the cold myocardial ischemia-reperfusion injury (IRI) in the rats through PI3K/Akt signaling pathway.Methods:Sixty-four health male Sprague-Dawley (SD) rats were randomly divided into cold IRI group,SFN group,LY (LY294002) + cold IRI group,and LY+SFN group (n=16).The allogeneic heterotopic heart transplantation model was established by donor heart into recipient abdomen.The myocardium tissue was taken 24 h after reperfusion for the detection of histological changes using HE staining.The expression levels of Akt,p-Akt,Bax and Bcl-2 proteins were detected by immunohistochemistry and Western boltting methods.Results:The morphological results showed that the myocardium tissue damage was serious in cold IRI group and LY+cold IRI group,it was light in SFN group;the myocardium tissue damage of the rats in SFN+ LY group was ranged between cold IRI group and SFN group.Compared with IRI group,the expression levels of p-Akt protein and Bcl-2 protein in SFN group were increased (P<0.05),and the expression level of Bax protein was decreased (P<0.05).After treatment of blockage LY294002,compared with LY-+-cold IRI group,the expression level of p-Akt protein in LY-+-SFN group was not statistically significant (P>0.05),the expression level of Bcl2 protein was increased (P<0.05),the expression levels of Bax protein was decreased),and the ratio of Bcl-2/Bax was also increased (P<0.05).Conclusion:SFN may attenuate cold IRI of heart transplantation through PI3K/Akt signaling pathway in the rats.
9.Establishment of air-pouch bladder cancer model and investigation of its feasibility for evaluating the effect of intravesical therapy
Pengyu GUO ; Li PENG ; Lu WANG ; Ziyin CHEN ; Jiuwei CHEN ; Wanhai XU
Chinese Journal of Urology 2020;41(9):698-702
Objective:To establish an air-pouch bladder cancer (APBCa) model and investigate whether it could be a new animal model to evaluate the efficacy of intravesical therapy through chemotherapeutics and BCG instillation.Methods:Filtered sterile air was injected subcutaneously into the backs of BALB/c Nude mice to create a 2.5 cm×3.5 cm air pouch. After 24 hours, human bladder cancer cells EJ were seeded on the inner face of the pouch wall to establish APBCa of human cancer (H-APBCa). Gemcitabine instillation was used to investigate whether chemotherapy could inhibit tumor growth in the H-APBCa model, and Tunel staining was used to verify the apoptosis of tumor cells 20-day treatment. Filtered sterile air was injected subcutaneously into the backs of C57BL/6 mice to create a 2.5 cm×3.5 cm air pouch. After 24 hours, mice bladder cancer cells MB49 were seeded on the inner face of the pouch wall to establish APBCa with intact immunity (I-APBCa). BCG instillation was used to investigate whether BCG could inhibit tumor growth in the APBCa model. Immunofluorescence was used to verify the infiltration of immune cells after 20-day treatment.Results:H-APBCa and I-APBCa mice models could be established by immune deficiency and intact mice. At day 20, chemotherapeutic instillation therapy could inhibit tumor growth (781.02±241.02 vs. 1213.88±214.02 mm 3, P<0.05) by inducing tumor cell apoptosis with statistically significant differences (77.33±4.63 vs. 14.67±2.60, P<0.05). BCG instillation was able to inhibit tumor growth (645.02±156.63 vs. 948.84±221.76, P<0.05) by increasing CD80 + macrophage (49.67±7.57 vs. 16.33±5.69, P<0.05) and T cells in the tumor with statistically significant differences (18.00±3.46 vs. 4.67±1.45, P<0.05). Conclusions:APBCa model could evaluate the efficacy of drug instillation and was expected to be a new animal model for studying drug for intravesical therapy.
10.YKL-40 at admission predicts stroke-associated pneumonia and outcome in patients with acute ischemic stroke
Guomei SHI ; Xiaorong WANG ; Wu XU ; Minwang GUO ; Meng WANG ; Pengyu GONG ; Junshan ZHOU ; Rujuan ZHOU
International Journal of Cerebrovascular Diseases 2022;30(8):569-576
Objective:To investigate the predictive value of YKL-40 at admission on stroke-associated pneumonia (SAP) and poor outcome in patients with acute ischemic stroke (AIS).Methods:Patients with AIS admitted to Taixing People’s Hospital from February 2020 to March 2021 were enrolled prospectively. The poor outcome was defined as 3-6 points on the modified Rankin Scale at 90 d after onset. Multivariate logistic regression analysis was used to determine the independent predictors of SAP and poor outcome, and the predictive value of serum YKL-40 on SAP and poor outcome was evaluated by receiver operating characteristic (ROC) curve. Results:A total of 377 patients with AIS were enrolled. The median serum YKL-40 was 127.16 μg/L. One hundred and four patients (27.6%) had SAP, and 126 (33.4%) had poor outcomes at 90 d after onset. Multivariate logistic regression analysis showed that after adjusting for confounding factors, YKL-40 was the independent predictors of SAP (odds ratio [ OR] 1.005, 95% confidence interval [ CI] 1.003-1.008; P=0.001) and poor outcome at 90 d ( OR 1.009, 95% CI 1.006-1.011; P=0.001). The ROC curve analysis showed that the area under the curve of YKL-40 for predicting SAP was 0.769 (95% CI 0.713-0.824; P<0.001), the best cutoff value was 168.70 μg/L, and the sensitivity and specificity were 71.2% and 75.1% respectively; the area under the curve of YKL-40 for predicting poor outcome at 90 d was 0.787 (95% CI 0.735-0.840; P<0.001), the best cutoff value was 195.56 μg/L, and the sensitivity and specificity were 68.3% and 84.1% respectively. Conclusion:Higher serum YKL-40 at admission has a good predictive value for SAP and poor outcome at 90 d in patients with AIS.