1.The role of Twist on cisplatin resistance of Hela cervical cancer cells in hypoxia microenvironment and its mechanism
Peng KOU ; Qin XU ; Lihua YANG
Journal of Medical Postgraduates 2015;(8):815-819
Objective Drug resistance is a major problem for the successful chemotherapeutic treatment and prognosis of cervi -cal cancer .The article investigated the role of Twist on cisplatin resistance of Hela cervical cancer cells in hypoxia microenvironment and its possible mechanism to provide an experimental basis to improve the therapeutic efficacy for cervical cancer . Methods Hela Cells were divided into two groups, normal oxygen group A (no CoCl2) and hypoxia group A (addition of 150μmol/L CoCl2 medium).6 hours later, different degrees(10-3, 10-4, 10-5, 10-6, 10-7 mol/L) of cisplatin (10 -3 , 10 -4 , 10 -5 , 10 -6 , 10 -7 mol/L) were added into two groups .24 hours later , MTT were added to measure IC 50 and growth inhibition ratio .CoCl2 were used to mimic hypoxia environ-ment.The working concentration and the treatment duration of Cispla-tin was optimized by MTT method.The expressions of Twist and MDR1 in normal oxygen group , cisplatin group , hypoxic group B and hypoxic cisplatin group B were determined by immunofluores-cence and western blot . Results The IC50 values were 10 -5.3 mol/L and 10 -4.5 mol/L of cisplatin respectively in normal oxygen group and hypoxia group, and there was significantly difference between them (P<0.05).The optimized working concentration and work time of cisplatin was 10 -5 mol/L and 24 h.Compare to the normal oxygen group (13.08 ±2.39, 29.57 ±12.80), the expres-sions of Twist (20.81 ±2.07, 24.25 ±4.51, 33.14 ±4.24) and MDR1 (35.26 ±8.41, 60.13 ±22.32, 76.00 ±9.96) was signifi-cantly higher than those in other three groups and which were the highest in hypoxic cisplatin group (P<0.05).There were significant positive correlations among them in hypoxic group and hypoxic cisplatin group (r =0.686,P <0.05;r =0.546,P <0.05). Conclusion The hypoxia microenvironment may be related to the cisplatin resistance of Hela cervical cancer cells .The possible mechanism is hypoxia activates Twist and upregulates MDR 1 expression , resulting in cisplatin resistance of Hela cells .
2.Risk factors of schistosomiasis cirrhosis esophageal varices bleeding
Qin WANG ; Peng CHEN ; Banghe XU ; Hongxia ZHU
Chinese Journal of Schistosomiasis Control 2014;(2):226-227
Objective To explore the risk factors of schistosomiasis cirrhosis esophageal varices hemorrhage. Methods A total of 113 cases of schistosomiasis cirrhosis esophageal varices hemorrhage and 128 schistosomiasis cirrhosis esophageal varices patients without hemorrhage were selected and their relevant factors were analyzed statistically. Results There were significant differences between the two groups in the prothrombin time,portal vein diameter,degree of esophageal varices,and varicose vein tumor-like lesions(P<0.01). Conclusion The risk factors of schistosomiasis cirrhosis esophageal varices hemorrhage are the prothrombin time,portal vein diameter,degree of esophageal varices,and varicose vein tumor-like lesions.
3.Effects and mechanism of hypoxia on the growth and invasion of human cervical cancer Hela cells
Peng KOU ; Qin XU ; Jianjun LIU ; Jing YANG ; Lihua YANG
The Journal of Practical Medicine 2014;(4):522-524
Objective To investigate the effects and mechanism of hypoxia on the growth and invasion of human cervical cancer Hela cells. Methods Hypoxia microenvironment was mimicked by cobalt chloride (CoCl2), in which the growth of Hela cells were determined by MTT method, the migration ability was detected by scratch test, and the expressions of HIF-1αand Vimentin protein were measured by immunofluorescence. Results The growth of Hela cells treated with 150 μmol/L of CoCl2 for 6 h was promoted. The migration ability in hypoxia group at 48 h was significantly higher than that in ordinary oxygen group. The expressions of HIF-lαand Vimentin in Hela cells in hypoxia group were significantly higher than those in ordinary oxygen group. There was significantly positive correlation between the expressions of HIF-1α and Vimentin (P < 0.05). Conclusion Hypoxia could promote the expression of HIF-lα, which then could up-regulate the expression of Vimentin , and finally enhanced the ability of invasion and metastasis of Hela cells.
4.A preliminary study on the influence of low-iron environment on serum osteocalcin and bone micro-structure in ovariectomized mice
Chao GAO ; Qin SHI ; Peng JIA ; Zengli ZHANG ; Youjia XU
Chinese Journal of Endocrinology and Metabolism 2012;(12):967-971
Objective To investigate the changes of serum osteocalcin and bone micro-structure in ovariectomized mice exposed to low-iron environment.Methods Twenty-four 12-week-old female C57BL/6 mice were divided equally into sham operation (SHAM) group,model(OVX) group,and low iron(OVX+DFO) group.In low-iron group,deferoxamine(DFO) was injected 3 times per week for 5 weeks after operation ; the other groups were injected with the same dose of 0.9% normal saline for 5 weeks.The serum,left femur,uterus were harvested after five weeks of treatment.The serum osteocalcin and ferritin levels were measured by ELISA kit,the weight of the uterus was recorded by analytical balance.A high resolution micro-CT was used to scan the left femur for cortical bone and cancellous bone analysis.Results (1) The serum osteocalcin and serum ferritin levels in low-iron group were significantly lower than those in the other 2 groups (P<0.01) ; (2) Compared with the sham group and ovx group,there were significant decrease of the BMD、BV/TV and Tb.N,but increase of Tb.Th and Tb.Sp in low-iron group (P<0.01).Conclusion A certain dose of DFO (30 mg/kg) can decrease the serum ferritin levels as well as the bone formation index in ovariectomized mice.
5.Analysis of risk factors of acute respiratory distress syndrome secondary to severe multiple trauma
Yanming QIN ; Peng WANG ; Xuanxuan XU ; Zheng LI
Chinese Critical Care Medicine 2021;33(3):299-304
Objective:To explore the risk factors of acute respiratory distress syndrome (ARDS) secondary to severe multiple trauma and the role of clinical guidance.Methods:The clinical data of 115 patients with severe multiple trauma admitted to the trauma center of Zhenjiang First People's Hospital from December 2017 to September 2020 were retrospectively analyzed. According to whether ARDS occurred within 1 week of the disease course, the patients were divided into ARDS group and non-ARDS group. The basic post-traumatic data, initial treatment measures (within 24 hours), pathophysiology, stress metabolism, and post-traumatic complications of the two groups of patients were selected for univariate analysis, the statistically different indicators of univariate analysis were incorporated into the multivariate Logistic regression analysis to screen out independent high-risk factors that affect the occurrence of ARDS in patients with severe multiple trauma, and a receiver operating characteristic curve (ROC curve) was drawn to analyze the effects of each risk factor on the occurrence of ARDS.Results:Among 115 patients, there were 45 casesin the ARDS group and 70 cases in the non-ARDS group. Compared with the non-ARDS group, the patients in the ARDS group were older (years: 57.45±15.37 vs. 45.68±12.70), and the proportion of patients combined with moderate-severe chest trauma, traumatic brain injury (TBI), shock, and massive blood transfusion were higher (71.11% vs. 31.43%, 44.44% vs. 28.57%, 80.00% vs. 67.14%, 46.67% vs. 27.14%). In the ARDS group, procalcitonin [PCT (μg/L):29.73±6.08 vs. 12.45±2.12], thrombomodulin [TM (ng/L): 83.43±16.34 vs. 37.66±14.64], blood glucose (mmol/L:17.2±5.0 vs. 10.3±2.4), triacylglycerol [TG (mmol/L): 3.77±0.57 vs. 2.22±0.63], interleukin-6 [IL-6 (ng/L):38.97±10.79 vs. 25.98±5.40], tumor necrosis factor-α [TNF-α (ng/L): 48.78±13.99 vs. 35.30±13.03], intra-abdominal pressure [mmHg (1 mmHg = 0.133 kPa): 25.21±3.59 vs. 11.98±4.91], serum creatinine [SCr (μmol/L):180.45±42.35 vs. 132.17±49.36] and blood urea nitrogen [BUN (mmol/L): 13.83±4.97 vs. 8.80±4.32] were significantly higher than those in the non-ARDS group; the proportion of patients with crystal infusion volume ≥ 3 000 mL(26.67% vs. 34.29%) and platelet count [PLT (×10 9/L): 72.67±7.96 vs. 127.99±17.65] and the levels of plasma glutathione peroxidase [GSH-Px (kU/L): 87.15±27.81 vs. 161.15±17.94], plasma superoxide dismutase [SOD (kU/L):92.65±32.67 vs. 125.58±38.96] were significantly lower than those in the non-ARDS group, the differences were statistically significant (all P < 0.05). Multivariate Logistic regression analysis showed that 11 indicators such as age, combined moderate-severe chest trauma, combined TBI, massive blood transfusion, PCT, TM, blood glucose, TNF-α, plasma GSH-Px, intra-abdominal pressure and SCr were independent risk factors that could predict ARDS secondary to severe multiple trauma, the odds ratio ( OR) and 95% confidence interval (95% CI) were 1.201 (1.035-1.165), 3.414 (1.217-8.876), 2.889 (1.124-8.109), 3.134 (1.322-9.261), 1.467 (1.096-2.307), 2.428 (0.024-0.973), 5.787 (1.246-9.642), 1.106 (0.949-5.108), 7.450 (1.587-10.261), 3.144 (1.217-8.876), 1.051 (1.002-1.542) respectively, the P valueswere 0.008, 0.024, 0.044, 0.017, 0.018, 0.045, 0.026, 0.037, 0.005, 0.029, 0.033 respectively. ROC curve analysis showed that plasma GSH-Px had a higher predictive value for ARDS secondary to severe multiple trauma, the area underROC curve (AUC) = 0.873, 95% CI was 0.798-0.928, P = 0.000, when the best cut-off value at 72.22 kU/L, its sensitivitywas 86.7%, specificity was 75.7%, positive predictive value was 69.6%, and negative predictive value was 89.8%. The Logistic regression model established by 11 independent high-risk factors had an accuracy rate of 81.74% in predicting ARDS secondary to severe multiple trauma, which had a good guiding significance for predicting ARDS. Conclusion:Our study showed that there are many risk factors for ARDS secondary to severe multiple trauma, involvingbasic post-traumatic data, initial treatment measures, pathophysiology, stress metabolism, post-traumatic complications, etc. Early identification and intervention may be beneficial to improve the success rate of treatment for such patients.
6.Construction and prokaryotic expression of His-tagged expression vector of human IP-10 and identification of its activity
Ziyun SHAO ; Zhifeng LIU ; Yi PENG ; Jia XU ; Qinghe QIN ; Peng DENG ; Yong JIANG
Chinese Journal of Pathophysiology 1989;0(05):-
AIM: To construct prokaryotic expression vector of His-tagged human IP-10 for further study of its biological function in the inflammatory response. METHODS: The coding sequence of IP-10 lacking signal peptide was amplified from human lung cDNA library by polymerase chain reaction (PCR) and the fragment was cloned into pET-14b plasmid for the construction of His-tagged fusion protein expressing vector, pET-14b/IP-10. After being identified by enzyme digestion and sequencing, the recombinant vector was transformed into a strain of E. coli, BL21 (DE_3). The expression of His-tagged fusion protein was induced with IPTG and purified with Ni+-NTA affinity chromatography. Then the chemotactic activity of IP-10 was determined by transwell migration assay on THP-1 cells. RESULTS: The construction of pET-14b/IP-10 recombinant vector was proved by enzyme digestion and sequencing. The fusion protein IP-10, which was purified by a routine Ni+ affinity method, had an activity on the induction of cell migration of THP-1. CONCLUSION: We successfully construct IP-10 fusion protein expressing vector and get the fusion protein with high bioactivity, which provides essential materials for the future studies on IP-10.
7.NIBP regulates colon cancer cell express MMP2 and MMP9 via NF-κB signaling pathways
Chunyan XU ; Jiean HUANG ; Mengbin QIN ; Peng PENG ; Mengzi LIANG ; Qian LI ; Chujie CHEN
The Journal of Practical Medicine 2015;(11):1752-1755
Objective To observe after the NIBP (NIK and IKK beta binding protein) gene transfected into colon cancer cell lines HT29, the migration of cells and expression of p65, MMP2, MMP9 mRNA and proteins. Methods The experimental group: divided into without transfection HT29 cell (HT29 group) and transfection no-load HT29 cell (HT29-NC group) and transfection NIBP HT29 cell (HT29-NIBP steady group). Using the Transwell test to detect cell migration ability. Q-PCR method to detect the mRNA expressions of NIBP , p65, MMP2, MMP9. Western Blot method to detect the expressions of NIBP, p65, phosphorylation p65 (p-p65) proteins. The method of ELISA was used to detect the secretion of matrix metalloproteinase (MMP)-2, MMP-9. Results The high expression of NIBP may enhance the migration ability of colon cancer cell lines HT29 , increasing the expression of p-p65, MMP2, MMP9 mRNA and proteins (P < 0.05). Conclusion NIBP potently enhances colon cancer HT29 cell migration and invasion. Activation of NF-κB signaling pathways resulted in up-regulation of MMP-2 and MMP-9 maybe one of its molecular mechanisms.
8.Strengthen the management of outside projects of general hospital and improve the level of scientific research management
Baohua FENG ; Peng QIN ; Pengjun ZHANG ; Miao MIAO ; Lingling YU ; Yang XU ; Yuxuan PENG
Chinese Journal of Medical Science Research Management 2016;29(6):472-476
Objective The number of awarded provincial-level research projects and their budget had been declining year by year.We analyzed the data of provincial-level research projects in a general hospital in last ten years to be able to describe the distribution of research projects,the subject crossing situation,the distributions of subject and age structure,degree,title of the project leaders.We tried to find out the problems of them and to provide evidence for policy making.Methods Based on the scientific research management software,the data of the provincial-level research projects can be collected from 2005 to 2014.To Analyze the distributions of subject and structure of age,degree,professional title of the projects leaders by SPSS software.Results In general,the number and funds of provincial-level projects were unstable in the nearly ten years.Many of them focused mainly on basic departments and key subject.Young and middle-aged professionals were the backbone of the hospital team and talents with a high degree and title were the main force in the successful provincial-level projects.Conclusions According to the distribution trend of the projects,the hospital should deliver a carefully targeted set of actions.According to the distribution trend of the subject departments,the hospital should strengthen cooperation among departments and promote the integration of subjects.Strengthen the construction of talent team,pay attention to the cultivation of young talents.Strengthen scientific research management,To provide services for provincial level projects management.
9.Establishment of Quality Standard System for Gui Erbai Gel Based on a Method of A System to Multiple Evaluation
Xianshun XU ; Hongen QIN ; Sibo ZHANG ; Yongfang XIA ; Lei PENG ; Yuancui XU
China Pharmacist 2017;20(9):1559-1563
Objective:To establish the quality standard system of Gui Erbai gel based on a method ofa system to multiple evalu-ation and discuss the feasibility of the method used for the quality standard for traditional Chinese medicine. Methods:TLC identifi-cation of Gui Erbai gel was established by one thin layer system. An HPLC method was used to detect 6 active components in Gui Erbai gel. Results:Five active components in the gel could be identified by one thin layers system simultaneously with clear spots and good separation. Six active components in the gel could be determined by the same HPLC system with high accuracy. The average content of podophylotoxin,quercetin,kaempferol,imperatorin,dictamnine and rutin is as follows 0. 154,0. 052,0. 138,0. 051,0. 060,0. 048 mg· g-1 . RSD<3%. Conclusion:The established method based on a system to multiple evaluation can be used for the quality standard establishment for Gui Erbai gel with the properties of promising feasibility, simple operation, low cost, high accuracy and good stabili-ty.
10.Effect of Qingguang'an Ⅱ on expression of PAX6, Ngn1 and Ngn2 mRNA of rats with chronic high intraocular pressure
Ya-Sha, ZHOU ; Jian, XU ; Yue, LIU ; Jun, PENG ; Yi-Jing, YANG ; Gen-Yan, QIN ; Qing-Hua, PENG
International Eye Science 2017;17(9):1631-1634
AIM:To remark the effect of Qingguang'an Ⅱ on expression of PAX6, Ngn1, and Ngn2 mRNA of rats with chronic high intraocular pressure.METHODS:Totally 40 male SD rats were randomly divided into 6 groups, that was:A:blank group, B:model group, C:Qingguang'an Ⅱ low dose group, D:Qingguang'an Ⅱ moderate dose group, E:Qingguang'an Ⅱ high dose group, F:Yimaikang disket group.B, C, D, E, F groups of experimental rats were established the model of chronic high intraocular pressure (IOP) by cauterizing of superficial scleral vein.Animal model was established successfully by using monitoring IOP consistently keep above 25mmHg for 8wk as cut-off criterion.Tissues of Eyes were obtained after intragastric administration for 2wk and 4wk.The expressions of PAX6, Ngn1, and Ngn2 mRNA were investigated by Real-time PCR.RESULTS:At the time-point of 2wk, PAX6, Ngn1, and Ngn2 mRNA in group B were statistically expressed in lower level comparing with other groups (P<0.05).Moreover, at the time-point of 4wk, PAX6, Ngn1, and Ngn2 mRNA in group C, D and E were statistically expressed in higher level comparing with group F (P<0.05).Besides, PAX6, Ngn1, and Ngn2 mRNA in group C and D were statistically expressed in lower level comparing with group E (P<0.05).PAX6, Ngn1, and Ngn2 mRNA in group C and D were expressed in similar level(P>0.05).CONCLUSION:In summar, Qingguang'an Ⅱ and Yimaikang disket can remarkably increase the expressions of PAX6, Ngn1, and Ngn2, which suggest protecting the optic nerve of rats caused by chronic high IOP.What's more, this study indicated that, in the protection of optic nerve of rats with chronic high IOP, the high dose of Qingguang'an Ⅱ at the time-point of 4wk was the better choice.