1.Effect of low-expressed MK on proliferation, migration and angiogenesis of HUVECs cells in human breast cancer cell line MDA-MB-231
Dongmei LIU ; Yongping WU ; Qingling WANG
Chinese Journal of Clinical and Experimental Pathology 2015;(7):729-733
Purpose To observe the effect of MK on proliferation, migration and angiogenesis of human umbilical vein endothelial HU-VECs cells and to explore the role of MK in tumor angiogenesis. Methods siRNA targeting MK was used to downregulate MK expres-sion in human breast cancer cell line MDA-MB-231. Then, the experiment was divided into three groups: the untreated group, the empty vector transfection group and MK gene interference group. CCK-8 assay was used to detect the endothelial cells proliferation, tr-answell method was for detection of endothelial cell migration numbers, and matrigel in vitro small tube formation assay was used to sur-vey the state of tube formation. Expression of MK in siRNA transfection was identified by RT-PCR and Western blot. Results The MK gene interference group showed lower cell proliferation activity, less number of migration of cells and tube formation than the other two groups (P<0. 05). Conclusion Low-expressed MK in human breast cancer cell line MDA-MB-231 can inhibit proliferation, mi-gration and angiogenesis of endothelial cells, which shows that MK may play an important role in tumor angiogenesis.
2.QCC: a kind of quality management for reducing re-washing rate of suction aspirator tip
Lingling ZHUANG ; Qian LIU ; Qingling WU
China Medical Equipment 2017;14(8):129-132
Objective:To discuss the application effect of quality management of quality control circle (QCC) for reducing re-washing rate of suction aspirator tip in the quality management of medical instrument of hospital.Methods: Through established QCC group to designate one topic, to investigate its current situation and to protocol project plan of quality management. The qualified rate of first washing of 904 suction aspirator tips was analyzed before control management of QCC was implemented, and then series of QCC methods including fishbone diagram were applied to analyze and confirm main factors. The protocol countermeasure, depend on above information, achieved improvement for washing quality of 856 suction aspirator tips after the quality management of QCC was implemented in practice. The re-washing rates of washing quality of suction aspirator tip between before and after QCC was applied were compared.Results: The qualified rate of first washing of 904 suction aspirator tips before the quality management of QCC was implemented was 65.7%, while it was 97.6% for 856 suction aspirator tips after quality management of QCC was implemented. The washing qualified rate of suction aspirator tip after quality management of QCC was applied was significantly higher than that before it was applied (x2=8.429,P<0.05), and the result has achieved the aim that continuous improved quality.Conclusion: The application of quality management of QCC can decrease re-washing rate of suction aspirator tip, and contribute to decrease cost, to increase work efficiency, to strengthen the attention and degree of involvement of staff for work quality. Therefore, the management can fully exert potential of member of quality management of QCC, and enhance washing quality of suction aspirator tips.
3.Clinical observation of sufentanil and midazolam assisted epidural anesthesia for cholecystectomy
Kan YANG ; Xiaodong PENG ; Zhonghou XU ; Qingling WU
Chinese Journal of Primary Medicine and Pharmacy 2011;18(3):329-331
Objective To investigate the effect of sufentanil and midazolam assisted epidural anesthesia for cholecystectomy on sedative effects,visceral pulling reaction and the functions of respiratory and circulatory system.MethodsFourty patients underwent cholecystectomy received informed consent, were randomly allocated to two groups:group A and group B( n =20 each). When the level of epidural block is appropriate,auxiliary medicine infusion was given 5min before the skin incision. In group A,first injection was sufentanil 0. lμg · kg-1 and midazolam 0. 02mg · kg-1 in 1-2 min, then continuous infusion of sufentanil and midazolam was given at the rate of 0. 2μg ·kg-1 · h-1 and 0. 04μg · kg-1 · h-1 respectively,and the infusion was stopped at the time of wound closure. Group B received slow intravenous injection of pethidine 1mg · kg-1 and droperidol 0. 5mg · kg-1. We recorded the differences of SBP、DBP、SpO2 、HR、Ransay score and visceral pulling reaction classification. Results SBP、DBP、HR after anesthesis,were decreased in different degree in the above groups,but there was no statistically significant difference( P >0. 05). According to Ramsay score,in group A there were 3 cases in grade Ⅱ、8 cases in Ⅲ and 9 cases in Ⅳ ,while in group B there were respectively 4 cases in Ⅱ、13 cases in Ⅲ and 3 cases inⅣ. By comparision,the differences had statistical significance ( u = 3.75, P < 0. 05 ); Group A according to visceral pulling reaction classification (0- Ⅲ ), were 9 cases in grade 0,9 cases in Ⅰ , 1 cases in Ⅱ and 1 cases in Ⅲ, while in group B there were respectively 4,5,7 and 4 cases. By comparision, the differences had statistical significance(u = 4. 01,P < 0. 01 ). Conclusion In the cholecystectomy,sufentanil and midazolam assisted epidural anesthesia,could improve the level of sedation ,prevent visceral pulling reaction ,and had a minor interference to respiratory and circulatory function.
4.The effect of total mesorectal excision for low rectal cancer
Zhongxue SU ; Hongjun LIU ; Liming LIN ; Qingling MU ; Taihuang WU
Chinese Journal of General Surgery 2001;0(10):-
Objective To investigate the therapeutic effect of total mesorectal excision(TME) for low rectal cancer.Methods One hundred nineteen patients with rectal cancer,located on an average within 8 cm of the anal verge,were included in the study.Fifty-four patients underwent traditional operation,and sixty-five patients underwent total mesorectal excision.Operation time,loss of blood at operation and local recurrence rates were compared between the two groups.Results The average operation time and blood loss were 118 minutes,and 100mL respectively in TME group,and they were 182 minutes,and 340 ml respectively in traditional operation group.There were significant differences between them(all P0.05).Conclusions In TME group,operation time was shorter,operative blood loss was less,and local recurrence rate was lower.TME should be applicated for patients with low rectal cancer.
5.Study of MK promoting breast cancer cell MDA-MB-231 angiogenesis through EPCR/PAR1 pathway in vitro
Qingling WANG ; Dongmei LIU ; Zhengjie HAN ; Yongping WU
Chinese Journal of Clinical and Experimental Pathology 2015;(9):961-965
Purpose To observe the effects of midkine ( MK) on human breast cancer cell line MDA-MB-231 angiogenesis in vitro, and to explore its mechanism. Method shRNA interference was performed to silence the expression of MK in MDA-MB-231 cells, and Western blot was used to identify the expression of MK and EPCR. After MK and EPCR knockdown, or treated with anti protease-activated receptor 1 (PAR1) antibody, the culture medium of MDA-MB-231 cells were collected and the conditioned medium were pre-pared. Human umbilical vein endothelial cells ( HUVECs) were cultured with conditioned medium, and the endothelial cells prolifera-tion was detected by CCK-8 assay, cell migration was detected by transwell method, vasculogenic activity was assessed by Matrigel-based tube formation assay. Results After knockdown of MK, the protein level of EPCR was decreased in MDA-MB-231 cells. Com-pared with control, knockdown of MK and EPCR decreased the proliferation, migration and angiogenesis ability of HUVECs significant-ly (P<0. 05), and the effect of EPCR knockdown group was stronger than MK knockdown group (P<0. 05). After treated with anti-PAR1 antibody, the proliferation, migration and angiogenesis ability of HUVECs were decreased compared with control and EPCR knockdown group (P<0. 05). Conclusion MK promotes human breast cancer cell line MDA-MB-231 angiogenesis through EPCR /PAR1 signaling pathway in vitro.
6.Expression of p27 in Rectal Carcinoma and Modulatory Relation Between Transforming Growth Factor-?1 and p27
Changqing JING ; Leping LI ; Zhao LI ; Taihuang WU ; Qingling MU ;
Chinese Journal of Bases and Clinics in General Surgery 2004;0(01):-
Objective To detect p27 expression in rectal carcinoma and serum transforming growth factor ? 1 (TGF ? 1) level in these patients, and to elucidate the modulatory effect of serum TGF ?1 on p27 expression in rectal carcinoma. Methods Expression of p27 was measured in 37 cases of rectal carcinoma, 22 of rectal adenoma and 19 of normal control specimens by immunohistochemical staining using antibodies to p27. Serum level of TGF ?1 was measured in these patients by enzyme linked immunosorbent assay (ELISA) method. Results p27 protein was expressed in normal rectal tissue, rectal adenoma and rectal carcinoma, and the positive rate was 89.47%, 90.91% and 64.87%, respectively. The positive rate of p27 in rectal carcinoma was significantly lower than that of normal rectal tissue and rectal adenoma ( P =0.025). p27 was mainly located in nucleolus of normal rectal tissue and rectal adenoma, and the positive rate of p27 in cytoplasm of rectal carcinoma was higher than that of normal and rectal adenoma. The positives rates of serum TGF ?1 in normal group, rectal adenoma group and rectal carcinoma group were 21.05%, 27.27% and 51.35%( P =0.045),respectively. The expression of p27 related to histological differentiation, lymph node metastasis and infiltration depth. Serum level of TGF ?1 related to lymph node metastasis, infiltrated depth and CEA level. The positive rate of p27 in TGF ?1 negative group and positive group was 88.89% and 42.11%(Mantel Haenszel ? 2=6.755, P =0.009), respectively. Conclusion TGF ?1 may be useful in assessment of malignance and prognosis of rectal carcinoma. TGF ?1 can down regulate p27 expression in rectal carcinoma.
7.Proteomics analysis of the apoptotic effects of hepatitis B virus large envelope protein on HepG2 cells
Dali ZHENG ; Qingling HUANG ; Yunli WU ; Jingyin LIN ; Xu LIN
Chinese Journal of Microbiology and Immunology 2010;30(12):1085-1091
Objective To assess the effects of hepatitis B virus(HBV) large envelope protein (LHB) on the apoptosis of HepG2 cells and explore the possible mechanism by proteomic approaches.Methods LHB gene was cloned into pShuttle-IRES-hrGFP-1,and the recombinant adenovirus either barboring LHB(Ad-LHB) or empty vector(Ad-GFP) were separately generated.Annexin V-FITC apoptosis detection kit,JC-1 mitochondrial membrane potential assay kit and propidium iodide(PI) staining kit were employed combined with flow cytometry to detect the apoptotic cells infected with Ad-LHB or control of Ad-GFP.The cellular proteins were collected after infection of HepG2 cells by Ad-LHBs or Ad-GFP,and a total of 600 μg proteins were submitted to two-dimensional gel electrophoresis(2-DE) and stained with R350.The gel images were captured by ImageScanner Ⅱ Imaging System,the differentially expressed proteins were identified by ImageMaster 2D Platinum analysis software and picked up by Ettan Spot Picker.After enzyme digestion,the protein samples were analyzed by MALDI-TOF-TOF MS.Results HepG2 cells infected with Ad-LHB were much more prone to apoptosis.There were thirty nine differentially expressed proteins were determined by 2-DE between HepG2 cells infected with Ad-LHB and Ad-GFP,and they were identified ultimately and categorized into thirty three kinds of proteins by MALDI-TOF-TOF MS.Among these proteins,nine were found to be closely related to cell apoptosis,in which CAPN2,eIF3K and PPP2CB were higher expressed in Ad-LHB infected HepG2 cells,and SERPINH1,LASP1,PRDX1,DHRS2,LDHA and PS-MA4 were lower expressed in Ad-LHB infected HepG2 cells.Conclusion LHB could induce apoptosis of HepG2 cells,and several apoptosis-related proteins participated in this process.
8.The evaluation of Survivin protein expression for the prognosis of postoperative patients with hepatocellular carcinoma
Hong CHANG ; Taihuang WU ; Qingling MU ; Leping LI ; Xiaohua PAN ; Changqing JING
Chinese Journal of General Surgery 2000;0(12):-
Objective To evaluate the correlation between the expression of Survivin、VEGF and MMP 2 and the prognosis of patients with hepatocellular carcinoma Methods Expressions of Survivin、 VEGF and MMP 2 were detected by immunohistochemical S P method in 67 postoperative HCC cases Twelve candidate factors associated with long term survival were analyzed by Kaplan Meier Log rank estimation A multivariative Survival analysis of these individual variables was undertaken using the cumulative survival rate by the COX proportional hazards model Results A multivariative analysis determined three independent significant factors influencing overall cumulative survival including Survivin, VEGF, liver function (Child grade) The three prognostic factors predicted an increased risk of death from HCC Conclusion Survivin, VEGF, liver function, particularly the Child Pugh classification are the most significant prognostic factors for HCC patients undergoing hepatic resection
9.IGF-Ⅱ inhibits adriamycin-induced apoptosis of HepG2
Ming LIU ; Hong CHANG ; Guojun WU ; Qingling MU ; Jian XU ; Chengkun QIN
Chinese Journal of General Surgery 2008;23(4):282-284
Objective To investigate the effects of IGF-Ⅱ on adriamycin-induced apoptosis of hepatocellular carcinoma cell line(HepG2).Methods Cultured HepG2 cells were divided into:①group A:control group;②group B:ADM group(200 ng/ml);③group C:2 ng/ml IGF-Ⅱ+200 ns/ml ADM group;④group D:20 ng/ml IGF-Ⅱ+200 ng/ml ADM group;⑤group E:200 ng/ml IGF-Ⅱ+200 ng/ml ADM group. After HepG2 were treated for 48 h,MTT colorimetry was performed to determine the cell viability,Flow cytometry was used to detect the cell apoptosis rate,and Weastern-blot was performed to evaluate the expression of survivin. Results The cell viability of group A、B、C、D、E was respectively 0.568±0.025、0.201±0.020、0.232±0.027、0.268±0.013 and 0.304±0.019;The cell apoptosis rate of group A、B、C、D;E was respectively 6.9%±1.3%、35.4%±2.1%、31.2%±2.2%、26.4%±1.7%and 21.7%±1.9%;Survivinβ-actin of group A、B、C、D、E was respectively 0.527±0.039、0.147±0.081、0.311±0.069、0.421±0.033 and 0.469±0.031.The cell viability in IGF-Ⅱ+ADM group was better than ADM group(P<0.01),the cell apoptosis rate in IGF-Ⅱ+ADM group was significantly lower than ADM group(P<0.01),the expression level of survivin in IGF-Ⅱ +ADM group was significantly higher than ADM group(P<0.01). Conclusions IGF-Ⅱ inhibits ADM-induced apoptosis of HepG2 cells,probably by up-regulating cell's survivin expression.
10.Relationship of adiponectin and visfatin with fetus intrauterine growth
Lixin SHANG ; Qingling TANG ; Jing WANG ; Fang ZHANG ; Nan WU ; Shuhe WANG ; Ping LI
Chinese Journal of Obstetrics and Gynecology 2009;44(4):246-248
Objective To explore the correlation between adipocyte factors(adiponectin and visfatin)and fetus intrauterine growth.Methods Enzyme immunoassay was used to measure the adiponeetin and visfatin levels in maternal and umbilical serum from 14 women with fetal growth restriction (FGR group),14 women with maerosomia(macrosomia group)and 14 normal pregnant women(control group).The correlations of cord serum adiponectin and visfatin with matemal serum adiponectin and visfatin were analyzed.Results (1)Serum visfatin levels in FGR mothers[(41.4±5.5)]μg/L were significantly higher than that in control women[(34.7±4.9)μg/L and macrosomia mothers[(37.3±4.4)μg/L;P<0.01,P<0.05].Serum adiponectin levels in maerosomia mothers [(4.1±1.3)mg/L]were significantly lower than that in control women [(6.6±1.5)mg/L]and FGR mothers[(6.4±1.3)mg/L;P<0.01].(2)Serum visfatinin levels in FGR babies[(58.1±7.6)μg/L] were significantly increased than that in control newborns[(42.6±7.8)μg/L]and macrosomia babies[(48.5±9.1)μg/L;P<0.01,P<0.05].Serum adiponeetin levels in macrosomia babies[(6.5±1.3)mg/L]were significantly decreased than that in control newborns[(7.7±1.5)mg/L]and FGR babies[(7.7±1.0)mg/L;P<0.05,P<0.05].(3)Maternal serum visfatin levels were positively correlated with umbilical serum visfatin levels(r=0.720.P<0.01).Umbilical serum adiponectin levels were higher than that in maternal serum,but there were no relationship between them(r=0.301,P>0.05).Conclusion The changes of visfatin and adiponeetin levels may be related to the oceurrenee of FGR and fetal macrosomia.