1.Effect of β-elemene Combined with Radiotherapy on the Expression of KU70 mRNA and Apoptosis of Lung Adenocarcinoma Cell Line A549
Zhuo ZHANG ; Lijuan ZOU ; Yingying TIAN
Chinese Journal of Clinical Oncology 2010;37(4):184-186
Objective: To study the effect of β-elemene combined with radiotherapy on the expression of KU70 mRNA and apoptosis of lung adenocarcinoma cell line A549. Methods: A549 cells were divided into the control group (c), irridation group (IR), β-elemene group (0.1×IC50 and 0.2×IC50 I.e. 10 μg/mL and 20pg/mL β-elemene) and β-elemene combined with irridation group (0.1×IC50 + IR and 0.2×IC50 + IR). IC50 was obtained through MTT method and cell survival rate was analyzed by colony formation test. Cell cycle and apoptosis were detected with flow cytometry. The expression level of KU70 mRNA was detected by RT-PCR. Results: MTT assay showed that IC50 value of A549 cells was 120 μg/mL. The number of cell clones and survival rate of β-elemene groups were declined significantly. The radiosensitivity of A549 cells can be enhanced by β-elemene. The flow cytometry confirmed that the ratio of G_2/M and apoptosis were significantly in-creased under the effect of β-elemene, statistically different from those in the control group (P <0.05). The expression level of KU70 mRNA in β-elemene with radiotherapy group was declined significantly, statistically different from those in the con-trol group (P<0.05). Cendusion: The rediosensitization effect of β-elemene on A549 cells is associated with induction of cell apoptosis and down-regulation of KU70 mRNA expression.
2.Effect of elemene on radiosensitivity of A549 cells and its possible molecular mechanism
Zhuo ZHANG ; Lijuan ZOU ; Yingying TIAN
Chinese Journal of Radiological Medicine and Protection 2011;31(5):519-522
Objective To investigate the effect of elemene on the radiosensitivity of A549 cells and its possible molecular mechanism.Methods The effect of radiosensitivity was detected by colony forming assay.The protein expressions of DNA-PKcs,Bcl-2 and P53 were detected with Western blot.The correlation between the protein expression of DNA-PKcs and Bcl-2,DNA-PKcs and P53 was analyzed.Results Elemene had radiosensitizing effect on A549 cells,with the SERDo and SERDq 1.54 ± 0.20 and 1.43±0.15,respectively for 10 μg/ml elemene,and 1.63 ±0.32 and 1.75 ±0.19,respectively for 20 μg/ml elemene.Compared with irradiation group,the expression of DNA-PKcs was reduced significantly in 10,20 μg/ml elemene combined with radiation group ( t =7.52,8.33,P < 0.05 ),so was for Bcl-2(t =10.74,11.33,P <0.05).The expression of P53 protein increased significantly (t =-9.25,7.66,P <0.05).There was a remarkable negative correlation between the expression of DNA-PKcs and P53(r =-0.569,P <0.05),and a remarkable positive correlation between DNA-PKcs and Bcl-2 (r =0.755,P <0.05 ).Conclusions Elemene has radiosensitizing effect on A549 cells,which might be related to down-regulation of DNA-PKcs gene expression,up-regulation of P53 and down-regulation of Bcl-2.
3.Screening radiosensitizing-related genes mediated by element in lung adenocarcinoma A549 cells by using gene chip
Zhuo ZHANG ; Lijuan ZOU ; Yingying TIAN
Chinese Journal of Radiological Medicine and Protection 2011;31(3):264-268
Objective To screen radiosensitizing-related genes mediated by elemene in lung adenocarcinoma A549 cells by using gene chip. Methods MTT test was used to calculate the IC50 of elemene. ①The effect of radiosensitivity was detected by colony forming assay. A549 cells were divided into 2 groups: radiation group and radiation + elemene group. Oligonucleotide chip was used to screen the gene expression changes of A549 cells from these 2 groups. The up-regulated gene Egr-1 and the down-regulated gene CyclinDl were selected to undergo RT-PCR so as to confirm the reliability of the result. Results MTT test showed the elemene inhibited the proliferation of the A549 cells dose-dependently. The IC50 value of elemene on the A549 cells was 120 mg/L. ②10 mg/L elemene had radiosensiting effect on A549 cells.The values of SERDO and SERDq obtained from the survival curve were (1.54±0. 20) and (1.43±0. 15 )respectively. Gene chip screened 122 differentially-expressed genes, including 89 up-regulated genes and33 down-regulated genes. ③These altered genes could be related to cell structure, substance metabolism,cell proliferation, cell differentiation, signal transduction, material transport, DNA repair, apoptosis,immune response and so forth. The RT-PCR results of Egr-1 and Cyclin D1 were consistent with the genenchip analysis.Conclusions The mechanism of elemene enhancing the radiosensitivity of lung adenoearcinoma A549 cells is the result of participation and collaboration of multiple genes. Further study of the newly-discovered differentially-expressed gene helps find out new radiosensitizational targets of elemene.
4.Determination of 9 Residual Organic Solvents in Blonanserin by Headspace Gas Chromatography
Lijuan PEI ; Chen TIAN ; Ranran QI ; Zhuo ZHANG ; Ye WANG
China Pharmacy 2017;28(12):1699-1702
OBJECTIVE:To establish a method for the determination of 9 residual organic solvents in blonanserin as methanol, alcohol,isopropyl alcohol,acetonitrile,dichloromethane,hexane,ethyl acetate,tetrahydrofuran and methylbenzene. METHODS:Headspace gas chromatography was adopted. The determination was performed on DB-624 capillary column using temperature pro-gramming. The inlet temperature was 150 ℃,and flame ionization detector was used with temperature of 250 ℃. High purity nitro-gen was used as carrier gas with flow rate of 2.8 mL/min. The split ratio was 1:1,and headspace sample size was 1 mL. Head-space heating temperature was 90 ℃,and equilibration time was 35 min. RESULTS:The linear ranges of methanol,alcohol,iso-propyl alcohol,acetonitrile,dichloromethane,hexane,ethyl acetate,tetrahydrofuran and methylbenzene were 6-1500 μg/mL(r=0.9998),10-2500 μg/mL(r=0.9999),10-2500 μg/mL(r=0.9998),0.82-205 μg/mL(r=0.9994),1.2-300 μg/mL(r=0.9995), 0.58-145 μg/mL(r=0.9994),10-2500 μg/mL(r=0.9999),1.44-360 μg/mL(r=0.9996),1.78-445 μg/mL(r=0.9995),respec-tively. The limits of quantitation were 17.71,6.02,3.17,7.45,1.53,0.69,0.93,1.01,0.22 μg/mL;the limits of detection were 5.89,1.90,1.05,2.48,0.51,0.23,0.31,0.33,0.07 μg/mL,respectively. RSDs of precision and stability tests were all lower than 3.0%,and isopropanol was found in repeatability test (RSD=2.1%). The average recoveries ranged 96.67%-102.66%(RSD=1.9%,n=9),96.00%-101.83%(RSD=1.9%,n=9),97.17%-101.50%(RSD=1.4%,n=9),96.97%-102.44%(RSD=2.2%,n=9),95.83%-103.33%(RSD=2.5%,n=9),95.83%-100.28%(RSD=1.9%,n=9),98.17%-101.25%(RSD=1.0%,n=9),96.55%-102.30%(RSD=1.9%,n=9),96.30%-102.22%(RSD=1.8%,n=9),respectively. CONCLUSIONS:The method is simple,rapid,accurate and suitable for simultaneous determination of 9 residual organic solvents in blonanserin as methanol,alco-hol,isopropyl alcohol,acetonitrile,dichloromethane,hexane,ethyl acetate,tetrahydrofuran and methylbenzene.
5.Clinical analysis of 29 cases of fetal digestive tract malformation diagnosed prenatally by ultrasound
Na ZHUO ; Qing DUAN ; Hui ZHANG ; Jing TIAN ; Tong SUN
Tianjin Medical Journal 2015;(9):1054-1055
Objective To investigate the clinical significance of prenatal ultrasound examination in the diagnosis of fe?tal digestive tract development. Methods Twenty-nine cases of congenital digestive tract malformation were examined in according to the different characteristics of their different fetal ultrasound images. Results There were 11 cases with non-magenblase or less magenblase (37.93%), 4 cases with combination of multiple malformations, and 9 cases with combination of amniotic fluid in the 29 cases. There were 7 cases (24.14%) with dilatation of intestine and intestinal vesicles, in which 3 with multiple malformations and 3 with polyhydramnios. There were 8 cases (27.58%) with double bubbles, in which 1 case with multiple malformations and 7 cases with amniotic fluid. Conclusion The prenatal ultrasound examination in 30 to 32 weeks of pregnancy is very valuable in diagnosis of fetal digestive tract development, which is worthy of clinical application.
6.Investigation and Intervention of Vision-related Quality of Life in Patients with Glaucoma
Rongjiang LUO ; Yehong ZHUO ; Shaorui LIU ; Zhen TIAN ; Yan ZHANG
Chinese Journal of Rehabilitation Theory and Practice 2009;15(3):261-262
Objective To investigate the vision-related quality of life (VRQL) in patients with glaucoma and to explore the intervention. Methods 260 patients with primary open angle glaucoma (POAG) were investigated with questionnaire of VRQL and self-evaluation before and after health education. 100 health adults with similar age and vision was selected as controls. Results There were significant decrease on VRQL and self-evaluation in patients with glaucoma. Systematic health education can significantly improve their VRQL and self-evaluation. Conclusion The in patients with Glaucoma may result in decrease of VRQL, and systemic health education can improve the quality of life.
7.Recent research advances on markers, isolation and purification of mouse hematopoietic stem cells.
Journal of Experimental Hematology 2012;20(1):196-199
Hematopoietic stem cells (HSC), the well-characterized adult stem cells both in the markers and function, show tremendous therapeutic potential. However, the level of hematopoietic stem cells in bone marrow is very low, which makes it difficult to work with. Based on cell surface markers and dye staining, HSC can be isolated from bone marrow and peripheral blood by using magnetic-activated cell separation and fluorescence-activated cell sorting. Over 40 years of research, many surface markers have been identified to purify mouse HSC, and CD34(-)LSK cells are regarded as the mostly used enrichment of HSC in mouse bone marrow. The purpose of this review is to provide insight into the advance of these markers, and isolation and purification of HSC.
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Flow Cytometry
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Hematopoietic Stem Cells
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cytology
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8.Endoscopic esophageal submucosal tunnel resection of gastric fundus-cardiac tumors originating from muscularis propria
Zhifeng ZHAO ; Shuren MA ; Ning ZHANG ; Zhuo YANG ; Zhaojie GONG ; Yanan SUN ; Haoyang TIAN
Chinese Journal of Digestive Endoscopy 2012;29(9):506-509
ObjectiveTo investigate the clinical value of endoscopic esophageal submucosal tunnel resection of gastric fundus-cardiac tumors originating from muscularis propria.Methods Clinical date of 18 patients with gastric fundus-cardiac submucosal tumors originating from muscularis propria who underwent endoscopic esophageal submucosal tunnel resection from January 2011 to December 2011 were retrospectively collected and analyzed.ResultsAll lesions were successfully and completely resected in 18 patients,with sizes ranging from 0.7 cm to 7.2 cm,mean (2.43 ± 1.91 ) cm.Pneumoretroperitoneum,pneumomediastinum and pneumohypoderma occured during the procedure in 2 cases,but spontaneously resolved in 3 days.Fever with increased WBC within 24 h after the procedure occurred in one patient,and was cured in two days with antibiotics.There were no severe complications including bleeding,perforation or death.All patients could have liquid diet 3 days later after the operation.Follow-up endoscopy at 1 week after the operation showed a healing of esophageal incision.ConclusionEndoscopic esophageal submucosal tunnel resection is a safe and effective method for gastric fundus-cardiac submucosal tumors originating from the muscularis propria,lessening the difficulty of traditional endoscopic resection.
9.Radical nephrectomy and thrombectomy for Mayo clinic stage Ⅲ tumor thrombus:a surgical technique and clinical experience
Zhuo LIU ; Lulin MA ; Xiaojun TIAN ; Guoliang WANG ; Xiaofei HOU ; Shudong ZHANG ; Shaohui DENG
Journal of Peking University(Health Sciences) 2017;49(4):597-602
Objective: To evaluate the effectiveness and safety of radical nephrectomy and inferior vena cava thrombectomy in the treatment of patients with Mayo Ⅲ tumor thrombus, and to introduce our experience and surgical technique.Methods: The clinical data of 8 patients with Mayo Ⅲ tumor thrombus from October 2014 to September 2016 were analyzed retrospectively.Of the 8 patients, 3 were male and 5 were female.The average age was (50.8±18.7) years (18 to 77 years).The average body mass index (BMI) was (22.7±4.4) kg/m2 (15.2 to 30.8 kg/m2).Imaging suggested the right renal tumor in all the 8 cases.The average tumor size was (7.9±2.5) cm.Open radical nephrectomy and inferior vena cava thrombectomy was conducted in 5 cases and laparoscopic surgery in 3 cases, and 1 case was converted to open surgery.Results: All the 8 surgeries were completed successfully with no death case.The average surgery time was (370.3±101.6) min, ranging from 272-567 min.The average vena cava blocked time was (41.0±12.1) min, ranging from 17-55 min.The blood loss volume was (1 181.3±915.7) mL, ranging from 200-3 000 mL.During the operation, 5 cases were infused with suspended red blood cells, the amount of blood transfusion was 800-2 000 mL.3 cases were infused of plasma with 400-1 000 mL.The average hospital stay was 9-23 d, with an average of (14.1±4.0) d.In the 8 patients, 4 cases underwent inferior vena cava wall resection because of invasion by tumor thrombus.Preoperative serum creatinine was 60-101 μmol/L, with an average of (76.4±15.3) μmol/L.Serum creatinine 1 week after the operation was 74-127 μmol/L, with an average of (100.8±21.1) μmol/L.Pathological diagnosis showed 6 cases of clear cell carcinoma, 1 case of papillary carcinoma type Ⅱ, and 1 case of Ewing''s sarcoma.Among the 8 patients, early postoperative complications occurred in 5 cases.Postoperative complications were graded as level Ⅱ, according to the Clavien classifications.The 8 cases were followed up for 2 to 24 months with an average of 11.3 months.There was 1 patient who suffered from lung metastasis.Conclusion: Our initial clinical results show that radical nephrectomy and inferior vena cava thrombectomy is safe and effective for patients with Mayo Ⅲ tumor thrombus.The wide extension of grade Ⅲ vein tumor thrombus leads to the difficulty of operation technique.Sufficient preoperative preparation, rich operative experience and skills can improve the safety of operation.
10.Protective Roll of Atorvastatin on Cardiac Remodeling Induced by Pressure Overload in Experimental Mice With its Mechanism
Di ZHAO ; Wei WANG ; Xiaojuan ZHANG ; Weixing GUO ; Tian ZHAO ; Hongju JIANG ; Zhuo ZHAO
Chinese Circulation Journal 2015;(11):1090-1095
Objective: To observe the protective roll of atorvastatin on post-operative cardiac remodeling induced by transverse aortic constriction (TAC) in experimental mice with its possible mechanism.
Methods: A total of 48 C57BL/6 mice were randomly divided into 4 groups: Sham group, TAC group, TAC + valsartan group and TAC + atorvastatin group,n=12 in each group. Myocardial hypertrophy model was successfully established at 4 weeks after the operation, and then the animals were further treated by normal saline, valsartan 5mg/kg and atorvastatin 10mg/kg respectively for 8 weeks. Left ventricular anterior wall thickness at diastole (LVAWd), left ventricular posterior wall thickness at diastole (LVPWd), LVEF and left ventricular fractional shortening (FS) were examined by echocardiography, cardiac hypertrophy indexes were calculated. Protein expression of NF-κB was detected by Western blot analysis, cardiac tissue hydroxyproline (Hyp) level was measured by alkaline hydrolysis, serum levels of TNF-α, IL-1β were determined by ELISA, cardiac collagen deposition was identiifed by HE and Masson staining.
Results: Compared with Sham group, TAC group had increased LVAWd, LVPWd, cardiac hypertrophy indexes and increased area of cardiac fibrosis, allP<0.01; elevated protein expressions of NF-κB, Hyp, TNF-α, IL-1β, all P<0.01. Compared with TAC group, TAC + valsartan group and TAC + atorvastatin group presented improved cardiac hypertrophy indexes, decreased LVAWd, LVPWd and decreased area of cardiac ifbrosis, allP<0.01; reduced protein expressions of NF-κB, Hyp, TNF-α, IL-1β, allP<0.01.
Conclusion: Atorvastatin had protective roll on myocardial hypertrophy induced by pressure overload in experimental mice, which might be related to its anti-inlfammatory effect.