1.Experimental study on the killing effects of oxymatrine on human colon cancer cell line SW1116
Jian ZOU ; Zhihua RAN ; Qi XU
Chinese Journal of Digestion 2001;0(04):-
Objective To evaluate the killing effects of oxymatrine(OM) on human colon cancer cell line SW1116 and the mechanism of its anti-neoplastic effect. Methods Methyl thiazolyl tetrazolium analysis, flow cytometry , polymerase chain reaction(PCR)-enzyme linked immunosorbent assay (ELISA) and RT-PCR were used to detect the killing effects of OM and its influence on cell cycle dis- tribution , telomerase activity, expressions of human telomerase reverse transcriptase (hTERT), c-myc ,p53 and mad1 genes in SW1116 cells. Results OM exhibited dose-dependent killing effects on SW1116 cells and induced the increase of G1/G0-phase cells and decrease of S-phase cells. It was found that OM could supress the telomerase activity of SW1116 cells, and the effects were dose- and time-dependent. After OM administration, the expression of hTERT gene in SW1116 cells was decreased, those of p53 and mad1 genes were increased, and the expression of c-myc gene had no marked changes. Conclusions OM has dose-dependent killing effects on SW1116 cells.The anti-neoplastic activity of OM might be due to the inhibition of telomerase activity by means of its influence on hTERT and the up-stream regulation genes.
2.The value of combined detection of NT-proBNP,CA-125 and Hs-CRP for the diagnosis of chronic heart failure
Guizhong ZHU ; Li SUN ; Zhihua QI
Chongqing Medicine 2014;(24):3195-3198
Objective To investigate the changes of serum N-terminal proBNP (NT-proBNP) ,carbohydrate antigen 125(CA-125) and high sensitive CRP(Hs-CRP) levels in patients with chronic heart failure(CHF) ,to evaluate the severity of CHF using the three markers alone or in combination .Methods A total of 62 patients with CHF(CHF group) from our hospital were select-ed .Among them ,13 cases were in NYHA class Ⅱ ,27 cases were in class Ⅲ and 22 cases were in class Ⅳ .The control group in-cluded 54 patients without CHF .Serum levels of NT-proBNP ,CA-125 and Hs-CRP were detected in CHF group and control group . The left ventricular ejection fraction(LVEF) were detected by echocardiography .Results The serum levels of NT-proBNP ,CA-125 and Hs-CRP of CHF group before treatment were significantly higher than those of control group (P<0 .01) ,and the serum levels increased with the degree of severity of heart failure .There were statistically significant difference among patients in NYHA classⅡ ,class Ⅲ and class Ⅳ (P< 0 .01) .There was a high correlation between NT-proBNP ,CA-125 and LVEF (NT-proBNP vs . LVEF ,r=0 .934 ,P<0 .01 ;CA-125 vs .LVEF ,r=0 .878 ,P<0 .01) ,a low correlation between Hs-CRP and LVEF(r=0 .437 ,P<0 .05) .Compared with the single marker detection ,the combined detection of NT-proBNP ,CA-125 and Hs-CRP increased the AUC .Conclusion The combined detection of NT-proBNP ,CA-125 and Hs-CRP can increase the value of the observed severity of heart failure .
3.Effect of telomerase-antisense DNA on apoptosis of hepatoma cells induced by arsenic trioxide
Zhihua DENG ; Ziyan HAN ; Qi WANG ; Shuisheng SHI
Chinese Journal of Cancer Biotherapy 2006;0(06):-
Objective: To observe the effect of telomerase-antisense DNA on apoptosis of hepatoma cells induced by arsenic trioxide, in an effort to look for a new anti-hepatic cancer agent with high efficiency, low cytotoxicity. Methods: We designed and synthesized a 20nt telomerase-antisense DNA targeting telomerase template and observed its influence on the telomerase activity of hepatoma cells. H-E staining, flow cytometry, and DNA agarose electrophoresis were used to study the preventive effect of telomerase-antisense DNA on hepatoma cells apoptosis induced by arsenic trioxide. Flow cytometry was used to detect the expression of Fas, Fas-L, and bcl-2. Results: Telomerase-antisense DNA (5 ?mol/L) effectively inhibited the telomerase activity of hepatoma cells after 24 hours (P
4.Construction of cell line with overexpression of rat serotonin1A receptor
Zhihua LI ; Bojuan WU ; Wanjun SONG ; Yi DANG ; Xiaoyong QI
Chinese Journal of Anesthesiology 2011;31(1):74-77
Objective To establish a cell line with overexpression of rat serotonin1A receptor (5-HT1AR).Methods Human neuroblastoma cells-SH-SY5Y were donated by cancer institute attached to the 4 th Affiliated Hospital, Hebei Medical University. Total RNA was extracted from brain tissues of male SD rats and rat 5-HT1A R was obtained by RT-PCR. Plasmid pc-DNA3. 1/hisC containing the rat 5-HT1AR (pc-DNA3.1/hisC-Rat-5-HT1AR)was constructed and transfected into SH-SY5Y cells. The transfected cells were isolated by G418 selection and SH-SY5Y-Rat-5-HT1A R cells were obtained. Expression of 5-HT1A R was detected by Western blot analysis. Cell viability was evaluated by MTT assay. SH-SY5Y-Rat-5-HT1AR cells were further observed for 5-HT1AR by immuno-fluorescence staining. Results Plasmid pc-DNA3. 1/hisC-Rat-5-HT1AR was successfully constructed by linking Rat-5-HT1A R with pc-DNA3.1/hisC and transfected into SH-SY5Y. The SH-SY5Y-Rat-5-HT1A R cells were more slender than SH-SY5Y cells with less and longer processes. MTT showed that the viability of SH-SY5Y-Rat-5-HT1A R cells was much lower than SH-SY5Y. Rat 5-HT1A R was expressed efficiently on the membrane of SH-SY5Y-Rat-5-HT1A R cells. Conclusion A cell line with overexpress of rat 5-HT1A R is successfully established.
5.Trends of colorectal cancer incidence in Xuhui District of Shanghai (2001-2011)
Qi YANG ; Zhihua GU ; Chunmin LIU ; Haiyan GU ; Hongwei NI
Chinese Journal of General Practitioners 2013;(3):181-184
Objective To investigate the trends of colorectal cancer incidence in Shanghai Xuhui District from 2001 to 2011.Methods Total 3042 cases of colon cancer and 1958 cases of rectal cancer were collected from the Shanghai Cancer Reporting System.The results were age-adjusted according the census data of 2000 as the standardized incidence.Results The standardized incidence rate of colorectal cancer in Xuhui District rose from the 21.83/100 000 in 2001 to 27.35/100 000 in 2006 and increased by 25.29% ; from 2009 to 2011 the incidence rate was stabilized.The ratio of colon to rectal cancer was 1.55:1,and the incidence of colon cancer was significantly higher than rectal cancer.The incidence rate of male and feinale colorectal cancer increased with age,especially at > 45 y group accounting for 98.70% and 98.37% of total incidence rate respectively.The ratio of male to female in colon cancer incidence rate was 0.98∶ 1,while that of rectal cancer was1.25∶ 1.Conclusion The incidence of colorectal cancer in Xuhui District from the last 11 years is significantly higher than of national level.More effective preventive measures should be taken.
6.Determination the Equilibrium Solubility and Apparent Oil/Water Partition Coefficient of Phloridzin by RP-HPLC
Zhihua QI ; Shuyun GUO ; Xiuling TANG ; Xiaoye LI ; Qingwei WANG
Herald of Medicine 2015;(10):1329-1331,1332
Objective To establish a reversed-phase high performance liquid chromatograph ( RP-HPLC ) method for determination of equilibrium solubility and oil/water partition coeficient of phloridzin in different solvents. Methods A RP-HPLC method was established to detect the concentration of phloridzin in water and different organic solvents. The partition coefficients in the n-octanol-water/buffer solution systems of phloridzin were determined by shaking flask method. The Inertsil ODS-3 (4.6 mm×150 mm, 5μm) column was used and the detection wavelength was 284 nm.The flow rate was 1.0 mL??min-1, and acetonitrile-0.05% phosphoric acid(30??70)was used as mobile phase. Results The equilibrium solubility of phloridzin was 2.07 mg??mL-1 in water and 838.63 mg??mL-1 in methanol at 25 ℃.A good linear relationship of phloridzin was obtained within the range of 0.054 9-1.098 0 μg.The regression equation was Y=2 152.9X+7.26 (r=0.999 9).The solubility values of phloridzin were higher in ethanol and propylene glycol than in other solvents. Conclusion RP-HPLC method is simple, quick and accurate for the determination of phloridzin.Phloridzin was almost insoluble in petroleum ether and poorly soluble in water.The equilibrium solubility is higher in methanol than in other solvents. The apparent distribution coefficient of phloridzin varies significantly with pH under the alkaline conditions but less in the acidic solution.
7.Calibrating the working standard for rhG-CSF
Qian JIA ; Xingjun ZHOU ; Xuehong TIAN ; Yuling FAN ; Xiaobo QI ; Zhihua ZHANG
Chinese Journal of Biochemical Pharmaceutics 2001;22(2):81-84
Purpose Both the working standard and corresponding sample(GRAN75)were first calibrated by WHO international standard for G-CSF.Methods MTT method by NFS-60 cells was used. The results were calculated by (4,4)method.Results Three batchs working standards were prepared,two batchs were freeze-dry and the prescription was same as WHO international standard for G-CSF,one batch was injection and the prescription was same as corresponding sample(GRAN75).The biological potency and the FL% of average potency were 3.062×106 、4.276×106IU/ampoule、1.635×107IU/ml and 5.529%、4.291%、4.244% for working standard, and 1.880×107IU/ml and 5.175% for corresponding sample,respectively.Conclusion The working standard which calibrated could be used as working standard in the measurement of rhG-CSF biological activity.
8.Diagnosis and treatment of infantile congenital subglottic hemangioma
Lan CHENG ; Qi HUANG ; Hao WU ; Jun YANG ; Li CHEN ; Zhihua ZHANG
Journal of Clinical Otorhinolaryngology Head and Neck Surgery 2009;(15):693-696
Objective: To discuss the diagnosis and treatment of infantile congenital subglottic hemangioma. Method:Nine patients with recurrent stridor,feeding difficulties and laryngeal obstruction were diagnosed as con-genital subglottic hemangioma with bronchia endoscopy. The lesions were unilateral in 7 cases and bilateral in 2 ca-ses. Eight cases were treated by intralesional Pingyangmycin injection after tracheotomy. Patients were discharged with metal trachea and were followed up endoscopically 2 weeks thereafter. A second treatment would be required if the tumor reduced in size but not disappeared completely. Plugging tubes and extubation would be done when the tumor disappeared completely. One case with unilateral lesion underwent power-assisted tumor resection with-out tracheotomy. Result: Hemangioma vanished completely in 6 patients with unilateral subglottic hemangioma who were treated with single stage procedure. Tumor size was found reduced significantly after one procedure and van-ished completely after a second procedure in 2 cases with bilateral subglottic hemangioma. All of the 8 cases were extubated successfully. The patient recovered thoroughly in two weeks after power-assisted tumor resection. Con-clusion: Intralesional Pingyangmycin injection after tracheotomy can relief the airway obstruction quickly, shorten the nature course of hemangioma, shorten the period of wearing trachea, has no complication such as subglottic stenosis and is thought to be a safe and effective therapy for large size hemangioma. Power-assisted tumor resec-tion under good total anesthesia without tracheotomy combined with well hemostasis is effective, less injure, and suitable for small size hemangioma.
9.Expression of delta like ligand 4 in pancreatic cancer and its relationship with angiogenesis
Guifang XU ; Weijie ZHANG ; Qi SUN ; Jun YANG ; Zhihua ZHOU ; Haibin ZHAO ; Xiaoping ZOU
Chinese Journal of Pancreatology 2012;12(3):164-166
Objective To detect the expression of delta-like ligand 4 (DLL4) and its relation with the angiogenesis in pancreatic cancer.Methods SP immunohistochemistry method was used to detect the expression of DLL4 in pancreatic cancer and para-cancerous tissue.The microvessel density (MVD) was calculated with CD34 staining for microvessel endothelium and the correlation between expression of DLL4,MVD and clinicopathological parameters of pancreatic carcinoma was investigated.Results The expression of DLL4 in pancreatic cancer was significantly higher than that in para-cancerous normal pancreatic tissue (68.3% vs 20%,P <0.01 ).The over-expression of DLL4 in pancreatic cancer was positively related to the differentiation degree,TNM staging,metastasis and invasion (P <0.05),but not related to the location,tumor size and histological types.The MVD in pancreatic cancer tissues was significantly higher than that in para-cancerous normal tissues (34.9 ± 13.2 vs 18.9 ±2.2,P <0.01).MVD was correlated with the differentiation degree,TNM staging,metastasis and invasion (P <0.05 ),but not with the tumor size and location,histological types.MVD in pancreatic cancer expressing DLL4 was significantly higher than that in pancreatic cancer not expressing DLL4 ( 38.8± 10.7vs 29.0 ± 15.2,P =0.038 ),and the expression of DLL4 and MVD were positively correlated ( r =0.669,P < 0.05 ).Conclusions DLL4 expression may play an important role in pancreatic carcinogenesis by promoting angiogenesis and may be associated with tumor invasion and metastasis.
10.Study on determination method of the rhG-CSF biological activity
Qian JIA ; Xingjun ZHOU ; Xuehong TIAN ; Yuling FAN ; Xiaobo QI ; Zhihua ZHANG
Chinese Journal of Biochemical Pharmaceutics 2001;22(1):25-26
Purpose The determination method of rhG-CSF bio logical activity by NFS-60 cells was studied in accordance with WHO internation al standard for G-CSF.Methods MTT method was adopted.R esults The chomosone number of NFS-60 cell was 39, about 1×105 cells/ml to 1×107 cells/ml,the dying color showed gradient when the NFS-60 c ells was dyed by MTT,the method(4,4) was adopted in the determination of rhG-CS F biological activity,the average FL% of potency was 13.560% for single exprimen t,and the CV of inter-assay and intra-assay was lower than 10% and 10.109%,respectively.Conclusion method(4,4) can be us ed in the determination of rhG-CSF biological activity,and the results can guid e the study and the manufacture of rhG-CSF effectively.