3.Effects of HMG-CoA Reductase Inhibitors on Proliferation and Growth of Gastric Cancer Cells in Vitro
Xiao-Yun DING ; Ding-Guo LI ; Han-Ming LU ; Qin-Fang XU
Chinese Journal of Cancer 2001;20(5):474-476
Objective: This study was designed to investigate the effects of HMG-CoA reductase inhibitors on proliferation and growth of gastric cancer cells. Methods: MTT colorimetric assay and flow cytometry were used to measure the proliferative activity and the cell cycle progression of gastric cancer MNK450 cells. Results: Gastric cancer cells treated with Simvastatin had reductions in proliferative activity, S-phase fraction and proliferative index in a dose dependent manner with statistical significance at the concentrations of more than 1 μ mol/L as compared with normal control. The inhibitory rate ranged from 8.0% to 51.5 % when cells were treated with Simvastatin at the concentrations of 0.2 -100 μ mol/L for 72 hours. Inhibition induced by Simvastatin was reversed by simultaneous addition of mevalonic acid at the concentation of 10 mmol/L. Gastric cancer cells were arrested by Simvastafin in G1 at the concentations of 1-20 μ mol/L. The fraction of S-phase cells was reduced from 32.1% to 15.0% after exposure to 20 μ mol/L Simvastatin for 36 hours. The addition of 10 mmol/L mevalonic acid overcome G1 arrest induced by 5 μ mol/L Simvastatin. Otherwise, the percentage of cells in G2 plus M2 remained rather constant throughout the treatment. Meanwhile, Pravastatin had no significant effect on the proliferatiive activity of gastric cancer MNK450 cells. Conclusion: Simvastatin could inhibit the proliferation and growth of gastric cancer cells, which suggests that Simvastatin has the effect of antitumor.
4.Proteins structure changes of COL4A5 gene point mutations and structure-phenotype relations in Alport syndrome.
Yun-Feng WANG ; Jie DING ; Fang WANG ; Ding-Fang BU
Chinese Journal of Pediatrics 2004;42(8):589-594
OBJECTIVEAlport syndrome (AS) is a progressive hereditary nephritis presented with hematuria and renal failure, frequently associated with sensorineural deafness and ocular lesions. So far, more than 300 gene mutations in AS have been identified which provides a better way to analyze the association between genotype and phenotype. It is hard to understand all the phenotype according to the gene mutations, because the structure and function changes of the relevant protein, alpha5(IV) chain, encoded by mutated COL4A5 gene are rare to know. This study aimed to detect the proteins structure encoded by COL4A5 gene with different missense mutations and to analyze the effect of gene mutations on the secondary structure of alpha5(IV) chain and structure-phenotype relations.
METHODSTwo X-linked AS patients with different missense mutations (g.3246G > T resulting in p.G1015V and g.3290G > A resulting in p.G1030S, respectively) diagnosed by clinical manifestations, family history and skin or renal biopsy examinations, as well as a control were included in this study. The fragments of cDNA with the two mutations, respectively, and that of corresponding cDNA from the control were expressed in E. coli. The secondary structure of the recombinant polypeptides were analyzed by using circular dichroism (CD) spectroscopy.
RESULTSCD spectra of the control exhibited a negative peak near 200 nm whereas that of the patient 1 exhibited a negative peak near 220 nm. Furthermore, the magnitude of the negative peak of patient 1 decreased from -9000 deg x cm2 x dmol(-1) to -4000 deg x cm2 x dmol(-1) as compared with that of the control. CD spectra of the patient 2 were slightly changed with the negative peak remaining near 220 nm but the magnitude increasing from -9000 deg x cm2 x dmol(-1) to -11000 deg x cm2 x dmol(-1) as compared with that of the control. In addition, the secondary structure of the control polypeptide was mainly composed of beta-sheet and random coil without alpha-helix, whereas that of the patient 1 presented 12.9% alpha-helix. Although the secondary structure of polypeptide of the patient 2 was also mainly composed of beta-sheet and random coil, the composition of beta-sheet reduced and random coil increased.
CONCLUSIONAlthough the glycine substitutions existed in the same domain of alpha5(IV) chain, the patient 1 with the severe AS phenotype and g.3246G > T mutation, and patient 2 with the mild AS phenotype and g.3290G > A mutation were revealed with different secondary structures of alpha5(IV) chain. Moreover, the secondary structure changes of alpha5(IV) chain were consistent with their corresponding phenotype severity.
Collagen Type IV ; genetics ; Humans ; Mutation, Missense ; Nephritis, Hereditary ; genetics ; Phenotype ; Point Mutation ; Protein Structure, Secondary ; genetics ; Spectrum Analysis
6.Measurement of the mRNA level of Polo-like kinase 1 in bronchoscopic bioptic specimens by realtime quantitative PCR
Ke-Jing YING ; Fang-Chun SHAO ; Bi-Yun YU ; Jin-Min WU ; Jia-Yi DING ;
Chinese Journal of Laboratory Medicine 2003;0(09):-
0.05)and the TNM staging (P=0.55).A mild elevated compared other pathological classification was found in small cell lung cancer (0.191?0.275).Conclusions The results showed that RFQ-PCR was suitable for measurement of the mRNA level of PLKI in bronchoscopic bioptic specimens.This study suggest elevated expression of PLK1 might play a important role in development of lung cancer,so that PLK1 might be a potential tumor marker for Lung cancers.Advanced studies will be needed to clarify that PLKI mRNA level do not relate to TNM staging and pathological classification.
7.Spatial distribution of wall shear stress in the common carotid artery of type 2 diabetes mellitus patients
Zhifang HUANG ; Ming CHEN ; Yuhui ZHANG ; Yun DONG ; Ying WEI ; Fang DING ; Qiuyun LIU
Chinese Journal of Ultrasonography 2014;23(5):393-397
Objective To investigate the wall shear stress(WSS) in the common carotid artery of type 2 diabetes mellitus(DM) patients,and analyzed the spatial distribution of WSS by using quantitative visualization of blood flow shear stress analysis software.Methods Eighteen male type 2 DM subjects were enrolled as DM group and 18 age-matched healthy subjects were selected as control group.None of the participants was hypertensive,hyperlipidemic or a cigarette smoker.Intimal-medial thickness (IMT),number and size of plaques in the common carotid artery were evaluated by high-resolution echo-Doppler.Color Doppler flow images of common carotid arteries in the two groups were extracted from DICOM files.WSS in the common carotid arteries was calculated by shear stress visualization quantitative analysis software,and the corresponding spatial distribution maps of WSS were designed.Results WSS of the common carotid arteries in the control group were ranged from 4 to 14 dyne/cm2.WSS of the common carotid arteries in the DM group were ranged from 2 to 8 dyne/cm2.Compared to mean WSS value [(6.96 ± 1.17)dyne/cm2] of common carotid arteries in the control group,mean WSS value [(3.14 ± 0.79)dyne/cm2] of common carotid arteries in the DM group was significantly lower (t =9.380,P =0.000).Six diabetic participants had a plaque in one carotid artery and no lesions in the contralateral carotid.Among these subjects,mean WSS was significantly lower in the side with lesion (t =7.324,P =0.000).Therefore,IMT of common carotid arteries in participants was significantly inversely related to WSS (r =-0.76,P <0.01).Conclusions The common carotid arteries of DM patients are more prone to atherosclerosis which is associated with reduction of WSS.The hemodynamic profile might represent an additional factor contributing to the increased prevalence and severity of carotid atherosclerosis in diabetic patients compared with general population.
9.Clinical trial on treatment of acute cerebral infarction with TCM treatment according to syndrome differentiation combining Western medicine by staging.
Ding-Fang CAI ; Yun-Ke YANG ; Xi-Xi GU
Chinese Journal of Integrated Traditional and Western Medicine 2007;27(9):789-792
OBJECTIVETo observe the curative effect of integrated Chinese and Western medicine on acute cerebral infarction (ACI).
METHODSTwo hundred and seventy-nine ACI patients were assigned to two groups. The control group (140 cases) was treated with Western medicine by staging and the treated group (139 cases) was given TCM therapy according to syndrome differentiation on the basis of Western medicine. The end point was set at the 90th day of the administration. The curative effect was assessed by Chinese stroke scale (CSS), National Institutes of health stroke scale score (NIHSS), Rankin scale and Barthel index (BI).
RESULTSAt the end point of the trial, the total effective rate was 73.38% in the treated group and 61.43% in the control group, the former was superior to the latter (P<0.05). There were 66 cases (47.14%) in the control group and 80 cases (57.55%) in the treated group with improvement rate of NIHSS > or =40%, 65 cases (46.43%) with their Rankin scale within 0-2 grade in the control group and 78 cases (56.12%) in the treated group, 60 cases (42.86%) in the control group and 71 cases (51.08%) in the treated group with BI > or =85, 61 cases (43.57%) in the control group and 72 cases (51.80%) in the treated group with improvement rate of CSS > or = 46%, comparison between them showed significant differences (P < 0.05).
CONCLUSIONTCM therapy accord-ing to syndrome differentiation combined with Western medicine by staging shows better curative efficacy on ACI.
Acute Disease ; Adult ; Aged ; Anticoagulants ; therapeutic use ; Cerebral Infarction ; diagnosis ; drug therapy ; Diagnosis, Differential ; Drug Therapy, Combination ; Drugs, Chinese Herbal ; therapeutic use ; Female ; Humans ; Male ; Medicine, Chinese Traditional ; Middle Aged ; Phytotherapy ; Syndrome ; Treatment Outcome