1. The study on epithelial-mesenchymal transition induced by X-ray radiation in human colorectal carcinoma SW480 cells and its potential mechanism
Tumor 2014;34(12):1115-1119
Objective: To investigate the effect of X-ray irradiation on epithelial-mesenchymal transition (MET) in human colorectal cancer SW480 cells and its involved potential signaling pathway. Methods: SW480 cells were irradiated with different doses of X-ray (0, 2, 4, 6 and 8 Gy). The migration and invasion abilities of SW480 cells after irradiation were detected by Transwell method. The expression levels of E-cadherin (E-cad), vimentin, K-ras and Smad3 mRNAs and proteins in SW480 cells were detected by real-time fluorogenic quantitative-PCR and Western blotting, respectively. Results: The migration and invasion abilities of SW480 cells after irradiation with X-ray were increased as compared with that of the control cells (no irradiation) (P < 0.05). The mRNA and protein expression levels of E-cad and K-ras were significantly decreased while the expression levels of vimentin mRNA and protein were significantly increased after irradiation as compared with those of the control cells (all P < 0.05). The expression level of Smad3 mRNA and protein in transforming growth factor-p1 (TGF-p1)/Smad3 signaling pathway were also increased (both P < 0.05) after irradiation. Conclusion: The results of this study support that X-ray irradiation can induce EMT of human colorectal cancer SW480 cells, and this effect may be related to the activation of TGF-β1/Smad3 signaling pathway.
2.The minimum local analgesic concentration of epidural bupivacaine with hydromorphone for labour
Lei LIN ; Chun CHEN ; Qibin KE ; Heqing TANG ; Wei FANG ; Jun HOU
Chongqing Medicine 2015;(6):772-774
Objective To investigate the minimum local analgesic concentration(MLAC)of epidural hydromorphone combined with bupivacaine for analgesia during the first stage of labor by establishing clinical model.Methods Sixty four labouring parturi-ents at 3-7 cm cervical dilation who requested epidural analgesia were allocated to one of two groups.After a lumbar epidural cath-eter was placed,study participants received 15 mL bupivacaine(n=30),bupivacaine with hydromorphone 200 μg(n=30).The con-centration of bupivacaine was determined by the response of the previous patient using 0 - 100 mm visual analog pain scores, with≤30 mm within 30 min defined as effective.Results Four women were excluded,leaving 30 patients in each of the two groups for analysis.The MLAC of bupivacaine alone was 0.103%(95% CI :0.094%-0.113%).The addition of hydromorphone at doses of 200 μg resulted in significant reduction(P <0.05)in the MLAC of bupivacaine to 0.044%(95% CI :0.034%-0.053%),the difference was significant (P <0.05).There was no significant difference in Bromage scores,HR,BP,FHR,UC and adverse reac-tion (P >0.05).Conclusion The study showed a significant reduction in the MLAC of bupivacaine by hydromorphone.Hydromor-phone could be safely used in epidural analgesia for labor.
3.Clinical analysis of 56 patients with relapsing polychondritis
Xu-Hua SHI ; Jin-Mei SU ; Zhi-Ke CHEN ; Feng-Chun ZHANG ; Fu-Lin TANG ;
Chinese Journal of General Practitioners 2005;0(10):-
Objective To investigate clinical characteristics of relapsing polychondritis(RP)and to improve early recognition for it.Methods Clinical and laboratory data of 56 patients with RP were analyzed retrospectively.Results Ratio of number of male patients to female ones was 1.2.Age at onset was(46?11)years(ranging from 27 to 71)and average interval between onset and diagnosis was(21? 35)months,(8?6),(16?31)and(29?37)months for patients initial onset with auricle,respiratory tract and joints involved,respectively.Site involved included airway in 40 patients(71.4%),auricle in 32 (57.1%),joints in 32(57.1%),eyes in 27(48.2%),nasal chondritis in 25(44.6%)and inner ear in 13(23.2%).At initial stage of the course,17 patients were misdiagnosed as respiratory infection (30.4%),nine as perichondritis(16.1%),six as pulmonary tuberculosis(10.7%),five as rheumatoid arthritis(8.9%).Seven of 40 patients with airway involvement received metallic stents for their tracheobronchial stenosis.Four patients whose condition never improved after regular therapy all had respiratory involvement.Conclusions Patients of RP with initial onset at non-auricle,non-nasal sites tended to be misdiagnosed.Prevalence of airway involvement was not so low with a poor prognosis in patients of RP.
4.Clinical on molecular basis of atrial fibrosis in patients with atrial fibrillation investigation.
Dan KE ; Chun-xuan XU ; Ya-zhou LIN ; Jian-cheng ZHANG ; Lin CHEN ; Li-fang LIN ; Xi-zhong HU
Chinese Journal of Cardiology 2005;33(5):459-463
OBJECTIVETo determine the molecular mechanisms involved in atrial fibrosis which occurs in patients with atrial fibrillation (AF) and to investigate their effects on the initiation and maintenance of AF.
METHODSThe right atrial tissue samples were taken from 73 patients with rheumatic heart disease who underwent heart valve replacement surgery. 34 patients had no history of AF (sinus rhythm group), 9 patients had paroxysmal AF and 30 patients had persistent AF. The mRNA content of collagen type I, collagen type III, MMP-2, TIMP-1, TIMP-2, TIMP-3 and TIMP-4 was measured by semi-quantitative reverse transcription-polymerase chain reaction (RT-PCR) and normalized to beta-actin or GAPDH.
RESULTSCompared to sinus rhythm group, the mRNA of collagen type I and MMP-2 increased significantly in the persistent AF group (all, P < 0.01), followed by the paroxysmal AF group (all, P < 0.05). The mRNA of collagen type III was slightly higher in both AF groups than in the sinus rhythm group, but the differences were not statistically significant (P > 0.05). The mRNA of TIMP-1, TIMP-2 and TIMP-3 was down-regulated in the persistent AF group (all, P < 0.01, respectively), however, the trends of reduction did not reach statistical significance in the paroxysmal AF group (P > 0.05). The mRNA of TIMP-4 remained compatible in each group. The mRNA of collagen type I was significantly correlated with left atrial dimension (r = 0.336, P = 0.004) and AF duration (r = 0.339, P = 0.003). The mRNA of MMP-2 was significantly correlated with the mRNA of TIMP-2 (r = -0.326, P = 0.006), the mRNA of collagen type I (r = 0.322, P = 0.006), left atrial dimension (r = 0.300, P = 0.011) and AF duration (r = 0.300, P = 0.010).
CONCLUSIONThe increased level of collagen type I associated with selective downregulation of TIMP-2 and upregulation of MMP-2 gene expression in atrium could be one of the molecular mechanisms of atrial fibrosis during atrial fibrillation, which correlates with the initiation and maintenance of AF.
Adolescent ; Adult ; Atrial Fibrillation ; metabolism ; pathology ; Collagen Type I ; genetics ; Female ; Fibrosis ; Humans ; Male ; Matrix Metalloproteinase 2 ; genetics ; Middle Aged ; Myocardium ; pathology ; RNA, Messenger ; analysis ; Tissue Inhibitor of Metalloproteinase-2 ; genetics ; Tissue Inhibitor of Metalloproteinases ; genetics
5.Study on the genotoxicity of exhausts of diesel engine with ethanol-diesel blending fuel.
Ke-ming LIU ; Chun-hua WANG ; Lei ZHOU ; Ming-yue ZHANG ; Chong-lin SONG ; Guo-liang FAN ; Peng LIU
Chinese Journal of Industrial Hygiene and Occupational Diseases 2005;23(1):42-44
OBJECTIVETo study the genotoxicity of components of diesel engine exhausts with ethanol-diesel blending fuel. To provide scientific arguments to find more economical and less polluted fuels.
METHODSAmes test, comet assay and GC-MS technique were used to test the genotoxicity and 16 kinds of PAHs on diesel engine exhausts with different proportions of ethanol (E0, E5, E10, E20).
RESULTSBoth Ames test and comet assay were positive. It shows that diesel engine exhausts can lead to mutation and DNA damage, especially in pure diesel oil. But the content of 16 kinds of PAHs and DNA damage level decreased in exhausts of E5. With the increase of ethanol proportion in diesel oil, the content of 16 kinds of PAHs and DNA damage level increased.
CONCLUSIONCompared with pure diesel oil and high proportion of ethanol fuel, E5 can reduce the genotoxicity and the brake specific exhausts of PAHs.
Air Pollutants ; toxicity ; Air Pollution ; Carbon Monoxide ; Comet Assay ; DNA Damage ; Ethanol ; toxicity ; Gasoline ; toxicity ; Mutagenicity Tests ; Particulate Matter ; Vehicle Emissions ; toxicity
6.Variance of cost-effectiveness when treat different acute myocardial infarction with different pattern.
Ming FANG ; Heng YE ; Hong-Ke ZENG ; Xin LAI ; Chun-Bo CHEN ; Kairan HE ; Wei-Feng ZHAN ; Xiao-Jun LIN ;
Chinese Journal of Emergency Medicine 2006;0(12):-
Objective To investigate the variance of cost-effectiveness when treat acute myocardial infaretion of different severe extents with different pattern.Methods Acute myocardial infarction patients were selected from emergency eommand center of Guangzhou from October 2003 to December 2005.These patients wew assigned by the center to First-Class Hospitals at Grade 3 and First-class Hospital at Grade 2,and were followed up after 6 months after post-discharge.Cost in hospital and mortality in hospital were registered.The health of all patients were quantificated using SF-36.According to the assigned hospitals,the patients were divided into single infarction group and complex infarction group.Cost in hospital,mortality in hospital,short-term quality of life were compared between the them.Results Compared with and First-Class Hospital at Grade 2 (101 cases),the single infarction patients in First-Class Hospitals at Grade 3 had higber costs in hospital (P=0.016),better society function,affection role,mental health and health status (P
7.Kiss-1 gene expression after radiation and its association with proliferation and apoptosis in colorectal cancer cells.
Shao-qin CHEN ; Ming-mei TU ; Qi-bao DAI ; Su-yong LIN ; Chun-lin KE
Chinese Journal of Gastrointestinal Surgery 2012;15(5):508-511
OBJECTIVETo investigate the change of expression level of metastasis suppressor gene Kiss-1 in the colorectal cancer cell line SW480 after radiation, and to determine its association with the proliferation and apoptosis of SW480 cells.
METHODSSW480 cells were divided into control group (0 Gy) and study groups (2, 4, 6, 8 Gy). Cells in the study groups were irradiated by 6-MV X-ray radiation for 48 hours. Immunohistochemistry and real-time PCR methods were used to investigate the influence of radiation on Kiss-1 gene expression of SW480. Colony formation assay was used to detect the proliferation of SW480. Flow cytometry-Annexin- V/PI assay was used to observe the change of the apoptosis rate.
RESULTSCompared with the control group, Kiss-1 protein expression increased after radiation of 6, 8 Gy (P<0.05), but no significant changes were observed after radiation of 2, 4 Gy(P>0.05). Kiss-1 gene mRNA level increased after radiation of 2, 4, 6 Gy, while no obvious change was observed for 8 Gy radiation. The apoptosis rates increased for 4, 6, 8 Gy radiation(P<0.05), however, there was no significant difference for 2 Gy radiation (P<0.05).
CONCLUSIONRadiation may increase Kiss-1 gene expression in SW480 cells, which results in decreases proliferation and increases apoptosis in residual surviving cells.
Apoptosis ; Cell Line, Tumor ; Cell Proliferation ; Colorectal Neoplasms ; metabolism ; pathology ; Humans ; Kisspeptins ; genetics ; metabolism ; radiation effects ; RNA, Messenger ; genetics ; X-Rays
8.A modified cytogenetic study for multiple myeloma.
You-wen QIN ; Dao-lin WEI ; Chun WANG ; Shi-ke YAN ; Yan-rong GAO ; Qi CAI
Chinese Journal of Medical Genetics 2006;23(2):213-215
OBJECTIVETo evaluate the effect of modified culture method used to cytogenetic analysis and the clinically significance of chromosomal abnormalities to multiple myeloma (MM).
METHODSMononuclear cells were isolated from bone marrow aspirate of 20 MM patients; and then cultured for 3 days without any cytokines, and 6 days in the presence of IL-6 (10 ng/mL) and GM-CSF (30 ng/mL) before RHG banding analysis; the remained part of aspirates were treated directly. Eight cases of iron deficiency anemia were taken as control.
RESULTSThe experiment was failure in 2 cases because of blood clot, and another 2 cases could be analyzed only by direct method due to inadequate cells. The karyotype abnormalities were found from 4 cases of 16 available patients. Of them, three cases had complex karyotypes. The abnormalities were detected after 6 days culture with addition of cytokines. No abnormalities were detected from those groups of directly analysis and 3 day culture. Meantime, the clinical data showed that the patients with cytogenetic abnormalities were in stage III, and had a high percentage of MM cells (25%-56%) in their bone marrow, and also poor responses to prior chemotherapy. No cytogenetic abnormalities were found from control individuals in all groups.
CONCLUSIONExtended culture in the presence of cytokines could improve the efficiency of cytogenetic analysis to MM. Complex karyotype was common cytogenetic abnormalities in MM patients with poor response to chemotherapy.
Aged ; Chromosome Aberrations ; Cytogenetic Analysis ; Cytokines ; metabolism ; Female ; Humans ; Karyotyping ; Male ; Middle Aged ; Multiple Myeloma ; genetics ; pathology
9.Detection of residual diphenylmethane diisocyanate monomer in a hemodialysis catheter
Yuan-Li HUANG ; Xue SUN ; Lin-Nan KE ; Chun-Ren WANG
Chinese Journal of Tissue Engineering Research 2018;22(6):927-931
BACKGROUND:In the current quality control file or technical standards of the hemodialysis catheter,the indicators of the component contents and detection methods of the residual diphenylmethane diisocyanate (MDI) monomer are undefined.To ensure the safety and effectiveness of these products,we should try to establish and improve the quality standards.OBJECTIVE:To establish a method for determination of the residual MDI monomer in a hemodialysis catheter by gas chromatography (GC),and to analyze the bio-security of the MDI.METHODS:Samples collected in the hemodialysis catheter were heated to reflux with ethyl acetate and the residual MDI content was analyzed by the GC.The GC separation was performed on a DB-5 MS column (30 mx0.25 mm),the temperature of which rose by program.The initial temperature was 60 ℃,maintained for 5 minutes,rose to 280 ℃ with a rate of 15 ℃/min,and maintained for 6 minutes.The temperature of the Injector and FID detector was both 280 ℃.Carrier gas was 99.999% nitrogen.RESULTS AND CONCLUSION:The linearity was achieved in the range of 4.970-99.40 mg/L (r=0.999 64) for MDI.The mean recovery rate was 100.9% with the relative standard deviation of 3.2% (n=6).The residue of MDI monomer in the three batches of samples was lower than the tolerable exposure.Therefore,it is a sensitive,rapid,accurate,specific method that can be used for the quality control of the residual MDI monomer in the hemodialysis catheter.
10.Simultaneous determination of 18 different amino acids in in vitro fertiIization medium by ultra-high performance liquid chromatography-tandem mass-spectrometry
Yuan-Li HUANG ; Lin-Nan KE ; Chang-Kun LI ; Chun-Ren WANG
Chinese Journal of Tissue Engineering Research 2018;22(18):2915-2920
BACKGROUND: At present, in the quality control file or technical standards of in vitro fertilization medium, indicators for component contents and detection methods have not yet been defined. To ensure the safety and effectiveness of these products, we should try to establish and improve the quality standards. OBJECTIVE: To establish a method for simultaneously testing 18 kinds of amino acids by ultra-high performance liquid chromatography-tandem mass-spectrometry (UHPLC-MSMS), and to analyze the difference in the content of different amino acids in the medium for different uses. METHODS: Using the UHPLC-MSMS, we detected the indicator components of 18 different amino acids in the in vitro fertilization medium. These amino acids included glycine, leucine, methionine, tyrosine, histidine, threonine, alanine, isoleucine, tryptophan, cystine, lysine, aspartic acid, valine, phenylalanine, valine, serine, glutamic acid, arginine. The UHPLC separation was performed on a SUPELCO Discovery HS F5-3 column (15 cm×2.1 mm, 3 μm) in a gradient elute mode with acetonitrile and water (both containing 0.1% formic acid) as the mobile phase at a flow rate of 0.35 mL/min. The column temperature was 40 ℃. MS detection was performed with multiple-reaction monitoring mode using negative electro spray ionization. RESULTS AND CONCLUSION: The linearity was achieved in the range of 0.25-12.5 mmol/L for the 18 different amino acids. The average recovery rate of these amino acids ranged 86.3% to 125.3%. The relative standard deviation of the precision experiment and the repeatability experiment was less than 4.7%. These findings indicate that this is a sensitive, rapid, accurate, and specific method that can be used for the quality control of in vitro fertilization medium.