1.Effect of hexavalent chromium-induced ribosomal DNA copy number variation on DNA damage response in various cell lines
WU Fan ; FENG Lingfang ; CHEN Junfei ; JIANG Zhaoqiang ; GONG Xiaoxue ; QIN Yao ; LOU Jianlin
Journal of Preventive Medicine 2023;35(5):374-379
Objective:
To investigate the effect of ribosomal DNA (rDNA) copy number variation caused by hexavalent chromium exposure on DNA damage response in different cell lines, so as to provide insights into the involvement of hexavalent chromium-induced rDNA copy number variation in DNA damage responses.
:
Methods Human lung epithelial BEAS-2B cells and human embryonic lung MRC-5 cells were treated with 2 μmol/L potassium dichromate for 24 hours, and then cells were transferred to fresh media for further incubation, while cells treated with the same volume of phosphate buffer solution served as controls. Cells treated with potassium dichromate for 24 hours, and 3 and 7 days post-detoxification, were harvested, and rDNA copy number was quantified in cells using a quantitative fluorescent real-time PCR assay. Cell cycle, apoptosis and DNA damage were detected using a Muse cell analyzer, and the DNA damage was evaluated with the proportion of ataxia telangiectasia-mutated (ATM) gene activation, proportion of double-strand DNA breaks and the percentage of the H2A.X variant histone phosphorylatio.
:
Results The 45S and 5S rDNA copy numbers of were significantly higher in MRC-5 cells than in BEAS-2B cells [(1.54±0.26) vs. (1.02±0.18), P<0.05; (6.97±1.07) vs. (3.00±0.15), P<0.05]. The 45S rDNA copy number was lower in MRC-5 cells 3 days post-detoxification (0.80±0.04) than in controls (P<0.05), and was higher in BEAS-2B cells 3 days post-detoxification (1.43±0.07) than in controls (P<0.05) . G0/G1 phase arrest was found in MRC-5 cells 24 hours post-treatment, and the apoptotic rates were significantly higher in MRC-5 cells 3 and 7 days post-detoxification than in controls [(11.53±1.53)%, (18.33±0.70)% vs. (3.53±0.93)%, P<0.05]. The overall apoptotic rates 24 hours post-treatment and 3 days post-detoxification [(2.80±0.17)%, (3.33±0.57)% vs. (1.53±0.61)%, P<0.05], proportion of ATM gene activation 3 days post-detoxification [(3.37±0.67%) vs. (1.18±0.22)%, P<0.05], proportion of double-strand DNA breaks 3 days post-detoxification [(4.45±0.85)% vs. (0.97±0.21)%, P<0.05] and percentage of the H2A.X variant histone phosphorylation 3 days post-detoxification [(1.68±0.56)% vs. (0.29±0.06)%, P<0.05] in BEAS-2B cells were higher than in controls.
Conclusions
Hexavalent chromium-induced rDNA copy number variation affects DNA damage response in different cell lines. A stronger DNA damage response is found in BEAS-2B cells with a low rDNA copy number, and a relative stable response is observed in MRC-5 cells with a high rDNA copy number.
2.Effect of exposure to chrysotile on ribosomal DNA copy number variation and DNA damage response
Jiaqi LIU ; Lingfang FENG ; Junfei CHEN ; Hailing XIA ; Zhaoqiang JIANG ; Fan WU ; Xiaoxue GONG ; Jianlin LOU
Journal of Preventive Medicine 2022;34(6):547-554
Objective:
To investigate the effect of chrysotile exposure on ribosomal DNA (rDNA) copy number and DNA damage response, so as to provide insights into the mechanism of asbestos-induced carcinogenesis.
Methods:
Human pleural mesothelial MeT-5A cells were treated with chrysotile suspensions at doses of 1.25, 2.5 and 5 μg/cm2 (low-, medium-, high-dose group), while PBS served as controls. MeT-5A cells were harvested 6, 24, 48 and 72 h post-treatment, and the rDNA copy numbers and the BIRC5, HRAS, GINS4 and RRM2 mRNA expression were determined using a quantitative real-time PCR (qPCR) assay. The apoptosis of MeT-5A cells and DNA damage were detected using Muse cell analyzer. The rDNA copy numbers, DNA damage responses and BIRC5, HRAS, GINS4 and RRM2 mRNA expression were compared in MeT-5A cells treated with different doses of chrysotile suspensions.
Results:
There were significant differences in 45S rDNA copy numbers among low-, medium-, high-dose groups and the control groups 6, 48 and 72 h post-treatment with chrysotile suspensions, and significantly lower 45S rDNA copy numbers were measured in low-, medium- and high-dose groups than in the control group 6 h post-treatment, while significantly higher 45S rDNA copy numbers were found in the high-dose group than in low- and medium-dose groups 48 and 72 h post-treatment (all P<0.05). There were significant differences in 5S rDNA copy numbers among low-, medium-, high-dose groups and the control groups 24, 48 and 72 h post-treatment with chrysotile suspensions, and significantly lower 5S rDNA copy numbers were measured in medium- and high-dose groups than in the control group 24 and 48 h post-treatment, while significantly lower 5S rDNA copy numbers were found in medium- and high-dose groups than in the low-dose group 24, 72 h post-treatment (all P<0.05). There were significant differences in the overall apoptotic rate of MeT-5A cells among groups at different time points, and the overall apoptotic rate of MeT-5A cells were significantly higher in medium- and high-dose groups than in the control group (all P<0.05), with late-stage apoptosis predominantly detected. There were significant differences in the rates of ATM activation and DNA double-strand break in MeT-5A cells among groups 72 h post-treatment, and higher rates of ATM activation and DNA double-strand break were measured in medium- and high-dose groups than in the control group (all P<0.05). In addition, there were significant differences in the relative mRNA expression of BIRC5, HRAS, GINS4 and RRM2 genes among groups 24 and 48 h post-treatment, and significantly lower BIRC5, HRAS, GINS4 and RRM2 mRNA expression was quantified in medium- and high-dose groups than in the control group (all P<0.05).
Conclusion
Exposure to chrysotile may induce rDNA copy number variations and altered expression of nucleolar proteins in human pleural mesothelial cells, which may be involved in the regulation of DNA damage responses.
3.MSCT appearances of lung lacerating inj ury
Junfei FAN ; Minling WANG ; Youxue XU ; Jiawen QUAN ; Qiancheng SHEN ; Rongbiao LI ; Baoting HUANG
Journal of Practical Radiology 2016;32(12):1861-1863,1875
Objective To explore MSCT appearances of lung lacerating injury.Methods The MSCT findings of lung lacerating injury in 31 patients were analyzed retrospective.Results The lung lacerating injury of the 31 cases with 67 lesions in total was found,18 of whom were located on the back side of lung near the pleura,11 of whom had solitary lesion and 20 of whom had multiple ones. The MSCT findings included lung cavity in 9 eases,liquid airbag cavity in 1 7 and lung hematoma in 5 .The pulmonary contusion with different degrees was found in all 3 1 cases.Dynamic observation showed the cavities and hematoma could be transformed into each other.Conclusion MSCT is the best method for diagnosis and observation of lung lacerating injury and helpful for the guide of clinical treatment.
4.Cerebral blood stream status observation of cervical dizziness treatmen with traction and heating acupuncture
Peiwu FAN ; Junfei WANG ; Xufang PAN ; Ming LIU ; Zuo WANG ; Ting YAO
Chinese Journal of Primary Medicine and Pharmacy 2006;0(12):-
Objective To explore the improve mechanism of the cerebral blood stream state in patients with cervical dizziness(CD) after treatment combined with traction and heating acupuncture.Methods The results of transcranial Doppler(TCD) were compared in 53 patients with CD being treated with traction and heating acupuncture.Results The results showed that in 53 CD cases after treatment the velocity of the blood flow of double middle cerebral arteries,posterior cerebral arteries and right anterior cerebral artery were decreased remarkablely,besides this left anterior cerebral artery.The TCD examined results got near to normal value of contrast group(P
5.MSCTfindingsofextrarenalappendageinvolvementwithchronicurinarytractobstruction
Youxue XU ; Junfei FAN ; Minling WANG
Journal of Practical Radiology 2019;35(4):598-601
Objective Toinvestigatetheincidenceofchronicrenalobstructiveexternalappendageinvolvementandtheresultsof MSCTdetection.Methods MSCTdataof120patientswithchronicurinarytractobstructionwereretrospectivelyanalyzed.Results Among120patients,75caseswerefoundwiththeextrarenalappendageinjury(62.5%).Theinjuryoftheextrarenalappendageincluded thickeningofrenalcapsule(8.00%),effusionofrenalsubcapsular(14.67%),thickeningofbridgingseptaoftheperinephricspace (100.00%),lamellarshadowoffatlayerintheperirenalspace(36.00%),thickeningofrenalfascia(96.00%),pararenalspaceeffusion (88.00%),thickeningperitoneal(76.00%),pseudocyst(1.33%),lamellarshadowoffatlayerinextraperitoneal(22.67%)andnapes(6.67%), increasedintraperitonealfatdensity(32.00%)andperitonealeffusion(4.00%).Conclusion MSCTcanclearlydemonstrateextentand rangeoftheperinephricspaceandpararenalspaceinvolvementinchronicurinarytractobstruction.CTscanplayanimportantrolein chronicurinarytractobstruction.
6.Ribosomal DNA copy number variation in peripheral blood and its influencing factors among patients with pneumoconiosis
GONG Xiaoxue ; FENG Lingfang ; CHEN Junfei ; FU Hao ; JIANG Zhaoqiang ; LIU Shuang ; DONG Xiaowen ; WU Fan ; LOU Jianlin
Journal of Preventive Medicine 2024;36(2):101-104
Objective:
To explore the changes in ribosomal DNA copy number in peripheral blood among patients with pneumoconiosis and its influencing factors, so as to provide insights into prevention and treatment of pneumoconiosis.
Methods:
Eighty-eight patients with pneumoconiosis who visited a designated hospital and 71 community residents with no history of pneumoconiosis or dust exposure were selected as the pneumoconiosis group and control group, and age, smoking history, drinking history and cumulative years of exposure to dust were collected through questionnaire surveys. The copy number of 45S rDNA and 5S rDNA was detected using real-time fluorescence quantitative PCR, and the differences between the two groups were compared. Factors affecting the copy number of 45S rDNA and 5S rDNA were identified by a multiple linear regression model.
Results:
The pneumoconiosis group had a median age of 56.00 (interquartile range, 15.25) and a mean cumulative dust exposure duration of (12.40±8.08) years, with 56.82% smoking and 62.50% drinking. The control group had a median age of 64.00 (interquartile range, 37.00) years, with 32.39% smoking and 26.76% drinking. The median copy number of 45S rDNA in the pneumoconiosis group was 1.29 (interquartile range, 0.59), which was lower than 2.10 (interquartile range, 1.88) in the control group; the median copy number of 5S rDNA in the pneumoconiosis group was 5.33 (interquartile range, 0.85), which was higher than 4.66 (1.34) in the control group (both P<0.05). Multiple linear regression analysis identified age (β=-0.034) and pneumoconiosis (β=-1.595) as factors affecting 45S rDNA copy number, age (β=-0.013) as a factor affecting 5S rDNA copy number, and age (β=0.018) as a factor affecting 5S rDNA copy number in the pneumoconiosis group (all P<0.05).
Conclusions
Compared with community residents with no history of pneumoconiosis or dust exposure, the copy number of 45S rDNA in peripheral blood among patients with pneumoconiosis is reduced and the copy number of 5S rDNA is increased.
7.MSCT findings of pulmonary tuberculoma
Junfei FAN ; Wujiang YU ; Youxue XU ; Minling WANG
Journal of Practical Radiology 2018;34(4):526-528
Objective To explore MSCT findings of pulmonary tuberculoma.Methods MSCT data of 62 patients with pulmonary tuberculoma confirmed by clinical and pathological results were analyzed retrospectively.Results Of the 62 patients,64 lesions were detected with single in 60 cases and multiple in 2 cases.The MSCT signs showed as follows:circumscribed round,oval,egg-like or irregular nodules or masses,size ranging from 2-3 cm,cavity in 31 lesions,calcification in 25 lesions,pleural indentation sign in 45 lesions and pleural tail sign in 1 2 lesions.Conclusion The MSCT features of pulmonary tuberculoma are relatively specific such as the multiple patch calcifications within mass,calcification of the whole mass and semilunar cavity on the lateral side of the hilum.The MSCT is a valuable examination for the accurate diagnosis of pulmonary tuberculoma.