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Objective:To study the influence of circular RNA hsa_circZDHHC21_004 on the proliferation of human small intestinal epithelial cells HIEC-6 after 60Co γ-rays exposure. Methods:HIEC-6 cells were exposed to 60Co γ-rays at 0, 5, 10, and 15 Gy with a dose rate of 1 Gy/min. The expression level of hsa_circZDHHC21_004 in the irradiated HIEC-6 cell was detected. Hsa_circZDHHC21_004 was knocked-down to investigate the influences of hsa_circZDHHC21_004 on the proliferation of irradiated HIEC-6 cells by CCK-8 assay and colony formation assay. Results:The expression level of hsa_circZDHHC21_004 in HIEC-6 cells was upregulated by (1.00±0.24), (1.34±0.28), (1.85±0.31), and (2.80±0.64) times of control after 0, 5, 10, and 15 Gy irradiation, respectively and there were significant difference between 10 or 15 Gy group and 0 Gy group ( F=10.86, P=0.008). Knockdown of hsa_circZDHHC21_004 significantly increased the proliferation rate of HIEC-6 cells at 24, 48, and 72 h after 10 Gy irradiation compared with non-irradiated control ( t=-6.25, -5.83, -7.75, P < 0.001). Under 2 and 5 Gy irradiation, the clone formation rates of the hsa_circZDHHC21_004 knockdown cells were significantly higher than those of the control ( t=-7.45, -8.83, P<0.01). Conclusions:Hsa_circZDHHC21_004 is increased after irradiation and influenced the proliferation of irradiated HIEC-6 cells.
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Purpose: To explore the neuroprotective effects of Lutongkeli (LTKL) in traumatic brain injury (TBI) and detect the related mechanism. Methods: TBI model was established with LTKL administration (2 and 4 g/kg/d, p.o.). Motor function of rats was examined by Rotarod test. Nissl staining was used to show neuron morphology. Furthermore, the disease-medicine common targets were obtained with the network pharmacology and analyzed with Kyoto Encyclopedia of Genes and Genomes. Lastly, the predicted targets were validated by real-time polymerase chain reaction. Results: After LTKL administration, neural behavior was significantly improved, and the number of spared neurons in brain was largely increased. Moreover, 68 bioactive compounds were identified, corresponding to 148 LTKL targets; 2,855 genes were closely associated with TBI, of which 87 overlapped with the LTKL targets and were considered to be therapeutically relevant. Functional enrichment analysis suggested LTKL exerted its pharmacological effects in TBI by modulating multiple pathways including apoptosis, inflammation, etc. Lastly, we found LTKL administration could increase the mRNA level of Bcl-2 and decrease the expression of Bax and caspase-3. Conclusions: This study reported the neuroprotective effect of LTKL against TBI is accompanied with anti-apoptosis mechanism, which provides a scientific explanation for the clinical application of LTKL in the treatment of TBI.
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Animals , Male , Rats , Apoptosis/drug effects , Neuroprotective Agents/administration & dosage , Brain Injuries, Traumatic/therapy , Rats, Sprague-Dawley , Medicine, Chinese TraditionalABSTRACT
This paper reports the diagnosis and treatment of a pregnant woman complicated with myotonic dystrophy (DM) type 1(DM1). The pregnant woman complained of occasional dull pain in the precardiac region accompanied by night sweats in the first trimester and a mild increase in myocardial enzymes was observed. Symptoms were improved after oral medication followed by no regular reexamination. While in the third trimester, abnormal myocardial enzyme, liver enzyme, and urinary protein were found and echocardiography and abdomen ultrasound suggested multiple organic lesions involving the heart and spleen. A girl was born through cesarean section at 36 +5 weeks of gestation due to maternal heart failure, with Apgar scores of 10 at 1 and 5 minutes. The patient developed acute respiratory failure, ventricular bigeminy, and diarrhea after surgery. Neurology consultation found incomplete buried eyelids in both eyes, weak bulging cheeks, and weak upper extremity muscle strength. Muscle biopsy confirmed the diagnosis of DM and genetic testing showed abnormal amplification of non-coding region [CTG] n in myotonic dystrophy protein kinase gene in chromosome 19q13.3, which indicated DM1. The mother recovered well at nine months postpartum. Further genetic analysis of pedigree and offspring is recommended.
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Objective:To explore the characteristics of lipid metabolism in rat plasma after total body irradiation(TBI) in order to provide scientific evidence of radiation biomarkers.Methods:For the non-targeted lipidomics study, 50 SD rats were divided into 6 groups and irradiated with 0, 1, 2, 3, 5 or 8 Gy 60Co γ-rays, respectively. For the targeted lipidomics study, 25 rats were divided into 5 groups and irradiated with 0, 0.5, 2.5, 4 or 6 Gy. Venous blood samples were collected and plasma were separated 4 h after TBI. Radiation-sensitive lipids were screened and their concentrations were determined. Receiver operating characteristic curve (ROC) and dose-response were analyzed. Results:A total of 15 radiation differential lipids were screened out based on non-targeted lipidomics study and 7 of them were identified as radiosensitive lipids by targeted lipidomics analysis. The ROC of radiosensitive lipids distinguished area under curve (AUC) of samples in 0 Gy group and > 0 Gy group, < 2 Gy group and ≥ 2 Gy group were all > 0.75. The AUC values were increased to 0.96 and 0.94 after the panel of radiation sensitive lipids ROC analysis. The concentrations of LysoPC(18: 2), LysoPC(22: 0), PC(18: 0/18: 2), PE(18: 2/16: 0) and PE(18: 2/18: 0) decreased with irradiation dose within 0-6 Gy.Conclusions:A total of 7 plasma radiosensitive lipids in rat plasma were identified 4 h after TBI, and the panel of them could be used for specific dose classification. Five of the lipids had good dose-response relationship.
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Objective:To explore the influence of dose-rate on radiation-induced gene expression in human peripheral blood.Methods:Human peripheral blood ex vivo was irradiated with 0, 1, 2, 4 and 6 Gy of 60Co γ-rays with different dose-rates of 0.2, 1 and 2 Gy/min. Human blood cells were harvested at 24 h post-irradiation. Following RNA isolation, the mRNA expression levels ofCDKN1A, MDM2, PCNA, FDXR, GADD45A, PHPT1, ASTN2, TNFSF4, POLH, GDF-15 and PPM1D were measured by quantitative real-time PCR. The stepwise regression method was used to establish the gene combination models. Results:The relative mRNA expression levels of 11 genes significantly increased in a dose-dependent manner within the dose range of 0-6 Gy with three dose-rates of irradiation ( R2=0.744-0.998, P< 0.05). Following the exposure to 2 Gy(0.2 Gy/min) 60Co γ-rays, the expression levels of CDKN1A, FDXR, PHPT1 and TNFSF4 genes were significantly higher than that of the 1 and 2 Gy/min groups ( t=3.73, 5.73, 2.44, 2.77, 3.53, 2.68, 2.43, 2.05, P< 0.05). With 6 Gy irradiation, the changes of radiation-induced PPM1D expression level under a dose rate of 2 Gy/min were significantly higher than other two dose-rates( t=3.82, 2.54, P< 0.05). The combined expression model at different dose rates was composed of 2-3 genes, and the R2values of regression equations were 0.976, 0.964 and 0.951, respectively. Conclusions:In a certain range, the dose-rate may affect the changes of radiation-induced gene expression in human peripheral blood.
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Objective:To explore the feasibility of the optimized cytokinesis-block (CB) assay on radiation-induced nucleoplasmic bridge (NPB), and to provide a scientific basis for the application of NPB in biological dose estimation.Methods:Human peripheral blood in vitro was irradiated with 2 Gy 60Co γ-rays at a dose rate of 1 Gy/min (0 Gy control group). According to the culture time after irradiation, blood samples were divided into group 48, 56, 68 and 72 h. Cytochalasin-B (Cyt-B) with a concentration of 6 μg/ml was added into the samples at 28 h and harvested at 48, 56, 68 and 72 h after irradiation, respectively. On the other hand, the blood samples were treated with different concentration of Cyt-B i. e., 0.6, 1, 2, 6 and 10 μg/ml at the beginning of culture (0 h) and harvested at 68 h after irradiation. The proportion of mononucleated, binucleated and multinucleated cells, radiation-induced NPB and micronucleus (MN) frequencies were analyzed. Results:The nuclear division index (NDI) and proportion of binucleated cells at 2 Gy and 0 Gy had tendency of increasing with cell culture time. NPB frequencies (0.023 0-0.033 0/cell) and MN frequencies had no significantly difference ( P> 0.05). With the increase of Cyt-B concentration, NDI and the proportion of binucleated cells in group 2 Gy and 0 Gy also increased, but NPB frequencies (0.023 0-0.047 0/cell) had no significant difference ( P> 0.05). MN frequencies of group 10 μg/ml were significantly lower than that of group 6 μg/ml ( U=2.74, P< 0.01). Conclusions:Cell culture time and Cyt-B concentration had no significant influence on radiation-induced NPB frequencies, suggesting that NPB could be obtained by appropriately reducing cell culture time and Cyt-B could be added into blood samples at the beginning of culture. But this protocol reduced the number of cells for further analysis, and thus its feasibility for dose estimation still need to be studied.
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Objective:To screen radiosensitive lipid metabolites in rat small intestine and analyze their metabolic pathways, in order to provide scientific basis for radiation enteropathy biomarkers.Methods:The total body irradiation of 60Co γ rays was performed to rats with different doses of 0, 1, 2, 3, 5 and 8 Gy. The changes of lipids in small intestine were studied by targeted lipidomics method based on liquid chromatography coupled mass spectrometry (LC-MS). Results:Fifteen lipids in small intestine were screened as radiosensitive metabolites at 3 d after irradiation, including 4 up-regulated lipids and 11 down-regulated lipids( t=-6.395, 5.998, 5.836, -5.503, -5.449, -5.422, 4.841, 4.802, 4.621, 4.457, 4.426, 4.373, 4.110, 3.945, 3.902, P< 0.05 and FDR < 0.05). The metabolic pathways of sphingolipid, glycerophosphoplipid were significantly enriched. Four phosphatidyl serines (PS)increased while 1 phosphatidic acid(PA), 2 sphingomyelins(SM) and 4 fatty acids(FA)decreased in a good dose-response manner( R2> 0.80, P< 0.05), which were more potential radiation enteropathy biomarkers. Conclusions:Lipid metabolites in rat small intestine were significantly changed after the rat was total body irradiated with 60Co γ-rays.Eleven lipids with good dose-response relationship were more potential to be radiation enteropathy biomarkers.
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OBJECTIVE: To observe the effect of low dose long-term chronic irradiation on chromosome aberration and micronucleus in peripheral blood lymphocytes of interventional radiology workers.METHODS: A total of 100 interventional radiology workers in a grade A tertiary hospital of Henan Province were selected as the interventional group using a convenient sampling method, and 78 healthy individuals without radiation exposure were selected as the control group. The incidence of chromosomal aberration and micronucleus of peripheral blood lymphocytes in individuals of these two groups were investigated. RESULTS: The frequencies of cells with acentric fragment(ace), dicentric chromosome(dic), reciprocal translocation(t), chromosome type aberration and total aberration in the interventional group were higher than those in the control group, and the differences were statistically significant(0.55% vs 0.30%, 0.25% vs 0.03%, 0.38% vs 0.01%, 1.21% vs 0.34%, 1.37% vs 0.54%, all P<0.01). In the interventional group, the frequency of cell with chromosome type aberration was higher than that of chromatid type aberration(1.21% vs 0.16%, P<0.01).The frequency of cell with unstable chromosome aberration was higher than that of stable chromosome aberration(0.80% vs 0.41%, P<0.01), and the frequency of cell with chromosomal type aberrant in the interventional group ranked from high to low in order of ace, t, dic and deletion(P<0.01). The incidence of micronuclear cells and micronucleus in the interventional group were higher than those in the control group(0.86‰ vs 0.40‰, 0.89‰ vs 0.41‰, all P<0.01). The incidence of micronuclear cells and micronucleus of female interventional radiology workers were higher than those of male workers(1.12‰ vs 0.62‰, 1.19‰ vs 0.62‰, all P<0.01).CONCLUSION: Low dose and long-term chronic irradiation exposure can affect both chromosome aberration and micronucleus of interventional radiology workers. The main chromosomal aberrations were non-stable chromosomal aberrations ace, dic and stable chromosomal aberrations t.
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PURPOSE@#To evaluate the impact of five different tooth preparation designs on the marginal and internal fit discrepancies of cobalt-chromium (CoCr) crowns produced by computer-aided designing (CAD) and selective laser melting (SLM) processes. @*MATERIALS AND METHODS@#Five preparation data were constructed, after which design crowns were obtained. Actual crowns were fabricated using an SLM process. After the data of actual crowns were obtained with structural light scanning, intaglio surfaces of the design crown and actual crown were virtually superimposed on the preparation. The fit-discrepancies were displayed with colors, while the root means square was calculated and analyzed with one-way analysis of variance (ANOVA), Tukey’s test or Kruskal-Wallis test (α =.05). @*RESULTS@#The marginal or internal color-coded images in the five design groups were not identical. The shoulder-lip and sharp line angle groups in the CAD or SLM process had larger marginal or internal fit discrepancies compared to other groups (P < .05). In the CAD process, the mean marginal and internal fit discrepancies were 10.0 to 24.2 µm and 29.6 to 31.4 µm, respectively. After the CAD and SLM processes, the mean marginal and internal fit discrepancies were 18.4 to 40.9 µm and 39.1 to 47.1 µm, respectively. The SLM process itself resulted in a positive increase of the marginal (6.0 – 16.7 µm) and internal (9.0 – 15.7 µm) fit discrepancies. @*CONCLUSION@#The CAD and SLM processes affected the fit of CoCr crowns and varied based on the preparation designs. Typically, the shoulder-lip and sharp line angle designs had a more significant effect on crown fit. However, the differences between the design groups were relatively small, especially when compared to fit discrepancies observed clinically.
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Objective:To explore the advantage of dose estimation based on semi-automated dicentric (dic) scoring and the feasibility of reflecting a large scale radiation accident for population clinical triage by mean of the comparison between semi-automatic and manual detection.Methods:Human peripheral blood samples from two healthy volunteers were irradiated by 60Co γ-rays at the doses of 0, 0.5, 1, 2, 3, 4, 5 and 6 Gy with a dose rate of 0.27 Gy/min, and chromosome preparation was carried out using the conventional method. The dic and dic plus ring were analyzed automatically with the DCScore software and manually with the Ikaros software, respectively, in a high-throughput chromosome automatic scanning system. The dose-response curves were fitted with dic or dic plus ring per cell. Twelve standard samples of biodosimetry were used to validate the dose-response curves. Results:The numbers of dic or dic plus ring per cell by semi-automatic or manual detection increased with the exposure doses ( r=0.984, 0.972, 0.972, P<0.01). The yields of semi-automated dic or manually detected dic plus ring were well fitted with the correlation coefficients ( R2=0.998, 0.999, 0.999, P<0.01). When the exposure dose of the standard samples was more than 2 Gy, the relative deviation between actual and predict doses was within 21% using the dose-response curve based on automated dic before human verification and correction of dic (elimination of false positives and inclusion of true positives), and wiht ±10% after manual elimination of false positive dic. Bio-doses estimated from the dic detected by manual scoring were similar to the actual exposure doses with the exception of 0.7 Gy, but the efficiency of semi-automatic analysis of dic was increased by 6-times in bio-dose assessment. Conclusions:The semi-automated dic detection obviously improves the level and efficiency of biodosimetry analysis, and thus can meet the requirements of clinical classification diagnosis of medical emergency response to large-scale nuclear radiation events.
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Objective:To explore the influence factors of chromosomal aberration levels in radiation workers in hospitals.Methods:Two hundred and fourteen age- and sex- matched hospital radiation workers were recruited by stratified random sampling method. According to the job title, the individuals were divided into four groups including diagnostic radiology group ( n=57), radiotherapy group ( n=49), nuclear medicine group ( n=52) and interventional radiology group ( n=56). Chromosomal aberrations in peripheral blood lymphocytes from the subjects were measured using conventional cytogenetic analysis method, and the influence factors of chromosomal aberrations were analyzed. Results:There was significant difference in the frequencies of acentric fragment, translocation and total chromosome-type aberrations among the four groups ( χ2=9.906, 19.965, 32.824, P<0.05), and the rates of aberrations were significantly higher in the interventional radiology group and the nuclear medicine groups than those in the diagnostic radiology (interventional group: χ2=4.711, 10.798, 10.845, P<0.05; nuclear medicine group: χ2=3.853, 7.674, 7.708, P<0.05) and the radiotherapy groups (interventional group: χ2=9.209, 9.772, 21.330, P<0.05; nuclear medicine group: χ2=8.010, 6.969, 10.812, P<0.05). The rates of translocation and total aberrations ( χ2=7.706, 6.667, P<0.05) and the frequencies of acentric fragment, translocation and total aberrations ( χ2=12.263, 15.360, 21.478, P<0.01) were dependent on the length of service and the dose among different groups. The rates of translocation and total aberrations significantly increased along with exposure doses ( r=0.347, 0.263, P<0.01). Poisson regression analysis indicated that the job titles and annual effective dose partly affected the levels of chromosomal aberrations[ IRR=1.797 (nuclear medicine group), 2.136 (interventional group) and 1.422 (0.5-1 mSv group); P<0.05]. Conclusions:The frequencies of chromosomal aberrations in the radiation workers of interventional and nuclear medicine groups remain higher levels in hospital, thus it is necessary to strengthen the radiation protection on these radiation workers.
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Objective:To explore the effects of ultraviolet B (UVB) on the premature senescence of human immortalized keratinocytes HaCaT cells and the possible underlying molecular mechanism.Methods:HaCaT cells were exposed with UVB of different doses (20, 50, 80 and 100 mJ/cm 2). At 72 h after exposure, cellular morphology was observed by Giemsa staining, cell proliferation was detected by clone formation assay, and the proportion of premature senescence cells was detected by β-galactosidase staining. The number change of lysosomes was detected by Lyso-Tracker Red fluorescence probe at 24, 48 and 72 h after exposure. Cell migration was measured by scratch test at 24 h and 48 h after exposure. The protein expressions of p53 and p16 related to premature senescence were detected by Western blot assay at 72 h after exposure. Results:After UVB exposure, HaCaT cells showed a premature senescence phenotype. At 72 h after exposure, the cell volume increased ( F=115.18, P<0.05), the cell proliferation ability decreased ( F=410.32, P<0.05), the activity of β-galactosidase increased ( F=16.31, P<0.05), and the expressions of P53 and P16 increased. In addition, the number of lysosomes increased at 24, 48, and 72 h after exposure ( F=17.65, 38.36, 13.66, P<0.05), and cell migration capacity was inhibited at 24 and 48 h after exposure ( F=8.21, 11.48, P<0.05). Conclusions:UVB exposure can induce premature senescence of HaCaT cells by increasing the expression of p53 and p16 proteins.
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Objective:To investigate the feasibility of dose estimation based on the dose-response curves established by different types of dicentrics (dic).Methods:Human peripheral blood samples were irradiated by 60Co γ-rays at the doses of 0, 0.5, 1, 2, 3, 4, 5 and 6 Gy with the dose rate of 0.27 Gy/min, and chromosome preparation was carried out. The different types of dics were analyzed. The dose-response curves were fitted with software of Chromosomal Aberration Calculation Software (CABAS). Two samples were used to validate the dose-response curves. Results:The aberration rates of different types of dics increased with the exposure doses ( R2=0.886-0.943, P<0.01). The proportion of typical and single-ended dic components accounted for more than 92% of all types of dic, while the proportion of short-range dic and double-ended dic components was less than 4% at each dose point. The dose-response curves based on different types of dics were established ( R2=0.998, P<0.01). There was no significant difference in the doses estimated by four different dose-response curves ( P>0.05). The relative deviations between actual and predict doses were less than 13.08% by using dose-response curve based on typical dics. Conclusions:Dose-response curves based on different types of dics have the feasibility of dose estimation.
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Objective:To screen the indicators of retrospective dose estimation, based on 5 cytogenetic methods to assess the victim followed-up at 4 year after 192Ir radiation accident in Nanjing. Methods:The chromosome aberration (dic + r) assay, cytokinesis block micronucleus (MN) and nucleoplasmic bridge (NPB) assay, fluorescence in situ hybridization (FISH)-based and G banding-based translocation analysis were used to retrospective biological dose estimation. Results:The estimated doses of FISH-based and G banding -based analysis were 1.45 and 1.21 Gy respectively, which was similar to the biological dose estimated short time after the accident. However, the estimated doses by chromosome aberration, micronucleus and nucleoplasmic bridge method were 0.56, 0.45 and 0.41 Gy respectively, which were lower than the corresponding biodose. Correction factors were used to adjust the biodose.Conclusions:In the 4th years after exposure, the estimated biological doses by FISH-based and G banding-based translocation were consistent with the biodose.Therefore, the two methods were suitable for retrospective dose estimation, while correction factors should be considered in chromosome aberration method for retrospective dose estimation.
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Objective To study the effects of sex,age,length of service,type of work and annual effective radiation dose on nucleoplasmic bridge (NPB) in the peripheral blood lymphocytes of radiation workers.Methods The peripheral blood samples of 100 radiation workers in Henan province were collected and the NPB in peripheral blood lymphocytes were measured by CBMN assay.The frequencies of NPB formation and NPB-containing cells were calculated,and the effects of various factors on NPB incidence were analyzed statistically.Results The NPB frequency in radiation workers was higher than that in healthy people (Z=-8.123,P＜0.01).Except for sex,the factors of age,length of service,type of work and annual effective dose had significant influences on NPB (x2=7.202-45.571,P＜0.05).Conclusions NPB reflects the effect of low-dose long-term chronic irradiation on the occupational radiation workers.
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Objective@#To study the effects of sex, age, length of service, type of work and annual effective radiation dose on nucleoplasmic bridge (NPB) in the peripheral blood lymphocytes of radiation workers.@*Methods@#The peripheral blood samples of 100 radiation workers in Henan province were collected and the NPB in peripheral blood lymphocytes were measured by CBMN assay. The frequencies of NPB formation and NPB-containing cells were calculated, and the effects of various factors on NPB incidence were analyzed statistically.@*Results@#The NPB frequency in radiation workers was higher than that in healthy people (Z=-8.123, P<0.01). Except for sex, the factors of age, length of service, type of work and annual effective dose had significant influences on NPB (χ2= 7.202-45.571, P<0.05).@*Conclusions@#NPB reflects the effect of low-dose long-term chronic irradiation on the occupational radiation workers.
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Objective To investigate the effect of continual care on postoperative medication compliance of patients with osteoporotic vertebral compression fractures (OVCF) after vertebroplasty.Methods A retrospective case control study was performed to analyze the clinical data of 150 patients with OVCF who underwent vertebroplasty in Guizhou People's Hospital from January 2016 to May 2017.There were 38 males and 112 females,aged 47-88 years,with an average of 67.5 years.Seventy-five patients were given continual care such as telephone follow-up and home visit (continual care group).Seventy-five patients were given routine health education (routine care group) when they were discharged from hospital.Visual analogue scale (VAS) and Oswestry dysfunction index (ODI) before operation,1 day,1,3,6 and 12 months after operation,as well as medication compliance and vertebral re-fracture at 1,3,6 and 12 months after operation were compared between the two groups.Results The preoperative VAS of the routine care group was 6 (6-7),1 (0-1),1 (1-3),2 (1-3) and 3 (2-5) points at 1 day and 1,3,6 and 12 months after operation,respectively.The preoperative ODI was 21 (18-27),0 (0-0),2 (1-4),5 (3-7),7 (5-10),10 (7-14) points at 1 day,1,3,6 and 12 months after operation.In the continual care group,VAS was 7 (6-7) points before operation,0 (0-1),1 (0-1),1 (0-2) and 2 (1-3) points at 1 day,1,3,6 and 12 months after operation,respectively.ODI of the continual care group was 18 (22-28) points before operation,0 (0-1),2 (0-4),4 (1-5),4 (3-6) and 6 (4-9) at 1 day,1,3,6 and 12 months after operation.The VAS and ODI of the two groups were lower than those before operation,and the scores of the continual care group were lower than those of the routine care group at 1,3,6 and 12 months after operation (P ＜ 0.05).The medication compliance rates of continual care group were 93％,89％,91％ and 84％ at 1,3,6 and 12 months after operation,while those of routine care group were 44％,40％,47％ and 40％ respectively (P ＜0.05).The incidence of vertebral re-fracture was 1％,1％,3％ and 3％ in continual care group and 3％,5％,5％ and 7％ in routine care group at 1,3,6 and 12 months after operation respectively (P ＜ 0.05).Conclusion Continual care can improve the medication compliance of OVCF patients after treatment with vertebroplasty,relieve pain,improve the quality of life,and reduce the incidence of vertebral re-fracture,which is worthy of clinical promotion.
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Objective To investigate the damage mechanism of typeⅡalveolar epithelial cells (AECⅡ) after hyperoxia exposure by proteomics. Methods The primary AECⅡ of preterm Sprague-Dawley (SD) rats were divided into normoxia and hyperoxia groups, and cultured in room air (21% O2) or hyperoxia (95% O2) condition, respectively. The cell morphology change was observed under an inverted contrast microscope; the protein expressions of Bcl-2 and caspase-3 were detected by Western Blot to ensure a successful model. Total protein in AECⅡ was collected, and mass spectrometry-based tandem mass tag (TMT)-labeled quantitative proteomics were used to detect the change of protein profile. Proteins with changes greater than 1.5-fold and P < 0.05 were considered differentially expressed, and bioinformatics analysis was performed. According to the proteomic results, AECⅡ were divided into three groups:normoxia group, hyperoxia group and hyperoxia+MW167 group (γ-secretase inhibitor MW167 was added to culture medium 30 minutes before they were placed into the chamber). The cell viability was detected by the cell proliferation and toxicity kit (CCK-8), and the expressions of Hes1, Bax mRNA were detected by real-time fluorescence quantitative reverse transcription-polymerase chain reaction (qRT-PCR). Results ① The cells in the normoxia group proliferated and prolonged significantly, and the cytoplasmic particulate matter was abundant. In the hyperoxia group, nucleus pyknosis and cytoplasmic particulate matter decreased significantly. Compared with the normoxia group, the expression of caspase-3 in the hyperoxia group was significantly increased, and the expression of Bcl-2 was significantly decreased (caspase-3/GAPDH: 1.352±0.086 vs. 0.769±0.080, Bcl-2/GAPDH: 0.614±0.060 vs. 1.361±0.078, both P < 0.01).② A total of 162 differentially expressed proteins were identified between normoxia and hyperoxia groups, the proteins up-regulated by hyperoxia were commonly associated with response processes to various stimuli, and located in the extracellular region; the proteins down-regulated by hyperoxia were commonly associated with synthesis of substances, and located in the cellular matrix. KEGG Pathway analyses suggested that metabolism by cytochrome P450, oxidative phosphorylation, and Notch signaling pathway were associated with the mechanism of hyperoxia injury on AECⅡ.③Compared with the normoxia group, the viability of cells in the hyperoxia group was significantly decreased, and the expressions of Hes1 and Bax mRNA were significantly increased [cell viability (A value): 0.060±0.003 vs. 1.058± 0.017, Hes1 mRNA (2-ΔΔCt): 2.235±0.606 vs. 1.144±0.107, Bax mRNA (2-ΔΔCt): 2.210±0.240 vs. 1.084±0.096, all P < 0.05]. Compared with the hyperoxia group, the viability of cells in the hyperoxia+MW167 group was significantly increased, and the expressions of Hes1 and Bax mRNA were significantly decreased [cell viability (A value): 0.271±0.025 vs. 0.060±0.003, Hes1 mRNA (2-ΔΔCt): 0.489±0.046 vs. 2.235±0.606, Bax mRNA (2-ΔΔCt): 1.289±0.041 vs. 2.210±0.240, all P < 0.05]. Conclusion The mechanism of hyperoxia injury on AECⅡ may be related to the metabolism by cytochrome P450, oxidative phosphorylation and activation of Notch signaling pathway.
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OBJECTIVE: To explore the dose-effect of X-ray irradiation and chromosome aberrations in human peripheral blood cells,and establish a dose-response curve of dicentric and ring(dic+r)aberration induced by X-ray irradiation.METHODS: Human peripheral blood samples were collected from three healthy individuals and were exposed to X-ray at the doses of 0.00,0.25,0.50,0.75,1.00,2.00,3.00,4.00 and 5.00 Gy in vitro.The dose rate was 1.158 Gy/min.The blood cells were harvested after routine culture,and the chromosome preparation was carried out.The dicentrics and rings in metaphase cells were counted under microscope,and a dose-response curve was fitted by using the software of CABAS.Dose estimation was performed according to the curve from two blind samples.RESULTS: Aberration of dic+r increased with irradiation doses in the range of 0.00-5.00 Gy(P<0.01).The dose-response relationship followed a linear-quadratic equation:■,where■ is the yield of dic+r,and D is the irradiated dose.The estimated doses of the two blind samples were in accordance with the actual doses.CONCLUSION: The dose-response curve and mathematical model of chromosome aberration following exposure to 0.00-5.00 Gy X-ray irradiation is established in this study provide a reliable method for the accurate dose estimation.
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Objective To investigate the clinical significance of serum ferritin (SF) detection in myelodysplastic syndromes (MDS).Methods A total of 42 cases of MDS without blood transfusion who were admitted to General Hospital of Huainan Eastern Hospital Group from January 2010 to July 2017 were collected.There were 27 males and 15 females,aged 46-90 years old with a median age of 72 years old.In 42 cases of MDS,10 cases had refractory anemia (RA),4 cases had refractory anemia with ring increased iron bead young cell (RARS),8 cases had refractory blood cells decreased with more dysplasia (RCMD),and 20 cases had refractory anemia with primitive cells increased (RAEB).According to the International Prognostic Scoring System,22 cases had low risk,and 20 cases had high risk.At the same time,40 healthy checkers were chosen as healthy control group.SF levels were measured and compared between the MDS patients and healthy controls.Results The SF level in the MDS group was significantly higher than that in the healthy control group [(479±267) ng/ml vs.(153±56) ng/ml],and the difference was statistically significant (t =7.379,P ＜ 0.05).The difference in SF levels among the patients with different subtypes of MDS was statistically significant (F =4.81,P =0.006).Among them,the RA group had the lowest SF level,while the RAEB group had the highest,and the difference between the two groups was statistically significant [(346±257) ng/ml vs.(818±443) ng/ml,P =0.005].The SF level in the low-risk group was lower than that in the high-risk group [(456±238) ng/ml vs.(788±467) ng/ml],and the difference was statistically significant (t =2.867,P =0.008).Conclusion Patients with MDS show a significant increase in SF level,and high-risk patients are associated with iron overload,and SF may be an auxiliary indicator for judging the prognosis of MDS.