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Objective:To investigate the protective effect of lactoferrin(Lf) on lung injury in mice exposed to irradiation.Methods:C57BL/6 J mice were randomly divided into control group, 15 Gy irradiation group (IR group) and lactoferrin combined 15 Gy irradiation group (Lf+ IR group), with 5 mice in each group. The mice in the Lf+ 15 Gy group drank lactoferrin solution (10 mg/ml) from 3 days before irradiation and contained the whole experiments. Then, single chest 15 Gyirradiation was performed both in the IR and Lf+ IR groups. The body weight and other characteristics were monitored during the experiment. The mice were killed at day 14 after irradiation. The lung histopathology was observed by HE staining. Serum inflammatory cytokine such as HMGB1, TNF-α, IL-1β and IL-6 was determined by ELISA method . The expression of inflammatory related protein in lung tissue including HMGB1, TLR4, MyD88 and NF-κB were performed by immune histochemistry and Western blot method.Results:Compared with the control group, lung weight was significantly increased ( t=3.20, P<0.05), pulmonary hyperemia and inflammatory cell infiltration was observed in the IR group. Exposure also significantly increased serum level of TNF-α[(291.80±5.49) vs.(332.25±22.18)pg/ml]( t=3.07, P<0.05), up-regulated the expression of inflammatory related protein in lung tissue ( t=4.04, 4.78, 3.77, 6.14, P<0.05). Lactoferrin intervention (Lf+ IR group) significantly decreased lung weight ( t=2.18, P<0.05), alleviated histopathologic changes, decrease serum levels of HMGB1, TNF-α and IL-1β ( t=4.67, 2.97, 3.49, P<0.05). On the other hand, lactoferrin intervention decreased the positive cell number of HMGB1 and NF-κB, and down-regulated the protein expression of HMGB1, TLR4, MyD88 and NF-κB in lung tissues, with significant difference with the IR group ( t=8.06, 9.80, 3.07, 5.56, P<0.05). Conclusions:Lactoferrin plays the protective effect of radiation-induced lung injury through the downregulation of inflammatory response, such as HMGB1/TLR4/MyD88/NF-κB signaling pathway.
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Objective To investigate the effect of Zhubei Dingchuan pill on clinical symptoms and inflammatory mediators in serum and induced sputum of patients with chronic bronchitis. Methods 80 cases of patients with chronic bronchitis treated in the Department of Respiration in our hospital from January 2016 to January 2017 were selected and were randomly divided into the control group and the observation group, with 40 cases in each group. The control group was given symptomatic support treatment and the observation group was additionally given Zhubei Dingchuan pills on the basis of the control group. The changes of inflammatory mediators levels in serum and induced sputum were measured before and after treatment between the two groups, and the clinical efficacy was observed in the two groups. Results After treatment, the levels of interleukin-6 (IL-6), tumor necrosis factor-α (TNF-α) and high-sensitivity C reactive protein (hs-CRP) in serum or induced sputum in the observation group were significantly lower than those in the control group (P<0.05). The effective rate of treatment in the observation group was significantly higher than that in the control group (95.0％ vs. 82.5％) (P<0.05). Conclusion Zhubei Dingchuan pill can effectively relieve the clinical symptoms in patients with chronic bronchitis, and its pharmacological mechanism is related to the decrease of levels serum and airway inflammatory mediators.
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Objective To compare the analgesic effect and incidences of adverse events of different sufentanil concentration regimens in postoperative intravenous patient-controlled analgesia(PCA).Methods We retrospectively analyzed the clinical data of 6231 patients undergoing elective general anesthesia using sufentanil as postoperative intravenous PCA regimen in Peking Union Medical College Hospital from January 2004 to December 2016.These patients were subcategorized into 4 groups according to the sufentanil concentration regimens:0.4 μg/ml(SF4,n=1421),0.6 μg/ml(SF6,n=2489),0.8 μg/ml(SF8,n=1326),and 1.0 μg/ml(SF10,n=995).Total drug consumption within 48 h after surgery,analgesic effect,and incidences of adverse events were compared among these four groups.Results The cohort consisted of 2874 males(46.1%)and 3357 females(53.9%)in the age group from 3 years to 91 years(median:52.5 years).The postoperative 48 h sufentanil consumption was significantly different among these four groups in terms of volume(χ=87.316,P<0.001)and dosage(χ=20.261,P<0.001).Meanwhile,the VAS scores at rest and during activity on postoperative day 1(POD1)and POD2 showed no statistical significance among these four groups (both P>0.05).As for the adverse events,the sedation score in POD1(χ=9.042,P=0.029)and incidence of no bowl movement on both POD1(χ=7.855,P=0.012)and POD2(χ=5.635,P=0.044)were significantly different among groups,whereas the incidences of other adverse events showed no significant difference(all P>0.05).Conclusion In patients using intravenous sufentanil PCA as their postoperative analgesia regimen after general anesthesia,regimens with higher sufentanil concentrations may result in more adverse events such as sedation and no bowl movement without improving analgesic outcomes.
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Objective To investigate the diagnostic value of ultrasound elastography hardness score and area ratio to identify benign and malignant lesions.Methods 90 patients with breast lumps were selected and divided into the malignant and non -malignant group according to surgical pathology diagnosis,they were 40 cases (a total of 45 tumors)and 50 cases (a total of 57 tumors),two groups of patients were detected in ultrasound elastography,anal-ysis of imaging hardness ratings and area ratio and other information,to investigate the clinical diagnostic value. Results The ultrasound elastography of 50 cases with benign tumor were 0 -2 point,7 cases were 3 -4 point, 7 cases with malignant tumors were 0 -2 point,38 cases were 3 -4 point,the difference between the two groups were statistically significant (χ2 =28.55,P <0.05);The average area ratio of benign tumor was (1.01 ±0.27),malignant tumors was (2.28 ±1.68),the difference between the two groups was statistically significant(t =9.22,P =0.001);the sensitivity,specificity,accuracy of elastography hardness rating in diagnosis of malignant breast tumors were 87.72%,84.44%,86.27%,the area ratio method were 85.96%,86.67%,86.27%,joint inspection of the two groups were 96.49%,95.55%,96.08%,joint inspection had obvious advantages,the difference was statistically sig-nificant(χ2 =16.24,13.58,P <0.05).Conclusion Ultrasound elastography hardness rating combined area ratio has a higher accuracy rate for differentiating benign and malignant breast tumors.
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Objective To investigate the value of Color Doppler ultrasound in hemodynamic changes of non-alcoholic fatty liver disease.Methods 120 cases with non -alcoholic fatty liver disease were selected and divided into the mild group,moderate group and severe group.According to the degree of fatty infiltration of the liver taxono-my,40 cases in each group,hemodynamic changes of each group were detected in Color Doppler ultrasound equip-ment.Results Hepatic venous pulse waveform analysis and comparison constituted of the three groups and control group,the difference was statistically significant (χ2 =25.343,P <0.05).PPVV,MPVV,HARI of the three groups and control group were compared,the difference was statistically significant(F =34.67,51.03,31.06,all P <0.05).Conclusion Color Doppler ultrasound which is used to detect hemodynamic changes in non -alcoholic fatty liver disease had good clinical value,can assess disease progression in patients,can provide the basis for clinical treat-ment.
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Objective To investigate the biological functions of IncRNA-BG on the radiosensitivity of normal human bronchial epithelial cell line Beas-2B.Methods Three IncRNA-BG siRNAs were designed,synthesized and traasfected into Beas-2B cells via lipofectamine.The RNA transcription level of BG was detected by quantitative real time-PCR to confirm the siRNA transfection efficiency.The experiment was divided into control group,control siRNA transfected group,and BG transfected group.Cell survival was detected by clonogenic assay,and the cell cycle distribution was determined by flow cytometry assay.The γ-H2AX foci formation after irradiation was visualized via immunofluorescence.Western blot assay was performed to detect the protein expressions of RAD50,p-P53,KU70,KU80,MDM2,CDK2 and RB.Results BG-siRNA transfection significantly reduced the BG transcription level (t =8.32-15.29,P ＜0.05) and increased cell survival after irradiation at 0.5,1,2,4 and 6 Gy.Analyzed with the multi-target model,the SERD0 of Beas-2B cells and control siRNA transfected cells were calculated to be 0.80 and 0.82,respectively.In addition,BG-siRNA transfection enhanced radiation-induced cell cycle arrest at G2 phase so that,after 4 Gy irradiation,the cells in G2 phase was increased from (37.37 ±0.63) ％ of control siRNA cells to (64.19 ± 1.01) ％ (t =30.65,P ＜ 0.05).Meanwhile,the γ-H2AX foci of BG-siRNA transfected cells was decreased from 76 ± 1.78 per 100 cells to 59-± 3.49 per 100 cells (t =13.72,P ＜0.05).The expressions of DNA damage related proteins including KU70,KUS0,CDK2 and RB were increased,but the expressions of p-P53 and RAD50 were decreased.Conclusions LncRNA-BG could regulate the radiosensitivity of the normal human bronchial epithelial cells,probably through inducing cell cycle G2 phase arrest and promoting DNA damage repair after irradiation.
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Objective To investigate the effect of X-ray irradiation on the expression of free ubiquitin in the serum, small intestine, and heart tissues of C57 BL/6N mice. Methods The amount of free ubiquitin protein in the serum and tissue homogenates was analyzed quantitatively with ELISA and Western Blotting assay. The mRNA expressions of free ubiquitin in the tissues were detected by RT-PCR. Results At 24 or 48 h after radiation, the free ubiquitin level in the serum and small intestine tissue increased asymptotically with increasing of radiation dose (F=183?1, 435?3, P 0?05). Conclusions Because of the high expressions of free ubiquitin protein in the radiosensitive mice tissues, X-ray radiation could increase the concentration of free ubiquitin in serum. The changes of free ubiquitin may be related to cellular radiosensitivity and tissue injury.
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Objective To study the effect of sanguinarine on growth and radiosensitivity of human glioma A172 cells.Methods MTT assay was used to evaluate cell growth.Cell cycle analysis and reactive oxygen species(ROS) burst were performed by flow cytometry assay.Annexin V/PI assay was used to detect cell apoptosis.Colony formation assay was used to detect the influence of sanguinarine on cell radiosensitivity.Results Exposure of A172 cells to sanguinarine led to dose-and time-dependent cytotoxicity with IC50 values of 4.8,3.9 and 3.2 μmol/L for 12,24 and 48 h,respectively.Furthermore,sanguinarine caused an arrest of S phase.After being treated with 5 μmol/L sanguinarine for 24 h,the ratios of early apoptosis,late apoptosis and necrosis were (60.01 ± 3.73)％,(2.70 ± 0.18)％ and (3.93 ± 0.76)％ respectively in A172 cells compared with the untreated control(t =55.28,8.32,9.51,P ＜0.05).An increased generation of ROS was found after treatment with sanguinarine,however,NAC inhibited the effect of sanguinarine.As analyzed with multi-target click model fitting curves,the SERD0 of sanguinarine-treated cells was 1.48.Conclusions Sanguinarine inhibits the A172 cells growth via apoptosis induction and ROS burst.Moreover,sanguinarine at a non-cytotoxicity dose increases cell sensitivity to X-rays.
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Objective To explore the influence of honokiol on growth,cell cycle and radiosensitivity in nasopharyngeal carcinoma cells,CNE-1 and CNE-2.Methods MTT assay and clonogenic assay were used to detect cell growth and survival respectively.Flow cytometry was employed to analyze cell cycle progression.Annexin V-FITC kit was used to detect cell apoptosis.Western blot assay was applied to examine protein expression.Results Honokiol signficantly inhibited proliferation of CNE-1 and CNE-2 cells in a dose and time dependent manner,the IC50 value was 2.84 and 2.68 μmol/L(24 h)and 2.50 and 2.20 μmol/L (48 h),respectively.After being treated with 2.5 μmol/L honokiol for 24 h,the ratios of early apoptosis,late apoptosis and necrosis were 24.53％,23.05％ and 7.13％ in CNE-1 cells compared with the control group(t =-41.17,-8.18,-6.08,P ＜0.05).The expression levels of pro-apoptotic proteins Caspase-3 and Bax were significantly increased to 2.31 and 1.89 times (t =-15.92,-17.15,P ＜ 0.05),4.43 and 1.85 times (t =-29.39,-13.47,P ＜ 0.05).Simultaneously the expression level of anti-apoptotic protein Bcl-2 was reduced by 2.22 and 2.74 times(t =26.94,66.14,P ＜ 0.05) as compared with controls after being treated with 4 and 3 μmol/L honokiol.Additionally,honokiol at lower doses signicantly enhanced the senstivity of CNE-1 and CNE-2 cells to X-ray irradiation.The SER was 1.41 and 1.88 in CNE-1 and CNE-2 cells.3 Gy irradiation of X-rays increased the proportion at G2/M state in both cell lines (t =-14.96,-19.26,P ＜ 0.05).Honokiol reduced the G2/M cell cycle arrest induced by irradiation significantly (t =7.65,4.98,P ＜ 0.05).Simultaneously,cyclin B1 protein expression obviously elevated (t =-33.07,-73.49,P ＜ 0.05).Conclusions Honokiol is a potent inhibitor of nasopharyngeal carcinoma cell growth by inducing cell apoptosis and necrosis and works as a radiosensitizer by disrupting G2/M cell cycle checkpoint.
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Objective To investigate the effect of ethanol on radiosensitivity of human breast cancer MCF-7 cells.Methods Human breast cancer MCF-7 cells were divided into four groups including control group,ethanol treatment group,X-ray exposed group,and ethanol combined with X-ray group.Clonogenic assay was used to determine cell survival.Flow cytometry was employed to analyze cell cycle progression.Annexin V-FITC kit was used to determine cell apoptosis induction.Results Ethanol(50 and 100 mmol/L,50 h)had no influence on MCF-7 cell growth( t =0.82,1.15,P ＞0.05 ).The radiosensitivity of MCF-7 cells was reduced when the cells were pretreated with 50 mmol/L ethanol (t =4.15,P ＜0.05)and 100 mmol/L ethanol ( t =10.28,P ＜ 0.05 ) for 2 h. Compared with irradiation with X-ray alone,ethanol treatment decreased G2/M phase arrest(t =7.18,P ＜0.05) and sub-G1population(an indicator of apoptosis induction) ( t =5.39,P ＜ 0.05).A decrease of advanced and early apoptosis in the cells pretreated with ethanol was also confirmed by Annexin V-FITC apoptosis assay( t =4.86,7.59,P ＜ 0.05 ).Conclusions Ethanol causes radioresistance in human breast cancer MCF-7 cells,where the decreases of radiation-induced G2/M phase arrest and apoptosis may be involved.
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Objective To explore the effect of low calorie metabolism on the survival of HeLa cells exposed to X-rays,and the influence of starvation on the antioxidative factors in the blood of rats after irradiation.Methods MTT method was used to evaluate the impact of different concentration glucose on the proliferation of HeLa cells.Colony formation assay was employed to detect the influence of glucose ( 1,5,10 and 25 mmol/L) on radiosensitivity of HeLa cells. Flow cytometry assay was used to analyze distribution of cell cycle and apoptosis.60 male SD rats were randomly divided into 6 groups with 10 rats each.Rats in every two groups were fed ad libitum,fasted for 24 h and fasted for 48 h,respectively.Rats in one group of each approach were respectively exposed to whole-body X-rays at 11 Gy. At 2 h after irradiation,all of rats were sacrificed and their venous blood was collected.Elisa kits were used to detect superoxide dismutase (SOD) and total antioxidant capacity (T-AOC).Results An increased viability was observed in HeLa cells treated with the glucose at low concentration ( ＜25 mmol/L),while HeLa cell growth was inhibited by glucose at doses of ＞25 mmol/L. Relevant to cells treated with 1 mmoL/L glucose,SERs (sensitive enhancement ratio) in cells exposed to 5,10 and 25 mmol/L glucose were 1.07,1.10 and 1.23,respectively. A reduction of G2/M and S arrests and apeptosis caused by 6 Gy X-ray irradiation were observed [(49.68 ±1.88)％ and (35.54±1.45)％ at G2/M phase,(16.88 ±1.22)％ and (10.23 ±1.65)％ atS phase,t=10.42,5.61,P＜0.05]and in the cells treated with 1 mmol/L glucose compared with cells treated with 25 mmol/L glucose [ ( 25.50 ± 0.95 ) ％ and (7.56 ± 1.07 ) ％,t =21.72,P ＜0.05 ].Without irradiation,calorie restriction exhibited a negligible influence on SOD and T-AOC in rats.However,after 11 Gy irradiation,compared with rats fed ad libitum,the levels of SOD and T-AOC were significantly increased in rats with calorie restriction ( t =40.32,42.78, P ＜ 0.05 ).Conclusions Calorie restriction has a certain radioprotective effect in vivo and in vitro.
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Objective To investigate the effect of EGCG on the radiosensitivity of human nasopharyngeal carcinoma CNE-1 cells.Methods CNE-1 cells were divided into four groups:control,EGCG treatment,UVC or X-ray exposure,and EGCG combined with UVC or X-rays.After treatment with different concentrations of EGCG for 24,48 and 72 h and UVC or X-rays,cell growth was determined with MTT assay,cell survival was measured with clonogenic assay,cell cycle was deteeted with flow cytometry,cell apoptosis was detected by the annexin V-FITC cell apoptosis kit,and protein expression was assayed by Westem blot.Results EGCG inhibited cell growth in a dose-and time-dependent manner(r =0.817and 0.364).Compared with UVC or X-ray irradiation alone,the radiosensitivity of CNE-1 cells was enhanced by 2 h pre-treatment of 50 μmol/L EGCG,which disrupted S phase arrest caused by UVC( t =18.68,P ＜ 0.05 ) and increased the population of S and G2/M arrest caused by X-rays ( t =7.11 and 6.99,P ＜0.05 ).UVC could cause a significant increase of sub-G1 population( t =6.67,P ＜ 0.05 ) and Annexin V-FITC assay indicated apoptosis was further elevated by EGCG ( t =10.28,P ＜ 0.05 ).However,no significant induction of apoptosis was observed in the cells either irradiated with X-rays alone or combinationly treated with EGCG and X-rays.The combination treatment of EGCG and UVC significantly increased the expression of Bax and Caspase-3 proteins,but failed to affect Bcl-2 protein expression.Conclusions EGCG enhances the growth inhibition of CNE-1 cells caused by UVC or X-rays,which is relevant to apoptosis induction or cell cycle arrest.
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Objective To explore the effect of ionizing radiation and cisplatin on the expression of Transducer of erbB2,1 ( TOB1 ) in human lung adenocareinoma SPCA-1 and LTEP-α-2 cell lines.Methods SPCA-1 and/or LTEP-α-2 cells were respectively irradiated with 2,4,6,8,and 10 Gy X-rays generated by a linear accelerator (with the source skin distance of 100 cm and dose rate of 200 cGy/min).The cell molecular samples were subtracted at 6,12,18,and 24 h after 6 Gy irradiation.For X-rays and cisplatin combination treatment,cells were divided into radiation group (6 Gy X-rays),cisplatin group (20 mol/L cisplatin),and combination group (6 Gy X-rays + 20 mol/L cisplatin).SPCA-1 and LTEP-α-2 cells without any treatment were used as control group.The relative levels of TOB1 mRNA and protein expression were detected by rigorously controlled semi-quantitative RT-PCR and Western blot analysis with quantitation of the mRNA/protein bands by densitometry.Results The expression level of TOB1 was significantly increased in both mRNA and protein level after radiation exposure (t =8.25-24.48,P ＜0.05).For time-dependent induction of TOB1,the expression was significantly increased 6 h after X-rays exposure (t =14.23-15.82,P ＜ 0.05 ).More significant increase of TOB1 expression was identified after the combination treatment of X-rays and cisplatin,compared to that of the radiation and/or cisplatin treatment alone (t =11.21-13.67,P ＜0.05).Conclusions lonizing radiation and cisplatin could upregulate the expression of TOB1 in lung cancer cells in both mRNA and protein levels.TOB1 may be a molecular target for ionizing radiation and cisplatin treatment in lung cancer and it may have potential implication in evaluating the curative efficiency of chemo-and radio-therapy.
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Objective To study the impacts of berberine on the growth, migration and radiosensitivity in human breast cancer cells.Methods MTT assay was used to evaluate cell growth.In vitro scratch migration assay was used to determine cell migration.Annexin V assay was used to detect cell apoptosis.The distribution of cell cycle was evaluated by flow cytometry assay.Colony formation assay was used to detect the influence of berberine on cell radiosensitivity. Western blot assay was employed to measure protein expression.Results Berberine inhibited cell growth and migration in two human breast cancer cell lines, MCF-7 and MDA-MB-231, in a dose-and time-dependent manner. Furthermore,berberine resulted in a cell cycle G0/G1 arrest.Compared with control,the early apoptosis in MDA-MB-231 and MCF-7 cells treated with 40 pμmol/L of berberine was as high as 86.6％ and 66.6％ (t =8.79,10.32,P ＜ 0.01 ),respectively. Berberine caused a dose-dependent increase in Bax and Caspase-3 protein expressions,but did not change Cyclin D1 protein expression,while suppressed the expressions of Cyclin B1 and Bcl-2 protein. As analyzed with multi-target click model fitting curves,the SERD0 of berberine-treated cells were 1.12 and 1.22 for MDA-MB-231 and MCF-7cells respectively at the dose D0 of X-rays.Conclusions The berberine inhibited the growth and migration of breast cancer cells via apoptosis induction and cell cycle arrest.Moreover,berberine increases cell sensitivity to X-ray irradiation.
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Objective To study the effect of sanguinarine on the growth and radiosensitivity of ovarian cancer SK-OV-3 cells.Methods Cell growth was determined by MTT and clonogenic assay.Cell cycle analysis was performed by flow cytometry assay.The cell apoptosis was analyzed by Annexin V/PI assay.Results Sanguinarine inhibited SK-OV-3 cell growth in a dose-and time-dependent fashion and its IC50 values were 3.02 and 1.11 μmol/L at 24 and 48 h,respectively. Sanguinarine also significantly triggered a sub-G1 peak,an indicator of apoptosis,and caused a G0/G1 arrest.Furthermore,the cell apoptosis induced by X-irradiation was significantly increased at 6 Gy when the cells were pre-treated with sanguinarine,in which the early apoptotic population increased from 10.28％ to 43.28％ (t =19.41,P ＜0.01 ) and the late apoptotic population increased from 20.26％ to 30.80％ ( t =8.78,P ＜ 0.01 ).The multi-target click model was used to fit survival curves and the SER of sanguinarine treatment approached to 1.625 at the dose of D0. Conclusions Sanguinarine could inhibit SK-OV-3 cell growth by inducing apoptosis and cell cycle arrest and enhance cell radiosensitivity at low doses.
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Objective To investigate the probability of high mobility group box 1 protein as a biological dosimeter of ionizing radiation. Methods The medium of cultured human fibroblast cell line GM was collected 24 h after exposed to 60Co γ-rays, and HMGB1 protein concentration was detected by ELISA assay, and the dose-effect curve was then fitted. The levels of HMGB1 protein was also detected after exposed to 4 Gy irradiation at 24,48 and 72 h. Results The level of HMGB1 protein in culture medium was increased in a dose-dependant way, and the dose-effect curve of HMGB1 level for 24 h after irradiation fitted the linear model y = 0.5655 + 0.0358x (r = 0. 9339) . After exposure to 4 Gy irradiation, the levels of HMGB1 protein in the culture medium were also increased time-dependently.Conclusion The release of HMGB1 protein to neoplasm is reactivated by irradiation in GM cell. It is necessary to further the studies on HMGB1 as biological dosimeter for radiation injury protection and therapy.