1.Study on the mechanism of curcumin on the apoptosis and radiosensitivity of human chordoma cell line CM-319
Jie YAO ; Li QI ; Xuejian WU
Chinese Journal of Orthopaedics 2021;41(1):33-42
Objective:To investigate the effects of curcumin on apoptosis and radiosensitivity of human chordoma cell line CM-319 and its mechanism.Methods:CM-319 cells were cultured in vitro and divided into curcumin 0 μmol/L group, curcumin 5 μmol/L group, curcumin 10 μmol/L group, curcumin 20 μmol/L group, si-NC group, si-UHRF1 group, curcumin+pcDNA group, curcumin+pcDNA-UHRF1 group.Flow cytometry was usedtodetect cell apoptosis, Western Blot was used to detectthe protein expression of Bcl-2, Bax, Cleaved Caspase-3 and UHRF1, qRT-PCR was used to detect the expression of UHRF1 mRNA, and clone formationexperiment was used to detect the sensitivity of CM-319 cells to radiation.Results:Compared with the curcumin 0 μmol/L group, the apoptosis rate of CM-319 cellsinthe curcumin 5 μmol/L group, curcumin 10 μmol/L group and curcumin 20 μmol/L groupincreased 12.61%±1.57%, 18.42%±1.54%, 29.37%±2.65% vs 7.24%±0.73% ( t=9.305, 19.680, 24.153; all P<0.05), and the protein expression of Bcl-2 decreased 0.62±0.06, 0.51±0.05, 0.33±0.03 vs 0.76±0.07 ( t=4.556, 8.719, 16.939; all P<0.05), while the protein expression of Bax 0.41±0.04, 0.55±0.05, 0.69±0.06 vs 0.26±0.03 ( t=9.000, 14.920, 19.230; all P<0.05) and Cleaved Caspase-3 0.37±0.04, 0.49±0.05, 0.62±0.06 vs 0.24±0.03 ( t=7.800, 12.862, 16.994; all P<0.05) increased, and the expression of UHRF1 mRNA decreased 0.82±0.08, 0.67±0.07, 0.42±0.04 vs 1.01±0.09 ( t=4.734, 8.946, 17.972; all P<0.05), the protein expression of UHRF1 decreased 0.61±0.06, 0.48±0.05, 0.31±0.03 vs 0.83±0.08 ( t=6.600, 11.130, 18.258; all P<0.05), and were dose-dependent, and the sensitization ratios were 1.433, 1.708, and 2.183, respectively. Compared with the si-NC group, the protein expression of UHRF1in CM-319 cells in the si-UHRF1 group decreased 0.35±0.04 vs 0.79±0.08 ( t=14.758, P<0.05), the apoptosis rate of CM-319 cells increased 21.49%±2.11% vs 8.24%±0.83% ( t=17.531, P<0.05), the protein expression of Bcl-2 decreased 0.34±0.03 vs 0.71±0.07 ( t=14.575, P<0.05), while the protein expression of Bax 0.62±0.06 vs 0.25±0.03 ( t=16.547, P<0.05) and Cleaved Caspase-3 0.58±0.05 vs 0.23±0.03 ( t=18.007, P<0.05) increased, and the sensitization ratio was 1.727. Compared with the curcumin+pcDNA group, the apoptosis rate of CM-319 cells in the curcumin+pcDNA-UHRF1 groupreduced 13.59%±1.25% vs 28.31%±3.01% ( t=13.549, P<0.05), the protein expression of Bcl-2 increased 0.63±0.06 vs 0.31±0.03 ( t=14.311, P<0.05), while the protein expression of Bax 0.42±0.04 vs 0.71±0.07 ( t=10.791, P<0.05) and Cleaved Caspase-3 0.38±0.04 vs 0.65±0.05 ( t=12.650, P<0.05) decreased, and the sensitization ratio was 0.539. Conclusion:Curcumin can induce apoptosis of CM-319 cells and enhance the radiosensitivity of CM-319 cells, , and the mechanism is related to the down-regulation of UHRF1 expression.
2.Diagnostic significance of urethral sphincter electromyography and external anal sphincter electromyography in patients with multiple system atrophy
Feng QIU ; Xiaokun QI ; Liping LI ; Jianguo LIU ; Wei YAO
Chinese Journal of Neurology 2012;45(3):188-191
Objective To assess the significance of urethral sphincter electromyography (US-EMG) and external anal sphincter electromyography (EAS-EMG) for the diagnosis of multiple system atrophy (MSA).Methods US-EMG and EAS-EMG were performed in 9 patients who were diagnosed as MSA.Duration,motor unit action potentials amplitude,polyphasicity,as well as recruited pattern and amplitude during powerful contraction were recorded and analyzed.Results Among 9 patients who were diagnosed as MSA,7 cases showed neural injury by both US-EMG and EAS-EMG.There was significant difference of electromyographic findings between US-EMG group and EAS-EMG group (average volatility (μV):1063.44 ±499.92 vs 634.89 ±265.07; polyphasic wave:0(0,20% ) vs 57% (28%,63% ) ; t =2.567,P=0.033;t =2.833,P=0.012).Conclusions Although US-EMG may be difficult to perform,US-EMG may have the same specificity as EAS-EMG for the diagnosis of MSA,especially for the diagnosis of MSA patients only with urination disorders,who are involved in Onuf neclear according to some of the abnormal indexes.
3.The diagnostic value of urethral sphincter electromyography in patients with multiple system atrophy
Xiaokun QI ; Liping LI ; Wei YAO ; Jianguo LIU ; Feng QIU
Chinese Journal of Internal Medicine 2012;(12):975-977
Objective To analyze diagnostic evaluation of urethral sphincter electromyography (US-EMGs) for patients with multiple system atrophy (MSA).Methods Totally 15 patients who were diagnosed as MSA were examined as treatment group while 17 non-MSA patients were examined as controls.US-EMGs were performed in the both groups.Spontaneous activities when relax,parameters of motor unit potentials(MUPs) mean duration and amplitude,percentage of polyphasic ware,satellite potential,recruitment potentials and amplitude when strong contraction were recorded and analyzed.Results USEMGs changes of various abnormalities were found in 13 cases (86.7%) in MSA group.There were significant differences of electromyographic findings between the MSA group and control group including MUPs mean duration[(12.79 ±3.18)ms vs (9.49 ± 1.51)ms] and amplitude[(828.53 ±459.89) μV vs (378.76 ± 152.26) μV] as well as recruitment potentials [(11.47 ± 21.55) % vs (8.23 ± 10.74) %] and amplitude [(2.19 ± 1.24) mV vs (0.75 ± 0.42) mV] when strong contraction (all P values < 0.05).Conclusions There is certain value of US-EMGs for the diagnosis of MSA.It could be used as a routine electrophysiological method for the patients who are suspected of MSA.It could be a supplement of externalanal sphincter electromyography.
4.Expression of E-cadherin and β-catenin in primary colorectal carcinoma and metastatic lymph nodes and it's clinical significance
Jianzhi CHEN ; Xuan LI ; Qi YAO ; Xiuhuai CAO
Chinese Journal of Primary Medicine and Pharmacy 2011;18(15):2022-2024
ObjectiveTo investigate the expression of E-cadherin and β-catenin in the primary colorectal carcinoma and metastatic lymph nodes and it's clinical significance. MethodsImmunohistochemical analysis of Ecadherin and β-catenin was performed in colorectal mucosa( n =20 ,group A) ,colorectal carcinoma without metastasis ( n = 25, group B) and colorectal carcinoma with metastatic lymph nodes ( n = 39, group C) The expression of E-cadherin and β-catenin and the metastasis and prognosis of colorectal carcinoma were observed. ResultsThe strongly positive expression of E-cadherin in primaries of group C (5.1% ) was obviously lower than in group B ( U = 2. 149,P =0. 032) ,whereas no statistic differences of the expression of E-cadherin between primaries and metastases of group C were observed(P =0.186). There was no difference of the expression of β-catenin between group B and group C,the same of primary colorectal carcinoma and self control lymph nodes of group C(P =0.235 and P =0. 715). There was positive correlation between membranous down regulation of E-cadherin and β-catenin in group C( r = 0.387 ,P =0.024) ,but no relationship was found in group B( r = 0.080 ,P = 0.723 ). ConclusionThe decreased membranous expression of E-cadherin in the colorectal carcinoma was correlated with lymph nodes metastasis, and was obviously related to membranous down regulation of β-catenin. There was no relevance between the expression of β-catenin and the lymph nodes metastasis of colorectal carcinoma.
5.Expression and signiifcance of PLTP and MIF in mice with bronchopulmonary dysplasia
Guifang LI ; Baohai SHI ; Guo YAO ; Junli LIU ; Chunhua QI
Journal of Clinical Pediatrics 2013;(9):876-879
Objective To study the expression and signiifcance of phospholipid transfer protein (PLTP) and macrophage migration inhibitory factor (MIF) in mice with bronchopulmonary dysplasia (BPD). Methods Ninety-six 4-day-old mice were randomly divided into oxygen group and air group. Mice in oxygen group were exposed to a FiO2 of 65%, and mice in air group were exposed to air. On day 7, 14, 21 and 28, blood and lung tissue samples from 12 randomly selected mice in each group were obtained. The serum levels of MIF and PLTP were measured by ELISA assay. The morphological changes of lung tissue were ob-served with HE staining. Results The mice in oxygen group showed thickened lung parenchyma and obvious pulmonary ifbrosis. The radioactive alveolar count was signiifcantly lower in oxygen group than that in air group (P<0.01). PLTP level in air group was increased gradually from day 7 to day 21, and began to decrease on day 28. PLTP level in oxygen group was increased from day 7 to day 14, and decreased on day 21 and day 28. MIF level in air group did not change during the experiment. MIF level in oxygen group was signiifcantly increased from day 7 to day 21, and began to decrease on day 28. Conclusions MIF and PLTP may be good biomarkers for the diagnosis of BPD.
6.Epidemiological survey on bronchial asthma in remission stage of Liaoning province
Qi WANG ; Yinghai CUI ; Ming YAO ; Guoxin LI ; Xuefeng YU
International Journal of Traditional Chinese Medicine 2013;35(12):1057-1058
Objective To survey the epidemiological situation of asthma in remission stage in Liaoning province,and provide evidence for preventing and treating asthma.Methods 800 patients of asthma in the remission stage were investigated through the questionnaire investigation.Results Onset age of asthma in Liaoning province was 30~40 years old.The sex ratio between male and female was 1 ∶ 1.34.The number of staff was the highest (25.75%).Most of the course of asthma was 1~5 years (33.50%).The number of patients who often receive treatments was 59 (7.38%)and the number of patients with allergy history was 531 (66.38%).Conclusion Prevention,management and treatment of asthma should be strengthened.
7.Anti-apopttoic effect of resveratrol on free fatty acid-i nduced human hepatic L02 cells and its mechanisms
Qi YAO ; Meiling LI ; Mengyuan CHEN ; Jianguo DAI
Journal of Medical Postgraduates 2015;(8):804-808
Objective Resveratrol can improve nonalcoholic fatty liver disease , but its action mechanisms remain unclear . This study aimed to investigate the protective effect of resveratrol against the free fatty acid ( FFA)-induced apoptosis of human hepatic L02 cells and its possible mechanisms . Methods Human hepatic L02 cells were incubated with FFA and resveratrol for 24 hours.The prepared cells were divided into a blank control , an FFA ( 2 mmol/L) , and a resveratrol group ( 50 μmol/L resveratrol +2 mmol L/FFA).After treatment, we measured the triglyceride (TG), glutathi-one (GSH), and malonaldchyde (MDA) contents and caspase3 ac-tivity in the hepatocytes , determined the apoptosis of the cells by flow cytometry , and detected the protein expression of silent information regulator 1 (SIRT1) by Western blot as well as the mRNA expressions of catalase (CAT), Mn superoxide dismucase (MnSOD), Bcl-2, and Bax by qRT-PCR. Results The TG content and caspase 3 activity in the hepatocytes were significantly increased in the FFA ([3518.±64.2] μmol/L and [5.97 ±0.78] U/g) and the resveratrol group ([201.1 ±60.1] μmol/L and [3.60 ±0.73] U/g) as compared with those of the blank control ([40.2 ±7.4] μmol/L and [2.56 ±0.49] U/g) (both P<0.05), but the caspase3 ac-tivity was markedly decreased in the resveratrol group in comparison with that of the FFA group (P<0.05).Both early and late apop-tosis rates of the hepatocytes were remarkably higher in the FFA ([6.75 ±0.81]%and [8.52 ±0.54]%) and the resveratrol group ([4.94 ±0.44]%and [6.52 ±0.61]%) than those in the blank control ([3.38 ±0.33]% and [2.72 ±0.19]%) ( both P<0.05), with statistically significant differences between the former two groups (P<0.05).The resveratrol group showed significant differences in the GSH content ([100.2 ±8.8] nmol/g), the MDA level ([2.36 ±0.82] mg/g), and the relative expression of SIRT1 (0.84 ±0.04) from the FFA group ([73.8 ±13.1] nmol/g, [3.77 ±0.92] mg/g, and 0.61 ±0.07) and the control ([113.7 ±13.8] nmol/g, [1.85 ±0.41] mg/g, and 0.90 ±0.02) (all P<0.05).The resveratrol group also exhibited statistically significant differences in the relative expressions of the MnSOD , CAT, and Bax genes from the FFA and control groups (P<0.05). Conclusion Resveratrol attenuates FFA-induced apoptosis of human hepatic L 02 cells by activating SIRT1 and reducing the oxidative stress of hepatocytes .
8.Early effects of iodine excess on spleen cells of methallothionein Ⅰ/Ⅱ knockout mice
Lingyan WANG ; Na ZHANG ; Yongmei LI ; Qi DUAN ; Xiaomei YAO
Chinese Journal of Endemiology 2015;34(3):168-171
Objective To investigate the effects of iodine excess on spleen cell viability,lactate dehydrogenase (LDH) leakage,mitochondrial superoxide production and peroxiredoxin (Prx)3 expression in methallothionein Ⅰ / Ⅱ knockout (MT-Ⅰ / Ⅱ KO)mice.Methods Spleen cell suspensions were prepared from six to eight-week old and healthy male MT-Ⅰ / Ⅱ KO mice and wild type (WT) mice; the cell number was adjusted to 5 × 107/L and the cells were plated in 96-well plates (100 μl each well); the cells were exposed to various concentrations of KI (0,10-4,10-3,10-2 mol/L) and 10-3 mol/L H2O2,respectively,for two hours,and control group did not give KI nor H2O2.Cell viability was assayed by methyl thiazolyl tetrazolium (MTT) colorimetric method.Cell damage was detected by chemical colorimetric method.Mitochondrial superoxide production in the spleen cells was measured by flow cytometry.Western blotting technology was used to investigate the expression of Prx3.Results In both MT-Ⅰ/Ⅱ KO and WT mice,the differences of cell viability,LDH leakage,mitochondrial superoxideproduction and the expression of Prx3 of spleen cells among the treatment groups were statistically significant (F =357.92,71.03,130.36,10.36,179.58,26.92,187.43,and 7.16,all P < 0.05).Compared to the control group [(100.00 ± 2.00)%,(100.00 ± 1.63)%,(3 202.22 ± 85.63),(3 161.51 ± 144.49)U/L,43.82 ± 1.56,38.60 ± 2.81,0.61 ± 0.09,0.50 ± 0.08],cell viability of 10-4,10-3,10-2 mol/L KI treatment and 10-3 mol/L H2O2 groups [(80.77 ± 1.86)%,(89.89 ± 2.90)%,(76.08 ± 1.92)%,(87.66 ± 1.74),(73.26 ± 1.86)%,(84.30 ± 2.23)%,(66.22 ± 1.71)%,(70.80 ± 1.49)%] was decreased (all P < 0.05); LDH leakage [(3 880.00 ± 190.62),(3 431.17 ± 170.45),(4 178.33 ± 170.43),(3 598.63 ± 189.09),(4 388.61 ± 123.79),(3 863.72 ± 195.64),(4 615.28 ± 196.17),(4 148.12 ± 195.81)U/L] was increased significantly (all P< 0.05); and mitochondrial superoxide production in the spleen cells (53.83 ± 3.22,47.03 ± 1.60,58.92 ± 4.00,50.48 ± 2.59,72.72 ± 2.14,68.53 ± 2.97,80.76 ± 4.11,75.26 ± 3.41) was increased significantly (all P < 0.05); Prx3 expressions in 10-3、10-2 mol/LKI and 10-3 mol/L H2O2 treatment groups (0.82 ± 0.12,0.65 ± 0.12,0.96 ± 0.15,0.73 ± 0.16,1.04 ± 0.13,0.85 ± 0.16) significantly increased (all P < 0.05),the differences of Prx3 expressions between 104 mol/L KI groups (0.73 ± 0.15,0.55 ± 0.09),and control groups were not statistically significant (all P > 0.05).In 104,10-3,10-2 mol/L KI and 10-3 mol/L H2O2 treatment groups,cell viability of MT-Ⅰ/Ⅱ KO mice spleen was lower than that of WT mice (t =6.47,10.93,9.30 and 4.96,all P < 0.05); LDH leakage was higher than that of WT mice (t =4.30,5.58,5.56 and 4.13,all P < 0.05); mitochondria superoxide production was higher than that of WT mice (t =4.64,4.33,2.80 and 2.52,all P < 0.05); Prx3 expression was higher than that of WT mice (t =2.54,2.37,2.59 and 2.27,all P < 0.05).Conclusions KI may decline the cell viability,increase the leakage of LDH and increase the production of mitochondrial superoxide production and Prx 3 expression,which are much more significant in MT-Ⅰ /Ⅱ KO mice,suggesting that MT Ⅰ /Ⅱ has some antioxidative effect in high concentration of iodide induced oxidative stress in the spleen.
9.Chinese Medicine and Western Medicine Prevention the Anaphylaxis of Ionic-type Radiography Agent in Brain Enhancement CT
Qi WANG ; Shengguo YIN ; Shuxin CHEN ; Li ZHANG ; Min YAO
Journal of Practical Radiology 2001;0(08):-
Objective To study Chinese medicine and Western medicine preventing the anaphylaxis of ionic-type radiography agent,to solve the anaphylaxis in brain enhancment CT.Methods Eight hundred and forty cases have been diagnosed in brain enhancement CT at our hospital,from Fed.1995 to May 2000.Results Chinese medicine,ginger,pinellia and hormone medicine were all useful in preventing the anaphylaxis of ionic-type radiography agent in brain enhancement CT.Conclusion Chinese medicine:ginger and pinellia could prevent the anaphylaxis of ionic-type radiography agent.
10.Research Progress of the Relationship between the Expression of TAR DNA-bind-ing Domain Protein 43 and Brain Injury
Yao YE ; Rubo LI ; Shiyu MA ; Xueting WEI ; Qi XU
Journal of Forensic Medicine 2017;33(3):289-292
T A R D N A-binding dom ain protein 43 (T D P-43) is a highly conserved and w idely expressed nuclear protein. N ow adays, the expression of T D P-43 can be found in m ost neurodegenerative diseases such as A lzheim er's disease, w hich m akes it becom e a neurodegenerative disease associated m arker pro-tein. From the current research status at hom eland and abroad, and around the relationship betw een the expression of T D P-43 and brain injury, this article em phatically probes into the specific expression and function of T D P-43 in acute and chronic brain injury based on the know ledge of its biological charac-teristics, w hich aim s to explore the feasibility for determ ining the cause of death and the injury and dis-ability situations by T D P-43 in forensic pathology.