1.Effect of epidermal growth factor receptor inhibitor AG1478 induces cell death in HeLa cells
Congling MA ; Yiping YIN ; Tingting MAO
The Journal of Practical Medicine 2017;33(2):202-205
Objective To investigate the biological features of epidermal growth factor receptor inhibitor AG1478 on cervical carcinoma cell. Methods The proliferation of HeLa cells under AG1478 stimulation was determined by CCK8 assay. The expression of EGFR downstream signaling protein and apoptotic relative protein were examined by Western blot and transcription of apotosis?related genes were measured by RT?qPCR in AG 1478 treated HeLa cells. Nuclear transport of phosphorylated ERK were measured through ICC assay. TUNEL assay was used to determine early stage of apoptosis. Results CCK8 assay showed that AG1478 inhibit the proliferation of HeLa cells and also block phosphorylated level of EGFR ,ERK and AKT. Furthermore ,nuclear transport of phosphorylated ERK upon EGF stimulation were blocked and pro?apoptotic proteins were up?regluated with activat ed cleaved Caspases. Conclusion AG1478 inhibited the proliferation and induced apoptosis in HeLa cells and it could be a potential therapeutic drug for cervical carcinoma.
2.Neuroprotective effect of PTEN inhibitor BPV on cerebral ischemia-reperfusion injury in rats and its mechanism
Lunlin MAO ; Donglin HAO ; Xiaowei MAO ; Tingting HUANG ; Bona WU
International Journal of Cerebrovascular Diseases 2015;(9):691-698
Objective To investigated the neuroprotective effect of PTEN inhibitor BPV on cerebral ischemia-reperfusion injury in rats and its mechanism. Methods Male Sprague-Dawley rats were used to induce a reperfusion model of middle cerebral artery occlusion for 1 h. During the reperfusion, the BPV solution (0. 2 mg/kg daily) or the equal volume of saline was injected intraperitonealy immediately. The neurological deficit scores were conducted at day 1, 3,5, and 7 after ischemia-reperfusion. At day 4, triphenyl tetrazolium chloride staining was used to assess cerebral infarction volume. Enzyme-linked immunosorbent assay was used to detect the levels of interleukin 10 (IL-10) and tumor necrosis factor α(TNF-α) in cortical ischemic border zones. Real-time quantitative polymerase chain reaction was used to detect the expression level of PTEN mRNA. Western blotting was used to detect the expression levels of PI3K, Akt, and p-GSK-3β. At day 7, Bielschowsky silver staining was used to detect the axonal distribution in the ischemic border zone of the striatum. Immunohistochemical staining was used to detect the expression of myelin basic protein (MBP). Results At day 4 after ischemia-reperfusion, the infarct volume (32. 27% ± 1. 71% vs. 45. 49% ± 2. 12% ; P < 0. 001), TNF-α concentration in the cortical ischemic border zones (134. 17 ± 10. 38 pg/ml vs. 264. 17 ± 24. 84 pg/ml; P < ), and PTEN mRNA level (1. 19 ± 0. 08 vs. 2. 50 ± 0. 06; P < 0. 001) in the rats of the BPV group were al significantly lower than those of the normal saline group. The IL-10 concentration (186. 83 ± 10. 83 pg/ml vs. 147. 83 ± 11. 62 pg/ml; P < 0. 001), and the expression levels of PI3K (0. 43 ± 0. 08 vs. 0. 26 ± 0. 06; P = 0. 004), Akt (0. 52 ± 0. 05 vs. 0. 40 ± 0. 04;P = 0. 001), and p-GSK-3β (0. 75 ± 0. 08 vs. 0. 38 ± 0. 06; P < 0. 001) were al significantly higher than those of the normal saline group. At day 7 after ischemia-reperfusion, the neurological deficit score (4. 83 ± 0. 41 vs. 6. 33 ± 0. 52; P < 0. 001) in the rats of the BPV group was significantly lower than that of the normal saline group. The axon densities in the ischemic border zones (35. 51% ± 2. 45% vs. 25. 31% ± 2. 79% ; P < 0. 001) and the expression level of MBP (32. 56% ± 3. 46% vs. 27. 81% ± 4. 18% ; P = 0. 037) were significantly higher than those of the normal saline group. Conclusions BPV has neuroprotective effect for cerebral ischemia-reperfusion injury in rats. Its mechanism may be associated with the up-regulation of PTEN downstream proteins PI3K, Akt and p-GSK-3β expression to regulate inflammatory mediators and reduce the inflammatory response.
3.Study on the expression of vimentin during the rat tongue carcinogenesis induced by 4NQO
Tingting DENG ; Hong TAN ; Liang MAO ; Minhai NIE
Chongqing Medicine 2016;45(16):2194-2197
Objective To study the expression of vimentin in the tongue mucosa carcinogenesis and to explore its significance in the invasion and metastasis of tongue squamous cell carcinoma .Methods The occurrence of tongue squamous cell carcinoma in rat was induced by means of 4NQO water solution ,and 56 cases in total were collected in the cancerous process ,including normal tongue mucosa ,epithelial hyperplasia ,mild dysplasia ,moderate dysplasia ,severe dysplasia and the tongue tissue specimen of squa‐mous cell carcinoma .The immunohistochemistry was used to evaluate protein expression and real‐time fluorescent quantitative PCR method was used to obtain the expression quantity of mRNA .Results In immunohistochemistry ,with the increase of degree of rat tongue mucosa dysplasia ,the positive rate of vimentin expression increases obviously .The difference between groups was statistical‐ly significant (χ2 =10 .685 ,P<0 .05) .Lesion groups compared with normal group ,their mRNA expression differences all hold sta‐tistical significance(P<0 .05);Mild dysplasia ,moderate dysplasia ,severe dysplasia and squamous cell carcinoma groups were com‐pared with epithelial hyperplasia group .The difference between squamous cell carcinoma group and epithelial hyperplasia group was statistically significant (P<0 .05) .mRNA expression of epithelial hyperplasia ,mild dysplasia ,moderate dysplasia and severe dys‐plasia were respectively 1 .22 times ,1 .28 times ,1 .29 times and 1 .42 times of that of the normal group .Conclusion During rat tongue carcinogenesis induced by 4NQO ,the expression of vimentin was increased with the increase of the degree of pathological change ,which is closely related to the invasion of tumor and could be regarded as a predictor of tongue squamous cell carcinoma .
4.MRI in the differential diagnosis of breast ductal cancer in situ
Chao YOU ; Yajia GU ; Weijun PENG ; Jian MAO ; Tingting JIANG
China Oncology 2014;(6):463-468
Background and purpose: Ductal carcinoma in situ (DCIS), is precursor lesions of invasive breast cancer, belongs to non-systemic ductal lesions, which is similar to other ductal lesions on imaging. This study aimed to investigate the differential diagnostic value of MRI in DCIS between DCIS with microinvasion (DCIS-MI) and breast intraductal papilloma (BIDP). Methods:All the cases were surgico-pathologically conifrmed. Twenty-four patients were DCIS, 9 patients were DCIS-MI, and 20 patients were BIDP. The MRI charateristics of DCIS, DCIS-MI and BIDP were analyzed and compared. Results:DCIS and DCIS-MI’s performance in the form of MRI is dififcult to differentiate (P<0.05). Compared with BIDP, the ductal and segmental enhancement, typeⅢtime-signal intensity curve (TIC), and the red pseudo-color image were associated with the DCIS. The local area enhancement, typeⅡTIC, and no-red pseudo-color image were associated with BIDP. Conclusion:MRI is a useful for differentiate between DCIS and BIDP, but is dififcult for DCIS and DCIS-MI.
5.Clinical application of transplantation of autologous fat with stromal vascular fraction for breast augmentation
Lijun ZHENG ; Qiming ZHAO ; Dongsheng MAO ; Tingting SI
Chinese Journal of Medical Aesthetics and Cosmetology 2015;21(6):331-333
Objective To explore the clinical effect of transplantation of autologous fat with stromal vascular fraction for breast augmentation.Methods From September 2012 to March 2014,15 people accepted breast augmentation by transplanting autologous fat with SVF under local anaesthesia.Three-dimensional computer tomography reconstruction (3D-CT reconstruction) was applied preoperatively and postoperatively,the data of the thickness of breast's subcutaneous tissue was compared to observe the breast contour improvement.Results Autologous fat was injected 2 times in 3 cases,and the rest were 1 time.6 months after operation,13 cases were satisfied with the results,and 2 cases were quite satisfied.3D-CT reconstruction was performed before operation and 6 months after operation;statistics showed that before operation the thickness of breast subcutaneous tissue of the right side was (11±4) mm,the left side was (11±6) mm;6 months after operation the right side was (14±3) mm,the left side was (16±3) mm (P>0.05).The breast was soft,no major complication such as nodules,cysts or calcification were seen.Conclusions The transplantation of SVF with autologous fat is a safe,effective procedure for breast augmentation.3D-CT reconstruction can show more details of breast structures,which is a more effective method to guide and evaluate the lipotrans plantation.
6.Effects of prenatal hypoxia on renin-angiotensin system in plasma and liver of offspring rats
Caiping MAO ; Tingting ZHONG ; Nan CHEN ; Shan JIANG ; Xiangwei ZHANG ; Li CAO
Chinese Pharmacological Bulletin 2016;32(7):975-979
Aim To study the effects of prenatal hy-poxia on the risk of fatty liver disease to search the drug targets .Methods Intrauterine hypoxia rats model was established .The bodies and livers of fetal rats of 21 days, and adult offspring rats of 5 months with and without anoxic treatment were all weighed .The liver index was calculated and the concentrations of renin-angiotensin system components in circulation system and livers of offspring rats were measured .Results The weight of the bodies , livers and index of liver weight to body weight ( liver index ) were significantly decreased in the PH group compared with the normal group.These differences disappeared in adulthood . However, the liver index of adult offsprings in the PH group after hypoxia stress for 7 days was significantly increased compared with that of adult offsprings in nor-mal group.There were no significant differences in the concentrations of AngⅠ, AngⅡ and ACE in plasma and livers between the two groups of fetal rats and the two groups of adult offspring rats separately .The con-centrations of AngⅡ in the livers of adult rats in PH group were significantly higher than those in normal group.The concentrations of AngⅠ in livers and the concentrations of AngⅡ in plasma and livers in the group treated with hypoxia stress for 7 days were signif-icantly higher than those without hypoxia stress .The concentrations of ACE in livers and the concentrations of AngⅡ in plasma and livers in PH adult offsprings were significantly higher than those of normal adult off-springs .Conclusion PH can induce the increase of RAS content in the livers of fetus and adult rats , RAS is more likely to be activated after hypoxia stimulation in the following adulthood .PH is a potential mecha-nism that mediates offspring susceptibility of fatty liver .
7.The changes of miR-126 and SPRED1 levels after transient ischemic attack and their effects on prognosis
Lidong DING ; Zhanghong XIAO ; Tingting ZHANG ; Huawu MAO ; Xiaofang HANG ; Xiaobo LU ; Qi FANG
Chinese Journal of Geriatrics 2017;36(8):858-863
Objective To investigate the effects of changes of miR-126 and spouty related EVH,domain containing proteinl(SPRED1) after transient ischemic attack(TIA)on prognostic value for pathogenesis of secondary cerebral infarction.Methods Retrospective analysis of the clinical data of 106 patients with TIA was performed.The expression levels of miR-126,SPRED1 and vascular endothelial growth factor(VEGF)in peripheral blood were detected at 3 h,6 h and 12 h after TIA onset respectively.The specificity and sensitivity of miR 126 and SPRED1 in the diagnosis of TIA were analyzed.The miR-126 and SPRED1 levels versus ABCD2 score were compared for evaluating their predictive value in the diagnosis of secondary cerebral infarction within 30 days after TIA onset.Results The miR-126 level was declined after TIA onset at 3 h(9.41±1.04),especially at 12 h(6.59 ±2.78),versus in healthy control (9.35±1.76)(t =-7.764,P=0.000).The SPRED1 level after TIA onset was increased at 3 h(58.05 ± 17.53)pg/L,12 h(82.64 ± 18.60)pg/L versus in healthy control(52.38 ± 13.24)pg/L(t=12.374,P =0.000).A closely negative correlation was found between levels of miR 126 and SPRED1 at 12 h point but not at 3 h and 6 h(r=-0.278,P=0.004).Both miR-126 and SPRED1 levels at 12 h after TIA were implied to sensitivity and specificity evaluation.Additionally,VEGF was significantly increased at 3 h (345.61 ± 76.76) pg/L,6 h (461.65 ±103.87)pg/L and 12 h (519.22 ± 103.55)pg/L after TIA onset as compared with healthy control (107.77± 26.04) pg/L(t =26.569,29.756,34.699,all P =0.000).The decrease of miR-126 and increase of SPRED1 at 12h after TIA indicated high incidences of cerebral infarction but their significance was less than ABCD2 score.Combination of miR 126,SPRED1 and ABCD2 score significantly improved the prediction for cerebral infarction(Z=2.105,P =0.035).Conclusions After the onset of TIA,levels of miR-126 and SPRED1 expression in combination of ABCD2 score can improve predictive value for cerebral infarction development.
8.Effect of Bcl-2 and PCNA expression in rat myoblast injury induced by hydrogen peroxide
Tingting MAO ; Yahua ZHENG ; Hongbo FANG ; Luji XIE ; Weidan ZHANG ; Liping JIANG
Chinese Journal of Pathophysiology 2017;33(5):935-938,943
AIM:To explore the role of Bcl-2 and PCNA expression in the injury of rat myoblasts induced by hydrogen peroxide (H2O2).METHODS:Rat myoblasts at growth phase were divided into 4 groups based on basic fibroblast growth factor (bFGF) and H2O2 levels:normal control group, bFGF group, model group (H2O2 group) and treatment group (bFGF+H2O2 group).The expression of Bcl-2 and Bax was observed by immunohistochemistry and fluorescence methods.The protein levels of Bax, Bcl-2 and PCNA were determined by Western blot.RESULTS:Compared with model group, both immunofuorescence and fluorescence in treatment group showed enhanced Bcl-2 and low expression of Bax.Furthermore, the results of Western blot showed up-regulated PCNA and Bcl-2 protein and decreased Bax expression in treatment group.CONCLUSION:Oxidative stress results in the pathologic changes of myoblasts, and the up-regulation of Bcl-2 and PCNA may attenuate myoblast injury.
9.Analysis of renal glucose threshold and related factors in patients with type 2 diabetes mellitus
Jinfu SHEN ; Zhuoqun WANG ; Shuangshuang FENG ; Mao LI ; Juan LI ; Tingting GAO ; Jingjing KANG ; Xingpo MA ; Min NIU
Chinese Journal of Endocrinology and Metabolism 2021;37(1):34-38
Objective:To investigate the value of renal glucose threshold and related factors in patients with type 2 diabetes mellitus.Methods:According to the cut-off point of normal renal glucose threshold(RT G 8.9-10 mmol/L), 107 patients with type 2 diabetes mellitus hospitalized in the Endocrinology Department of our hospital were divided into three groups: high RT G group(RT G>10 mmol/L), medium RT G group(8.9 mmol/L≤RT G≤10 mmol/L), and low RT G group(RT G<8.9 mmol/L). The clinical data and biochemical characteristics of each group were collected and analyzed. Results:The proportions of patients with high, medium, and low RT G of type 2 diabetes mellitus were 56%, 29%, and 15%, respectively. There were significant differences in RT G value, age, course of disease, body mass index(BMI), fasting plasma glucose(FPG), HbA 1C, total cholesterol(TC), serum creatinine, mean blood glucose(MBG), and 24-hour urine glucose between high and medium RT G groups. RT G, gender, BMI, FPG, HbA 1C, TC, and MBG in patients with high RT G group were different from those in low RT G group. Only RT G revealed a difference between medium and low RT G groups. Correlation analysis showed that RT G was positively correlated with gender, age, BMI, HbA 1C, TC, and low density lipoprotein-cholesterol(LDL-C). Regression analysis showed that BMI, HbA 1C, and LDL-C were the related factors affecting the RT G of patients with type 2 diabetes. Conclusion:There is a larger proportion of patients with high RT G in type 2 diabetes mellitus. Their BMI, HbA 1C, and LDL-C are associated with RT G in the patients with type 2 diabetes mellitus.
10.The fork head box M1 effects on human colon cancer cells malignant phenotype
Xiaobei MAO ; Xiaobei LIU ; Kai XU ; Xiaoyuan CHU ; Hongju YU ; Lijun XUE ; Yanan CHEN ; Lili REN ; Tingting DAI ; Longbang CHEN
Journal of Medical Postgraduates 2014;(6):582-586
Objective The invasion and metastasis of colon cancer often leads to treatment failure and mortality in patients . Our research is to investigate the influence of FoxM 1 to malignant human colon cancer line . Methods In two human colon cancer lines, the protein and mRNA expression levels of FoxM 1 were analyzed with the application of RT-PCR and Western blot , from which high-expressed HT-29 and low-expressed HCT-116 were determined.The expression of FoxM1 was down-regulated by RNA interfering in HT-29 and up-regulated by constructing overexpression transgenic line in HCT-116.The proliferation of the above cells was assayed by healing method;while the metastasis and invasion ability were examined by Transwell chamber assay . Results Two colon cancer lines were selected with high-expression or low-expression of FoxM1 separately named HT-29 and HCT-116.Application of PEX-2-FoxM1 raised after HCT-116 cells express FoxM1, cell scratches in HCT-116 experimetal group ([70.92 ±1.48]%) compared with HCT-116 control group([16.92 ±4.05]%)and HCT-116 blank control group([16.66 ±2.63]%) will markedly enhance its capabil-ity of healing (P<0.05), Transwell Chambers in membrane cells in HCT-116 experimetal group (186.0 ±6.8) compared with HCT-116 control group(42.0 ±2.0) and HCT-116 blank control grou (37.0 ± 2.2)was increased (P<0.05).On the other hand, the applied pG-PH-shFoxM1 can reduce FoxM1 expression in HT-29 cell, cell scrat-ches healing ability in HT-29 experimetal group ( [ 10 .37 ± 3.86]%) compared with HT-29 control group([34.63 ±2.35]%)and HT-29 blank control group([67.36 ±2.61]%) decreased significantly (P<0.05), Transwell Chambers in membrane cells in HT-29 experimetal group (53.0 ±1.8)compared with HT-29 control group(95.0 ±2.2)and HT-29 blank control grou(118.0 ±4.0) was also reduced (P<0.05). Conclusion The expression of FoxM1 is in close relation to the invasion and metastasis of CRC .The fact that the siRNA interfering FoxM1 could effectively inhibit the proliferation, metastasis and invasion, suggesting FoxM1 could po-tentially be a new molecular target for inhibiting the proliferation of human colon cancer line .