1. ASPP2 gene silence promotes hepatocellular carcinoma cell proliferation via regulating autophagy under starvation
Academic Journal of Second Military Medical University 2013;34(3):266-270
Objective To study the effects of apoptosis-stimulating protein of ρ53-2 (ASPP2) gene on hepatocellular carcinoma (HCC) cell proliferation and apoptosis under starvation and the related mechanism. Methods Lentivirus encoding shRNA against ASPP2 was constructedto knockdown ASPP2 expression in hepatoma cell HepG2. Cell proliferation and apoptosis were observed by transmission electron microscopy, MTS analysis, flow cytometry analysis and morphologic changes. The influence of silencing ASPP2 gene on the proliferation and apoptosis under amino acid-starvation and serum-deprivation culture was observed; autophagy inhibitor 3-methyladenine (3-MA) was added in the experiment so as to detect the involvement of autophagy in the changes induced by ASPP2 down-regulation. Results Transmission electron microscopy showed cytoplasmic accumulation of autophagosomes when ASPP2 was knocked down under amino acid-starvation and serum- deprivation (P<0. 05), with increased GFP-LC3 dots (P<0. 05). MTS analysis showed that silence of ASPP2 gene greatly enhanced the proliferation of HepG2 cells (P<0. 05), which could be inhibited by addition of 3-MA (P<0. 05). Microscope observation showed that silence of ASPP2 gene promoted the anti-apoptotic ability of HepG2 cells, which was reversed by treatment with 3-MA. Early sign of apoptosis was observed in shASPP2 + 3-MA group. Annexin V_PI double staining showed that ASPP2 silence decreased the apoptotic rate ofHepG2 cells from (38±5)% to(15±4)% (P<0. 05), and 3-MA treatment increased it to (36 ±3)% (P<0. 05). Conclusion Down-regulation of ASPP2 expression may facilitate the survival and proliferation of HCC cells through activating autophagy under starvation.
2.Radiofrequency treatment of hemangioma of nasal cavity under nasal endoscopy
Jianping LIANG ; Dongyun LI ; Bei LIU
Journal of Clinical Otorhinolaryngology Head and Neck Surgery 2000;(10):456-457
Objective:To further understand the advantage of radiofrequency treatment of hemangioma ofnasal cavity under nasal endoscopy. Method:Fifteen cases with hemangioma of nasal cavity were treated withradiofrequency under nasal endoscopy. Result:The hemangioma of fifteen cases could be removed completely. Thecomplication was absent. None of them recured in six months to four years following up. Conclusion:This methodhas many advantages such as clear operation visual field,less hemorrhage and postoperative pain,no facial scar.The radiofrequency under endoscopy is valuable in treatment for hemangioma of nasal cavity.
3.The effect of colchicine on the proliferation of human orbital fibroblast in thyroid associated ophthalmopathy in vitro
Yan, LIU ; Bei, XU ; Xue-liang, XU
Chinese Journal of Experimental Ophthalmology 2013;(7):630-635
Background The orbital fibroblasts (OFs) in thyroid associated ophthalmopathy (TAO) play important roles in the proliferative and inflammatory response.Seeking the drug which inhibit OFs growth is of a vital significance for the prevention and treatment of TAO.Research documented that colchicine has an anti-fibrosis effect.But its influence on OFs of TAO patient is few known.Objective This study was to investigate the effect of colchicine on growth and apoptosis of OFs in vitro.Methods The retroobital connective tissue was obtained form 3 TAO patients and cultured using explant method.OFs were passaged and identified by immunochemistry,and 3-8 genetaions of cells were used in the study.Colchicine at the concentrations of 1 × 10-8,1 × 10-7,1 × 10-6,1 × 10-5,1 ×10-4 mol/L was added into the RPMI 1640 with 10% fetal bovine serum(FBS) to incubated the cells for 24,48 and 72 hours respectively,and only RPMI 1640 was used to culture the cells as the control group.Cell counting kit-8 (CCK-8)was used to detect the absorbance value (A450) of OFs for the evaluatuion of OFs and the inhibitory rate of colchicine to OFs.The colchicine of 1 ×10-6,1 ×10-5,1 × 10-4 mol/L was added into the culture medium for 48 hours,and then the apoptotic rate of the cells and the cell percentage in various cellular cycle was assayed by flow cytometry(FCM).The expression of transforming growth factor-β (TGF-β)in the cells was detected by immunochemistry to assess the influence of colchicine on the serection of the cells.Results Cultured cells showed the spindle-like in shape and the cell number was significantly increased with the incubation time.After incubated with 1 × 10-4,1 × 10 5,1 × 10-6,1 ×10-7,1 × 10-8 mol/L colchicines,the A450 values were gradually reduced with the increase of the concentrations of colchicine(F ion =62.004,P<0.05),and significant differences were found between different contrations of colchicine groups(all P<0.05).Aslo,gradually declined A450 values of the cells were seen with the lapse of culture time among the groups(Ftime =459.582,P<0.05).The inbitory rate of colchicine to the cells was elevated with the increase of concentrations.The apoptotic rates of the cells were (1.73 ± 0.15) %,(21.04 ± 4.56) %,(31.84 ±6.21)%and(35.32±5.56)% in the control group and 1 × 10-6,1 × 10-5,1 × 10 4 mol/L colchicine groups respectively,with statistically significant difference among the 4 groups (F =83.905,P<0.05).With the increase of concentrations of colchicines,the cell percentage in G2 +M phase lessened gradually,showing significant difference among the control group and the 1 × 10-6,1 × 10-5,1 × 10-4 mol/L colchicine groups (F =20.443,P<0.05).The expression of the TGF-β in the cells was (97.60± 2.09) % in the control group,and that in the 1 × 10-4 mol/L colchicine group was (44.43 ± 3.96) %,presenting a significant difference between them (t =65.330,P < 0.05).Conclusions Colchicine can induce apoptosis of OFs and inhibit the prolilferation of OFs in a time-and dose-dependent manner probably by decreasing the TGF-β secretion
4.Relationship between serum IGF-Ⅱand peripheral neuropathy in type 2 diabetics.
Li LIANG ; Chengjiang LI ; Bei ZHANG
Chinese Journal of Practical Internal Medicine 2006;0(24):-
Objective To evaluate the possible relationship between IGF-Ⅱand peripheral neuropathy in type 2 diabetes.Methods Seventy-one type 2 diabetic patients without peripheral neuropathy(Group A),Seventy-seven patients with peripheral neuropathy(Group B)and thiry healthy subjects(Group C)were recruited.Serum IGF-Ⅱwas determined in all subjects.Other clinical parameters(fasting plasma glucose、fasting plasma insulin,et al)and nerve conduction velocity were determined in Group A and Group B.Results Group B had significantly lower levels of IGF-Ⅱ,compared to GroupA and Ⅲ(P
5.Determination the concentration of linezolid in Mueller hinto broth in vitro pharmacodynamic model by HPLC-UV
Bei-Bei LIANG ; Nai BAI ; Rui WANG
The Chinese Journal of Clinical Pharmacology 2010;26(4):296-298
Objective To establish an analytical method for the determination the concentration of linezolid in vitro pharmacodynamic model.Methods Linezolid was precipitated from MH broth with hydrochloric acid.HPLC-UV method was carried on with a venusil XBP C_8(4.6 mm ×250 mm,5 μm)column.The mobile phase consisted of water-acetonitrile(80:20).The flow rate was 1.5 mL·min-1 and the detection was performed at 251 nm.Results The calibration curve was linear with in the range of 0.5-40.0 μLg·mL~(-1)(γ=0.9996).The absolute recoveries of linezolid at the concentrations of 2,10,40 μg·mL~(-1) were all higher than 90%,and the relative recoveries were from 96%to 98%.The RSDs of the within-day and between-day variations were lower than 2%and 4%,respectively.Conclusion This HPLC-UV method was simple,sensitive and accurate,and suitable for routine determination the concentration of linezolid for vitro pharmacodynamic study.
6.CD25 content variation with the responses of T helper cell to IL-2 induced proliferation
Fanqiang KONG ; Zhimou WANG ; Bei SUN ; Dongchun LIANG
Chinese Journal of Microbiology and Immunology 2012;32(5):430-435
Objective To elucidate the expression diversity of IL-2R subunits in T helper(Th) cells under different activation condition and clarify its relationship with the response to IL-2 induced proliferation.Methods In vitro cultured Th1 cells were activated by plate bound anti-mouse CD3 McAb.In both activated and inactivated Th1 cells the expression of different IL-2R subunit was evaluated by real-time PCR and fluorescence staining,3H incorporation assay was adopt to measure the proliferation of the cells in response to IL-2 treatment,IL-2 affinity was determined via I125 labeled IL-2.IL-2Rα siRNA transfection was applied to knockdown CD25 expression in activated Th1 cells and its effect was confirmed by Western blot,IL-2 induced proliferation in IL-2Rα siRNA transfected Th1 cells were subsequently detected.CD4 cells were isolated from na(i)ve BALB/c mice and were also stimulated by plate bound anti-CD3 McAb.Cells were harvested at different days posterior to the stimulation,CD25 content and IL-2 induced proliferation were determined by flow cytometry and 3H incorporation assay respectively.Results In comparison with inactivated cells,only the expression of CD25but not other IL-2R subunits was significantly increased in activated Th1 cells.In accordance with elevated CD25,IL-2 affinity was also increased in activated Th1 cells,however,which resulted in decreased cell proliferation in response to IL-2 treatment.In activated Th1 cells,when CD25 expression was differently knockdown by different-dosed IL-2Rα siRNA transfection,the highest IL-2 response was observed in the cells with partially CD25 depression.Forin vitro activated na(i)ve CD4 cells the elevated CD25 expression gradually decreased and the highest proliferation response was detected at day 8 post stimulation upon IL-2 treatment.Conclusion Although CD25 is necessary for IL-2 induced proliferation,however,the excessively expressed CD25 lead to lower proliferation response.Not fully activated Thl or CD4 cells,but partially activated cells with lightly increased CD25 content possessed highest proliferation rate in response to IL-2 treatment.
7.Surgical treatment of chronic pancreatitis and choice of procedure
Liang JI ; Bei SUN ; Hongchi JIANG ; Jie LIU ; Linfeng WU
Chinese Journal of Pancreatology 2014;14(1):30-33
Objective To analyze the results of surgical treatment on chronic pancreatitis(CP),and investigate how to choose the appropriate surgical procedure.Methods The clinical data of 54 patients with chronic pancreatitis who underwent surgery at the First Affiliated Hospital of Harbin Medical University from January 2007 to December 2011 were retrospectively analyzed.Results All the 54 patients underwent surgery,including 8 decompression and drainage procedure (Partington procedure) ; 13 resections (7 cases of pancreaticoduodenectomy,1 case of distal pancreatectomy,4 cases of distal pancreatectomy combined with splenectomy and 1 case of pancreaticoduodenectomy combined with distal pancreatectomy,respectively); 12hybrids (7 cases of Beger procedure and 5 cases of Frey procedure,respectively) and 21 other procedures (15 cases of pancreatic pseudocyst jejunostomy,4 cases of exploratory laparotomy combined with pancreatic tissue biopsy,2 cases of gastrojejunostomy combined with choledochojejunostomy,respectively).There were 4 cases of post-operative pathologic evidence of cancer.Twelve patients had postoperative complications and were cured with non-operative management.Forty-four patients (81.5%) were followed for 2 to 67 months,36 out of 42(85.7%) patients who suffered from abdominal pain had a persistent remission,there were one case of new on-set diabetes and no steatorrhea was reported.Conclusions For CP patients with surgical indications,the choice of procedure should be individualized for the purpose of preserving the endocrine and exocrine functions of pancreas,and taking effectiveness as well as safety into consideration.
9.The relationship between the direct signs of breast cancer and the metastasis in the mammography
Bei PEGN ; Hong LIANG ; Longbai MA ; Jun CHEN
Journal of Practical Radiology 2016;32(7):1036-1039
Objective To analyze the relationship between the direct signs of invasive ductal carcinoma and the metastasis in the mammography.Methods 21 7 patients with invasive ductal carcinoma in the mammography were divided into four groups:mass with calcification group,mass without calcification group,calcification without mass group and the group without both mass and calcification. The correlation between metastasis and the size and shape of mass or calcification was studied.Results The rate of metastasis in the group with mass and calcification was 56.31%,and the percentage of the skin and nipple depressed in this group was 38.83%,both of whom were highest (P =0.004 and P =0.043)among four groups.The rate of metastasis in mass group was higher than that in the group without mass (46.27% vs 18.75%,P =0.033).Meanwhile,the edge burr and lobulation of the mass didn’t exert remarkable influence on the rate of metastasis (46.70% vs 25.00% and 43.37% vs 52.00%,P =0.389 and P =0.348).The probability of me-tastasis in the calcification group was higher than that in noncalcification group (53.57% vs 34.29%,P =0.001 ).However,in the calcification group,the different shapes of calcification did not result in significant difference in the rate of metastasis (51.19% vs 60.71%, P =0.382).In the 201 patients with breast cancer mass,the size of mass did not correlate with metastasis [(81.04±1 1 9.45)mm3 vs (70.06±208.30)mm3 ,P =0.654].The masses with lobulation were bigger than others [(7 6 .5 0 ± 1 7 4 .1 3 )mm 3 vs (8 .3 9 ± 1 0 .2 7 )mm 3 ,P =0.000],while the masses with edge burr were smaller than those with smooth edges [(52.10 ±85.90)mm3 vs (144.75±304.13)mm3 ,P =0.038].There was no significant correlation between the size of mass and the calcification [(80.39 ± 126.62)mm3 vs (69.63±209.12)mm3 ,P =0.660].However,the masses with point calcification were larger than those with both point and worm calcifications [(92.79±137.21)mm3 vs (41.71±94.58)mm3 ,P =0.041].Conclusion The mass and calcification in the mammography are valuable for judging the malignant degree of invasive ductal carcinoma.
10.One case of pulsatile tinnitus and literatures review.
Tao WANG ; Jianping LIANG ; Bei LIU
Journal of Clinical Otorhinolaryngology Head and Neck Surgery 2013;27(10):497-500
To investigate the etiology.diagnosis and treatment of pulsatile tinnitus. One case with pulsatile tinnitus in our hospital admitted in May 2012 was reported and the relevant literatures were reviewed. The chief complaint when visited was pulsatile tinnitus. This case was treated by surgery. Pulsatile tinnitus was completely subsided without recurrence. Pulsatile tinnitus is an uncommon otologic symptom, which often presents a diagnostic and treatment dilemma to the otolaryngologist. The majority of patients with pulsatile tinnitus have a treatable cause. Failure to establish correct diagnosis may have disastrous consequences, because a potentially life threatening and underlying disorder may be present.
Adult
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Humans
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Tinnitus
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diagnosis
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etiology
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therapy