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Objective@#To investigate the effect of Δ40p53, an alternative spliced isoform of p53 lacking the N-ter minus, on the pro-apoptotic function of p53.@*Methods@#The wild-type p53 was ectopically expressed in HCT116-p53-/- (endogenous Δ40p53 expression), HCT116-p53+ /+ (wild-type p53) and H1299 (p53-null) cells by adenoviral delivery, while Δ40p53 plasmid were transfected into these cells to overexpress Δ40p53. The levels of Δ40p53 and p53 mRNA were detected by reverse transcription-polymerase chain reaction (RT-PCR) and quantitative PCR. The expression of related proteins was deter mined by Western blotting. The interaction of p53 and Δ40p53 was observed by co-immunoprecipitation assay. Calcein-AM/propidium iodide (PI) staining and flow cytometry were used to detect the apoptotic rate of tested cells in each group.@*Results@#HCT116-p53-/- cells expressed endogenous Δ40p53 isoform. Neither transcription nor protein expression of wild-type p53 was interfered by the increased expression of Δ40p53. Full length p53 and Δ40p53 could bind to each other. Calcein-AM/PI staining showed that the apoptotic rates of H1299-Control, HCT116-p53-/- -Control, H1299+ p53, HCT116-p53-/-+ p53, H1299+ oxaliplatin (Oxa), HCT116-p53-/-+ Oxa, H1299+ p53+ Oxa and HCT116-p53-/-+ p53+ Oxa groups were (2.50±0.47)%, (2.40±0.32)%, (5.20±0.58)%, (4.10±0.18)%, (22.40±1.73)%, (19.30±1.11)%, (29.90±1.15)% and (39.30±2.26)%, respectively. It was statistically significant between H1299+ p53+ Oxa and HCT116-p53-/-+ p53+ Oxa groups (t=3.721, P=0.0205). Moreover, the apoptotic rates of H1299-Control, H1299+ Δ40p53, H1299+ p53, H1299+ p53+ Δ40p53, H1299+ Oxa, H1299+ Δ40p53+ Oxa, H1299+ p53+ Oxa and H1299+ p53+ Δ40p53+ Oxa groups were (2.60±0.35)%, (2.20±0.17)%, (4.80±0.49)%, (4.90±1.10)%, (20.30±1.10)%, (19.60±1.45)%, (27.90±1.39)%, (35.20±1.43)%, respectively. Furthermore, flow cytometry assay showed that the apoptotic rates of above cells were (2.70±0.32)%, (2.20±0.24)%, (4.60±0.48)%, (3.90±0.67)%, (19.30±1.11)%, (17.70±0.66)%, (28.30±2.76)% and (37.50±1.51)%, respectively. H1299+ p53+ Δ40p53+ Oxa cells showed higher cell apoptosis than H1299+ p53+ Oxa cells (t=2.930, P=0.042).@*Conclusion@#Δ40p53 isoform can bind to full-length p53, and enhance its pro-apoptotic function in tumor cells.
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Background The incidence of myopia is gradually increasing,and how to choose a better corrective method of myopia for the best visual demand is very important.Objective This study was to compare visual quality of implantable collamer lens (ICL) implantation with femtosecond laser in situ keratomileusis (FSLASIK) for moderate and high myopia using double-pass optical quality analysis system (OQAS).Methods A non-randomized controlled clinical trail was performed.Fifty-two eyes with-4.00 to-9.00 D of 26 consecutive patients were included in NO.1 Hospital of Xi'an from January 2015 to January 2016.Twenty-four eyes of 12 patients with the corneal thickness ＜500 μm received ICL implantation as ICL group and 28 eyes of 14 patients which corneal thickness was ≥500 μm underwent FS-LASIK surgery as FS-LASIK group.The demography was matched between the two groups (all at P＞0.05).All the patients were followed-up for 3 months after surgery.The preoperative best corrected distance visual acuity (BCDVA),spherical equivalent (SE),postoperative uncorrected distance visual acuity (UCDVA),BCDVA and SE were examined and compared between two groups.The parameters from OQAS were evaluated and intergrouply compared,including the objective scattering inders (OSI),modulation transfer function (MTF) cut off frequency,Strehl ratio and OQAS values under the contrast of 100％,20％ and 9％ (OV100,OV20,OV9).Results The postoperative BCDVA was not significantly different from preoperative UCDVA in both ICL group and FS-LASIK group (-0.04±t0.10 vs.0.05±0.12;-0.07±0.12 vs.0.00±0.12) (t=3.128,2.358,both at P＞0.05).No statistically significant differnces were found in SE,UCDVA and BCDVA after operation between ICL group and FS-LASIK group (t =1.292,0.900,-0.653,all at P＞0.05).OQAS examination showed that MTF cut off,OSI,Strehl ratio,OV100 were not significantly different after operation between ICL group and FS-LASIK group (t=-2.032,-1.440,-0.224,all at P＞0.05).The postoperative OV20 and OV9 were 0.82±0.14 and 0.80±0.21 in the ICL group,which were significantly higher than those in the FS-LASIK group(0.59±0.15 and 0.47±0.13) (t =4.105,4.702,both at P＜0.05).Conclusions Both ICL implantation and FS-LASIK provide good optical and visual quality for moderate to high myopic eyes,and ICL appears to have a better visual quality in comparison with FS-LASIK under the contrasts of 20％ and 9％.
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<p><b>OBJECTIVE</b>To summarize the treatment status of gastric gastrointestinal stromal tumor (GIST) in Shandong province,by analyzing the clinicopathological features and prognostic factors.</p><p><b>METHODS</b>Clinicopathological and follow-up data of 1 165 patients with gastric GIST between January 2000 and December 2013 from 23 tertiary referral hospitals in Shandong Province were collected to establish a database. The risk stratification of all cases was performed according to the National Institutes of Health(NIH) criteria proposed in 2008. Kaplan-Meier method was used to calculate the survival rate. Log-rank test and Cox regression model were used for univariate and multivariate prognostic analyses.</p><p><b>RESULTS</b>Among 1 165 cases of gastric GIST, 557 were male and 608 were female. The median age of onset was 60 (range 15-89) years. Primary tumors were located in the gastric fundus and cardia in 623 cases(53.5%), gastric body in 346 cases(29.7%), gastric antrum in 196 cases(16.8%). All the cases underwent resection of tumors, including endoscopic resection (n=106), local resection (n=589), subtotal gastrectomy(n=399), and total gastrectomy(n=72). Based on the NIH risk stratification, there were 256 cases (22.0%) at very low risk, 435 (37.3%) at low risk, 251 cases (21.5%) at intermediate risk, and 223 cases (19.1%) at high risk. A total of 1 116 cases(95.8%) were followed up and the median follow-up period was 40 (range, 1-60) months. During the period, 337 patients relapsed and the median time to recurrence was 34 (range 1-60) months. The 1-, 3-, and 5-year survival rates were 98.6%, 86.1% and 73.4%, respectively. The 5-year survival rates of patients at very low, low, intermediate, and high risk were 93.1%, 85.8%, 63.0% and 42.3% respectively, with a statistically significant difference (P=0.000). Multivariate analysis showed that primary tumor site (RR=0.580, 95%CI:0.402-0.835), tumor size (RR=0.450, 95%CI:0.266-0.760), intraoperative tumor rupture(RR=0.557, 95%CI:0.336-0.924), risk classification (RR=0.309, 95%CI:0.164-0.580) and the use of imatinib after surgery (RR=1.993, 95%CI:1.350-2.922) were independent prognostic factors.</p><p><b>CONCLUSIONS</b>The choice of surgical procedure for gastric GIST patients should be based on tumor size. All the routine procedures including endoscopic resection, local excision, subtotal gastrectomy and total gastrectomy can obtain satisfactory curative outcomes. NIH classification has a high value for the prediction of prognosis. Primary tumor site, tumor size, intraoperative tumor rupture, risk stratification and postoperative use of imatinib are independent prognostic factors in gastric GIST patients.</p>
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<p><b>OBJECTIVE</b>To observe the impacts of eye acupuncture on neurological deficit and Barthel index in the patients of infarction hemiplegia and explore its function mechanism.</p><p><b>METHODS</b>Ninety-six patients of infarction hemiplegia were randomized into an observation group and a control group, 48 cases in each one. In the control group, the routine western medicines such as thrombolysis and antiplatelet aggregation were used. In the observation group, on the basis of the treatment as the control group, eye acupuncture was added atandareas bilaterally, once a day, 5 times a week. Separately, before treatment and after 2 weeks' treatment the score changes of the modified Edinburgh Scandinavia stroke scale (MESSS) and the activity of daily life scale (ADL, Barthel index, BI) were observed and the efficacy was compared between the two groups. The plasma endothelin was determined and compared before and after treatment in the two groups.</p><p><b>RESULTS</b>After treatment, the effective rate was 93.8% (45/48) in the observation group and was 79.2% (38/48) in the control group. The effective rate in the observation group was higher apparently than that in the control group (<0.05). The scores of neurological deficit were (13.29±1.45) and (18.24±1.33) in the observation group and control group respectively after treatment, which all lower apparently than (28.44±1.45) and (28.14±1.89) before treatment (both<0.05). Additionally, the difference was significant between the two groups after treatment (<0.05). The scores of Barthel index were (82.33±1.56) and (63.34±2.14) in the observation group and control group respectively, which all higher apparently than (38.53±1.54) and (38.14±2.56) before treatment (both<0.05), and the difference was significant between the two groups after treatment (<0.05). The levels of plasma endothelin were (54.55±11.48)ng/L and (62.44±9.88)ng/L in the observation group and the control group after treatment respectively, which were all lower apparently than (78.24±9.25)ng/L and (78.14±10.78)ng/L before treatment (both<0.05). Additionally, the difference was significant between the two groups after treatment (<0.05).</p><p><b>CONCLUSIONS</b>Eye acupuncture effectively improves the neurological deficit and Brathel index in the patients of infarction hemiplegia and comprehensively improves the efficacy. The effect mechanism is possibly relevant with reducing plasma endothelin.</p>
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BACKGROUND:Since damage control theory system was founded, this theory in the orthopedics has been applied gradualy, especialy in elderly hip fracture surgery that reduces the negative impacts due to inflammatory responses. OBJECTIVE:To explore whether flurbiprofen axetil can reduce inflammatory responses in rats with hip fractures based on the damage control theory. METHODS: Forty-nine healthy Sprague-Dawley rats weighing 250-300 g were randomly divided into four groups:control group (n=7), immediate internal fixation group (n=14), flurbiprofen axetil group (n=14), damage control group (n=14). Rats in the control group moved freely in the cages. Rats in the other three groups were intraperitonealy injected with composite anesthetics to make unilateral hip fracture models, and then respectively given internal fixation immediately after fracture, flurbiprofen axetil injection and delayed internal fixation, and delayed internal fixation. Levels of serum C-reactive protein, interleukin-6 and tumor necrosis factor-α were determined and analyzed before fixation, immediately after internal fixation and at 4, 8, 12, 24, 48 hours after internal fixation in different groups. RESULTS AND CONCLUSION:Postoperative serum levels of C-reactive protein, interleukin-6, tumor necrosis factor-αwere al increased in different groups. The level of C-reactive protein reached the peak at 24 hours after internal fixation. Flurbiprofen axetil injection had no significant influence on the level of C-reactive protein in rats with delayed internal fixation (P=0.51). Interleukin-6 levels were stil increased at 48 hours after internal fixation, but flurbiprofen axetil reduced the level of interleukin-6 significantly in rats with delayed internal fixation (P < 0.01). The tumor necrosis factor-α level peaked at 4 hours after internal fixation, and flurbiprofen axetil injection could significantly reduce the level of tumor necrosis factor-α in rats with delayed internal fixation (P < 0.01). These findings indicate that flurbiprofen axetil as a new non-steroidal anti-inflammatory drug can reduce the inflammatory response in rats with hip fractures after internal fixation, and also can aleviate the inflammatory response of rats undergoing delayed operation under the guidance of damage control theory.
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BACKGROUND:Bone marrow mesenchymal stem cels are crucial for bone and cartilage development and regeneration at a celular level. Insufficient quantity and functional impairment of bone marrow mesenchymal stem cels is widely considered to be one of osteoarthritis causes. OBJECTIVE: To explore the relationship between the functional status of bone marrow mesenchymal stem cels and disease progression in osteoarthritis patients.METHODS: Thirty patients with osteoarthritis were enroled from July 2013 to October 2014, and divided into control, mild osteoarthritis, and severe osteoarthritis groups, with 10 cases in each group. 5 mL bone marrow from the femur or tibia was extracted from each patient to isolate and culture bone marrow mesenchymal stem cels. Proliferation ability of cels at passage 3 was detected using cel counting kit-8; toluidine blue staining was performed at 14 days after chondrogenic induction; real-time PCR was used to detect the mRNA expression of Aggrecan and Col2A1 in the control group after chondrogenic induction. RESULTS AND CONCLUSION:Afterin vitro culture, bone marrow mesenchymal stem cels grew adherently in polygonal and fusiform shape with multiple processes at uniform size. The cytoplasm contained larger particles and the nuclei were ovoid. Most of cels were in cel division phase. The proliferation ability was strongest in the control group and weakest in the severe osteoarthritis group. Cels from the three groups were al at plateau phase after 1 week culture. At 14 days after chondrogenic induction, the cels were polygonal and quasi-circular, and purple metachromatic granules distributed outside of the cytoplasm. The expression of Aggrecan and Col2A1 in the control group displayed an overexpression trend. These findings indicate that the functional status of bone marrow mesenchymal stem cels from osteoarthritis patients is negatively correlated with the severity of disease, which can influence the disease progression in osteoarthritis patients.
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Taking the ICU We-chat platform of China Medical University Affiliated Shengjing Hospital as an example, the paper in-troduces its operation mode, analyzes its application effect, advantages and potential problems, points out that We-chat platform is ad-vantageous to improving nursing management quality, it could be a new way of nursing management.
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BACKGROUND:In recent years, many reports have focused on inflammatory cytokines, growth factors and mechanical loads affecting the cartilage and subchondral regeneration, but there is a lack of comprehensive understanding about the mechanism of osteoarthritis. OBJECTIVE: To explore the correlation between function status of bone marrow mesenchymal stem cels and disease progression in patients with osteoarthritis. METHODS:Femoral bone marrow was extracted from patients with femoral neck fractures (control group), mild (mild group) and severe (severe group) osteoarthritis to isolate and culture bone marrow mesenchymal stem cels. Cel counting kit-8 was used to detect the proliferative ability of bone marrow mesenchymal stem cels from different patient groups, and passage 3 bone marrow mesenchymal stem cels were subject to 2-week chondrogenic induction folowed by toluidine blue staining. RESULTS AND CONCLUSION: Bone marrow mesenchymal stem cels were isolated and cultured from the femoral bone marrow of different groups. The proliferative ability of cels in the control group was significantly higher than that in the mild and severe groups. After chondrogenic induction, bone marrow mesenchymal stem cels varied obviously in the morphology that was from fusiform to qusi-circular or polygon, the percentage of nucleoplasm became smaler, and cels were positive for toluidine blue staining. The number of chondrocytes generated in the severe group was less than that in the control group, but there was no great difference in cel morphology. These findings indicate that the occurrence of osteoarthritis is negatively correlated with the functional status of autologous bone marrow mesenchymal stem cels.
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BACKGROUND:Numerous studies confirmed that compared with deltopectoral approach, deltoid-splitting approach for proximal humeral fractures has good therapeutic effects, but precise superiority remains poorly understood. OBJECTIVE:To compare the therapeutic effects of deltopectoral approach and smal incision deltoid-splitting approach in the repair of proximal humeral fractures using meta-analysis. METHODS:We retrieved MEDLINE, Embase, the Cochrane library, Wanfang Database and PubMed by computer for articles on control ed trials of deltopectoral approach and smal incision deltoid-splitting approach in the repair of proximal humeral fractures published from 2010 to 2014. Neer score, operation time, intraoperative blood loss, incision length, length of hospital stay, Visual Analogue Scale score, fracture healing time, and Constant score were used as evaluation indexes of Meta analysis. RevMan 5.2 software was used for analysis. RESULTS AND CONCLUSION:Eventual y nine articles were included, total y 721 patients, published from 2010 to 2014 years. There were eight in Chinese, and one in English. Meta-analysis results showed that compared with deltopectoral approach, deltoid-splitting approach in the treatment of proximal humeral fractures could shorten operation time, reduce intraoperative blood loss, shorten length of incision, elevate Neer score and Constant score (P<0.05). No significant difference in fracture healing time, hospitalization days, and Visual Analogue Scale score was detected between the two methods of surgical approach. These data indicated that compared with deltopectoral approach, deltoid-splitting approach in the repair of proximal humeral fracture has certain superiority. Deltoid-splitting approach can be firstly selected in the permit of hospital conditions.
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<p><b>OBJECTIVE</b>To investigate the role of apoptosis stimulating protein 2 of p53 (ASPP2) phosphorylation status in the regulation of ASPP2-p53 apoptotic pathway activity.</p><p><b>METHODS</b>Cells were individually transfected with green fluorescent protein (GFP)-encoding vector, constitutively non-phosphorylatable ASPP2 mutant-ASPP2 (Am)-encoding vector, and wild type ASPP2 (Aw)-encoding vector) plasmids, respectively, to make them overexpressing phosphorylated and non-phosphorylated ASPP2 proteins, respectively. Cell apoptosis was induced by oxaliplatin. The apoptosis rate of cells was determined by flow cytometry after staining with FITC-conjugated annexin V and PI. ASPP2 protein level and its phosphorylation status were observed by Western blot. The interaction between ASPP2 and p53 was observed by immunoprecipitation assay.</p><p><b>RESULTS</b>Oxaliplatin induced cell apoptosis and caused phosphorylation of ASPP2 at ser92/ser361 in the HCT116 cells. The apoptosis rate of Aw and Am plasmids-transfected cells were (3.8 ± 1.0)% and (3.9 ± 1.2)% respectively, statistically with a non-significant difference (P > 0.05) in comparison with that of the GFP plasmid-transfected cells [(4.0 ± 0.8)%]. After oxaliplatin treatment, the apoptosis rate of Aw plasmid-transfected cells was (46.7 ± 3.9)%, significantly higher than that of the Am and GFP plasmid-transfected cells [(40.1 ± 10.2)% and (37.1 ± 6.9)%, respectively, P < 0.05], however, there was no statistically significant difference (P > 0.05) between Am and GFP plasmid-transfected cells. These results indicate that phosphorylated ASPP2 promoted the oxaliplatin-induced apoptosis of HCT116 cells through a p53-dependent pathway. Phosphorylation status of ASPP2 influenced its binding activity to p53.</p><p><b>CONCLUSION</b>Phosphorylation status of ASPP2 modulates p53 apoptotic function in oxaliplatin-induced apoptosis of colorectal cancer HCT116 cells.</p>
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Humans , Apoptosis , Apoptosis Regulatory Proteins , Metabolism , Colorectal Neoplasms , Metabolism , HCT116 Cells , Organoplatinum Compounds , Phosphorylation , Tumor Suppressor Protein p53 , Genetics , MetabolismABSTRACT
Objective To investigate the role of apoptosis stimulating protein 2 of p53 (ASPP2)in the apoptosis,cell cycle and autophagy of starvation-induced colorectal cancer HCT116 p53 +/+ (p53 wild-type) cell line.Methods Six groups were included:(1) control group; (2) green fluorescent protein adenovirus (rAd-GFP) infection group; (3)ASPP2 adenovirus (rAd-ASPP2) infection group; (4)starvation group; (5)rAd-GFP + starvation group; (6) rAd-ASPP2 + starvation group.HCT116 cells were infected with ASPP2 adenovirus (rAd-ASPP2),resulting ASPP2 gene over-expression.The apoptosis,autophagy and cell cycle changes were induced by culturing with serum-free medium for 24 h.Apoptosis was evaluated by Calcein/PI uptaking test,and autophagy was observed by counting the red fluorescent protein autophagy plasmid CFP-Lc3 which was transfected into cytoplasm.Cell cycle was detected by flow cytometry.Statistical analysis was performed by one-way analysis of variance (ANOVA).Results Over-expressed ASPP2 was found to significantly promote starvation-induced HCT116 apoptosis and autophagy.The cell apoptosis rate in rAd-GFP + starvation group was 10.00％ ± 1.42％,and 18.44％ ±2.06％ in rAd-ASPP2 + starvation group(q =9.548,P =0.000).The cell autophagy rate in rAd-GFP+ starvation group and rAd-ASPP2 + starvation group was 35.00％ ± 5.34％ and 57.61％ ± 6.06％ respectively(q =7.657,P =0.000).Over-expressed ASPP2 accelerated HCT116 G2/M arrest under starvation,but resulted in both G0/G1 and G2/M arrest without starvation.Conclusion These results suggest that ASPP2 can promote starvation-induced HCT116 p53 +/+ cells apoptosis and autophagy,and affect the cell cycle.
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Objective To investigate the role of ASPP2 (apoptosis stimulating protein 2 of p53,ASPP2) in starvation-induced autophagy and apoptosis of colorectal cancer HCT116 p53-/-(p53 gene deletion) cell line.Methods The study included three experiment groups:green fluorescent protein adenovirus (rAd-GFP) infection group,autophagy inhibitor LY294002 treatment group and ASPP2 adenovirus (rAd-ASPP2) infection group.Celluar autophagy and apoptosis were induced by coculturing with serum-free medium for 0 h,24 h,48 h.Apoptosis level was detected by Calcein/PI uptaking test.Autophagy level was observed under the fluorescence microscope via transfection with cerise fluorescent protein autophagy plasmid CFP-Lc3.Results In control group,starvation for 24 hours significantly promoted autophagy of HCT116 cells (0 h:1.04 ±0.24; 24 h:12.17 ±0.86,P ＜0.05),while apoptosis was not increased (0 h:2.01％ ±0.06％; 24 h:3.23％ ±0.34％,P ＞0.05).With 48 h starvation,autophagy(0 h:1.04 ±0.24; 48 h:21.09 ±3.32) and apoptosis(0 h:2.01％ ±0.06％ ; 48 h:30.20％ ±3.18％)of HCT116 increased (P ＜ 0.05).With the use of LY294002 apoptosis induced by 24 h starvation significantly increased (rAd-GFP group:3.23％ ± 0.34％ ; LY294002 group:15.68％ ± 1.24％,P ＜0.01),but aopotosis under 48 h starvation decreased (rAd-GFP group:30.20％ ± 3.18％; LY294002group:25.44％ ± 3.01％,P ＜ 0.05).With ASPP2 transfection,autophagy under 24 h starvation significantly declined (rAd-GFP group:12.17 ± 0.86,ASPP2 group:1.45 ± 0.45,P ＜ 0.01),and apoptosis increased(rAd-GFP group:3.23％ ± 0.34％ ; ASPP2 group:10.45％ ± 0.81％,P ＜ 0.05).Both autophagy (rAd-GFP group:21.09 ± 3.32; ASPP2 group:29.93 ± 3.48) and apoptosis (rAd-GFP group:30.20％ ±3.18％ ; ASPP2 group:36.72％ ±2.74％) were higher than that in controls under 48 h starvation (P ＜ 0.05).Conclusions ASPP2 probably promotes apoptosis of colorectal cancer cells by two-way regulated autophagy.
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P53 protein plays a crucial role in inhibition tumor development.It will accumulate in cells in the condition of DNA damage,oncogenes activation or stress.As a nuclear transcription factor,P53 can transactivate the genes which correlate with apoptosis,cell cycle control and other procedures. Current research reveals P53 outside the nucleus can induce apoptosis and inhibit autophagy which contributes to its function of tumor suppression.The study of the extranuclear function of P53 is beneficial to further understanding the mechanism of P53 in the genesis and development of tumor.