MF2ResU-Net: a multi-feature fusion deep learning architecture for retinal blood vessel segmentation / 数字中医药（英文）

Objective@#For computer-aided Chinese medical diagnosis and aiming at the problem of insufficient segmentation, a novel multi-level method based on the multi-scale fusion residual neural network (MF2ResU-Net) model is proposed.@*Methods@#To obtain refined features of retinal blood vessels, three cascade connected U-Net networks are employed. To deal with the problem of difference between the parts of encoder and decoder, in MF2ResU-Net, shortcut connections are used to combine the encoder and decoder layers in the blocks. To refine the feature of segmentation, atrous spatial pyramid pooling (ASPP) is embedded to achieve multi-scale features for the final segmentation networks.@*Results@#The MF2ResU-Net was superior to the existing methods on the criteria of sensitivity (Sen), specificity (Spe), accuracy (ACC), and area under curve (AUC), the values of which are 0.8013 and 0.8102, 0.9842 and 0.9809, 0.9700 and 0.9776, and 0.9797 and 0.9837, respectively for DRIVE and CHASE DB1. The results of experiments demonstrated the effectiveness and robustness of the model in the segmentation of complex curvature and small blood vessels.@*Conclusion@#Based on residual connections and multi-feature fusion, the proposed method can obtain accurate segmentation of retinal blood vessels by refining the segmentation features, which can provide another diagnosis method for computer-aided Chinese medical diagnosis.

Research on the expression and regulation of ASPP2 and its methylationin human gastric carcinoma cell / 中华内分泌外科杂志

Objective To investigate the relationship between the expression of ASPP2 mRNA and the methylation of ASPP2 gene in gastric cancer cells,to observe the inhibitory effect of 5-Aza-CdR on the growth of gastric cancer cells,to observe the effect of demethylation on the expression of ASPP2 mRNA and the methylation of ASPP2 gene in gastric cancer cells,and to explore the molecular mechanism of gastric cancer.Methods Real-time PCR was used to detect the expression of ASPP2 mRNA in two gastric cancer cells and normal gastric epithelial cells.BSP was used to detect the methylation of ASPP2 gene in two gastric cancer cells and normal gastric epithelial cells.CCK-8 was used to detect the growth inhibition rate of gastric cancer cells treated with 5-Aza-CdR of different concentrations,then they were used to detect expression of ASPP2 mRNA and the methylation of ASPP2 gene in gastric cancer cells again after the demethylation.Results ① The expression of ASPP2 mRNA in MKN-45 cells was significantly lower than that in GES-1 cells(P＜0.01).The expression of ASPP2 mRNA in MGC-803 cells was significantly lower than that in GES-1 cells (P＜0.01).There was no significant difference in MGC-803 cells and MKN-45 cells(P＞0.05).② The methylation rate of ASPP2 in MKN-45 cells was significantly higher than that in GES-1 cells (P＜0.01).The methylation rate of ASPP2 in MGC-803 cells was not significantly different from that in GES-1 cells (P＞0.05).The methylation rate of ASPP2 in MKN-45 cells was significantly higher than that in MGC-803 cells (P＜0.01).③ At the same time,the growth inhibition rate of each 5-Aza-CdR concentration group increased as the drug concentration depended.4.The expression of ASPP2 mRNA in MKN-45 cells was significantly higher than that before treatment (P＜0.01),the expression of ASPP2 mRNA in MGC-803 cells was not significantly different from that before treatment(P＞0.05).The methylation rate of ASPP2 in MKN-45 cells was significantly lower than that before treatment (P＜0.01).The methylation rate of ASPP2 in MGC-803 cells was not significantly different from that before treatment (P＞0.05).Conclusion ① Abnormal hypermethylation of ASPP2 gene in MKN-45 cells may be a molecular mechanism of decreased ASPP2 mRNA expression.② 5-Aza-CdR can inhibit the growth of MKN-45 and MGC-803 cells,and it can enhance the expression of ASPP2 mRNA in MKN-45 cells.Reversal of methylation in the promoter region of ASPP2 gene is the possible mechanism.③ Abnormal hypermethylation of the promoter region of ASPP2 gene may lead to silencing of mRNA expression that may be associated with gastric cancer.

Expression of ASPP2 and P16INK4a and its relationship with apoptosis in esophageal carcinoma / 实用医学杂志

Objective To study the expression of ASPP2 and P16INK4a in esophageal carcinoma and their relationship to the apoptosis and related clinicopathological characteristics. Methods Immunohistochemistry S-P method was used to examine the expression of ASPP2 and P16INK4a in the pathological specimens of 112 esophageal carcinoma cases and 31 cases of normal esophageal mucosa. TUNEL was also employed to detect the rate of apopto-sis in 37 esophageal carcinoma cases and 12 cases of normal esophageal mucosa. Results The difference between the expression of ASPP2 and P16INK4a in esophageal carcinoma and normal esophageal mucosa was statistically signif-icant(P < 0.05). Their abnormal expressions were all related to lymph node metastasis and differentiation degree (P < 0.05). The difference between the positive rate of apoptosis in esophageal carcinoma and normal esophageal mucosa was statistically significant(P < 0.05). The abnormal expression of ASPP2 and P16INK4a was all related to apoptosis in esophageal carcinoma. Conclusions The different expression of ASPP2 and P16INK4a may cooperatively play a role in differentiation degree ,lymph nodes metastasis and apoptosis in esophageal carcinoma. Co-examina-tion of them may be useful for the diagnosis and guiding the clinical treatment in esophageal carcinoma.

Expression of ASPP2 and P16INK4a and its relationship with apoptosis in esophageal carcinoma / 实用医学杂志

Objective To study the expression of ASPP2 and P16INK4a in esophageal carcinoma and their relationship to the apoptosis and related clinicopathological characteristics. Methods Immunohistochemistry S-P method was used to examine the expression of ASPP2 and P16INK4a in the pathological specimens of 112 esophageal carcinoma cases and 31 cases of normal esophageal mucosa. TUNEL was also employed to detect the rate of apopto-sis in 37 esophageal carcinoma cases and 12 cases of normal esophageal mucosa. Results The difference between the expression of ASPP2 and P16INK4a in esophageal carcinoma and normal esophageal mucosa was statistically signif-icant(P < 0.05). Their abnormal expressions were all related to lymph node metastasis and differentiation degree (P < 0.05). The difference between the positive rate of apoptosis in esophageal carcinoma and normal esophageal mucosa was statistically significant(P < 0.05). The abnormal expression of ASPP2 and P16INK4a was all related to apoptosis in esophageal carcinoma. Conclusions The different expression of ASPP2 and P16INK4a may cooperatively play a role in differentiation degree ,lymph nodes metastasis and apoptosis in esophageal carcinoma. Co-examina-tion of them may be useful for the diagnosis and guiding the clinical treatment in esophageal carcinoma.

Quality control of human ASPP2 recombinant adenovirus / 中国药理学通报

Aim Toanalyzethekeyqualityandestablishmeth-ods for essential quality control of human recombinant ASPP2 adenovirus.Methods TheviralstructuralgeneofE2Bandtar-get gene of ASPP2 were identified by PCR;The number of virus particles was measured by UV-SDS methods;Infectious titer was determined by TCID50 assay;Target protein of ASPP2 was ob-served by Western blot assay;The biological effects of recombi-nant adenovirus on liver cancer cells were evaluated by MTT as-say;A549 cells were used to check replication of the competent adenovirus(RCA)by the observation of the cytopathic effect. Results PCRanalysisofE2BandASPP2wasinconsistent with theoretical values;Particle numbers of virus were 5. 6 × 1012 VP/mL,infectious titer was 2 ×1011 IU/mL and specific activity was 3. 5%;ASPP2 protein expression could be detected when cells were infected with virus for 24 h;Growth inhibition of liver cancer cells could be found by adding recombinant ASPP 2 adenovirus;The level of RCA was less than 1 RCA/3. 0 ×1010 VP,in line with the standards of China Food and Drug Adminis-tration(SFDA).Conclusion Thequalitycontrolmethodswere established aiming at key characters of human recombinant AS-PP2 adenovirus,which may provide foundations for its quality standard and future applications.

ASPP2 gene silence promotes hepatocellular carcinoma cell proliferation via regulating autophagy under starvation / 第二军医大学学报

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.

Progress on aberrant methylation of INK4a/ARF and ASPP genes in tumor / 基础医学与临床

Many tumorigenesis in human are closely linked to the functional loss of P53. The tumor-suppressive functions of P53 are abrogated by mutations, but in 50% of all tumors it remains wild type. In these cases, loss of P53 tumor-suppressive function mainly results from the abnormal methylation of 5' CpG islands of tumor suppressor genes such as INK4a, ARF and ASPP. Restoring to normal methylation state of these genes might provide a new strategy for tumor therapy.

Progress on aberrant methylation of INK4a/ARF and ASPP genes in tumor / 基础医学与临床

Many tumorigenesis in human are closely linked to the functional loss of P53. The tumor-suppressive functions of P53 are abrogated by mutations,but in 50% of all tumors it remains wild type. In these cases,loss of P53 tumor-suppressive function mainly results from the abnormal methylation of 5' CpG islands of tumor suppressor genes such as INK4a,ARF and ASPP. Restoring to normal methylation state of these genes might provide a new strategy for tumor therapy.

Construction and effect identification of RNAi eukaryotic expression vectors of iASPP / 第三军医大学学报

Objective To construct the RNAi eukaryotic vector of inhibitory member of the ASPP family(iASPP) gene and observe the interfering effect in Jurkat cell line after the vector transfection.Methods The specific siRNA sequence was designed according to the iASPP sequence in GenBank.The sequence was cloned into PGCsilencer~(TM) H1/Neo/GFP and then sequence analysis was performed.The recombinant plasmid was transfected into Jurkat cell by liposome.The iASPP expression was analyzed by RT-PCR and cell apoptosis was detected by FCM.Results The sequence of templet and specific siRNA was correct by sequence analysis.The iASPP expression in Jurkat cells decreased and the apoptosis rate increased from 11.81% to 33.15% after RNAi transfection.Conclusion The RNAi eukaryotic vector PGCsilencer~(TM)H1/Neo/GFP/RNAi was constructed successfully.The RNAi inhibitory effect on the Jurkat cells is confirmed.The successful construction of iASPP RNAi makes it possible to further study the interaction between iASPP and p53.