Objective To explore the early diagnostic value of Golgi membrane protein 73 (GP73),alpha-fetoprotein (AFP) and alpha-fetoprotein-L3 (AFP-L3) in patients with high risk of primary hepatic carcinoma (PHC).Methods Sixty-four cases of PHC were selected as the PHC group,60 cases of liver cirrhosis(LC) as the LC group,53 cases of hepatitis as the hepatitis group and 51 healthy checked-up people as the control group.Enzyme-linked immunosorbent assay (ELISA) was used to detect the serum level of GP73 in all the cases.AFP-L3 was isolated by using affinity micro centrifugal column,AFP and AFP-L3 were detected with chemiluminescent immunoassay and then the proportion of AFP-L3 was calculated.Results The positive rate of serum GP73,AFP and AFP-L3 in PHC group was significantly higher than that in LC group and hepatitis group [78.1％ (50/64)vs.25.0％ (15/60),17.0％ (9/53);48.4％ (31/64) vs.31.7％ (19/60),22.6％(12/53) ;53.1％(34/64) vs.30.0％(18/60),20.8％(11/53)] (P ＜ 0.05),In control group,GP73,AFP,AFP-L3 was no positive.The levels of GP73,AFP and AFP-L3 in PHC group were significantly higher than those in LC group,hepatitis group and control group [(245.69 ± 89.18)μ g/L vs.(116.37 ±38.52),(97.29 ± 24.58),(23.48 ±9.12) μ g/L; (403.27 ± 128.46) μg/L vs.(75.62 ± 19.35),(66.49 ± 15.14),(3.46 ± 1.02) μg/L; (15.64 ±3.19)％ vs.(5.24 ± 1.15),(4.21 ± 0.96),(2.95 ±0.73)％] (P ＜0.05).The levels of GP73,AFP in LC group and hepatitis group were significantly higher than those in control group (P ＜ 0.05).The levels of GP73,AFP and AFP-L3 had no statistically significant difference between LC group and hepatitis group (P ＞ 0.05).Sensitivity and accuracy of three combined detection for PHC was 96.9％(62/64),91.7％(209/228),significantly higher than that of AFP,AFP-L3 single detection (P ＜ 0.05).GP73 single detection and any two combined detection was no significant difference in sensitivity and accuracy,compared with three combined detection (P ＞ 0.05).The levels of GP73 in PHC patients with different age,gender,serum level of AFP,TNM stage and tumor diameter had no statistically significant difference (P ＞ 0.05).The levels of GP73 in PHC patients with positive HBsAg,extrahepatic metastases and LC had significant difference (P ＜ 0.05).Conclusions The diagnosis value of GP73 is evidently higher than AFP and AFP-L3 for PHC,and combined determination is superior to single marker.Combined determination enhances the degree of precision in populations with high risk of PHC diagnosis.

Objective To synthesize novel gold nanoparticles of GAL-PEG-GNPs,study its radiation effect on hepatocellular carcinoma cells HepG2 cells in vitro,and investigate the underlying mechanisms.Methods GAL-PEG-GNPs were synthesized and characterized successfully.HepG2 cells were divided into three groups of control,GNPs and GAL-PEG-GNPs.The cytotoxicities of these compounds were tested by the CCK-8 assay and their IC50 values of HepG2 cells were calculated.Cell uptake of nanoparticles was detected by TEM and ICP-MS.The radiosensitization effect of nanoparticles was tested by the colony formation assay.Cell cycle distribution was detected by FCM.The expressions of CAT,SOD,and total GSH were detected with a microplate reader,and the expressions of apoptosis-related proteins were tested by Western blot.Results The GNPs and GAL-PEG-GNPs had absorption peaks at 520 and 530 nm,respectively,and their diameters were (22.6-±2.12) and (32.0 ± 1.41) nm detected by ICP-MS.The GAL-PEG-GNPs and GNPs had similar cytotoxicity profiles (P ＞ 0.05),while GAL-PEG-GNPs could be more effectively uptaken by HepG2 cells than GNPs.The sensitive enhancement ratio (SER) of GNPs and GAL-PEG-GNPs to HepG2 cells were 1.46 和 1.95,respectively.The percentage of cells at phase of G2/M in HepG2 population treated with GNP was higher than that of untreated cells (t =14.20,P ＜0.05).The protein expressions of Cytochrome C,Bax,Caspase-3,and Caspase-9 were upregulated while Bcl-2 expression was down-regulated in the cells treated with GNPs/radiation or GAL-PEGGNPs/radiation.The expressions of CAT,SOD and total GSH in the GNP treated groups were significantly decreased compared with the control group(t =12.34,29.39,12.85,P ＜ 0.05).Conclusions GALPEG-GNPs has obvious radiosensitization effect on HepG2 cells,which is related to the induction of cell apoptosis and the generation of free radicals.

Objective To study the radiosensitizing effect of nano-gold nano-silver particles in hepatocellular carcinoma cells (HepG2) in vitro and the possible mechanisms.Methods MTT assay and clonogenic assay were performed to determine the killing effect of nano-gold and nano-silver particles in HepG2 cells.Flow-cytometry was used to measure cell apoptosis and cell cycle distribution.Western blotting was used to measure the expression of Caspase-3,Bax and Bcl-2.ELIASA was used to determine the content of catalase (CAT),superoxide dismutase (SOD),and total glutathione (GSH).Results Nano-gold and nano-silver particles inhibited the proliferation of HepG2 cells with IC50 of 6.51 μg/ml and 2.47 μg/ml,respectively.Nano-gold and nano-silver particles significantly enhanced the radiosensitivity of HepG2 cells.Obtained by Dq,the SER of 1/5 IC50 nano-gold and nano-silver particles were 1.37 and 1.48,and 1/10IC50 with 1.11 and 1.09.Nano-gold and nano-silver particles increased the expression of Caspase-3 and Bax and reduced the exprcssion of Bcl-2.CAT,SOD and total GSH were significantly reduced.Conclusions Nano-gold and nano-silver particles can enhance the radiation sensitivity of HepG2 cells.Specific sensitizing mechanism may be the activation of the mitochondrial apoptosis pathway and the induction of reactive oxygen species in apoptotic pathways,which ultimately induces apoptosis.

A novel unsaturated polyphosphoester (UPPE) was devised in our previous research, which is a kind of promising scaffold for improving bone regeneration. However, the polymerization process of UPPE scaffolds was unfavorable, which may adversely affect the bioactivity of osteoinductive molecules added if necessary, such as recombinant human bone morphogenetic protein-2 (rhBMP2). The purpose of this study was to build a kind of optimal scaffold named UPPE-PLGA-rhBMP2 (UPB) and to investigate the bioactivity of rhBMP2 in this scaffold. Furthermore, the cytotoxicity and biocompatibility of UPB scaffold was assessed in vitro. A W1/O/W2 method was used to fabricate PLGA-rhBMP2 microspheres, and then the microspheres were added to UPPE for synthesizing UPB scaffold. The morphological characters of PLGA-rhBMP2 microspheres and UPB scaffolds were observed under the scanning electron microscopy and laser scanning confocal microscopy. The cumulative release of UPB scaffolds was detected by using ELISA. The cytotoxicity and biocompatibility of UPB scaffolds were evaluated through examining the adsorption and apoptosis of bone marrow stromal cells (bMSCs) seeded on the surface of UPB scaffolds. The bioactivity of rhBMP2 in UPB scaffolds was assessed through measuring the alkaline phosphates (ALP) activity in bMSCs seeded. The results showed that UPB scaffolds sequentially exhibited burst and sustained release of rhBMP2. The cytotoxicity was greatly reduced when the scaffolds were immersed in buffer solution for 2 h. bMSCs attached and grew on the surface of soaked UPB scaffolds, exerting well biocompatibility. The ALP activity of bMSCs seeded was significantly enhanced, indicating that the bioactivity of rhBMP2 remained and still took effect after the unfavorable polymerization process of scaffolds. It was concluded that UPB scaffolds have low cytotoxicity, good biocompatibility and preserve bioactivity of rhBMP2. UPB scaffolds are promising in improving bone regeneration.

A novel unsaturated polyphosphoester (UPPE) was devised in our previous research, which is a kind of promising scaffold for improving bone regeneration. However, the polymerization process of UPPE scaffolds was unfavorable, which may adversely affect the bioactivity of osteoinductive molecules added if necessary, such as recombinant human bone morphogenetic protein-2 (rhBMP2). The purpose of this study was to build a kind of optimal scaffold named UPPE-PLGA-rhBMP2 (UPB) and to investigate the bioactivity of rhBMP2 in this scaffold. Furthermore, the cytotoxicity and biocompatibility of UPB scaffold was assessed in vitro. A W1/O/W2 method was used to fabricate PLGA-rhBMP2 microspheres, and then the microspheres were added to UPPE for synthesizing UPB scaffold. The morphological characters of PLGA-rhBMP2 microspheres and UPB scaffolds were observed under the scanning electron microscopy and laser scanning confocal microscopy. The cumulative release of UPB scaffolds was detected by using ELISA. The cytotoxicity and biocompatibility of UPB scaffolds were evaluated through examining the adsorption and apoptosis of bone marrow stromal cells (bMSCs) seeded on the surface of UPB scaffolds. The bioactivity of rhBMP2 in UPB scaffolds was assessed through measuring the alkaline phosphates (ALP) activity in bMSCs seeded. The results showed that UPB scaffolds sequentially exhibited burst and sustained release of rhBMP2. The cytotoxicity was greatly reduced when the scaffolds were immersed in buffer solution for 2 h. bMSCs attached and grew on the surface of soaked UPB scaffolds, exerting well biocompatibility. The ALP activity of bMSCs seeded was significantly enhanced, indicating that the bioactivity of rhBMP2 remained and still took effect after the unfavorable polymerization process of scaffolds. It was concluded that UPB scaffolds have low cytotoxicity, good biocompatibility and preserve bioactivity of rhBMP2. UPB scaffolds are promising in improving bone regeneration.
Bone Morphogenetic Protein 2 ; chemistry ; pharmacology ; Bone Regeneration ; drug effects ; Humans ; Lactic Acid ; chemistry ; pharmacology ; Phosphatidylinositol Phosphates ; chemistry ; pharmacology ; Polyglycolic Acid ; chemistry ; pharmacology ; Tissue Scaffolds

Percutaneous imaging-guided cryoablation (PICA) is a recently developed technique,which applies extreme hypothermia to destroy tumors guided with CT,MRI or ultrasound images.Compared with other oncologic cryoablation therapies,PICA has some advantages,such as treating under local anesthesia,superior monitoring capability on multimodal imaging,less intraoperative and postoperative pain and preservation of tissue collagenous architecture.In recent years,PICA has been used in treatment of tumors in lung,liver,kidney,breast,prostate and so on.In this article,the principle of cryoablation and the application progresses of PICA in oncotherapy were reviewed.

Objective To evaluate the value of dynamic contrast-enhanced MRI (DCE-MRI)in diagnosing benign and malignant solitary pulmonary nodule (SPN)with Meta-analysis.Methods The databases including PubMed,EBSCO,Cochrane Library,Ovid, CBM,VIP,Wan Fang Database and CNKI were searched to select the literatures about the diagnosis of SPN with DCE-MRI.Inclusion criteria were established,according to the validity criteria for diagnostic research published by the Cochrane Methods Group on Screening and Diagnostic Tests .Methodological quality was assessed by using the quality assessment of diagnostic studies (QUADAS-2)instrument.Meta-analysis was performed by Stata 12.0 and Meta-Disc 1.4.According to the results of heterogeneity of the included articles,proper effect model was selected to calculate the pooled sensitivity,specificity,positive likelihood ration,negative likelihood ration,and diagnostic odds ratio.Summary receiver operating characteristic (SROC)curve was drawed and the area under the curve (AUC)was calculated.Results A total of 17 studies involving 1255 lesions were included.The results of Meta-analysis showed the pooled sensitivity,specificity,positive likelihood ration,negative likelihood ration,and diagnostic odds ratio were 0.95,0.81,4.9,0.06 and 85,respectively.The AUC of SROC was 0.97.Conclusion DCE-MRI has relatively high sensitivity and specificity in the differential diagnosis of benign and malignant SPN.

Objective To investigate the effect of exosomes secreted from human lung adenocarcinoma A549 cells under hypoxic or normoxic conditions on the radiosensitivity and invasiveness of normoxia cells.Methods A549 cells were cultured in hypoxic (1％ O2) and normoxic (21％ O2) conditions,respectively.The exosomes (N-EXO and H-EXO) secreted from normoxic or hypoxic A549 cells were collected by ultracentrifugation and its number was measured using a NanoSight detector.The appearance and size distribution of exosomes were observed by a scanning electron microscopy.The exosomal marker protein CD63 was measured by Western blot.The proliferation of cells exposed to X-rays under hypoxic or normoxic conditions were detected by CCK8 assay.The cell uptake situation of exosomes labeled with PKH67 was observed by a fluorescence microscopy.Cell migration and invasiveness were detected by a cell scratch test and transwell assay.The expression of matrix metalloproteinase 2 (MMP2) and MMP9 was detected by ELISA.Cellular radioresistance effect of exosomes was evaluated by a colony formation assay.Results The NanoSight measurement showed the number of exosomes in cell culture medium was increased after hypoxia treatment.The H-EXO and N-EXO showed typical ring cake shape.The size distribution of H-EXO was mainly between 30 nm and 200 nm,smaller than that of N-EXO (50-220 nm).Western blot assay showed that CD63 was expressed in both H-EXO and N-EXO.At 4 and 6 days after 2 Gy X-rays irradiation,cell proliferation rate of hypoxia A549 cells was significantly higher than that of normoxia cells.The green fluorescent marker of exosomes,PKH67,was distributed inside of the cell.Cell scratch test showed that the width of H-EXO group was much smaller than that of N-EXO group at 12,24 and 48 hours after exosomes treatment (t =2.96,6.76,3.35,P ＜ 0.05).Transwell assay showed that the number of transmembrane cells in the H-EXO group was more than that in the N-EXO group and the control group (t =4.84,7.88,P ＜ 0.O1).The expression levels of MMP2 (t =4.70,3.21,P＜0.05) and MMP9 (t =5.61,3.76,P＜0.05) in the supernatant of H-EXO group were significantly higher than those in the control and N-EXO groups.Cell survival assay showed that the D0 values of control,N-EXO and H-EXO group were 2.614,2.552 and 4.50 respectively,indicating that H-EXO could enhance radioresistance of A549 cells significantly.Conclusions This study finds that the number of exosomes released from A549 cells was increased under hypoxic condition but its size becomes smaller than that under normoxia.Hypoxic exosomes can promote the migration of normoxia cells andenhance cell radioresistance as well.

Objective:To explore the effectiveness of a hysteroscopic endometrial lesion diagnosis model de-veloped based on deep learning(DL)algorithm combined with gradient-weighted class activation mapping(Grad-CAM)visualization technology.Methods:303 hysteroscopy videos(4781 images)of 291 patients who un-derwent hysteroscopy examination in the Department of Gynecology,Renmin Hospital of Wuhan University from June 1,2021 to December 31,2022 were selected.The dataset was divided into a training set(3703 images)and a test set(1078 images)by weight sampling method.After the training set was used for model learning and train-ing,two model architectures,residual neural network(ResNet18)and efficient neural network(EfficientNet-B0),were selected to verify the model in the test set by five-class and two-class classification tasks,respectively.Tak-ing histopathology as the gold standard,the diagnostic efficacy was evaluated to select the optimal model,and the Grad-CAM layer was embedded in the optimal model to output hysteroscopy images of Grad-CAM.Results:①In the five-class classification tasks,the accuracy of EfficientNet-B0 model(93.23％)was higher than that of Res-Net18 model(84.23％);the area under the curve(AUC)of EfficientNet-B0 model in the diagnosis of five disea-ses,including atypical endometrial hyperplasia,endometrial polyps,endometrial cancer,endometrial atypical hy-perplasia,and submucous myoma,was slightly higher than that of ResNet18 model,and the AUC of both models was almost above 0.980.②In the binary classification task of accuracy and the evaluation of specificity,the two models were similar,both above 93.00％,and the sensitivity of EfficientNet-B0 model(91.14％)was significantly better than that of ResNet18 model(77.22％).③EfficientNet-B0 model combined with Grad-CAM algorithm could identify the abnormal areas in the image.After biopsy and pathological examination,it was confirmed that about 95％of the marked areas in the model's output heatmap were lesion areas.Conclusions:The hysteroscopy di-agnostic model developed by EfficientNet-B0 model combined with Grad-CAM has high diagnostic accuracy,sen-sitivity,and specificity,and has application value in the diagnosis of endometrial lesions.

Objective:To explore the effectiveness of a hysteroscopic endometrial lesion diagnosis model de-veloped based on deep learning(DL)algorithm combined with gradient-weighted class activation mapping(Grad-CAM)visualization technology.Methods:303 hysteroscopy videos(4781 images)of 291 patients who un-derwent hysteroscopy examination in the Department of Gynecology,Renmin Hospital of Wuhan University from June 1,2021 to December 31,2022 were selected.The dataset was divided into a training set(3703 images)and a test set(1078 images)by weight sampling method.After the training set was used for model learning and train-ing,two model architectures,residual neural network(ResNet18)and efficient neural network(EfficientNet-B0),were selected to verify the model in the test set by five-class and two-class classification tasks,respectively.Tak-ing histopathology as the gold standard,the diagnostic efficacy was evaluated to select the optimal model,and the Grad-CAM layer was embedded in the optimal model to output hysteroscopy images of Grad-CAM.Results:①In the five-class classification tasks,the accuracy of EfficientNet-B0 model(93.23％)was higher than that of Res-Net18 model(84.23％);the area under the curve(AUC)of EfficientNet-B0 model in the diagnosis of five disea-ses,including atypical endometrial hyperplasia,endometrial polyps,endometrial cancer,endometrial atypical hy-perplasia,and submucous myoma,was slightly higher than that of ResNet18 model,and the AUC of both models was almost above 0.980.②In the binary classification task of accuracy and the evaluation of specificity,the two models were similar,both above 93.00％,and the sensitivity of EfficientNet-B0 model(91.14％)was significantly better than that of ResNet18 model(77.22％).③EfficientNet-B0 model combined with Grad-CAM algorithm could identify the abnormal areas in the image.After biopsy and pathological examination,it was confirmed that about 95％of the marked areas in the model's output heatmap were lesion areas.Conclusions:The hysteroscopy di-agnostic model developed by EfficientNet-B0 model combined with Grad-CAM has high diagnostic accuracy,sen-sitivity,and specificity,and has application value in the diagnosis of endometrial lesions.