1.Effects of cripto siRNA on liver metastasis of colorectal cancer cell in nude mice
Ximing ZHONG ; Yu FAN ; Yongjing ZHOU ; Jian CHEN ; Gengjin LIN
Fudan University Journal of Medical Sciences 2010;37(2):202-206
Objective To study the effects of cripto on migration, invasion, and liver metastasis of colorectal cancer cell. Methods After human colorectal cancer cell line SW480 was transfected by cripto small interfering RNA (siRNA), the mRNA and protein level were determined by Real-time RT-PCR and Western blot, respectively. The migration and invasion ability were evaluated by wound-healing assay and boyden chamber model, respectively. Thirty nude mice model of liver metastasis from colorectal cancer was established by splenectomy. Results The siRNA could down-regulate the level of mRNA and protein of cripto in a dose- and time-dependent manner. Suppression of cripto expression could inhibit migration and invasion ability of human colorectal cancer cell in vitro. The metastastic rate and tumor nodules were lower in transfection with cripto siRNA than in two control groups in vivo. Conclusions Cripto gene might play an important role in regulation of liver metastasis from colorectal carcinoma cell, and suppression of cripto gene by siRNA can inhibit liver metastasis of colorectal cancer.
2.Clinical significance of extracellular matrix and hepatic ultramicrostructural changes in patients with mild chronic hepatitis B
Wei LIN ; Chenwei PAN ; Lu ZHUGE ; Yi ZHENG ; Guangyao ZHOU ; Zhouxi FANG ; Ximing Lü ; Linxiang JIN
Chinese Journal of Clinical Infectious Diseases 2011;04(1):29-32
Objective To investigate the correlations of extracellular matrix and hepatic ultramicrostructural changes with clinical manifestations in patients with mild chronic hepatitis B (CHB).Methods Patients with chronic HBV infections were enrolled and were divided into mild CHB group (n=66) and HBV carrier group (n=10).Serum samples were collected from patients, and serum HBV markers, HBV DNA load and liver fibrosis indexes were measured.All subjects received liver biopsy, and the tissue samples were observed by light microscope and electron microscope.T test and χ2 test were performed for measurement data and enumeration data, respectively.Spearman test was used for ranked data.Results The differences on ALT and AST levels between mild CHB group and HBV carrier group were significant (t=12.42, 7.06, P<0.05), but there was no significant difference on HBV DNA load between two groups (t=0.24, P > 0.05).Serum liver fibrosis indexes (hyaluronic acid, type Ⅲ collagen,type Ⅳ collagen and laminin protein) in mild CHB group were not significantly higher than those in HBV carrier group (t=0.45, 0.95, 0.76 and 1.21, P >0.05).In mild CHB group, there were 33 patients with ≥G2 and ≥S2, but in HBV carrier group were only 2 patients (χ2=4.17, P < 0.05).Seventeen patients in mild CHB group were with S3-4, while that was not observed in HBV carrier group (χ2=4.75, P <0.05).In mild CHB group, hepatic ultramicrostrutural changes on fat storing cell, collagen protein and portal area were correlated with fibrosis grades, and the correlation coefficients were 0.351, 0.675 and 0.301, respectively (P=0.004, 0.000 and 0.014).Conclusion Electron microscope is of higher sensitivity than light microscope in observing hepatic ultramicrostructural changes, which is effective in evaluating the severity of mild CHB.
3.Preparation of the mixed serum calibrators for C-reactive protein
Yazhe FANG ; Huaian MA ; Nan GUO ; Baoping LI ; Ying LIN ; Ximing YANG
Chinese Journal of Laboratory Medicine 2014;(11):832-835
Objective To prepare serum calibrators for CRP measurement.Methods Fresh serum without infectious diseases , hemolysis, lipemia and choloplania were collected and divided into 3 groups, low, medium, and high, according to the CRP concentration.Each serum pool was mixed , filtered, sterilized and aliquoted.The materials were tested for homogeneity and stability.The values of the CRP was assigned by particle enhanced immunonephelpmetry , and calibrated with international reference materials.The expanded uncertainty was evaluated.Results The materials were tested to be homogeneous (Ubb﹤Ur, P>0.05) with Ubb values being 0, 0.125, 0, Ur values being 0.046, 0.213, 0.785, and F values being 0.803, 1.686, 0.966 in CL, CM, CH groups respectively.Stability study, where F values are 0.609, 0.259, and 1.557 at 22-25℃, 1.217, 4.583, and 0.893 at 2-8℃(P>0.05), showed that the materials were stable for at least 3 days at 22-25 ℃or 30 days at 2-8 ℃, respectively.The certified values of the 3 levels materials for CRP were ( 2.64 ±0.14 ) , ( 31.17 ±0.63 ) , ( 73.85 ±1.74 ) mg/L, respectively.Conclusion The calibrators prepared for serum CRP measurement were homogeneous , stable and accurately assigned and can be used to calibrate the CRP measure system.
4.The immunity induced by recombinant spike proteins of SARS coronavirus in Balb/c mice.
Li, QIN ; Ximing, WANG ; Shaoting, WU ; Shishan, YUAN ; Dana, HUANG ; Mingjun, LEI ; Huirong, PAN ; Qiping, LIN
Journal of Huazhong University of Science and Technology (Medical Sciences) 2007;27(3):221-4
The immune effect of two recombinant protein fragments of spike protein in severe acute respiratory syndrome coronavirus (SARS CoV) was investigated in Balb/c mice. Two partial spike gene fragments S1 (322 1464 bp) and S2 (2170 2814 bp) of SARS coronavirus were amplified by RT-PCR, and cloned into pET-23a prokaryotic expression vector, then transformed into competent Escherichia E. coli BL21 (DE3)(pLysS) respectively. Recombinant proteins were expressed and purified by Ni2+ immobilized metal ion affinity chromatography. The purified proteins mixed with complete Freund adjuvant were injected into Balb/c mice three times at a two-week interval. High titer antibody was detected in the serum of immunized Balb/c mice, and mice immunized with S1 protein produced high titer IgG1, IgG2a, IgG2b and IgG3, while those immunized with S2 protein produced high titer IgG1, IgG2a, but lower titer IgG2b and IgG3. Serum IFN-concentration was increased significantly but the concentrations of Il-2, IL-4 and IL-10 had no significant change. And a marked increase was observed in the number of spleen CD8+ T cells. The results showed that recombinant proteins of SARS coronavirus spike protein induced hormonal and cellular immune response in Balb/c mice.
5.Effect of silencing ACAT1 gene by RNA interference on proliferation, migration and invasion of human colon cancer cells
Xin CHEN ; Huiling LIANG ; Ximing XU ; Yiqiao ZHANG ; Neng CHEN ; Dandan LIN ; Ling FENG ; Yongbin ZHENG
The Journal of Practical Medicine 2017;33(7):1074-1077
Objective To evaluate the effect of silencing ACAT1 gene on colon cancer cells proliferation,migration,invasion and colon cancer development by using the small interference RNA (siRNA) in colon cancer cell line HT-29.Methods Acyl coenzyme A cholesterol acyltransferase 1 (ACAT1) gene was silenced in HT-29 cell lines using Hiperfect transfection reagent.The expression level of ACAT1 was detected by real time PCR.CFSE and transwell assays were used to evaluate the effect of ACAT1 gene interfering on cells proliferation,mi gration and invasion.Result ACAT1 mRNA expression decreased obviously after siRNA interference.Compared with pre-transfection,proliferation,migration and invasion of colon cancer cells have been significantly inhibited (P < 0.05).Conclusion ACAT1 gene interference reduced proliferation,migration and of invasion of HT29 cells,which provide a new potential target for colon cancer treatment.
6.The Immunity Induced by Recombinant Spike Proteins of SARS Coronavirus in Balb/c Mice
Li QIN ; Ximing WANG ; Shaoting WU ; Shishan YUAN ; Dana HUANG ; Mingjun LEI ; Huirong PAN ; Qiping LIN
Journal of Huazhong University of Science and Technology (Medical Sciences) 2007;27(3):221-224
The immune effect of two recombinant protein fragments of spike protein in severe acute respiratory syndrome coronavirus (SARS CoV) was investigated in Balb/c mice. Two partial spike gene fragments S1 (322-1464 bp) and S2 (2170-2814 bp) of SARS coronavirus were amplified by RT-PCR, and cloned into pET-23a prokaryotic expression vector, then transformed into competent Escherichia E.coli BL21 (DE3)(pLysS) respectively. Recombinant proteins were expressed and purified by Ni2+ immobilized metal ion affinity chromatography. The purified proteins mixed with complete Freund adjuvant were injected into Balb/c mice three times at a two-week interval. High titer antibody was detected in the serum of immunized Balb/c mice, and mice immunized with S1 protein produced high titer IgG1, IgG2a, IgG2b and IgG3, while those immunized with S2 protein produced high titer IgG1, IgG2a, but lower titer IgG2b and IgG3. Serum IFN-γ concentration was increased significantly but the concentrations of IL-2, IL-4 and IL-10 had no significant change. And a marked increase was observed in the number of spleen CD8+ T cells. The results showed that recombinant proteins of SARS coronavirns spike protein induced hormonal and cellular immune response in Balb/c mice.
7.IWR-1-endo affects the migration and proliferation of hepatocarcinoma cells by inhibiting the Wnt pathway
Lin XIONG ; Xiuyun ZHANG ; Xiaoyu ZHANG ; Yue LI ; Ximing XU
Journal of International Oncology 2021;48(12):711-715
Objective:To investigate the effects and mechanisms of Wnt pathway inhibitor IWR-1-endo on the biological behaviors of human hepatocarcinoma cell Huh7.Methods:Human hepatocellular carcinoma cell Huh7 was cultured in vitro, and Huh7 cells were treated with IWR-1-endo at different concentrations (0, 20, 40, 80, 160, 320 μmol/L). Scratch test was used to detect changes in cell migration ability at diffe-rent drug concentrations, plate cloning was used to detect changes in cell proliferation, Western blotting was used to detect changes in the expression of Wnt pathway related protein β-catenin, and immunofluorescence staining was used to detect the expression of β-catenin in cytoplasm and nucleus. Results:The results of the scratch test showed that the 24 h scratch healing rates of Huh7 cells treated with 0, 20, 40, 80, 160, 320 μmol/L IWR-1-endo were (20.55±0.05)%, (12.10±0.08)%, (9.36±0.10)%, (3.62±0.09)%, (0.62±0.04)% and (0.23±0.02)%, respectively, and there was a statistically significant difference ( F=230.87, P<0.001). Further pair comparison showed that there were statistically significant differences in 24 h scratch healing rates among different concentrations (all P<0.001). The 48 h scratch healing rates were (34.77±0.08)%, (17.69±0.05)%, (11.60±0.04)%, (5.68±0.07)%, (2.66±0.04)% and (1.75±0.02)%, respectively, and there was a statistically significant difference ( F=589.68, P<0.001). Further pair comparison showed that there were statistically significant differences in 48 h scratch healing rates among different concentrations (all P<0.001). After treatment with IWR-1-endo at the concentration of 0, 20, 40, 80, 160, 320 μmol/L, the clone formation rates of Huh7 cells were (61.67±0.21)%, (57.33±0.11)%, (50.00±0.25)%, (36.67±0.28)%, (23.33±0.12)% and (15.00±0.08)%, respectively, and there was a statistically significant difference ( F=403.56, P<0.001). Further pair comparison showed that there were statistically significant differences in clone formation rates among different concentrations (all P<0.001). After treatment with 0, 20, 40, 80, and 160 μmol/L IWR-1-endo for 24 h, the relative expression levels of β-catenin in Huh7 cells were 0.30±0.08, 0.25±0.07, 0.22±0.05, 0.15±0.01 and 0.06±0.02, respectively, and there was a statistically significant difference ( F=247.00, P<0.001). Compared with 0 μmol/L, the relative expression levels of β-catenin treated with 80 and 160 μmol/L had statistical significance ( P=0.014; P=0.008). Compared with 0 mol/L, immunofluorescence showed that the expressions of β-catenin in cytoplasm and nucleus were reduced after 80 μmol/L IWR-1-endo treatment. Conclusion:Wnt pathway inhibitor IWR-1-endo can inhibit the migration and proliferation of hepatocarcinoma cells Huh7 by inhibiting the activity of Wnt pathway. The above inhibitory effects are dose-dependent.
8.Correlation study of triglyceride-glucose index and plaque burden and characteristics of vulnerable plaque based on coronary CT angiography plaque quantitative analysis
Ruopeng WANG ; Hui GU ; Yan GAO ; Ying WANG ; Xinxin YU ; Fanfan CAI ; Lin GAO ; Ximing WANG
Chinese Journal of Radiology 2021;55(11):1153-1160
Objective:To explore the relationship between triglyceride-glucose (TyG) index with plaque components, plaque burden and characteristics of vulnerable plaque using coronary plaque analysis based on coronary artery computed tomography (CCTA).Methods:A total of 498 patients(male 296, female 202), the age ranged from 33 to 87 (63±9) years who underwent CCTA from January 2020 to September in Shandong Provincial Hospital Affiliated to Shandong First Medical University were included. The enrolled patients were divided into three groups according to the tertiles of TyG index: T 1 group (the lowest one-third), T 2 group (middle one-third) and T 3 group (the highest one-third). The plaque burden, volume and ratio of calcified, lipid and fibrous components of plaques as well as the incidence of vulnerable plaques were measured based on CCTA images. Chi-square test, ANOVA and Kruskal-Wallis test were used to compare whether the differences of indexes among the three groups were statistically significant. Multiple stepwise regression was used to analyze the influencing factors of coronary atherosclerotic plaque burden and multiple logistic regression was used to analyze the risk factors of CT high-risk plaque. Finally, ROC curve was used to evaluate the value of TyG index in the diagnosis of CT high-risk plaque, and the best diagnostic threshold of TyG index was determined. Results:The plaque burden, non-calcified plaque volume and ratio had positive relationship with TyG index ( P<0.001).TyG index was significantly correlated with plaque burden according to multiple stepwise regression analysis (regression coefficient 7.267, P<0.001). The results of CT vulnerable characteristics of plaques showed that positive remodeling, low-attenuation plaque sign and the incidence of vulnerable plaque increased with TyG index ( P<0.05). Multivariate Logistic regression analysis showed that TyG index was an independent risk factor for CT vulnerable plaque(OR=2.324,95 %CI 1.533-3.523, P<0.001). The cut-off value of TyG index that can predict vulnerable plaque was 8.43(sensitivity 77.24%, specificity 45.60%, AUC 0.645, P<0.001). Conclusions:With the increase of TyG index, the burden of coronary atherosclerosis plaque and the incidence of CT vulnerable plaque increased. TyG index is expected to be a simple and effective predictor of cardiovascular disease and adverse cardiovascular events.
9.The value of CT features in predicting the invasion and invasive degree of lung pure ground-glass nodules based on the new classification of lung tumor in 2021
Lin GAO ; Jing ZHANG ; Hui GU ; Bing KANG ; Xinxin YU ; Shuai ZHANG ; Yan GAO ; Fanfan CAI ; Ruopeng WANG ; Ximing WANG
Chinese Journal of Radiology 2022;56(6):616-622
Objective:To investigate the value of CT features in predicting the invasion and degree of invasiveness of lung pure ground-glass nodules (pGGN) in the new histological classification in 2021.Methods:A total of 281 patients (304 lesions) with pGGN confirmed by surgical pathology from December 2018 to January 2021 in Shandong Provincial Hospital Affiliated to Shandong First Medical University were retrospectively analyzed. According to the pathological types, the patients were divided into prodromal lesion group [atypical adenomatous hyperplasia (AAH) and adenocarcinoma in situ (AIS), 129 cases], minimally invasive group [minimally invasive adenocarcinoma (MIA), 116 cases] and invasive group [invasive adenocarcinoma (IAC), 59 cases]. Clinical data (age, gender, smoking history, family history of cancer), and CT parameters [shape, boundary, lobulation, burr, vacuolar sign, bronchial abnormality sign, internal vessel sign, pleural traction sign, longest diameter, shortest diameter, unenhanced CT value, contrast-enhanced CT value in arterial phase, contrast-enhanced CT values in venous phase, the degree of enhancement (ΔCT A-N, ΔCT V-N)] were recorded and measured. The ANOVA, Kruskal-Wallis H and χ 2 test were used to compare the differences among the three groups. Binary logistic regression analysis was used to evaluate the independent risk factors of nodular invasion [prodromal lesion and invasive lesion (MIA and IAC)] and the degree of nodular invasion (MIA and IAC), and receiver operating characteristic (ROC) curve analysis was performed for each parameter. Results:There were statistically significant differences in age, pGGN morphology, lobulation, vacuolar sign, bronchial abnormality sign, internal vascular sign, pleural traction sign, longest diameter, shortest diameter, unenhanced CT value, contrast-enhanced CT value in arterial phase, contrast-enhanced CT value in venous phase among the precursor lesion group, minimally invasive group and invasive group ( P<0.05). Binary logistic regression analysis showed that vacuole sign (OR=2.832, 95%CI 1.363-5.887, P=0.005), internal vascular sign (OR=3.021, 95%CI 1.909-4.779, P<0.001) and unenhanced CT value (OR=1.003, 95%CI 1.001-1.006, P=0.019) were independent risk factors for invasion. Lobulation (OR=5.739, 95%CI 2.735-12.042, P<0.001), internal vascular sign (OR=1.968, 95%CI 1.128-3.433, P=0.017) and unenhanced CT value (OR=1.004, 95%CI 1.001-1.008, P=0.012) were independent risk factors for the degree of invasiveness. ROC curve analysis showed that the efficiency of internal vascular sign was the highest in distinguishing precursor lesion and the invasive, the area under the curve (AUC) was 0.757, the sensitivity was 50.3%, the specificity was 89.8%. The efficiency of lobulation was the highest in distinguishing MIA and IAC (AUC=0.702), with a sensitivity of 61.0% and specificity of 79.3%. Conclusions:CT features are of certain value in predicting the invasion and degree of invasiveness of lung pGGN in the new histological classification in 2021, and internal vascular sign is more effective in predicting the invasion of lung pGGN. Lobulation can predict the degree of invasiveness of pGGN better.
10.Introduction of guidelines for iodine thyroid blocking
Chunhui CHANG ; Ximing FU ; Huifang CHEN ; Long YUAN ; Yuhan HOU ; Lin HE ; Cuiping LEI
Chinese Journal of Radiological Health 2021;30(3):253-257
During a radiological or nuclear emergency, iodine thyroid blocking is an urgent protection action to prevent or reduce the absorption of radioactive iodine by thyroid. Although potassium iodide (KI) administration is recommended by WHO guidelines for iodine prophylaxis following nuclear accidents and is also widely implemented in most national guidelines, the scientific evidence for the guidelines lacks as the guidelines are mostly based on expert opinions and recommendations. This paper introduces the development and revision of WHO guidelines for iodine thyroid blocking published in 1989 and 1999, as well as the revision methods and main recommendations of the guidelines published in 2017, which supports the use of iodine thyroid blocking during a radiological or nuclear emergency.