RESUMO
Objective To investigate the effect of Trichomonas vaginalis macrophage migration inhibitory factor (TvMIF) on THP-1 macrophages.. Methods Recombinant TvMIF protein was prokaryotic expressed and purified, and endotoxin was removed after identification. Following exposure to TvMIF at concentrations of 0, 1, 5, 10, 50 and 100 ng/mL, the cytotoxicity of the recombinant TvMIF protein to THP-1 macrophages was tested using cell counting kit (CCK)-8 assay, and the apoptosis of THP-1 macrophages and reactive oxygen species (ROS) were detected using flow cytometry. The relative expression of nucleotide-binding oligomerization domain-like receptor protein 3 (NLRP3), caspase-1, interleukin-1β (IL-1β) and IL-18 genes was quantified using real-time fluorescent quantitative PCR (qPCR) assay, and the expression of caspase-1, NLRP3, gasdermin D (GSDMD), gasdermin D N-terminal (GSDMD-NT) and pro-IL-1β proteins were determined using Western blotting assay. Results Sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) displayed successful expression and purification of the recombinant TvMIF protein with a molecular weight of 15.5 kDa, and the endotoxin activity assay showed the successful removal of endotoxin in the recombinant TvMIF protein (endotoxin concentration < 0.1 EU/mL), which was feasible for the subsequent studies on protein functions. Flow cytometry revealed that the recombinant TvMIF protein at a concentration of 10 ng/mL and less promoted the apoptosis of THP-1 macrophages, and the highest apoptotic rate of THP-1 macrophages was seen following exposure to the recombinant TvMIF protein at a concentration of 5 ng/mL, while the recombinant TvMIF protein at concentrations of 50 and100 ng/mL inhibited the apoptosis of THP-1 macrophages. Exposure to the recombinant TvMIF protein at a concentration 1 ng/mL resulted in increased ROS levels in THP-1 macrophages. qPCR assay quantified significantly elevated caspase-1, NLRP3, IL-18 and IL-1β expression in THP-1 macrophages 8 hours post-treatment with the recombinant TvMIF protein at a concentration 1 ng/mL, and Western blotting determined increased caspase-1, NLRP3, pro-IL-1β, GSDMD and GSDMD-NT protein expression in THP-1 macrophages following exposure to the recombinant TvMIF protein at a concentration 1 ng/mL. Pretreatment with MCC950 significantly reduced GSDMD and GSDMD-NT protein expression. Conclusions High-concentration recombinant TvMIF protein inhibits macrophage apoptosis, while low-concentration recombinant TvMIF protein activates NLRP3 inflammasome and promotes macrophage pyroptosis.
RESUMO
Objective:To investigate the clinical value of computer tomography (CT)-based three-section formula in identification of Siewert types of adenocarcinoma of esophagogastric junction (AEG).Methods:The retrospective and descriptive study was conducted. The clinicopatho-logical data of 62 AEG patients who were admitted to two medical centers, including 33 patients from Peking University Cancer Hospital and 29 patients from the First Affiliated Hospital of Amy Medical University, between January 2019 and January 2021 were collected. There were 53 males and 9 females, aged (66±9)years. All patients underwent CT examination to obtain the coronal and axial images and determine the upper and lower edges of the tumor and the esophagogastric junction, which were imported into the formula for Siewert classification. Observation indicators: (1) results of CT examination and pathological examination; (2) consistence of judgements for CT examination between doctors; (3) consistence of judgements between CT examination and patholo-gical examination. Results of pathological examination came from intraoperative surgical observa-tion and postoperative histopathological examination. Measurement data with normal distribution were represented as Mean± SD, and count data were described as absolute numbers or percentages. The consistency coefficient Kappa ( κ) was used to evaluate the consistency of diagnosis between resear-chers. The κ≥0.75 was regarded as excellent consistency, 0.40< κ<0.75 as good consistency, κ ≤0.40 as poor consistency. Wilcoxon rank sum test was used to evaluate the statistical difference between results of the CT-based three-section formula method and the pathological examination. Taking the results of histopathological examination as standard, the sensitivity, specifi-city, accuracy and 95% confidence interval of the CT-based three-section formula method were calculated. Results:(1) Results of CT examination and pathological examination: all the 62 patients underwent CT examination successfully to obtain the coronal and axial images and determine the upper and lower edges of the tumor and the esophagogastric junction, which were used for Siewert classifica-tion. There were 3 cases with AEG of Siewert type Ⅰ, 47 cases with AEG of Siewert type Ⅱ and 12 cases with AEG of Siewert type Ⅲ according to doctor's judgement from the Peking University Cancer Hospital, and there were 3 cases with AEG of Siewert type Ⅰ, 49 cases with AEG of Siewert type Ⅱ and 10 cases with AEG of Siewert type Ⅲ according to doctor's judgement from the First Affiliated Hospital of Amy Medical University, respectively. After arbitration, there were 2 cases with AEG of Siewert type Ⅰ, 48 cases with AEG of Siewert type Ⅱ and 12 cases with AEG of Siewert type Ⅲ determined by the CT based three-section formula. There were 7 cases of stage T1, 10 cases of stage T2, 24 cases of stage T3, 14 of stage T4a and 7 cases of stage T4b of pathological T staging. There were 2 cases with AEG of Siewert type Ⅰ, 48 cases with AEG of Siewert type Ⅱ and 12 cases with AEG of Siewert type Ⅲ determined by pathological examination. (2) Consistence of judgements for CT examination between doctors: the consistency of Siewert classification determined by CT-based three-section formula between 2 doctors was good ( κ=0.74, P<0.001). (3) Consistence of judgements between pathological examination and CT examination: with Siewert classification determined by pathological examination as reference, the accuracy of Siewert classification determined by CT based three-section formula was 90.3%( κ=0.73, P<0.001). The sensitivity and specificity of CT-based three-section formula were 66.7%(95% confidence interval as 20.8%-93.9%) and 100.0%(95% confidence interval as 93.9%-100.0%) for AEG of Siewert type Ⅰ, 97.7%(95% confidence interval as 88.2%-99.6%) and 72.2%(95% confidence interval as 49.1%-87.5%) for AEG of Siewert type Ⅱ, 73.3%(95% confidence interval as 48.0%-89.1%) and 97.9%(95% confidence interval as 88.9%-99.9%) for AEG of Siewert type Ⅲ, respectively. Conclusion:The CT-based three-section formula can be used for identification of Siewert types of AEG, with a high accuracy.
RESUMO
Objective:To investigate the efficacy of CT imaging features in evaluating occult peritoneal metastasis (OPM) of diffuse infiltrating gastric cancer (Borrmann Type Ⅳ).Methods:Totally 101 patients with locally advanced Borrmann type Ⅳ gastric cancer were retrospectively collected who were admitted to Peking University Cancer Hospital from March 2014 to March 2021. The patients were divided into OPM group (53 cases) and the non-OPM group (48 cases) according to the results of preoperative CT and laparoscopic exploration/peritoneal cytology examination. The pathological examination results were recorded, including the degree of histological differentiation and Lauren classification. The evaluation indicators included the tumor center position, the number of tumor-occupied portions, involved orientation, mucosal broadband sign, stratified enhancement, serosa invasion, increased density of peripheral fat tissue, and enlarged lymph nodes. The maximum thickness of the primary tumor, average CT value of the primary tumor (arterial phase, venous phase, and delayed phase), difference between venous phase and arterial phase, difference between delayed phase and venous phase, and pattern of the enhanced curve were recorded. The Mann-Whitney U or Chi-square test was used to compare the differences of pathological and CT features between two groups. The multivariate logistic regression was used to screen independent predictors and establish a nomogram. The receiver operating characteristic curve was used to evaluate the performance of the nomogram in predicting OPM, and the Hosmer-Lemeshow test was used to test the model′s goodness of fit. Results:There was statistical significance in the seven indicators between the OPM and non-OPM groups, including tumor-occupied portions of stomach, mucosal broadband sign, stratified enhancement, serosa invasion, increased density of peripheral fat tissue, the enhanced curve pattern and the degree of histological differentiation ( P<0.05). Among them, the degree of histological differentiation (OR=0.19, P=0.033), stratified enhancement (OR=7.02, P=0.005) and serosa invasion (OR=14.27, P<0.001) were independent predictors of OPM. The nomogram was established based on the three significant features. The area under the curve for predicting OPM was 0.826 (95%CI 0.745-0.908), the sensitivity was 0.566 and the specificity was 0.938. The Hosmer-Lemeshow test showed a good agreement between the OPM risk predicted by the nomogram and the actual risk ( P=0.525). Conclusions:The CT features of Borrmann type Ⅳgastric cancer complicated with OPM have specific characteristics. The diagnosis model based on the degree of histological differentiation, stratified enhancement, and serosa invasion had high efficacy in evaluating OPM.
RESUMO
Objective:To investigate the computed tomography (CT) features of adenocarcinoma of esophagogastric junction (AEG) after neoadjuvant chemotherapy.Methods:The retrospective and descriptive study was conducted. The clinicopathological data of 59 patients with AEG who underwent neoadjuvant chemotherapy in Peking University Cancer Hospital from February 2010 to November 2014 were collected. There were 51 males and 8 females, aged from 46 to 82 years, with a median age of 63 years. All the 59 patients underwent enhanced CT examination before and after neoadjuvant chemotherapy. Observation indicators: (1) pathological examination and neoadjuvant chemotherapy of patients with AEG; (2) results of CT examination in patients with AEG, including ① qualitative indicators of CT and ② quantitative indicators of CT. Measurement data with skewed distribution were represented as M( P25, P75) or M (range), and comparison between groups was analyzed using the Mann-Whitney U test. Count data were described as absolute numbers, and comparison between groups was analyzed by the chi-square test. Results:(1) Pathological examination and neoadjuvant chemotherapy of patients with AEG: of the 59 patients with AEG, high-differentiated adenocarcinoma was observed in 1 patient, moderate-differentiated adenocarcinoma in 40 patients, and low-differentiated adenocarcinoma in 18 patients. Effective response to neoadjuvant chemotherapy was observed in 13 patients, including 6 patients of pathological tumor regression grading (pTRG) 0 and 7 of pTRG 1; poor response was observed in 46 patients, including 12 patients of pTRG 2 and 34 patients of pTRG 3. (2) Results of CT examination in patients with AEG. ① Qualitative indicators of CT: for the 13 patients with effective response to neoadjuvant chemotherapy, 13 had the presence of ulcers, 5 had layered enhancement, 10 had infiltration of adventitia surface, and 2 had positive extramural venous invasion (EMVI) before neoadjuvant chemotherapy; after neoadjuvant chemotherapy, 13 had shallowed or disappeared ulcers, 7 patients had changed enhancement pattern, 3 had infiltration of adventitia surface, and 1 had positive EMVI. For the 46 patients with poor response to neoadjuvant chemotherapy, 28 had the presence of ulcers, 18 had layered enhancement, 37 had infiltration of adventitia surface, and 22 had positive EMVI before neoadjuvant chemotherapy; after neoadjuvant chemotherapy, 23 had shallowed or disappeared ulcers, 7 patients had changed layered enhancement pattern, 33 had infiltration of adventitia surface and 21 had positive EMVI, respectively. There was no significant difference in the layered enhancement or infiltration of adventitia surface before neoadjuvant chemotherapy between patients with different treatment response ( χ2=0.002, 0.000, P>0.05). There were significant differences in the presence of ulcers and positive EMVI before neoadjuvant chemotherapy between patients with different treatment response ( χ2=5.591, 4.421, P<0.05). After neoadjuvant chemotherapy, there were significant differences in the changes of layered enhancement pattern, infiltration of adventitia surface and positive EMVI between patients with different treatment response ( χ2=6.359, 10.090, 4.728, P<0.05); while there was no significant difference in the shallowed or disappeared ulcers between patients with different treatment response ( χ2=1.239, P>0.05). ② Quantitative indicators of CT: for the 13 patients with good response to neoadjuvant chemotherapy, the maximum tumor height, the maximum tumor area, enhanced CT value of the lesion before neoadjuvant chemotherapy were 1.37 cm(0.94 cm, 1.88 cm), 8.9 cm 2 (4.7 cm 2, 9.9 cm 2), 53 HU(47 HU, 63 HU), respectively. After neoadjuvant chemotherapy, the above indicators were 1.17 cm(0.79 cm, 1.29 cm), 4.4 cm 2(2.5 cm 2, 6.1 cm 2), 30 HU(25 HU, 53 HU), respectively. The change rates of the maximum tumor height, the maximum tumor area, and enhanced CT value of the lesion were -23%(-42%, 9%), -51%(-60 %, -21%), -44%(-51%, 19%), respectively. For the 46 patients with poor response to neoadjuvant chemotherapy, the maximum tumor height, the maximum tumor area, enhanced CT value of the lesion were 1.57 cm(1.21 cm, 1.96 cm), 9.4 cm 2(6.6 cm 2, 13.1 cm 2), 60 HU(53 HU, 66 HU) before neoadjuvant chemotherapy, respectively. After neoadjuvant chemotherapy, the above indicators were 1.16 cm(0.94 cm, 1.37 cm), 6.2 cm 2(4.8 cm 2, 8.1 cm 2), 55 HU(47 HU, 65 HU), respectively. The change rates of the maximum tumor height, the maximum tumor area, and enhanced CT value of the lesion were -27%(-38%, -9%), -33%(-47%, -12%), -9%(-22%, 9%), respectively. There was no significant difference in the maximum tumor height, the maximum tumor area, enhanced CT value of the lesion before neoadjuvant chemotherapy between patients with different treatment response ( Z=-1.372, -1.372, -1.331, P>0.05). There was no significant difference in the maximum tumor height after neoadjuvant chemotherapy between patients with different treatment response ( Z=-0.503, P>0.05), while there were significant differences in the maximum tumor area and CT value of the lesion ( Z=-2.743, -3.049, P<0.05). There was no significant difference in the change rate of the maximum tumor height or the maximum tumor area between patients with different treatment response ( Z=0.000, -1.481, P>0.05), while there was a significant difference in the change rate of CT value of the lesion ( Z=-3.231, P<0.05). Conclusion:Effective response of AEG to neoadjuvant chemotherapy was characterized by the changes in tumor layered enhancement pattern, reduction in the maximum tumor area, reduced CT value of the lesion, negative infiltration of adventitia surface, and negative EMVI.
RESUMO
The commonly used radiological examinations for gastrointestinal stromal tumor (GIST) include ultrasound, CT, magnetic resonance imaging and PET, which play important roles in the individualized treatment of GIST through the assistance of the detection, location, diagnose and differential diagnosis, biological behavior and risk evaluation, operation planning, therapeutic effects prediction and follow-up. Standardized predisposal, multi-planar reconstruction and multi-phase enhancement are of great significance in the detection of small GIST lesions. Differences in drainage vessels and enhancement characteristics of tumors are helpful in differentiating small intestinal GIST from pancreatic cancer, neuroendocrine tumors and other abdominal non-GIST tumors. Radiology can provide multiple indicators to qualitatively or quantitatively reflect the risk of GIST. Recent study found that the risk classification of GIST has correlation with necrosis, but not cystic degeneration. The risk stratification of GIST by dual-energy CT showed that CT value and standardized iodine concentration in arterial-venous and delayed phases of high-risk tumors were significantly higher than those of non-high-risk tumors. The correlation between tumor enhancement pattern and risk stratificationwas still controversial. Recently, the rise of radiomics and deep learning techniques provide new direction for risk assessment of GIST. Some studies used various models and parameters to predict the risk classifications of GIST and achieved AUC values above 0.85, and some results were also superior to conventional indicators and radiologists′ subjective judgment in the prediction of exon 11 mutations, mitotic counts and survivals. Recent developments in PET and diffusion-weighted MRI provide more precise functional quantitative indicators for the prediction and early evaluation of the response of GIST to targeted therapy. These studies are constantly expanding our field of exploration, and prompting us to stride toward the goal of individualized treatment of GIST.
RESUMO
The accurate judgement of the upper and lower borders of the adenocarcinoma of esophagogastric junction (AEG) by radiology can facilitate the decisions on surgical approach and staging criteria. X-ray double contrast radiography, CT and MRI are the common modalities. The accuracy of X-ray double contrast radiography in determining the invasion length of esophagus and the central point of gastric infiltration can be improved by standardized pretreatment, combination of multiple contrast methods such as double contrast and flow-coating procedure, and combination of multi-angle observations such as conventional frontal, left /right anterior oblique and supine right posterior oblique position. Abdominal enhanced CT is the imaging method recommended by clinical guidelines for the radiological examination of AEG. The relative position of the central point of the tumor from 2 cm line can be determined through the combination of measurement and formula calculation on multi-planar reconstructed CT images. The "three-layer four-type" classification can provide reference for the selection of abdominothoracic incision. The direct demonstration of the tumor extension can be achieved through the CT curved planar reconstruction by drawing lines along esophagus to stomach. The combination of multiple sequences of MRI is helpful to determine the extension of the lesions. In the future, more radiological studies are needed to establish criteria with high accuracy, repeatability and convenient operation,and to assist clinical evaluation of AEG invasion.