Objective To produce anti-thrombomodulin antibodies.Methods Using genetic immunization: Eukaryotic expression plasmid pcDNA3.1/TM(LEO),encoding all the extracellular domain of human thrombomodulin and signal peptides but lacking the transmembrance and cytoplasmic domains was constructed, which recombinant thrombomodulin was secreted soluble product. The plasmid was isolated from large-scale bacterial cultures by treatment with alkali and SDS, purified by precipitation with polyethylene Glycol (PEG). Recombinant plasmid was injected into tibial muscle of BALB/c mice. The productions of TM and anti-TM have been detected. Results The positive of RT-PCR and expressed TM identified the function of the recombinant plasmid. The pcDNA3.1/TM(LEO) induced higher titer of anti-TM. The antibody titer peaked between the 5th and 7th injection with a titer of 1∶8 000 detected by cell-ELISA coated with EVC-304. Specificity has been identified by western blot and immunohistochemistry.Conclusion The production of antibody through genetic immunization was a feasible method due to the difficulties in obtaining and purification of natural thrombomodulin.

Objective To study the feasibility and timing of early laparoscopic cholecystectomy (LC) for the patients with mild-to-moderate acute pancreatitis with gallbladder stone.Methods 26 patients with mild-to-moderate acute pancreatitis with gallbladder stone underwent early laparoscopic cholecystectomy.The patients were initially treated with conservative treatment.When the diagnosis was confirmed and the patients were stable,LC was carried out within 48 hours of hospitalization.The results were compared with a delayed group of patients who received delayed LC for acute pancreatitis.Results All the LC operations were successfully carried out.There were no significant differences between the two groups in alkaline phosphatase,alanine aminotransferase,aspartate aminotransferase,blood amylase and urine armylase on postoperative day 1,3,5,7.Patients who received early laparoscopic cholecystectomy had a shorter hospital stay.There were no significant differences between the two groups in the operation time,intraoperative blood loss,postoperative complication rates and recurrence of acute pancreatifis.Conclusion After CBD stones have been ruled out,it is safe and feasible to carry out early laparoscopic cholecystectomy within 48 hours of hospital admission for patients with mild-to-moderate acute pancreatitis with gallbladder stone.

Objective To study the value of the technique of "Push-pull traction-relax homingrepeatedly confirmed" in the prevention of bile duct injury in LC. Methods From March 2001-August 2009, we applied this technique in 4800 cases of LC. The technique of "Push-pull traction" showed the structures of in the Calot's triangle. The technique of "relax homing" was to restore the cystic duct,hepatic duct and common bile duct to their original anatomical positions. The technique of "repeatedly confirmed" repeatedly identified the positions of the cystic duct, the common hepatic duct and the common bile duct. Results There was no bile duct injury. Conversion to open surgery happened in 118patients due to difficulties in identifying the Calot's triangle structures, bile duct stones, gallbladder cancer, and gallbladder-duodenal fistula. Conclusions The "Push-pull traction-relax homing-repeatedly confirmed" technique could effectively prevent bile duct injury in LC. The method is simple, easy to master and worthy of promotion.

Objective To compare the clinical efficacy of anatomical hepatectomy (AR) and parenchymal-sparing hepatectomy (PSH) for Barcelona clinic liver cancer (BCLC) stage A hepatocellular carcinoma(HCC),and investigate its prognostic factors.Methods The propensity score matching and retrospective cohort study was conducted.The clinicopathological data of 269 patients with BCLC stage A HCC who were admitted to the First Affiliated Hospital of Nanjing Medical University from January 2009 to December 2017 were collected.There were 226 males and 43 females,aged from 23 to 84 years,with a median age of 56 years.All the 269 patients underwent radical resection and were confirmed as HCC using postoperative pathological examination.Of the 226 patients,146 undergoing AR and 123 undergoing PSH were allocated into the AR group and PSH group,respectively.Observation indicators:(1) the propensity score matching conditions and comparison of general data between groups after the propensity score matching;(2) intraoperative and postoperative situations;(3) follow-up and survival situations;(4) prognostic factors analysis.Patients were followed up by outpatient examination and telephone interview to detect survival once every 3 months within 1 year postoperatively,once every 6 months within 2-5 years postoperatively and once a year after 5 years postoperatively up to October 2018.The overall survival time was from surgery data to death or end of follow-up.The tumor-free survival time was from surgery date to time of tumor recurrence detected or end of follow-up without tumor recurrence.The propensity score matching was used to perform 1∶1 matching by nearest neighbor method.Count data were represented as absolute number,comparison between groups was analyzed using the chi-square test and McNemar test after propensity score matching.Measurement data with skewed distribution were represented as M (range),and comparison between groups was done using the Mann-Whitney U test and Wilcoxon signed rank sum test after propensity score matching.The survival rate and curve were respectively calculated and drawn by the Kaplan-Meier method,and Log-rank test was used for survival analysis.The COX proportional risk model was used for univariate and multivariate analysis.Results (1) The propensity score matching conditions and comparison of general data between groups after the propensity score matching:180 of 269 patients had successful matching,including 90 in each group.The maximum tumor diameter,cases with vascular embolism,cases of stage Ⅰ and Ⅱ (TNM staging) before matching were 5.0 cm (range,0.8-17.0 cm),42,97,99 in the AR group and 3.0 cm (range,1.0-17.0 cm),16,49,24 in the PSH group,respectively,with statistically significant differences between the two groups (Z =-4.277,x2 =9.803,6.664,P＜ 0.05).The above indices after matching were 4.0 cm (range,0.8-16.0 cm),15,70,68 in the AR group and 3.5 cm (range,1.0-17.0 cm),16,20,22 in the PSH group,with no statistically significant difference between the two groups (Z =-0.241,x2=0.039,0.124,P＞0.05).The confounding bias of maximum tumor diameter,vascular embolism and TNM staging were eliminated.(2) Intraoperative and postoperative situations:the operation time,volume of intraoperative blood loss,cases with intraoperative blood transfusion,cases with surgical margin ＜ 1 cm and ≥ 1 cm,cases with postoperative severe complications,duration of hospital stay,cases with postoperative tumor recurrence,cases with tumor recurrence within 2 years postoperatively,cases undergoing surgical treatment due to postoperative tumor recurrence,cases undergoing transcatheter arterial chemoemblization due to postoperative tumor recurrence after matching were 180 minutes (range,60-448 minutes),130 mL (range,30-6 000 mL),9,2,88,8,18 days (range,8-77 days),41,32,15,23 in the AR group,and 150 minutes (range,55-400 minutes),100 mL (range,50-3 000 mL),6,2,88,6,18 days (range,9-37 days),37,29,10,24 in the PSH group,respectively,showing no statistically significant difference between the two groups (Z =-1.987,-0.439,x2 =0.655,0.000,0.310,Z=-0.805,x2=0.362,0.223,0.816,0.624,P＞0.05).(3) Follow-up and survival situations:180 patients were followed up for 4-114 months,with a median time of 43 months.Forty of 180 patients died (21 in the AR group and 19 in the PSH group) and 78 had tumor recurrence (41 in the AR group and 37 in the PSH group).The 1-,3-,5-year overall survival rates and tumor-free survival rates were 92.0％,76.3％,71.8％ and 70.8％,53.0％,47.4％ in the AR group,92.3％,80.6％,62.0％ and 72.3％,56.4％,46.1％ in the PSH group,respectively,showing no statistically significant difference between the two groups (x2 =0.034,0.000,P＞0.05).Stratified analysis:of the AR group,the 1-,3-,5-year overall survival rates and median tumor-free survival rate were 95.3％,82.0％,82.0％ and 54.6％ in the patients with grade Ⅰ of preoperative albumin-bilirubin,100.0％,86.8％,86.8％ and 61.5％ in the patients with maximum tumor diameter ≤≤5 cm,91.3％,75.0％,69.7％ and 43.1％ in the patients with liver cirrhosis,89.9％,73.2％,66.6％ and 54.6％ in the patients with moderate-low differentiated tumor.Of the PSH group,the 1-,3-,5-year overall survival rates and median tumor-free survival rate were 90.9％,74.9％,63.0％ and 43.4％ in the patients with grade Ⅰ of preoperative albumin-bilirubin,98.2％,85.8％,61.7％ and 46.0％ in the patients with maximum tumor diameter ≤≤ 5 cm,98.0％,88.7％,70.0％ and 43.4％ in the patients with liver cirrhosis,90.7％,79.2％,59.0％ and 43.4％ in the patients with moderate-low differentiated tumor.There were no statistically significant difference in the 1-,3-,5-year overall survival rates between the two groups (x2 =1.892,1.320,0.732,0.002,P＞0.05) and a statistically significant difference in the tumor-free survival rate between the two groups (x2 =0.337,0.051,0.551,0.061,P ＞ 0.05).(4) Prognostic factors analysis.Results of univariate analysis showed that preoperative albumin-bilirubin grade,preoperative alpha fetoprotein (AFP),maximum tumor diameter,number of tumors,satellite lesion,vascular embolism,TNM staging,volume of intraoperative blood loss,postoperative severe complications were related factors affecting overall survival after radical resection for HCC (hazard ratio=1.762,1.001,1.139,1.955,2.561,2.495,2.766,1.000,2.599,95％ confidence interval:1.048-2.962,1.000-1.001,1.080-1.201,1.063-3.596,1.254-5.227,1.446-4.304,1.655-4.624,1.000-1.001,1.317-5.128,P＜0.05).Preoperative AST,positive HBsAg,preoperative AFP,maximum tumor diameter,vascular embolism,TNM staging,postoperative severe complications were related factors affecting tumor-free survival after radical resection for HCC (hazard ratio=1.004,1.594,1.000,1.065,2.203,2.132,1.775,95％ confidence interval:1.001-1.007,1.020-2.490,1.000-1.001,1.019-1.113,1.474-3.293,1.462-3.109,1.034-3.047,P＜0.05).Results of multivariate analysis showed that preoperative AFP,maximum tumor diameter,satellite lesion,postoperative severe complications were independent factors affecting overall survival after radical resection for HCC (hazard ratio =1.001,1.114,2.241,2.251,95％ confidence interval:1.000-1.001,1.033-1.202,1.003-5.008,1.100-4.607,P＜0.05).Positive HBsAg was an independent factor affecting tumor-free survival after radical resection for HCC (hazard =1.576,95％ confidence interval:0.987-2.516,P＜ 0.05).Conclusions There was no significant difference in long-term efficacy between AR and PSH in patients with BCLC stage A HCC.Preoperative AFP,maximum tumor diameter,number of tumors,satellite lesion,postoperative severe complications are independent factors affecting long-term survival of BCLC stage A patients after HCC radical resection.

Lung squamous cell carcinoma with intratracheal diffuse leukoplakia as the main manifestation is very rare in clinic. The clinical data of a patient with pulmonary squamous cell carcinoma with intratracheal diffuse leukoplakia admitted to the Affiliated Hospital of Jining Medical University in December 25, 2021 were retrospectively analyzed in order to improve the understanding of this special manifestation. The patient was a 73-year-old male with clinical manifestations of cough, sputum, and blood-stained sputum. Chest CT indicated patchy high-density shadow on the upper right lung, whole-course thickening of trachea and bronchial walls, and bronchoscopy showed diffuse trachea-bronchial mucosa congestion and edema, with a large number of leukoplakia on the surface. The clinical effect was stable after 2 cycles of chemotherapy. Lung squamous cell carcinoma accompanied by diffuse leukoplakia in the trachea is a rare presentation. Chest CT can show thickening of the tracheal and bronchial wall, while the lesion and tumor signs of the primary lesion are not obvious. Electronic bronchoscopy as soon as possible can avoid missed diagnosis.

Objective To investigate the expression of hepatocyte growth factor ( HGF ) and its relationship with microsatellite instability ( MSI) and their influence on survival in patients with colorectal cancer. Methods Immunohistochemistry ( IHC) was used to detect the expression of HGF and MSI in 98 specimens of colorectal cancer. Tumors lacking protein expression of any of the four mismatch repair genes (MLH1, PMS2, MSH2 or MSH6) were labelled as MSI, and the rest were considered as microsatellite stable ( MSS) . The associations between expression and clinicopathological factors were assessed using Chi?square tests. Kaplan?Meier curves, log?rank test, and Cox regression were used to analyze the association between biomarker expressions and overall survival. Results The incidence rate of MSI in 98 colorectal specimens was 32. 7%, and was statistically significantly correlated with the location of tumor and differentiation degree ( P<0.05) . The HGF?expression rate was 71.4%. The patients with an MSI tumor had a significantly higher HGF expression, compared with the patients with an MSS tumor (P=0.048). The 5?year survival rate of MSI group and MSS group were 39.8% and 58.7%, respectively (P=0.009). The 5?year survival rate of HGF?positive group and HGF?negative group were 46.2% and 67.9% ( P=0.035) . The multivariate analysis showed that lymphocytic infiltration, TMN stage, MSI and HGF are independent prognostic factors in colorectal cancer ( P<0. 05 for all ) . Conclusions HGF is highly expressed in colorectal cancer patients with microsatellite instability. Both microsatellite instability and HGF are independent factors affecting the prognosis in patient with colorectal cancer.

Objective To investigate the expression of hepatocyte growth factor ( HGF ) and its relationship with microsatellite instability ( MSI) and their influence on survival in patients with colorectal cancer. Methods Immunohistochemistry ( IHC) was used to detect the expression of HGF and MSI in 98 specimens of colorectal cancer. Tumors lacking protein expression of any of the four mismatch repair genes (MLH1, PMS2, MSH2 or MSH6) were labelled as MSI, and the rest were considered as microsatellite stable ( MSS) . The associations between expression and clinicopathological factors were assessed using Chi?square tests. Kaplan?Meier curves, log?rank test, and Cox regression were used to analyze the association between biomarker expressions and overall survival. Results The incidence rate of MSI in 98 colorectal specimens was 32. 7%, and was statistically significantly correlated with the location of tumor and differentiation degree ( P<0.05) . The HGF?expression rate was 71.4%. The patients with an MSI tumor had a significantly higher HGF expression, compared with the patients with an MSS tumor (P=0.048). The 5?year survival rate of MSI group and MSS group were 39.8% and 58.7%, respectively (P=0.009). The 5?year survival rate of HGF?positive group and HGF?negative group were 46.2% and 67.9% ( P=0.035) . The multivariate analysis showed that lymphocytic infiltration, TMN stage, MSI and HGF are independent prognostic factors in colorectal cancer ( P<0. 05 for all ) . Conclusions HGF is highly expressed in colorectal cancer patients with microsatellite instability. Both microsatellite instability and HGF are independent factors affecting the prognosis in patient with colorectal cancer.

To assess the background field removal method usually used in quantitative susceptibility mapping (QSM), and to analyze the cause of serious artifacts generated in the truncated -space division (TKD) method, this paper discusses a variety of background field removal methods and proposes an improved method to suppress the artifacts of susceptibility inversion. Firstly, we scanned phase images with the gradient echo sequence and then compared the quality and the speed of reconstructed images of sophisticated harmonic artifact reduction for phase data (SHARP), regularization enable of SHARP (RESHARP) and laplacian boundary value (LBV) methods. Secondly, we analyzed the reasons for reconstruction artifacts caused by the multiple truncations and discontinuity of the TKD method, and an improved TKD method was proposed by increasing threshold truncation range and improving data continuity. Finally, the result of susceptibility inversion from the improved and original TKD method was compared. The results show that the reconstruction of SHARP and RESHARP are very fast, but SHARP reconstruction artifacts are serious and the reconstruction precision is not high and implementation of RESHARP is complicated. The reconstruction speed of LBV method is slow, but the detail of the reconstructed image is prominent and the precision is high. In the QSM inversion methods, the reconstruction artifact of the original TKD method is serious, while the improved method obtains good artifact suppression image and good inversion result of artifact regions.
Algorithms ; Artifacts ; Brain ; Image Processing, Computer-Assisted ; Magnetic Resonance Imaging ; Phantoms, Imaging

BACKGROUND: With the popularization of chest computed tomography (CT) early screening for lung cancer, the detection rate of lung cancer combined with multiple primary malignancies (MPM) in other organs has been increasing. In this paper, the incidence, pathological characteristics, diagnosis and treatment characteristics and prognosis were discussed and analyzed to provide research basis for improving the clinical diagnosis and treatment of this disease. METHODS: From September 2011 to September 2015, a total of 5,570 patients with lung cancer were treated in Jiangsu Cancer Hospital. The clinicopathological characteristics of 61 patients with lung cancer combined with MPM in other organs were retrospectively analyzed. RESULTS: The incidence rate of lung cancer combined with MPM in other organs in this group was 1.1%, of which 15 cases were synchronous MPM (SMPM), 46 cases were metachronous MPM (MMPM). Colorectal cancer, breast cancer and thyroid cancer accounted for the top three of lung cancer combined with MPM in other organs. The overall 5-year survival rate was 39.3% and 71.4% of patients died from metastasis or recurrence of lung cancer. Multivariate analysis showed that the clinical stage of lung cancer patients, the order of occurrence of lung cancer and other tumors, the treatment status of patients with other organ tumors and the presence of epidermal growth factor receptor (EGFR) gene mutation were important factors for the survival of the patients. CONCLUSIONS The incidence rate of lung cancer combined with MPM in other organs is not uncommon. Lung cancer is the main cause of death compared with other organs tumors. Patients with advanced lung cancer, SMPM, lung cancer first, combined with tumor only receiving palliative treatment and without EGFR gene mutation had a poor prognosis.

BACKGROUND: Cyclic RNA (circRNA) is a new type of non-coding RNA (ncRNA) which is different from traditional linear RNA. More and more studies suggest that circRNA can be used as a biological marker of many malignant tumors and becomes a potential target for treatment. Therefore, searching for new molecular targets of lung adenocarcinoma from the circRNA will help to reveal the new mechanism of the occurrence and development of lung adenocarcinoma, and provide new ideas for clinical diagnosis and treatment. In this study, the biological function of circ_0007766, a highly expressed circRNA found in a screen of lung adenocarcinoma tissue, was verified and analyzed in vitro, so as to preliminarily explore the mechanism of circ_0007766 in promoting the proliferation of lung adenocarcinoma. METHODS: The expression level of circ_0007766 in lung adenocarcinoma cells was detected by qPCR. Then siRNA was used to knock down the expression of circ_0007766. The effects of knockdown of circ_0007766 on proliferation, cell cycle and apoptosis of lung adenocarcinoma cells were detected by CCK8, scratch test, PI staining and Annexin V/PI double staining. In addition, the biological mechanism of circ_0007766 in lung adenocarcinoma was preliminarily studied by qPCR and Western blots. RESULTS: The expression of circ_0007766 in lung adenocarcinoma cell lines was detected by qPCR. The expression of circ_0007766 was interfered in SPCA-1 cells. The proliferation and migration abilities of cells were inhibited. The cell cycle was arrested in G0/G1 phase, but the apoptosis was not affected. The deletion of circ_0007766 did not affect the expression of ERBB2, but influenced the mRNA and protein expression of Cyclin D1/Cyclin E1/CDK4. CONCLUSIONS In vitro functional studies have shown that circ_0007766 may promote the proliferation and migration of lung adenocarcinoma cells. Further molecular mechanism studies have found that circ_0007766 can up-regulate the expression of Cyclin D1/Cyclin E1/CDK4, which are the key proteins of cell cycle, and thus promote the malignant proliferation of lung adenocarcinoma. From the perspective of circRNA, this study will provide new clues for the pathogenesis, development and prognosis of lung adenocarcinoma, and provide new target for clinical treatment.
Adenocarcinoma of Lung ; pathology ; Apoptosis ; genetics ; Cell Cycle ; genetics ; Cell Line, Tumor ; Cell Proliferation ; genetics ; Cell-Free Nucleic Acids ; genetics ; Cyclin D1 ; genetics ; Cyclin E ; genetics ; Cyclin-Dependent Kinase 4 ; genetics ; Humans ; Oncogene Proteins ; genetics ; Up-Regulation ; genetics