Transcription factor ETV1-induced lncRNA MAFG-AS1 promotes migration, invasion, and epithelial-mesenchymal transition of pancreatic cancer cells by recruiting IGF2BP2 to stabilize ETV1 expression.

We investigated the mechanism of ETS-translocation variant 1 (ETV1)/lncRNA-MAFG-AS1 in pancreatic cancer (PC). MAFG-AS1 and ETV1 levels in PC cell lines and HPNE cells were determined using reverse transcription quantitative polymerase chain reaction (RT-qPCR) and Western blotting (WB). After transfection with sh-MAFG-AS1, PC cell invasion, migration, proliferation, and epithelial-mesenchymal transition (EMT)-related proteins were measured by 5-ethynyl-2'-deoxyuridine (EdU), Transwell assay, and WB. The binding between ETV1 and MAFG-AS1 was studied using dual-luciferase assay and chromatin immunoprecipitation. The interactions between MAFG-AS1, IGF2BP2, and ETV1 were tested. Combined experiments were further performed using sh-MAFG-AS1 and pcDNA-ETV1 simultaneously. ETV1/MAFG-AS1 was highly expressed in PC cells. Blocking MAFG-AS1 inhibited the malignant behaviors of PC cells. ETV1 induced MAFG-AS1 transcription in PC cells. MAFG-AS1 stabilized ETV1 mRNA by recruiting IGF2BP2. ETV1 overexpression partially antagonized the suppression of silencing MAFG-AS1 on PC cells. ETV1-induced MAFG-AS1 stabilized the ETV1 expression by recruiting IGF2BP2 and promoted PC cell migration, invasion, proliferation, and EMT.

The role of neoadjuvant conventional transarterial chemoembolization with radiofrequency ablation in the treatment of recurrent hepatocellular carcinoma after initial hepatectomy with microvascular invasion.

OBJECTIVES: Recurrent hepatocellular carcinoma (HCC) patients with microvascular invasion (MVI)-positive primary tumor is at high risk of re-recurrence while treated with radiofrequency ablation (RFA). We aimed to investigate whether neoadjuvant conventional transarterial chemoembolization (cTACE) was effective in reducing re-recurrence after RFA for recurrent HCC patients with MVI-positive primary tumors. METHODS: In this retrospective multicenter study, 468 patients with solitary small recurrent HCC (≤3.0cm) underwent RFA alone (n = 322) or with neoadjuvant cTACE (n = 146) between June 2007 and December 2017 were included. Overall survival (OS) and recurrence-free survival (RFS) were compared. RESULTS: The 1-, 5-year OS rates were 74.8%, 42.5% for RFA with neoadjuvant cTACE group, and 53.5%, 28.7% for RFA group (P < 0.001). The corresponding RFS rates were 51.7%, 24.4% for RFA with neoadjuvant cTACE group, and 36.1%, 9.3% for RFA group (P < 0.001). In subgroup analyses, the OS and RFS for neoadjuvant cTACE group were longer than those for RFA group no matter tumor size > 2cm (HR = 0.52, 95% CI: 0.36-0.77; HR = 0.49, 95% CI: 0.36-0.67) or not (HR = 0.53, 95% CI: 0.32-0.88; HR = 0.65, 95% CI: 0.42-0.98), or the time interval of recurrence from initial treatment ≤ 1 year (HR = 0.53, 95% CI: 0.36-0.77; HR = 0.70, 95% CI: 0.52-0.94) or not (HR = 0.56, 95% CI: 0.34-0.95; HR = 0.39, 95% CI: 0.25-0.62). Multivariable analyses showed that RFA alone (HR = 1.329, P = 0.031; HR = 1.764, P = 0.004) and interval of recurrence from initial treatment > 1 year(HR = 0.642, P = 0.001; HR = 0.298, P = 0.037) were independent prognostic factors of OS and RFS. CONCLUSIONS: Neoadjuvant cTACE could effectively reduce re-recurrence after RFA, and improve the long-term survivals for patients with solitary small recurrent HCC whose primary tumor was MVI-positive. Key pointsFor recurrent hepatocellular carcinoma (HCC) patients whose primary tumor was positive for microvascular invasion, neoadjuvant conventional transarterial chemoembolization (cTACE) with radiofrequency ablation (RFA) achieved better efficacy.Multivariable analyses showed that the interval of recurrence from initial treatment > 1 year and RFA alone were independent prognostic factors of overall survival and recurrence-free survival, respectively.

MAFG-AS1 aggravates the progression of pancreatic cancer by sponging miR-3196 to boost NFIX.

BACKGROUND: A host of researches have demonstrated the regulation of long non-coding RNAs (lncRNAs) in the progression of pancreatic cancers (PC). In this study, our main task was to analyze the function of MAF bZIP transcription factor G antisense RNA 1 (MAFG-AS1) in PC. METHODS: RT-qPCR measured gene expression. Functional experiments, including EdU assay, flow cytometry analysis, TUNEL assay and transwell assay, assessed the biological changes of PC cells. RNA pull down assay, luciferase reporter assay and RIP assay verified the interaction between RNAs. RESULTS: MAFG-AS1 was lowly expressed in normal pancreatic samples but up-regulated in PC tissues and cell lines. Besides, MAFG-AS1 silence suppressed cell proliferation and migration whereas promoted cell apoptosis in PC. Mechanism assays verified that miR-3196 could bind with MAFG-AS1. Moreover, miR-3196 was discovered to be lowly expressed in PC cell lines, and its overexpression inhibited PC cell growth and migration. Importantly, nuclear factor I X (NFIX), overexpressed in PC cell lines, was validated to be positively modulated by MAFG-AS1 through absorbing miR-3196. Moreover, overexpression of NFIX could countervail the restraining effects of MAFG-AS1 knockdown on the growth and migration of PC cells. CONCLUSION: MAFG-AS1 had an oncogenic function in the progression of PC via regulating miR-3196/NFIX pathway, and decreasing MAFG-AS1 expression could attenuate PC progression.

Hepatic resection versus transarterial chemoembolization in infiltrative hepatocellular carcinoma: A multicenter study.

BACKGROUND AND AIM: Prognosis of infiltrative hepatocellular carcinoma (iHCC) is poor, and the treatments selection based on efficacy is unclear. We performed this multicenter study to compare the efficacy of hepatic resection and transarterial chemoembolization (TACE) in treating patients with iHCC. METHODS: We retrospectively analyzed the overall survivals (OS) in 319 patients with iHCC who were initially treated by hepatic resection (n = 133) or TACE (n = 186) at four tertiary centers. Fifty-eight patients in the TACE group were assessed as resectable and compared with the hepatic resection group in subgroup analysis. A propensity score matched (PSM) analysis was performed to reduce selection bias. Cox regression was performed to identify significant factors associated with OS. RESULTS: The median OS time was significantly longer in the hepatic resection group than that in the TACE group, before and after PSM (before PSM, 17.5 vs 7.3 months, P < 0.0001; after PSM, 14.0 vs 7.3 months, P < 0.0001). The multivariable analysis indicated TACE as a risk factor of OS (hazard ratio = 2.233, 95% confidence interval = 1.492 to 3.341, P < 0.0001), as well as portal venous tumor thrombosis grades 3-4 and alpha fetal protein (AFP) > 400 ng/mL. In the subgroup analysis, the better efficacy of hepatic resection over TACE persisted regardless of the grade of portal venous tumor thrombosis and the level of AFP. As for resectable patients, hepatic resection still showed significant survival benefit (before PSM, 17.5 vs 11.2 months, P = 0.0013; after PSM, 14.0 vs 10.9 months, P = 0.0304). CONCLUSION: Hepatic resection might be the better choice for patients with iHCC due to its better survival benefit than TACE.

Relationships between adiponectin and matrix metalloproteinase-9 (MMP-9) serum levels and postoperative cognitive dysfunction in elderly patients after general anesthesia.

OBJECTIVE: The aim of this study was to evaluate the relationships between the serum levels of adiponectin (ADP) and matrix metalloproteinase-9 (MMP-9) and postoperative cognitive dysfunction (POCD) in elderly patients after general anesthesia. METHODS: The cognitive functions of 98 elderly patients who were scheduled to undergo selective hip replacement surgery under general anesthesia were assessed using the Montreal Cognitive Assessment (MoCA) 3 days before surgery and on postoperative Days 1, 2, 3, and 7. The serum levels of ADP and MMP-9 were determined at the same time points, and the presence of POCD on postoperative Day 3 was recorded. The patients were divided into a POCD group and non-POCD group. RESULTS: Postoperative cognitive dysfunction was observed in 28 patients (28.5 %). Serum MMP-9 levels significantly increased and serum ADP levels significantly decreased in the POCD group at each postoperative time point and in the non-POCD group on postoperative Days 1 and 2 compared to the presurgical levels. Serum MMP-9 levels were significantly higher and serum ADP levels were significantly lower in the POCD group compared with those in the non-POCD group at each time point. In the POCD patients, serum MMP-9 levels were significantly and negatively correlated and serum ADP levels were significantly and positively correlated with the MoCA scores. CONCLUSIONS: The increased serum MMP-9 levels and decreased serum ADP levels in elderly patients after general anesthesia might be involved in the POCD pathophysiological process.

[Experimental study on concentrations and pharmacokinetics of antibiotics in bile and evaluation of their microbicidal potential].

OBJECTIVE: To study the concentrations and pharmacokinetics of 6 different kinds of antibiotics in rabbit bile, and evaluate their microbicidal potential. METHODS: Thirty-six health rabbits were randomly divided into 6 groups, and each group was 6 rabbits. After anaesthesia, the common bile duct of rabbit was isolated and cumulated with a silicone tube. The rabbits were administered intravenously with the equal-effect dose of antibiotics. Bile (1.5 ml) was collected at different time points after administration, and the concentration of antibiotics of bile was assayed by high performance liquid chromatography. The bile drug concentration-time data were processed by software to figure out the pharmacokinetic parameters such as maximum concentration (C(max)), peak time (T(max)), half-life time (T(1/2)), clearance (CL) and apparent volume of distribution (VD). The bile antibiotics concentration contrasted to the minimum inhibitory concentration (MIC), and attained the bactericidal index (C(max)/MIC) and the time when the drug concentration exceeded the MIC (T(>MIC)). RESULTS: The C(max) and T1/2 of each antibiotic were as the followings: piperacillin (7 950 ± 3 023) mg/L and (1.97 ± 1.23) h, ceftriaxone (1 104 ± 248) mg/L and (3.14 ± 0.57) h, cefoperazone (5 215 ± 2 225) mg/L and (0.89 ± 0.13) h, meropenem (31.97 ± 12.44) mg/L and (0.36 ± 0.11) h, levofloxacin (66.3 ± 36.9) mg/L and (3.32 ± 2.57) h, metronidazole (28.2 ± 10.2) mg/L and (0.81 ± 0.33) h, respectively. Piperacillin/tazobactam and cefoperazone/sulbactam had the largest bactericidal index and the longest T(>MIC), and their bactericidal indexes were (62.1 ± 23.6) - (993.8 ± 377.9) and (164.8 ± 69.0) - (659.3 ± 275.9), their T(>MIC) were (6.00 ± 2.53) - (8.00 ± 0.00) h and (6.33 ± 1.97) - (8.00 ± 0.00) h. The bactericidal index and T(>MIC) of levofloxacin were the smallest, which were (2.1 ± 1.2) - (8.3 ± 4.6) and (0.54 ± 0.25) - (2.67 ± 1.03) h . Ceftriaxone and meropenem were as the medium, and their bactericidal indexes and T(>MIC) were (4.3 ± 1.0) - (69.2 ± 15.5) , (1.42 ± 0.65) - (8.00 ± 0.00) h and (2.0 ± 0.8) - (1 031.3 ± 401.4) , (0.29 ± 0.10) - (1.83 ± 0.26) h. The bactericidal index of metronidazole to anaerobic ranged from 7.4 to 294.9, and the T(>MIC) ranged from 1.88 to 5.00 h. CONCLUSIONS: The bile concentrations of six antibiotics all exceed their effective bactericidal concentrations. The concentration-time curves of piperacillin, cefoperazone, meropenem and metronidazole conformed to one-compartment model, and ceftriaxone and levofloxacin are conformed to two-compartment model. Piperacillin/tazobactam and cefoperazone/sulbactam have the largest bactericidal index and the longest T(>MIC), so they can be chosen as the first choice for the therapy of hepatobiliary infection.For the anaerobic, the microbicidal potential of metronidazole is high.

Exosomes derived from mycobacterium tuberculosis-infected MSCs induce a pro-inflammatory response of macrophages.

Tuberculosis (TB) is a common infectious disease caused by Mycobacterium tuberculosis (M.tb), and macrophages serve as the primary natural host of M.tb. Mesenchymal stem cells (MSCs)-derived exosomes play an essential role in inflammatory responses. This study aimed to determine the role of exosomes derived from M.tb-infected MSCs (Exo-MSCs-M.tb) on macrophages in vitro and in vivo and the underlying mechanisms. Here, we demonstrated that M.tb infection promoted the production of Exo-MSCs-M.tb, but did not influence MSCs proliferation. Exo-MSCs-M.tb were taken up by macrophages and then induced the pro-inflammatory response of macrophages through elevating the production of TNF-α, RANTES, and iNOS. Also, pro-inflammatory response induced by Exo-MSCs-M.tb displayed a time-dependent pattern in macrophages, in which the highest level of inflammatory response was observed at 72 hours post-infection of MSCs. In addition, the effect of Exo-MSCs-M.tb was mediated through TLR2/4 and MyD88 signaling pathways. Furthermore, Exo-MSCs-M.tb could induce the pro-inflammatory response in mice in vivo, and exosomes isolated from Exo-MSCs-M.tb-treated mice could also promote the pro-inflammatory response. Taken together, these results indicate that Exo-MSCs-M.tb induced the pro-inflammatory response of macrophages through TLRs signaling. This study provides new insight into the potential of MSCs-derived exosomes for the treatment of TB.

[A mathematical calculation model of replacement fluid and citric acid infusion in continuous veno-venous hemofiltration].

OBJECTIVE: To design a mathematical calculation model for better understanding and grasping the logical problem of replacement fluid and citric acid anticoagulant infusion in continuous veno-venous hemofiltration (CVVH). METHODS: (1) Parameter definition: A, B, and T were respectively called the main part of pre-replacement fluid, 5% sodium bicarbonate solution, and 4% sodium citrate infused before filter. And a and b were respectively called the main part of post-replacement fluid, and 5% sodium bicarbonate solution infused after filter. (2) Logic conversion: The liquid in back terminal (Z) was artificially divided into two parts. One (X) was the original residual plasma after filtration. The second (Y) was the part excluding the plasma, including the left part of pre-replacement fluid with sodium citrate, and the post-replacement fluid. (3) The mathematical formulas of liquid volume and electrolyte concentration at X, Y and Z in unit time were listed according to the principle of CVVH and the screening coefficient of filter for different substances. (4) The calculation formulas were entered into Excel form, and a mathematical calculation model was made, and a simulation calculation with examples was carried out. RESULTS: An Excel model was established by inserting the calculation formulas of volume, electrolyte, and total calcium at X, Y and Z. And it was found that the concentration of Na+, K+, Cl-, HCO3- at Y point remained unchanged only when A, B and (or) a, b was kept in same side and proportion even with the change of blood flow and other parameters without sodium citrate as anticoagulant. Once any of the parameters (such as blood flow, replacement fluid volume, etc.) were adjusted in other infusion methods (such as different ratios, different directions of the same year, etc.), the calculation results at Y would vary, and the electrolyte concentration at Z would change accordingly. A change of dilution model or parameter would result in the change of the electrolyte concentration at Y and Z with sodium citrate as anticoagulant. The concentration of total calcium scarcely changed no matter in what model and parameters. CONCLUSIONS: All kinds of infusion ways could be included in the Excel model. The infusion results of all kinds of infusion matching could be intuitively evaluated. It is helpful for the medical staff to make a logical analysis and risk prediction in CVVH.

[Preoperative evaluation of multi-slice spiral computed tomography angiography in laparoscopic radical operation for colorectal carcinoma].

OBJECTIVE: To investigate the clinical application of 256 multi-slice spiral computed tomography angiography (MSCTA) technique in the preoperative evaluation of mesenteric angiography in order to provide a reference to vessel anatomy and dissociation in laparoscopic radical operation for colorectal carcinoma. METHODS: Clinical data of 50 patients with colorectal cancer who underwent preoperative MSCTA+FDCT and laparoscopic curative operation at our hospital from October 2013 to March 2015 were collected (MSCTA group). The evaluation item was visualization of mesenteric artery, which was compared with the findings under laparoscopic surgery. Meanwhile, another 50 colorectal cancer patients undergoing laparoscopic radical operation by the same surgeon team without preoperative MSCTA examination were used as control(control group). Clinical data were compared between the two groups. RESULTS: MSCTA precisely and correctly demonstrated anatomy and variations of the mesenteric artery and relative nutrient vessel in carcinoma. The angiography reconstruction images were consistent with the visual anatomy and variation from laparoscopic findings, whose diagnostic conformity rate of 100%. As compared to control group, operative time was shorter [(195.0±23.2) minutes vs.(218.0±19.6) minutes, t=8.326, P=0.015], and blood loss was less[(168.1±18.8) ml vs. (206.5±14.3) ml, t=-19.369, P=0.002] in MSCTA group. Differences of number of harvested lymph node, postoperative complication morbidity, postoperative hospital stay and hospitalization cost were not significant between two groups(all P>0.05). CONCLUSION: Preoperative MSCTA can demonstrate anatomy and variations of the mesenteric artery precisely and correctly, thus it is beneficial to shorten the operation time and to reduce blood loss.

[Detection of interferon-induced transmembrane-1 gene expression for clinical diagnosis of colorectal cancer].

OBJECTIVE: To investigate the expression of the interferon-induced transmembrane-1 (IFITM1) gene in colorectal cancer (CRC) tissue and the serum anti-IFITM1 antibody responses of the patients and assess their value in clinical diagnosis of CRC. METHODS: Semi-quantitative RT-PCR was performed to detect IFITM1 mRNA expression in the specimens of normal colonic mucosa, CRC tissue, inflammatory polyps, adenomatous polyps, gastric cancer, esophageal carcinoma and liver cancer tissues. Serum samples were collected from the patients to detect anti-IFITM1 antibody responses using Western blotting. The clinicopathological features of the carcinoma expressing IFITM1 gene were analyzed. RESULTS: IFITM1 mRNA was expressed in 47.4 % (18/38) of the CRC specimens, a rate significantly higher than that in adenomatous polyps [15% (3/20)] and gastric cancer [4.8% (1/21)]; no obvious IFITM1 expression was found in normal colonic mucosa, inflammatory polyp, esophageal carcinoma or liver cancer tissues (P<0.001 or P<0.05). IFITM1 mRNA was strongly expressed in CRC at the expression level of 0.8048-/+0.2273, which was significantly higher than that in adenomatous polyps (0.4447-/+0.0989, P<0.001). No anti-IFITM1 antibody response was detected in healthy human sera, but in the CRC patients, the serum antibody response was detected at the rate of 36.8% (14/38), significantly higher than the rate of 9.5% (2/21) in gastric cancer (P<0.05). No antibody response was detected in esophageal carcinoma, liver cancer, inflammatory polyp or adenomatous polyps. Most of the IFITM1-expressing CRC had a diameter exceeding 5 cm, often invading the serous membrane with metastasis to the lymph nodes and the distant organs; these tumors were identified mostly as well-differentiated adenocarcinoma in Dukes stage C or D. CONCLUSION: IFITM1 gene may play an important role in the pathogenesis, development and metastasis of CRC, and may serve as a potential biomarker for clinical diagnosis of CRC.