N-linoleyltyrosine resisted the growth of non-small cell lung cancer cells via the regulation of CB1 and CB2 involvement of PI3K and ERK pathways.

Background: N-linoleyltyrosine (NITyr), one of the anandamide analogs, exerts activity via the endocannabinoid receptors (CB1 and CB2), which showed anti-tumor effects in various tumors. Therefore, we speculated that NITyr might show anti-non-small cell lung cancer (NSCLC) effects via the CB1 or CB2 receptor. The purpose of the investigation was to reveal the anti-tumor ability of NITyr on A549 cells and its mechanisms. Methods: The viability of A549 cells was measured by MTT assay, and the cell cycle and apoptosis were both examined by flow cytometry; in addition, cell migration was tested by wound healing assay. Apoptosis-related markers were measured by immunofluorescence. The downstream signaling pathways (PI3K, ERK, and JNK) of CB1 or CB2 were examined through Western blotting. The expressions of CB1 and CB2 were detected by immunofluorescence. Finally, the AutoDock software was used to validate the binding affinity between the targets, such as CB1 and CB2, with NITyr. Results: We found that NITyr inhibited cell viability, hindered the cell cycle, resulted in apoptosis, and inhibited migration. The CB1 inhibitor, AM251, and the CB2 inhibitor, AM630, weakened the aforementioned phenomenon. The immunofluorescence assay suggested that NITyr upregulated the expression of CB1 and CB2. Western blot analysis indicated that NITyr upregulated the expression of p-ERK, downregulated the expression of p-PI3K, and did not affect p-JNK expression. In conclusion, NITyr showed a role in inhibiting NSCLC through the activation of CB1 and CB2 receptors involved in PI3K and ERK pathways.

Laparoscopic versus ultrasound-guided transversus abdominis plane block for postoperative pain management in minimally invasive colorectal surgery: a meta-analysis protocol.

Introduction: Transversus abdominis plane block (TAPB) is now commonly administered for postoperative pain control and reduced opioid consumption in patients undergoing major colorectal surgeries, such as colorectal cancer, diverticular disease, and inflammatory bowel disease resection. However, there remain several controversies about the effectiveness and safety of laparoscopic TAPB compared to ultrasound-guided TAPB. Therefore, the aim of this study is to integrate both direct and indirect comparisons to identify a more effective and safer TAPB approach. Materials and methods: Systematic electronic literature surveillance will be performed in the PubMed, Embase, Cochrane Central Register of Controlled Trials (CENTRAL), and ClinicalTrials.gov databases for eligible studies through July 31, 2023. The Cochrane Risk of Bias version 2 (RoB 2) and Risk of Bias in Non-randomized Studies of Interventions (ROBINS-I) tools will be applied to scrutinize the methodological quality of the selected studies. The primary outcomes will include (1) opioid consumption at 24 hours postoperatively and (2) pain scores at 24 hours postoperatively both at rest and at coughing and movement according to the numerical rating scale (NRS). Additionally, the probability of TAPB-related adverse events, overall postoperative 30-day complications, postoperative 30-day ileus, postoperative 30-day surgical site infection, postoperative 7-day nausea and vomiting, and length of stay will be analyzed as secondary outcome measures. The findings will be assessed for robustness through subgroup analyses and sensitivity analyses. Data analyses will be performed using RevMan 5.4.1 and Stata 17.0. P value of less than 0.05 will be defined as statistically significant. The certainty of evidence will be examined via the Grading of Recommendations, Assessment, Development, and Evaluation (GRADE) working group approach. Ethics and dissemination: Owing to the nature of the secondary analysis of existing data, no ethical approval will be required. Our meta-analysis will summarize all the available evidence for the effectiveness and safety of TAPB approaches for minimally invasive colorectal surgery. High-quality peer-reviewed publications and presentations at international conferences will facilitate disseminating the results of this study, which are expected to inform future clinical trials and help anesthesiologists and surgeons determine the optimal tailored clinical practice for perioperative pain management. Systematic review registration: https://www.crd.york.ac.uk/PROSPERO/display_record.php?RecordID=281720, identifier (CRD42021281720).

Simultaneous determination of 22 fatty acids in total parenteral nutrition (TPN) components by gas chromatography-mass spectrometry (GC-MS).

Evaluating total parenteral nutrition (TPN) products for quality assurance and quality control is crucial due to the chemical complexity of its components. With the advent of exploring different approaches for analysing TPN components using tandem mass spectrometry techniques, there is still a need for a robust and reproducible method for industrial routine analyses. This study allows simple, simultaneous determination of 22 fatty acids (FAs) commonly found in TPN components using gas chromatography-mass spectrometry (GC-MS). Five different transesterification techniques were applied for the FA standards and the sodium methoxide in methanol-dimethyl carbonate method was selected due to its good methylation efficiency. Fatty acid methyl esters (FAMEs) were separated in gas chromatography using an HP-5MS UI column with helium as the carrier gas. Mass spectrometry was used to fragment and quantify FAMEs using electron ionization (EI) and selected ion monitoring (SIM) mode. The analytical method was evaluated using the guidelines from the US Food and Drug Agency (FDA) and European Medicines Agency (EMA) in compliance with the International Council for Harmonization (ICH) document Q2(R2). Correlation coefficients (R2) of the calibration curves for FAMEs were 0.99, except for C24:1 n-9 and C24:0, both R2 = 0.98. The limits of detection (LOD) and quantification (LOQ) were found to be 1.69 µg mL-1 and 5.14 µg mL-1, respectively. The linear range was from 3.10-179.9 µg mL-1 for most FAMEs, except for C18:1 n-7 (3.96-224.9 µg mL-1) and C18:1 n-9 (6.30-349.57 µg mL-1). The intra-day and inter-day precision coefficients of variance (CV) of the method were less than 11.10% and 11.30%, respectively. Freeze-thaw cycles and ambient temperature measurements were performed for assessing sample stability. The validated method was applied to analyse major TPN components-fish and olive oils, and an unidentified lipid sample. The presented GC-MS method is simple and robust in the identification and quantification of 22 fatty acids simultaneously in the tested TPN components.

Oncologic outcomes of intersphincteric resection versus abdominoperineal resection for lower rectal cancer: a systematic review and meta-analysis.

BACKGROUND: The efficacy of intersphincteric resection (ISR) surgery for patients with lower rectal cancer remains unclear compared to abdominoperineal resection (APR). The aim of this study is to compare the oncologic outcomes for lower rectal cancer patients after ISR and APR through a systematic review and meta-analysis. MATERIALS AND METHODS: A systematic electronic search of the Cochrane Library, PubMed, EMBASE, and MEDLINE was performed through January 12, 2022. The primary outcomes included 5-year disease-free survival (5y-DFS) and 5-year overall survival. Secondary outcomes included circumferential resection margin involvement, local recurrence, perioperative outcomes, and other long-term outcomes. The pooled odds ratios, mean difference, or hazard ratios (HRs) of each outcome measurement and their 95% CIs were calculated. RESULTS: A total of 20 nonrandomized controlled studies were included in the qualitative analysis, with 1217 patients who underwent ISR and 1135 patients who underwent APR. There was no significant difference in 5y-DFS (HR: 0.84, 95% CI: 0.55-1.29; P=0.43) and 5-year overall survival (HR: 0.93, 95% CI: 0.60-1.46; P=0.76) between the two groups. Using the results of five studies that reported matched T stage and tumor distance, we performed another pooled analysis. Compared to APR, the ISR group had equal 5y-DFS (HR: 0.76, 95% CI: 0.45-1.30; P=0.31) and 5y-LRFS (local recurrence-free survival) (HR: 0.72, 95% CI: 0.29-1.78; P=0.48). Meanwhile, ISR had equivalent local control as well as perioperative outcomes while significantly reducing the operative time (mean difference: -24.89, 95% CI: -45.21 to -4.57; P=0.02) compared to APR. CONCLUSIONS: Our results show that the long-term survival and safety of patients is not affected by ISR surgery, although this result needs to be carefully considered and requires further study due to the risk of bias and limited data.

Elucidation of CKAP4-remodeled cell mechanics in driving metastasis of bladder cancer through aptamer-based target discovery.

Metastasis contributes to the dismal prognosis of bladder cancer (BLCA). The mechanical status of the cell membrane is expected to mirror the ability of cell migration to promote cancer metastasis. However, the mechanical characteristics and underlying molecular profile associated with BLCA metastasis remain obscure. To study the unique cellular architecture and traits associated with cell migration, using a process called cell-based systematic evolution of ligands by exponential enrichment (cell-SELEX) we generated an aptamer-based molecular probe, termed spl3c, which identified cytoskeleton-associated protein 4 (CKAP4). CKAP4 was associated with tumor metastasis in BLCA, but we also found it to be a mechanical regulator of BLCA cells through the maintenance of a central-to-peripheral gradient of stiffness on the cell membrane. Notably, such mechanical traits were transportable through exosome-mediated intercellular CKAP4 trafficking, leading to significant enhancement of migration in recipient cells and, consequently, aggravating metastatic potential in vivo. Taken together, our study shows the robustness of this aptamer-based molecular tool for biomarker discovery, revealing the dominance of a CKAP4-induced central-to-peripheral gradient of membrane stiffness that benefits cell migration and delineating the role of exosomes in mediating mechanical signaling in BLCA metastasis.

Prior Distribution Estimation of Monitored Information in the Intensive Care Unit with the Hidden Markov Model and Decision Tree Methods.

In the intensive care unit, the monitored variables collected from sensors may have different behaviors among patients with different clinical basic information. Giving prior information of the monitored variables based on their specific basic information as soon as the patient is admitted will support the clinicians with better decisions during the surgery. Instead of black box models, the explainable hidden Markov model is proposed, which can estimate the possible distribution parameters of the monitored variables under different clinical basic information. A Student's t-test or correlation test is conducted further to test whether the parameters have a significant relationship with the basic variables. The specific relationship is explored by using a conditional inference tree, which is an explainable model giving deciding rules. Instead of point estimation, interval forecast is chosen as the performance metrics including coverage rate and relative interval width, which provide more reliable results. By applying the methods to an intensive care unit data set with more than 20 thousand patients, the model has good performance with an area under the ROC Curve value of 0.75, which means the hidden states can generally be correctly labelled. The significant test shows that only a few combinations of the basic and monitored variables are not significant under the 0.01 significant level. The tree model based on different quantile intervals provides different coverage and width combination choices. A coverage rate around 0.8 is suggested, which has a relative interval width of 0.77.

Case report and literature review: Small bowel intussusception due to solitary metachronous metastasis from renal cell carcinoma.

Introduction: Solitary metachronous small bowel metastasis from renal cell carcinoma (RCC) is rare. In contrast to idiopathic intussusception frequently occurring in children, adult intussusception is fairly uncommon and usually indicates a malignancy. Case presentation: We presented an 84-year-old man with small bowel intussusception and obstruction due to a solitary metachronous metastasis from RCC. Computed tomography with intravenous contrast revealed small bowel obstruction and a 4 × 4 cm intraluminal soft-tissue mass with moderate enhancement. During urgent exploratory laparotomy, a pedunculated tumor of the distal ileum was found to be the lead point of intussusception. Hence, reduction of the intestinal invagination and segmental resection of the ileum with functional end-to-end anastomosis were performed. Histological examination finally confirmed the diagnosis. The postoperative recovery was uneventful. The patient was discharged without any complications on postoperative day 6. Conclusion: The case report highlights the rarity of solitary metachronous small bowel metastases from RCC and suggests that life-long follow-up of RCC patients is critical due to its unpredictable behavior and the possibility of a long period of dormancy. Complete surgical resection remains the mainstay treatment for such patients.

Impaired bone marrow microenvironment and stem cells in transfusion-dependent beta-thalassemia.

Beta-thalassemia (BT) is a hereditary disease caused by abnormal hemoglobin synthesis with consequent ineffective erythropoiesis. Patients with thalassemia major are dependent on long-term blood transfusions with associated long-term complications such as iron overload (IO). This excess iron can result in tissue damage, impaired organ function, and increased morbidity. Growing evidence has demonstrated that IO contributes to impairment of the bone marrow (BM) microenvironment that largely impacts the function of BM mesenchymal stem cells, hematopoietic stem cells, and endothelial cells. In this article, we review recent progress in the understanding of iron metabolism and the perniciousness induced by IO. We highlight the importance of understanding the cross-talk between BM stem cells and the BM microenvironment, particularly the pathological effect of IO on BM stem cells and BT-associated complications. We also provide an update on recent novel therapies to cure transfusion-dependent beta-thalassemia and iron overload-induced complications for their future clinical application.

The regulation of NONO by USP11 via deubiquitination is linked to the proliferation of melanoma cells.

Ubiquitin-specific protease 11 (USP11) has been implicated in the regulation of DNA repair, apoptosis, signal transduction and cell cycle. It belongs to a USP subfamily of deubiquitinases. Although previous research has shown that USP11 overexpression is frequently found in melanoma and is correlated with a poor prognosis, the potential molecular mechanism of USP11 in melanoma remains indefinitive. Here, we report that USP11 and NONO colocalize and interact with each other in the nucleus of melanoma cells. As a result, the knockdown of USP11 decreases NONO levels. Whereas, overexpression of USP11 increases NONO levels in a dose-dependent manner. Furthermore, we reveal that USP11 protects NONO protein from proteasome-mediated degradation by removing poly-ubiquitin chains conjugated onto NONO. Functionally, USP11 mediated melanoma cell proliferation via the regulation of NONO levels because ablation of USP11 inhibits the proliferation which could be rescued by ectopic expression of NONO protein. Moreover, a significant positive correlation between USP11 and NONO concentrations was found in clinical melanoma samples. Collectively, these results demonstrate that USP11 is a new deubiquitinase of NONO and that the signalling axis of USP11-NONO is significantly involved in melanoma proliferation.

NONO and tumorigenesis: More than splicing.

The non-POU domain-containing octamer-binding protein NONO/p54nrb , which belongs to the Drosophila behaviour/human splicing (DBHS) family, is a multifunctional nuclear protein rarely functioning alone. Emerging solid evidences showed that NONO engages in almost every step of gene regulation, including but not limited to mRNA splicing, DNA unwinding, transcriptional regulation, nuclear retention of defective RNA and DNA repair. NONO is involved in many biological processes including cell proliferation, apoptosis, migration and DNA damage repair. Dysregulation of NONO has been found in many types of cancer. In this review, we summarize the current and fast-growing knowledge about the regulation of NONO, its biological function and implications in tumorigenesis and cancer progression. Overall, significant findings about the roles of NONO have been made, which might make NONO to be a new biomarker or/and a possible therapeutic target for cancers.

Deubiquitinase DUB3 Regulates Cell Cycle Progression via Stabilizing Cyclin A for Proliferation of Non-Small Cell Lung Cancer Cells.

The deubiquitinase DUB3 is frequently overexpressed in non-small cell lung cancer (NSCLC) and contributes to its malignant phenotype. However, the underlying molecular mechanism of DUB3 in NSCLC is largely unknown. In this study, we report that DUB3 regulates cell cycle progression by deubiquitinating cyclin A that links to proliferation of NSCLC cells. We found that knockdown of DUB3 decreases cyclin A levels, whereas overexpression of DUB3 strongly increases cyclin A levels. Mechanistically, DUB3 interacts with cyclin A, which removes the polyubiquitin chains conjugated onto cyclin A and stabilizes the cyclin A protein. Furthermore, we demonstrate that DUB3 regulates cell cycle progression by stabilizing cyclin A, because ablation of DUB3 arrests cell cycle from G0/G1 to S phase and the resulting effect can be rescued by introducing cyclin A into NSCLC cells. Functionally, we found that the effect of DUB3 on cyclin A mediates proliferation of NSCLC cells. Moreover, a significant correlation between DUB3 abundance and cyclin A expression levels were also found in NSCLC samples. Taken together, these results reveal that DUB3 functions as a novel cyclin A regulator through maintaining cyclin A stability, and that the DUB3-cyclin A signaling axis plays a critical role in cell cycle progression for proliferation of NSCLC.

A successful combined laparoscopic cholecystectomy and laparoscopic exploration of common bile duct for acute gangrenous cholecystitis and choledocholithiasis during pregnancy: A case report.

INTRODUCTION: Choledocholithiasis during pregnancy is relatively uncommon but it can be life-threatening. Therapeutic endoscopic retrograde cholangiopancreatography (ERCP) poses the risk of ionizing radiation exposure to the developing fetus. Other strategies are thus needed to tackle this problem. PRESENTATION OF CASE: A 38-year-old 8 weeks' pregnant lady presented with acute onset of right upper quadrant pain. Ultrasonography showed features of acute cholecystitis and choledocholithiasis, which was later confirmed by magnetic resonance cholangiogram. Emergency combined laparoscopic cholecystectomy (LC) and laparoscopic exploration of common bile duct (Lap ECBD) were performed. Lap ECBD through trans-cystic duct approach was adopted. The patient recovered uneventfully and was discharged on postoperative day 4. A healthy baby boy was delivered at 40 weeks' gestation without developmental problem. CONCLUSION: Lap ECBD avoided ionizing radiation for choledocholithiasis during pregnancy, and can combined with LC as an effective minimally invasive procedure for complicated gallstone disease.

Combined associating liver partition and portal vein ligation for staged hepatectomy (ALPPS) followed by left trisectionectomy and Whipple operation for PNET.

Pancreatic neuroendocrine tumor (PNET) is slow-growing, and account only for 2% of all pancreatic primary tumors. Surgical resection is still the only curative treatment for PNET patients. Unfortunately, most of PNETs was found with unresectable multiple liver metastases and extrahepatic metastasis as their characteristics of non-functional and asymptomatic. With advances in liver surgery in these years, especially combined associating liver partition and portal vein ligation for staged hepatectomy (ALPPS), provide a new curative surgical treatment for PNET with liver metastases patient. Here we report a PNET with multiple liver metastases case underwent ALPPS (followed by left trisectionectomy) and Whipple operation within one-stage.

Effect of short-chain fatty acids on triacylglycerol accumulation, lipid droplet formation and lipogenic gene expression in goat mammary epithelial cells.

Short-chain fatty acids (SCFAs) are the major energy sources for ruminants and are known to regulate various physiological functions in other species. However, their roles in ruminant milk fat metabolism are still unclear. In this study, goat mammary gland epithelial cells (GMECs) were treated with 3 mmol/L acetate, propionate or butyrate for 24 h to assess their effects on lipogenesis. Data revealed that the content of triacylglycerol (TAG) and lipid droplet formation were significantly stimulated by propionate and butyrate. The expression of FABP3, SCD1, PPARG, SREBP1, DGAT1, AGPAT6 and ADRP were upregulated by propionate and butyrate treatment. In contrast, the messenger RNA (mRNA) expression of FASN and LXRα was not affected by propionate, but reduced by butyrate. Acetate had no obvious effect on the content of TAG and lipid droplets but increased the mRNA expression of SCD1 and FABP3 in GMECs. Additionally, it was observed that propionate significantly increased the relative content of mono-unsaturated fatty acids (C18:1 and C16:1) at the expense of decreased saturated fatty acids (C16:0 and C18:0). Butyrate and acetate had no significant effect on fatty acid composition. Overall, the results from this work help enhance our understanding of the regulatory role of SCFAs on goat mammary cell lipid metabolism.

[Nanogold inhibits angiogenesis and growth of liver cancer: experiment with mice].

OBJECTIVE: To investigate the effects of nanogold in inhibition of angiogenesis and growth of liver cancer cells. METHODS: Nanogold was co-incubated with VEGF165 and VDGF121 respectively. Atomic force microscopy (AFM) was used to observe the changes of the form of the particles. Human umbilical vascular endothelial cells (HUVEC) were serum-starved for 24 hours, then co-cultured with VEGF165 + nanogold or VEGF121 + nanogold for 24 h. ATM was used to observe the ultrastructure of the cells. Another HUVEC were serum-starved for 24 hours and then cultured with VEGF165 (10 microg/L) 100 microl + nanogold 125, 250, and 500 nmol/L 100 microl respectively for 5 min. Then Western blotting was used to detect the phosphorylation protein of phospholipase C (PLC)-gamma1 on VEGFR-2. Hepatocellular cancer cells of the line H22 were injected subcutaneously into the right armpit of 20 Balb/c nude mice. When the size of transplanted tumor reached about 8 mm, the mice were divided into 2 equal groups: experimental group undergoing injection of nanogold into the tumor once a day for 8 days, and control group injected with normal saline. On day 14 the mice were sacrificed with the liver tumors taken out to measure the size and weight. The microvascular density (MVD) of tumor was determined by immunohistochemical staining. RESULTS: ATM showed that acted with VEGF165, the size of nanogold became over 30 nm. Treated with VEGF165 the HUVEC became larger with obvious pseudopodium. However, such changes were obviously milder in those HUVEC treated with nanogold + VEGF165. The PLC-gamma1 phosphorylation level VEGF receptor-2 was decreased along with the increase of the concentration of nanogold. The MVD of liver cancer tissue in the experimental group was 14.27 +/- 1.08, significantly lower than that of the control group [(23.52 +/- 1.36), P < 0.01]. The mean weight and volume of tumor of the experimental group were (1.39 +/- 0.08) g and (1.37 +/- 0.34) cm(3) respectively, both significantly lower than those of the control group [(2.47 +/- 0.15) g and (2.49 +/- 0.78) cm(3) respectively, both P < 0.05] with a tumor growth inhibition rate of 43.72%. CONCLUSION: Nanogold significantly inhibits the angiogenesis and growth of liver cancer cells with the possible mechanism that nanogold inhibits the VEGF165-induced signaling.