Behaviors and Mechanisms of Adsorption of MB and Cr(VI) by Geopolymer Microspheres under Single and Binary Systems.

Geopolymers show great potential in complex wastewater treatment to improve water quality. In this work, general geopolymers, porous geopolymers and geopolymer microspheres were prepared by the suspension curing method using three solid waste products, coal gangue, fly ash and blast furnace slag. The microstructure, morphology and surface functional groups of the geopolymers were studied by SEM, XRD, XRF, MIP, FTIR and XPS. It was found that the geopolymers possess good adsorption capacities for both organic and inorganic pollutants. With methylene blue and potassium dichromate as the representative pollutants, in order to obtain the best removal rate, the effects of the adsorbent type, dosage of adsorbent, concentration of methylene blue and potassium dichromate and pH on the adsorption process were studied in detail. The results showed that the adsorption efficiency of the geopolymers for methylene blue and potassium dichromate was in the order of general geopolymers < porous geopolymers < geopolymer microspheres, and the removal rates were up to 94.56% and 79.46%, respectively. Additionally, the competitive adsorption of methylene blue and potassium dichromate in a binary system was also studied. The mechanism study showed that the adsorption of methylene blue was mainly through pore diffusion, hydrogen bond formation and electrostatic adsorption, and the adsorption of potassium dichromate was mainly through pore diffusion and redox reaction. These findings demonstrate the potential of geopolymer microspheres in adsorbing organic and inorganic pollutants, and, through five cycles of experiments, it is demonstrated that MGP exhibits excellent recyclability.

An Improved Genetic Algorithm for the Recovery System of USVs Based on Stern Ramp Considering the Influence of Currents.

With the progression of marine exploration and exploitation, as well as the advancements in mechanical intelligence, the utilization of the unmanned surface vehicle (USV) and the design of their guidance system have become prominent areas of focus. However, the stern ramp recovery of the USV is still in its infancy due to its unique attitude requirements and automation design. Furthermore, few studies have addressed the impact of maritime disturbances, with most research limited to simulations. To enhance the efficiency and accuracy of stern ramp recovery, this paper presents the development and construction of a novel recovery system. By incorporating physical modeling of disturbance forces acting on USVs at sea, the practicality of the system is improved. Additionally, an optimized genetic algorithm is introduced in the navigation module to improve convergence rates and subsequently enhance recovery efficiency. A line-of-sight (LOS) algorithm based on average velocity is proposed in this paper to ensure the attainment of unique attitude requirements and to improve the effectiveness of stern chute recovery. This paper provides a detailed description of the independently designed USV hardware system. Moreover, simulations and practical experiments conducted using this experimental platform are presented, offering a new solution for the USV's stern ramp recovery.

Effect of postoperative application of esketamine on postoperative depression and postoperative analgesia in patients undergoing pancreatoduodenectomy: a randomized controlled trial protocol.

BACKGROUND: Pancreatoduodenectomy (PD) is traumatic, difficult to perform, and has a high incidence of postoperative complications and perioperative mortality. Postoperative complications and pain occur frequently and seriously affect the psychological status of patients. Esketamine, an N-methyl-D-aspartate (NMDA) receptor antagonist, has analgesic and antidepressant effects. In this study, we aim to investigate the effect of esketamine on postoperative depression and pain in patients undergoing PD. METHODS/DESIGN: This prospective, single-center, randomized control trial will include 80 patients who will undergo elective PD. The patients will be randomly assigned to two groups: the experimental group that will receive esketamine (n = 40) and the control group (n = 40). In the esketamine group, the analgesic pump will be connected immediately after surgery. A solution of esketamine 1.5 mg/kg + sufentanil 2 µg/kg, diluted to 150 mL, will be administered continuously for 72 h at the background infusion and impact doses of 1 mL/h and 2 mL/time, respectively; the locking time will be 10 min. The control group will receive sufentanil 2 µg/kg that will be administered as per the esketamine group. The primary outcome will be the Hamilton Depression Scale (HAMD-17) score on the third day post-surgery (POD3). Secondary study indicators will include (1) visual analog scale (VAS) score and HAMD-17 score prior to surgery, immediately after entering the postanesthesia care unit (PACU) and 1, 2, 3, 4, and 5 days after surgery; (2) Richmond Agitation-Sedation Scale (RASS) score at 1, 2, 3, 4, and 5 days after surgery; (3) consumed doses of sufentanil and esketamine after surgery; (4) postoperative analgesia pump effective press times, rescue analgesia times, and rescue drug dosage, recording the number of rescue analgesia and rescue drug dosage at 6, 24, 48, and 72 h after the patient enters the PACU; (5) postoperative complications and adverse events; (6) postoperative hospital stay; (7) concentrations of brain-derived neurotrophic factor (BDNP), 5-hydroxytryptamine (5-HT), tumor necrosis factor (TNF-α) and interleukin-6, at 1, 3, and, 5 days post-surgery; and (8) the patient survival rate at 6 and 12 months post-surgery. DISCUSSION: The study hypothesis is that the postoperative HAMD-17 and VAS scores, incidence of postoperative adverse reactions, and concentration of serum markers BDNP, 5-HT, TNF-α, and IL-6 in the experimental group will be lower than those in the control group. TRIAL REGISTRATION: ClinicalTrials.gov ChiCTR2200066303. Registered on November 30, 2022. PROTOCOL VERSION: 1.0.

Subcutaneous soft tissue metastases from esophageal squamous cell carcinoma with neuroendocrine differentiation: Case report and literature review.

Background: Esophageal squamous cell carcinoma is the predominant subtype of esophageal cancer in China and so differs from presentations in Western countries. Common metastatic locations of esophageal cancer include the liver, lung, bone, and brain. In contrast, metastases in subcutaneous soft tissue are exceedingly rare. Case presentation: We present the experience of a 57-year-old man with a complaint of hand and leg dysfunction on the right side. He had a past medical history of esophageal squamous cell carcinoma. Further imaging workup revealed a solitary brain metastasis, thickening of the esophageal wall, swollen lymph nodes in the mediastinum, and right adrenal gland metastasis. Gamma knife radiosurgery of the brain metastasis and intensity-modulated radiotherapy of the esophagus and lymph nodes were administered. After 1.5 months, he was admitted to our hospital again, and nodules were identified in the anterior abdominal wall and left posterior chest wall. Ultrasound, CT, and radical excision of the abdominal wall mass were undertaken and revealed metastatic squamous cell carcinoma with neuroendocrine differentiation. We administered immunotherapy followed by targeted therapy. A PET/CT scan was performed to identify other organ metastases; the scan revealed multiple areas of fluorodeoxyglucose uptake and foci in the esophagus, lung, liver, bone, and right adrenal gland; and in various lymph nodes. In addition, an intensely hypermetabolic lesion was localized in the left posterior thorax. Conclusion: This case highlights the diagnosis and treatment of uncommon metastases of esophageal squamous cell carcinoma. We hope that our clinical experience provides insights into these uncommon metastases.

Advances in targeted therapy in non-small cell lung cancer with actionable mutations and leptomeningeal metastasis.

WHAT IS KNOWN AND OBJECTIVE?: Leptomeningeal metastasis (LM) is a serious complication of advanced non-small cell lung cancer (NSCLC) that is diagnosed in approximately 3%-5% of patients. LM occurs more frequently in patients with NSCLC harbouring epidermal growth factor receptor (EGFR) mutations or anaplastic lymphoma kinase (ALK) rearrangements and is usually accompanied by a poor prognosis, with a median overall survival (OS) of several months if patients receive conventional treatments. However, tyrosine kinase inhibitor (TKI) therapy after LM diagnosis is an independent predictive factor for extended survival. Here, we aim to summarize the latest advances in targeted therapy for LM and provide patients with better treatment options. METHODS: By reviewing the recent progress of targeted therapy in NSCLC with LM, especially the efficacy of newer generation TKIs, we aim to provide clinicians with a reference to further optimize patient treatment plans. RESULTS AND DISCUSSION: Osimertinib was confirmed to have a several-fold higher CNS permeability than other EGFR-TKIs and was recommended as the preferred choice for patients with EGFR-positive LM whether or not they harboured the T790M mutation. Second-generation ALK-TKIs have a higher rate of intracranial response and can be positioned as front-line drugs in NSCLC with LM. However, the sequence in which ALK-TKIs are administered for effective disease control requires further evaluation. In addition, targeted therapy revealed a potential choice in patients with LM and rare mutations, such as ROS1 and BRAF. WHAT IS NEW AND CONCLUSIONS?: The development of therapeutic agents with greater CNS penetration is vital for the management of CNS metastasis from NSCLC, particularly in the EGFR-mutant and ALK-rearranged subtypes. Systemic therapy with newer generation TKIs is preferred as the initial intervention. This is because newer generation TKIs are designed to penetrate the blood-brain barrier and possess significantly higher intracranial activities. However, their further effectiveness is limited by inadequate blood-brain barrier penetration and acquired drug resistance. Further studies are needed to further understand the mechanisms underlying resistance to treatment.

Identification of m6A-Related lncRNAs Associated With Prognoses and Immune Responses in Acute Myeloid Leukemia.

Background: Acute myeloid leukemia (AML) remains the most common type of hematopoietic malignancy in adults and has an unfavorable outcome. Herein, we aimed to construct an N6-methylandenosine (m6A)-related long noncoding RNAs (lncRNAs) signature to accurately predict the prognosis of patients with AML using the data downloaded from The Cancer Genome Atlas (TCGA) database. Methods: The RNA-seq and clinical data were obtained from the TCGA AML cohort. First, Pearson correlation analysis was performed to identify the m6A-related lncRNAs. Next, univariate Cox regression analysis was used to determine the candidate lncRNAs with prognostic value. Then, feature selection was carried out by Least absolute shrinkage and selection operator (LASSO) analysis, and seven eligible m6A-related lncRNAs were included to construct the prognostic risk signature. Kaplan-Meier and receiver operating characteristic (ROC) curve analyses were performed to evaluate the predictive capacity of the risk signature both in the training and testing datasets. A nomogram was used to predict 1-year, 2-year, and 3-year overall survival (OS) of AML patients. Next, the expression levels of lncRNAs in the signature were validated in AML samples by qRT-PCR. Functional enrichment analyses were carried out to identify probable biological processes and cellular pathways. The ceRNA network was developed to explore the downstream targets and mechanisms of m6A-related lncRNAs in AML. Results: Seven m6A-related lncRNAs were identified as a prognostic signature. The low-risk group hold significantly prolonged OS. The nomogram showed excellent accuracy of the signature for predicting 1-year, 2-year and 3-year OS (AUC = 0.769, 0.820, and 0.800, respectively). Moreover, the risk scores were significantly correlated with enrichment in cancer hallmark- and malignancy-related pathways and immunotherapy response in AML patients. Conclusion: We developed and validated a novel risk signature with m6A-related lncRNAs which could predict prognosis accurately and reflect the immunotherapy response in AML patients.

Comparison of postoperative analgesic effects in response to either dexamethasone or dexmedetomidine as local anesthetic adjuvants: a systematic review and meta-analysis of randomized controlled trials.

This review compares the effects of peripheral dexamethasone and dexmedetomidine on postoperative analgesia. We included six randomized controlled trials (354 patients) through a systematic literature search. We found that analgesia duration was comparable between dexamethasone and dexmedetomidine (58.59 min, 95% CI (confidence interval), - 66.13, 183.31 min) with extreme heterogeneity. Secondary outcome was also compared and no significant difference was observed in sensory block onset and duration and motor block duration and also for postoperative nausea and vomiting. It is noteworthy that dexamethasone reduced analgesic consumption (fentanyl) by 29.12 mcg compared with dexmedetomidine. We performed subgroup analyses and found no significant difference between the following: (1) lidocaine vs ropivacaine (P = 0.28), (2) nerve block vs nerve block + general anesthesia (P = 0.47), and (3) upper limb surgery vs thoracoscopic pneumonectomy (P = 0.27). We applied trial sequential analysis to assess the risks of type I and II errors and concluded that the meta-analysis was insufficiently powered to answer the clinical question, and further analysis is needed to establish which adjuvant is better. In conclusion, we believe that existing research indicates that dexamethasone and dexmedetomidine have equivalent analgesic effects in peripheral nerve blocks.

Effect of Sodium Dodecyl Sulfonate on the Foam Stability and Adsorption Configuration of Dodecylamine at the Gas-Liquid Interface.

In this study, the effect of sodium dodecyl sulfonate (SDS) on the foam stability of dodecylamine (DDA) and on its adsorption configuration at the gas-liquid interface was investigated. Froth stability experiments, surface tension measurements, time-of-flight secondary-ion mass spectrometry measurements, and molecular dynamics simulation calculations were performed in this investigation. The results revealed that the foam stability of DDA solution was extremely strong, and the addition of SDS could decrease the foam stability when the concentration of DDA was less than a certain value. The decrease in foam stability could be ascribed to several reasons, namely, the big cross-sectional area of SDS at the gas-liquid interface and low adsorption capacity of surfactants at the gas-liquid interface, the high surface tension, the change in the double-layer structure, the small thickness of the gas-liquid interfacial layer, the weak interaction intensity between the head groups of the surfactants and the water molecules, the strong movement ability of the water molecules around the head groups, and the sparse and less upright arrangement configuration of molecules at the gas-liquid interface. These findings can greatly help in solving the strong foam stability problem in DDA flotation and provide a method for reducing foam stability.

Development of a CD63 Aptamer for Efficient Cancer Immunochemistry and Immunoaffinity-Based Exosome Isolation.

CD63, a member of transmembrane-4-superfamily of tetraspanin proteins and a highly N-glycosylated type III lysosomal membrane protein, is known to regulate malignancy of various types of cancers such as melanoma and breast cancer and serves as a potential marker for cancer detection. Recently, its important role as a classic exosome marker was also emphasized. In this work, via using a magnetic bead-based competitive SELEX (systematic evolution of ligands by exponential enrichment) procedure and introducing a 0.5M NaCl as elution buffer, we identified two DNA aptamers (CD63-1 and CD63-2) with high affinity and specificity to CD63 protein (Kd = 38.71nM and 78.43, respectively). Furthermore, CD63-1 was found to be efficient in binding CD63 positive cells, including breast cancer MDA-MB-231 cells and CD63-overexpressed HEK293T cells, with a medium binding affinity (Kd~ 100 nM) as assessed by flow cytometry. When immunostaining assay was performed using clinical breast cancer biopsy, the CD63-1 aptamer demonstrated a comparable diagnostic efficacy for CD63 positive breast cancer with commercial antibodies. After developing a magnetic bead-based exosome immunoaffinity separation system using CD63-1 aptamer, it was found that this bead-based system could effectively isolate exosomes from both MDA-MB-231 and HT29 cell culture medium. Importantly, the introduction of the NaCl elution in this work enabled the isolation of native exosomes via a simple 0.5M NaCl incubation step. Based on these results, we firmly believe that the developed aptamers could be useful towards efficient isolation of native state exosomes from clinical samples and various theranostic applications for CD63-positive cancers.

Structures of Pb-BHA Complexes Adsorbed on Scheelite Surface.

Previous studies have shown that Pb-BHA complexes (lead complexes of benzohydroxamic acid) have better collecting ability and can be used in flotation experiments with BHA acting as a collector and lead ions acting as activators. However, the structures of Pb-BHA complexes adsorbed on a mineral surface remain unclear. In this work, the adsorption behavior of Pb-BHA complexes on the scheelite surface was studied by flotation experiments and adsorption capacity measurements, and the structures of the adsorbed Pb-BHA complexes were determined using X-ray photoelectron spectroscopy (XPS) and time-of-flight secondary ion mass spectrometry (TOF-SIMS). The adsorption capacity results showed that more BHA was adsorbed on the scheelite surface in Pb-BHA flotation, and the XPS and TOF-SIMS analysis showed that the species of Pb-BHA complexes adsorbed on the scheelite surface were similar in activation flotation and Pb-BHA flotation. Therefore, the different contents of the complexes on the scheelite surface were responsible for the flotation behavior. XPS and TOF-SIMS showed that BHA combined with lead ions to form complexes with different structures, such as five- and four-membered ring structures. Structure fragment inference based on the measurements indicated that lead ions formed monomer complexes with two BHAs, and that lead hydroxide polymers with a certain degree of polymerization bonded with oxygen atoms in the complexes. The Pb-BHA complexes combine with oxygen atoms on the scheelite surface to form an adsorbate, rendering the surface hydrophobic.

Prognostic value of pre-treatment peripheral blood markers in pancreatic ductal adenocarcinoma and their association with S100A4 expression in tumor tissue.

The aims of the present study were to clarify the prognostic value of peripheral blood variables in patients with pancreatic ductal adenocarcinoma (PDAC), including the neutrophil-to-lymphocyte ratio (NLR), platelet-to-lymphocyte ratio (PLR) and lymphocyte-to-monocyte ratio (LMR), and to determine the association between these variables and S100 calcium-binding protein A4 (S100A4) expression in tumor tissue, which is another prognostic factor for PDAC. Patients with PDAC were recruited at the Tianjin Medical University Cancer Institute and Hospital (Tianjin, China) between December 2008 and December 2014. A retrospective analysis was performed based on the recorded pre-treatment hematological parameters and clinical data. The prognostic value of NLR, PLR and LMR was examined. The association between these variables and S100A4 tissue expression was analyzed. Descriptive statistics and χ2 analyses were used in the present study. The median overall survival (OS) time of patients with PDAC was 9 months (range, 1-32 months). Univariate analysis revealed that NLR, LMR, carbohydrate antigen 19-9, surgery, chemotherapy, stage at diagnosis, tumor grade and age significantly affected OS. Although PLR exhibited no significant effects on OS, NLR and LMR were independent prognostic factors according to the multivariate analysis. Unpaired Student's t-test revealed differences between S100A4 expression and NLR, PLR and LMR. The results of the present study indicated that low NLR and high LMR were associated with a favorable prognosis in patients with PDAC. As a simply obtained and widely available index at diagnosis, NLR and LMR may become a novel predictive and classifying marker for PDAC in the clinical setting.

miR-212 regulated by HIF-1α promotes the progression of pancreatic cancer.

MicroRNA-212 (miR-212) is dysregulated in numerous tissues and cancer types and serves a role in the progression of human cancer. However, the function and mechanism of miR-212 in the development of pancreatic ductal adenocarcinoma (PDAC) remain unknown, particularly in a hypoxic microenvironment. In the present study, miR-212 expression was observed to be significantly upregulated in PDAC tissues compared with normal tissues. Clinical data analysis indicated that miR-212 was positively associated with a large tumor size, Tumor-Node-Metastasis stage, lymph node metastasis and vessel invasion, and influenced the overall survival time. Notably, there was a positive association between the expression of hypoxia-inducible factor-1α (HIF-1α) and miR-212 in vivo and in vitro in hypoxic conditions. Mechanistically, HIF-1α bound directly to a hypoxia response element in the miR-212 promoter region and activated miR-212 expression in PDAC cells. Collectively, these results demonstrated that HIF-1α positively regulated miR-212 expression and resulted in PDAC progression.

Plasma protein profiling in patients undergoing coronary artery bypass grafting surgery and clinical significance.

This study was designed to identify the protein profiling in patients with triple vessel coronary artery disease (CAD) undergoing CABG, in order to detect CAD-related differential proteins in these patients. CABG patients with triple vessel disease with/without left main stenosis (n =160) were compared to normal coronary angiographic subjects (n =160). Plasma samples of 20 males and 20 females in each group were analyzed with iTRAQ technique. ELISA test was used to test the chosen proteins from iTRAQ results in plasma samples from a new cohort of the CABG group (n=120, male/femal=61/59) and control (n =120, male/female=60/60). iTRAQ detected 544 proteins with 35 up-regulated and 41 down-regulated (change fold > 1.2 or < 0.83, p < 0.05). Three proteins including platelet factor 4 (PF4), coagulation factor XIII B chain (F13B), and secreted frizzled-related protein 1 (sFRP1) were selected for validation by using ELISA that demonstrated significant up-regulation of PF4 and sFRP1 (p < 0.05). There was a positive correlation between these proteins and CAD (p < 0.05) and myocardial infarction history (p < 0.05). Thus, we for the first time have found 76 proteins differentially expressed in plasma of CABG patients. The thrombotic disease/inflammation progress-related protein PF4 and sFRP1, a member of the Wnt/fz signal-transduction pathway and related to myocardial repair, are significantly up-regulated in triple-vessel disease with/without left main stenosis. PF4 may be developed as a biomarker for the diagnosis of the severity of CAD requiring CABG procedure.

Recurrent cis-SAGe chimeric RNA, D2HGDH-GAL3ST2, in prostate cancer.

Neighboring genes transcribing in the same direction can form chimeric RNAs via cis-splicing (cis-SAGe). Previously, we reported 16 novel cis-SAGe chimeras in prostate cancer cell lines, and performed in silico validation on 14 pairs of normal and tumor samples from Chinese patients. However, whether these fusions exist in different populations, as well as their clinical implications, remains unclear. To investigate, we developed a bioinformatics pipeline using modified Spliced Transcripts Alignment to a Reference (STAR) to quantify these fusion RNAs simultaneously in silico. From RNA-Seq data of 100 paired normal and prostate cancer samples from TCGA, we find that most fusions are not specific to cancer. However, D2HGDH-GAL3ST2 is more frequently seen in cancer samples, and seems to be enriched in the African American group. Further validation with our own collection as well as from commercial sources did not detect this fusion RNA in 29 normal prostate samples, but in 19 of 93 prostate cancer samples. It is more frequently detected in late stage cancer, suggesting a role in cancer progression. Consistently, silencing this fusion resulted in dramatic reduction of cell proliferation rate and cell motility.

Overexpression of histone demethylase JMJD5 promotes metastasis and indicates a poor prognosis in breast cancer.

In this study, we showed the expression of JMJD5 was increased in breast cancer tissues and breast adenocarcinoma cell lines MCF-7 as well as triple negative breast cancer cell lines MDA-MB-231 compared with paired adjacent normal mammary tissues and normal mammary epithelial cell lines MCF-10A. The higher expression of JMJD5 was significantly corresponded with clinical stage, histological grade and lymph node metastasis. Overexpression of JMJD5 promoted cell invasion and induce EMT, while JMJD5 siRNA inhibits MDA-MB-231 cells invasion in vitro. Moreover, qChIP analysis revealed the Snail family proteins Snai1 was the direct target of JMJD5 in breast cancer cells. Luciferase reporter assays suggested that the overexpression of JMJD5 resulted in the activation of Snail1 promoter-driven luciferase reporter. The changes in the level of RNA and protein implied that the activation of Snail was the important mechanisms by which JMJD5 triggers metastasis. We also detected the higher expression of JMJD5 protein was an independent unfavorable biomarker for worse overall survival in breast cancer patients. Therefore, our results identified an important role for JMJD5 in breast cancer through the regulation of snail1.

Discovery of CTCF-sensitive Cis-spliced fusion RNAs between adjacent genes in human prostate cells.

Genes or their encoded products are not expected to mingle with each other unless in some disease situations. In cancer, a frequent mechanism that can produce gene fusions is chromosomal rearrangement. However, recent discoveries of RNA trans-splicing and cis-splicing between adjacent genes (cis-SAGe) support for other mechanisms in generating fusion RNAs. In our transcriptome analyses of 28 prostate normal and cancer samples, 30% fusion RNAs on average are the transcripts that contain exons belonging to same-strand neighboring genes. These fusion RNAs may be the products of cis-SAGe, which was previously thought to be rare. To validate this finding and to better understand the phenomenon, we used LNCaP, a prostate cell line as a model, and identified 16 additional cis-SAGe events by silencing transcription factor CTCF and paired-end RNA sequencing. About half of the fusions are expressed at a significant level compared to their parental genes. Silencing one of the in-frame fusions resulted in reduced cell motility. Most out-of-frame fusions are likely to function as non-coding RNAs. The majority of the 16 fusions are also detected in other prostate cell lines, as well as in the 14 clinical prostate normal and cancer pairs. By studying the features associated with these fusions, we developed a set of rules: 1) the parental genes are same-strand-neighboring genes; 2) the distance between the genes is within 30kb; 3) the 5' genes are actively transcribing; and 4) the chimeras tend to have the second-to-last exon in the 5' genes joined to the second exon in the 3' genes. We then randomly selected 20 neighboring genes in the genome, and detected four fusion events using these rules in prostate cancer and non-cancerous cells. These results suggest that splicing between neighboring gene transcripts is a rather frequent phenomenon, and it is not a feature unique to cancer cells.

The clinical significance and regulation mechanism of hypoxia-inducible factor-1 and miR-191 expression in pancreatic cancer.

The aim of study was to discuss the correlation and regulatory mechanism of HIF-1 and miR-191 expression in pancreatic tumor. The association between the miR-191 and the clinicopathologic characteristics and the prognosis of pancreatic cancer was further explored. After hypoxic cultured for 6 and 12 h, qRT-PCR and Western blot were practiced to analyze the miR-191 and HIF-1 expression of MIA PaCa-2 and Aspac1 cells. We regulated the HIF-1 expression via plasmid and siRNA transfection to observe the alteration of HIF-1 and miR-191 expression. ChIP sequencing identified the binding sites of HIF-1 and miR-191. Dual luciferase assays were practiced to verify the binding sites. Immunohistochemical staining was practiced to analyze the expression of HIF-1, while qRT-PCR were done for investigating miR-191 in tumor tissues. Then, the association between the expression of them and the clinicopathologic characteristics and prognosis of pancreatic cancer were analyzed. After hypoxic cultured 12 h, the expression of HIF-1 protein, HIF-1mRNA and miR-191 of MIA PaCa-2 and AsPC-1 cells increased significantly (P < 0.05). After HIF-1 overexpressing plasmid transfected to the MIA PaCa-2 and AsPC-1 cells for 48 h, the expression of HIF-1 protein, HIF-1mRNA, and miR-191 upregulated significantly (P < 0.05). While after transfected the MIA PaCa-2 cells by HIF-1 siRNA, the significant decreasing of HIF-1 protein, HIF-1mRNA, and miR-191 expression were observed (P < 0.05). ChIP sequencing showed the protein synthesis of HIF-1 increased in hypoxia situation. Only the HRE5 (-1,169 bp, ChIP4) were significantly brighter in hypoxia in comparing with normoxic cells. In dual luciferase assays, after pGL3-miR-191 and HIF-1 overexpressing plasmid co-transfect the MIAPaCa-2 cells for 48 h, its relative expression of bioluminescence was higher than those co-transfected by mutant miR-191 vectors and HIF-1 overexpressing plasmid or by pGL3-miR-191 and HIF-1 empty plasmid. The expression of miR-191 closely associated with the tumor size, pTNM stage, lymph node metastasis, and perineural invasion (P < 0.05). Patients with higher expression of miR-191 were a risk factor for prognosis of pancreatic cancers. Expression of HIF-1 in pancreatic cancer cells increased under the condition of chronic hypoxia, which may bind to HRE2 in 5'flanking region of miR-191 and initiate transcription of miR-191. Expression of miR-191 was significantly higher in pancreatic tumor tissues. The expression of miR-191 closely associated with the tumor size, pTNM stage, lymph node metastasis and perineural invasion and poor prognosis of pancreatic cancer.

The hypoxia-inducible factor-1 regulates the microRNA185 expression through binding to hypoxia response elements sequence 2.

This study aimed to investigate the interaction and regulatory mechanism of microRNA185 (miR185) and hypoxia-inducible factor-1 (HIF-1) in pancreatic cancer. The significance of miR185 on the clinicopathologic characteristics and prognosis was further explored. qRT-PCR and immunohistochemistry examined miR185 and HIF-1 expression of tumor tissues. Western blot analyzed HIF-1 protein expression. We regulated HIF-1 via transfection to observe the impact of HIF-1 on miR185 expression. ChIP sequencing and dual luciferase identified binding sites of HIF-1 and miR185. MiR185 expression was significantly higher in pancreatic tumors. MiR185 closely associated with tumor size, pTNM stage, lymph node, and perneural invasion. After hypoxic culture, both HIF-1 and miR185 expression of MiaPaCa2 and AsPc1 cells increased significantly. Up- or down-regulating HIF-1 expression via transfection leads to synchronous alteration of miR185. In ChIP sequencing, only the HRE2 (-938 bp) was significantly brighter under hypoxia among four hypoxia response elements' (HREs) sequence of miR185 promoter. After pGL3-miR185 and HIF-1 over-expressing plasmids co-transfect the MiaPaCa2 cells, its relative expression of bioluminescence increased. MiR185 expression was significantly higher in tumor tissues and closely associated with the clinical features of pancreatic cancer. Expression of HIF-1 in pancreatic cancer cells increased in hypoxia. HIF-1 may bind to HRE2 of miR185 and initiate its transcription.

Ascites metabolism measurement enhanced the diagnostic value and accuracy of prognostic evaluation in 18F-FDG PET/CT studies in malignant ascites patients.

OBJECTIVE: This study aimed to evaluate the role of ascites metabolism measurement in (18)F-fluorodeoxyglucose ((18)F-FDG) PET/computed tomography (CT) for auxiliary diagnosis and prognostic evaluation of malignant ascites. MATERIALS AND METHODS: This was a retrospective study on 55 patients, including 36 with malignant ascites and 19 with benign ascites of undetermined origin, before they underwent their first (18)F-FDG PET/CT scan. The χ(2) -test was used to compare the diagnostic efficiencies among (18)F-FDG PET/CT ascites metabolism measurement, tumor localization, and ascites cytology examination. The standard uptake values of ascites and of the normal liver were measured, respectively, and their ratio, denoted as T/NT, was calculated for each patient. The receiver-operating characteristic curve was used to analyze the diagnostic efficiency of the ascites T/NT, ascites carcinoembryonic antigen, ascites CA1(25), and ascites CA(199), and the linear regression was used to analyze the relationship between the ascites T/NT and the survival time of patients. RESULTS: The metabolic level of malignant ascites was high. The sensitivity and accuracy of ascites metabolism measurement were higher than those of ascites cytology examination (χ(2) =6.98, 4.58; all P's<0.05). The specificity of ascites metabolism measurement was higher than that of (18)F-FDG PET/CT tumor localization (χ(2) =5.70, P<0.05). The T/NT value of malignant ascites (0.68 ± 0.17) was higher than that of benign ascites (0.38 ± 0.10) (t=7.21, P<0.05). The area under the receiver-operating characteristic curve of ascites T/NT was larger than those of ascites carcinoembryonic antigen, CA(125), and CA(199). There was a negative correlation between the ascites T/NT and the survival of patients with malignant ascites (r=-0.647, P<0.01). CONCLUSION: Ascites metabolism measurement has an important auxiliary diagnostic value in (18)F-FDG PET/CT for ascites patients. The ascites T/NT may be a good index for prognostic evaluation of malignant ascites.