Impact of therapeutic plasmapheresis on the duration of organ failure in patients with hypertriglyceridemia-associated acute pancreatitis.

BACKGROUND: Plasmapheresis is widely used for severe hypertriglyceridemia-associated acute pancreatitis (HTG-AP) to remove excessive triglycerides from plasma. This study aimed to evaluate whether plasmapheresis could improve the duration of organ failure in HTG-AP patients. METHODS: We analyzed a cohort of patients from a multicenter, prospective, long-running registry (the PERFORM) collecting HTG-AP patients admitted to the study sites within 72 h from the onset of symptoms. This study was based on data collected from November 2020 to March 2023. Patients who had organ failure at enrollment were involved in the analyses. The primary outcome was time to organ failure resolution within 14 days. Multivariable Cox regression model was used to evaluate the association between plasmapheresis and time to organ failure resolution. Directed acyclic graph (DAG) was used to identify potential confounders. RESULTS: A total of 122 HTG-AP patients were included (median [IQR] sequential organ failure assessment (SOFA) score at enrollment, 3.00 [2.00-4.00]). Among the study patients, 46 underwent plasmapheresis, and 76 received medical treatment. The DAG revealed that baseline serum triglyceride, APACHE II score, respiratory failure, cardiovascular failure, and renal failure were potential confounders. After adjusting for the selected confounders, there was no significant difference in time to organ failure resolution between patients undergoing plasmapheresis and those receiving exclusive medical treatment (HR = 1.07; 95%CI 0.68-1.68; P = 0.777). Moreover, the use of plasmapheresis was associated with higher ICU requirements (97.8% [45/46] vs. 65.8% [50/76]; OR, 19.33; 95%CI 2.20 to 169.81; P = 0.008). CONCLUSIONS: In HTG-AP patients with early organ failure, plasmapheresis was not associated with accelerated organ failure resolution compared to medical treatment but may be associated with more ICU admissions. TRIAL REGISTRATION: The PERFORM study was registered in the Chinese Clinical Trial Registry (ChiCTR2000039541). Registered 30 October 2020.

Construction of an enteral nutrition evaluation system for critically ill patients based on the Delphi method.

BACKGROUND: This study aimed to construct an enteral nutrition evaluation system for critically ill patients using the Delphi method to direct the formulation of enteral nutrition support strategies and reduce interruption to enteral feeding. METHODS: We used domestic and foreign databases to obtain and analyze the literature and form "The Whole-Proceeding Enteral Nutrition Evaluation System for Critically Ill Patients." The Delphi method was used to conduct two rounds of expert opinion consultation, combined with the suggestions from the research group to finalize the nutrition evaluation content of the system. RESULTS: After two rounds of expert consultation, a nutrition evaluation system was formed around three dimensions: before the start, during, and after the end of nutritional support. The effective recovery rates of the two rounds of expert consultation were 90.0% (18/20) and 100.0% (18/18), respectively. Authority coefficients were 0.865 and 0.908, while Kendall coordination coefficients were 0.108 ( P < 0.05) and 0.115 ( P < 0.001), respectively. Finally, the full enteral nutrition evaluation system for critically ill patients was constructed based on the Delphi method, including three primary items and seven secondary and 28 tertiary indicators. CONCLUSION: The established "Whole-Proceeding Enteral Nutrition Evaluation System for Critically Ill Patients" has high consistency from expert opinions and reliability, which can provide a practical evaluation tool for the process of enteral nutrition for severe patients.

A retrospective analysis of the clinical efficacy of pancreatic duct stent implantation in the management of acute biliary pancreatitis requiring ERCP.

BACKGROUND: This study aimed to investigate the feasibility, effectiveness, and safety of pancreatic duct stenting in managing acute biliary pancreatitis (ABP) necessitating endoscopic retrograde cholangiopancreatography (ERCP). It further aimed to provide valuable insights for subsequent clinical diagnosis and treatment. METHODS: This research employs an observational retrospective case-control study design, encompassing patients with ABP who underwent ERCP at the hepatobiliary surgery department of the General Hospital of Ningxia Medical University between August 1, 2018, and December 31, 2020. A total of 229 cases were screened based on inclusion and exclusion criteria. Regardless of ABP severity, patients were categorized into the stent group (141) and the non-stent group (88). Changes in blood amylase (Amy), lipase (LIP), leukocyte count (WBC), total bilirubin (TBIL), alanine aminotransferase (ALT), hematocrit (HCT), and creatinine (CR) were compared between the two groups. Moreover, variables such as recovery time for oral feeding, hospitalization duration, hospitalization costs, local complications, systemic complications, and new organ failure were recorded to assess the therapeutic effect of pancreatic duct stenting. RESULTS: No significant differences were observed in gender, age, Acute Physiology and Chronic Health Evaluation (APACHE) II score, ABP severity grade, organ failure (OF), cholangitis, or biliary obstruction between the pancreatic stent and non-stent groups (P > 0.05). There was no significant difference in the incidence of complications related to acute pancreatitis between the two groups (P > 0.05). The median fasting and hospitalization times of patients in the stent group were significantly shorter than those in the non-stent group (P < 0.05). No significant differences between the groups were observed in hospitalization costs and in-hospital mortality (P > 0.05). There were no significant variations in white blood cell (WBC) count, TBIL, ALT, and creatinine (Cr) at admission, 72 h, and in the differences between the two groups (P > 0.05). The levels of Amy at admission and 72 h in the stent group were significantly higher than those in the non-stent group (P < 0.05). The differences in LIP and HCT in the stent group were considerably higher than in the non-stent group (P < 0.05). Although no significant differences were observed in mean Amy and LIP between the two groups (P > 0.05), the mean 72-h HCT in the stent group was 38.39% (95% confidence interval [CI] 37.82%-38.96%) was lower than that in the non-stent group (39.44%, 95% CI 38.70-40.17%) (P < 0.05). CONCLUSION: In the stent group, feeding time and hospital stay were significantly shorter than those in the non-stent group. No significant differences were observed between the two groups in the incidence of complications and mortality. The HCT value decreased more rapidly in the stent group. Early pancreatic stent implantation demonstrated the potential to shorten the eating and hospitalization duration of patients with ABP, facilitating their prompt recovery. TRIAL REGISTRATION: This study was registered as a single-center, retrospective case series (ChiCTR1800019734) at chictr.org.cn.

PABPC1 promotes cell proliferation and metastasis in pancreatic adenocarcinoma by regulating COL12A1 expression.

BACKGROUND: The expression of cytoplasmic poly (A) binding protein-1 (PABPC1) has been reported in multiple cancer types. This protein is known to modulate cancer progression. However, the effects of PABPC1 expression in pancreatic adenocarcinoma (PAAD) have not been investigated. Here, we investigate the regulatory targets and molecular mechanisms of PABPC1 in PAAD. METHODS: PABPC1 and collagen type XII α1 chain (COL12A1) expression in PAAD and their role in tumor prognosis and tumor stage were investigated using The Cancer Genome Atlas database analysis. After silencing PABPC1, messenger RNA sequencing and Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) analyses were performed. The expression of differentially expressed genes (DEGs), cell viability, apoptosis, and cell migration and invasion were explored using reverse transcription-quantitative polymerase chain reaction, Cell Counting Kit-8 assay, flow cytometry assay, and transwell assay, respectively. The relationship between PABPC1 and COL12A1 expression was assessed by Pearson's correlation analysis. The regulatory function of COL12A1 in PABPC1-affected BXPC3 cell behavior was studied after COL12A1 was overexpressed. RESULTS: PABPC1 and COL12A1 expression was upregulated in patients with PAAD and was linked to poor prognosis. Four hundred and seventy-four DEGs were observed in BXPC3 cells after PABPC1 silencing. GO and KEGG analyses revealed that the top 10 DEGs were enriched in cell adhesion pathways. Additionally, PABPC1 silencing inhibited cell viability, migration, and invasion and accelerated apoptosis in BXPC3 cells. PABPC1 silencing increased AZGP1 and ARHGAP30 expression and decreased CAV1 and COL12A1 expression in BXPC3 cells. PABPC1 positively mediated COL12A1 expression, whereas PABPC1 knockdown induced the inhibition of BXPC3 cell proliferation, migration, and invasion. CONCLUSION: The results of this study indicate that PABPC1 may function as a tumor promoter in PAAD, accelerating BXPC3 cell proliferation and metastasis by regulating COL12A1 expression.

Early Plasmapheresis Among Patients With Hypertriglyceridemia-Associated Acute Pancreatitis.

Importance: The incidence of hypertriglyceridemia-associated acute pancreatitis (HTG-AP) is increasing. Plasmapheresis is theoretically effective in removing triglyceride from plasma, but whether it confers clinical benefits is unclear. Objective: To assess the association between plasmapheresis and the incidence and duration of organ failure among patients with HTG-AP. Design, Setting, and Participants: This is an a priori analysis of data from a multicenter, prospective cohort study with patients enrolled from 28 sites across China. Patients with HTG-AP were admitted within 72 hours from the disease onset. The first patient was enrolled on November 7th, 2020, and the last on November 30th, 2021. The follow-up of the 300th patient was completed on January 30th, 2022. Data were analyzed from April to May 2022. Exposures: Receiving plasmapheresis. The choice of triglyceride-lowering therapies was at the discretion of the treating physicians. Main Outcomes and Measures: The primary outcome was organ failure-free days to 14 days of enrollment. Secondary outcomes included other measures for organ failure, intensive care unit (ICU) admission, duration of ICU and hospital stays, incidence of infected pancreatic necrosis, and 60-day mortality. Propensity score matching (PSM) and inverse probability of treatment weighting (IPTW) analyses were used to control potential confounders. Results: Overall, 267 patients with HTG-AP were enrolled (185 [69.3%] were male; median [IQR] age, 37 [31-43] years), among whom 211 underwent conventional medical treatment and 56 underwent plasmapheresis. PSM created 47 pairs of patients with balanced baseline characteristics. In the matched cohort, no difference was detected concerning organ failure-free days between patients undergoing plasmapheresis or not (median [IQR], 12.0 [8.0-14.0] vs 13.0 [8.0-14.0]; P = .94). Moreover, more patients in the plasmapheresis group required ICU admission (44 [93.6%] vs 24 [51.1%]; P < .001). The IPTW results conformed to the results from the PSM analysis. Conclusions and Relevance: In this large multicenter cohort study of patients with HTG-AP, plasmapheresis was commonly used to lower plasma triglyceride. However, after adjusting for confounders, plasmapheresis was not associated with the incidence and duration of organ failure, but with increased ICU requirements.

Distribution analysis of positive and negative pathogenic bacteria in patients with acute pancreatitis and the clinical characteristics and model prediction analysis of positive infection bacteria.

Background: There were bacteria in the early pancreatic juice culture of severe acute pancreatitis (SAP) patients, but during the clinical time, some patients showed more positive bacteria and some patients showed more negative bacteria. Many scholars have different test results, and further clinical research needs to be carried out to clarify this fact. To determine evidence of infection in the early stage of acute pancreatitis (AP) by pancreatic juice bacterial culture and provide a reference for the anti-infective therapy of AP. Methods: Patients with AP who underwent pancreatic juice bacterial culture in the Department of hepatobiliary surgery of the General Hospital of Ningxia Medical University from January 1, 2019 to June 30, 2020were reviewed. Endoscopic retrograde cholangiopancreatography (ERCP) was used to collect pancreatic juice, which was sent to the laboratory for culturing. The clinical data and bacterial culture results of the patients were then recorded and analyzed. According to the results of the pancreatic juice culture, the patients were divided into a positive bacterial culture group (n=64) and a negative bacterial culture group (n=92). It was compared the data results of two groups [age, gender, etiology, acute physiology and chronic health evaluation (APACHE) II score, cultured bacteria, complications, local complications, Balthazar computed tomography (CT) score, inflammatory factors, the use of antibiotics, drug sensitivity analysis results, and the patient's co-infection] and performed multivariate analysis to identify the clinically valuable indicators. Moreover, a receiver operating characteristic (ROC) curve was drawn to predict the model of positive pancreatic juice culture in AP. Results: The patients in the positive bacterial culture group and the negative bacterial culture group had statistically significant differences in gender, age, body mass index (BMI), amylase, white blood cell count and the two groups of patients were comparable. A total of 156 patients were included in the study and pathogenic bacteria were cultured in the pancreatic juice of 64 patients (41.03%) and 94 strains of bacteria were found (Gram-positive bacteria, 38.30%; Gram-negative bacteria, 58.51%; fungi, 3.19%). A history of ERCP and early pancreatic necrosis were independent influencing factors of positive pancreatic juice culture. The incidence of complications, APACHE II, and inflammatory factor levels of patients with positive pancreatic juice bacterial culture were significantly higher than those of negative pancreatic juice bacterial culture (P<0.05). Multivariate regression and the ROC curve of pancreatic infection showed that positive pancreatic and Balthazar CT score >7 on admission were independent risk factors of pancreatic. The area under the ROC curve of patients with later pancreatic infection was 0.863 [95% confidence interval (CI): 0.769-0.957], specificity was 65.30%, sensitivity was 90.50%, and the Youden index was 0.603. Conclusions: Bacterial culturing of pancreatic juice provides evidence of infection in the early stage of AP, which has certain significance for the anti-infective therapy of AP.

Nonalcoholic Fatty Liver Hepatocyte-Derived lncRNA MALAT1 Aggravates Pancreatic Cell Inflammation via the Inhibition of Autophagy by Upregulating YAP.

Background: Acute pancreatitis (AP) is one of the most common gastrointestinal disorders, which causes death with a high mortality rate of about 30%. The study aims to identify whether the nonalcoholic fatty liver disease (NAFLD)-derived lncRNA MALAT1 participates in the inflammation of pancreatic cell and its potential mechanism. Methods: The NAFLD cell model was constructed by treating HepG2 cells with FFA. The in vitro model of acute pancreatitis (AP) was established by the administration of caerulein on AR42J cells. MALAT1 and si-MALAT1 were transfected into pancreatic cells, and then exosomes were collected from the NAFLD cell model and then were cocultured with AR42J cells. Transmission electron microscopy was used to observe the morphology of exosomes. Oil Red O staining was applied to reveal the lipid deposition. The triglyceride, IL-6, and TNF-α levels were detected using ELISA. The MALAT1 level in exosomes was detected by qRT-PCR. The CD9, CD63, CD81, and CYP2E1, LC3II, and LC3I levels were detected by western blot. Results: MALAT1 was upregulated in NAFLD-derived exosomes and increased the levels of IL-6 and TNF-α in pancreatic cells. NAFLD-derived exosomes inhibited YAP phosphorylation, decreased the levels of IL-6 and TNF-α, and reduced the ratio of LC3II/LC3I protein in pancreatic cells. Silencing MALAT1 significantly returned the inhibitory effect of NAFLD on hippo-YAP pathway. YAP1 signal transduction inhibitor CA3 reversed the decrease of LC3II/LC3I expression and the increase of IL-6 and TNF-α levels induced by MALAT1 in the AP cell model. Conclusions: NAFLD-derived MALAT1 exacerbates pancreatic cell inflammation via inhibiting autophagy by upregulating YAP.

Stenting of the pancreatic duct in the early phase of acute pancreatitis: a retrospective study.

BACKGROUND: The effectiveness of pancreatic duct (PD) stenting in the early stages of acute pancreatitis (AP) remains controversial. This study aimed to investigate the efficacy and safety of PD stenting in the early stages of AP. METHODS: This is a retrospective cohort study. The clinical data of 131 patients with AP from 2018 to 2019 were analysed and divided into two groups: the study group (n = 46, PD stenting) and the control group (n = 85, standard treatment). RESULTS: There was a statistically significant reduction in pain relief, oral refeeding, hospitalization, and intensive care unit (ICU) stay in the study group compared with that of the control group (P < 0.05). There were no significant differences in the incidence of complications between the two groups. Further multivariate analysis of risk factors for new-onset organ failure showed that the control group (odds ratio [OR] (95% confidence interval [CI]): 6.533 (1.104-70.181)) and a higher level of haematocrit (HCT) at admission (HCT > 46.1%, OR (95%CI): 8.728 (1.264-116.767)) were independent risk factors. CONCLUSIONS: In the early phase of AP, PD stenting has the potential to reduce pain relief time, oral refeeding time, ICU stay time, and overall hospital stay time. This finding highlights a new route for the treatment of AP.

Early endoscopic transpapillary drainage through the minor papilla in the treatment of acute pancreatitis.

Background: To explore the feasibility, safety, and efficacy of endoscopic transpapillary drainage through the minor papilla in the treatment of acute pancreatitis (AP). Methods: We retrospectively evaluated the safety and efficacy of endoscopic transpapillary drainage via the minor papilla among AP patients who were treated in our hospital from September 2018 to March 2020. Results: The present study included 18 patients (12 males and 6 females). All patients successfully received endoscopic transpapillary drainage via the minor papilla and were discharged upon recovery. No patient died, received ICU treatment, or had endoscopic operation-related complications. Two patients (11.11%) received additional abdominal paracentesis due to local complications. Fifteen patients (83.33%) resumed oral feeding within 3 days. The postoperative 24-hour leukocyte level, APACHE II score, serum amylase level, and lipase level significantly decreased compared with those at admission. The median hospitalization stay was 5 (3.75-9) days. The median hospitalization cost was 25,123.82 (22,942.50-43,874.68) RMB. The patients were followed up at 6-24 months, during which 4 patients (22.22%) had recurrence. Two patients had recurrence after pancreatic duct removal and other 2 patients in the period of carrying ducts. Conclusions: Early endoscopic transpapillary drainage via the minor papilla in cases of difficult cannulation or stenting via the major papilla is safe and effective in the treatment of AP, and is worthy of further popularization.

Treatment of acute pancreatitis with early pancreatic stenting: a case series of 336 patients.

BACKGROUND: Pancreatic duct (PD) obstruction and hypertension may play a central role in the onset and progression of acute pancreatitis (AP). However, only a few studies have reported using pancreatic stenting to relieve PD obstruction in the early phase of AP, with conflicting results. Whether pancreatic stenting is effective in the early phase of AP remains unknown. We conducted this experiment in order to study the therapeutic efficacy and safety of pancreatic stenting in the early stage of AP. METHODS: We conducted a retrospective analysis of 336 AP patients from 2011 to 2018 who underwent pancreatic stenting within 48 hours of admission. RESULTS: A total of 330 (98.2%) patients underwent successful pancreatic stenting, of whom 23 (7.0%) had severe AP, 178 (53.9%) had moderately severe AP, and 129 (39.1%) had mild AP. Visible PD obstructive material was observed in 94 (28.5%) patients. The mean oral refeeding time since admission and length of hospital stay were 3.5±2.7 and 7.4±6.7 days, respectively. Procedure-related adverse events, in-hospital mortality, and local complication rates were 0.3%, 0.3%, and 7.6%, respectively. CONCLUSIONS: Early endoscopic pancreatic stenting in AP patients effectively shortened the fasting time and length of hospital stay and did not increase the risk of adverse events, death, or local complications. A further prospective randomized controlled clinical trial is currently underway to validate the safety and efficacy of this procedure.

miR­32­5p suppresses the proliferation and migration of pancreatic adenocarcinoma cells by targeting TLDC1.

Pancreatic adenocarcinoma (PAAD) is one of the most fatal types of cancer in humans. However, the molecular mechanisms underlying the migration and invasion abilities of PAAD cells remain unclear. The aim of the present study was to explore the regulatory roles of microRNA (miR)­32­5p in PAAD cells. miR­32­5p mimic and inhibitor were used to transfect the human PAAD AsPC­1 cell line to determine the role of miR­32­5p in cell proliferation and metastasis. The starBase database predicted the binding of miR­32­5p to the target gene TBC/LysM­associated domain containing 1 (TLDC1). Further analyses were performed to assess miR­32­5p and TLDC1 expression levels in healthy and PAAD tissues, as well as the association between miR­32­5p or TLDC1 expression and the prognosis of patients with PAAD. The interaction between miR­32­5p and TLDC1 was verified using the dual­luciferase reporter assay. miR­32­5p and TLDC1 expression levels were detected by reverse transcription­quantitative PCR and western blotting, respectively. The Cell Counting Kit­8 assay was utilised to assess cell proliferation, whereas the wound­healing and Transwell assays were conducted to assess cell migration and invasion, respectively. miR­32­5p expression levels were markedly lower in PAAD tissue compared with those in healthy tissue, and were significantly lower in PAAD cell lines compared with those in the human pancreatic duct cell line HPDE6, which corresponded with poor prognosis. miR­32­5p significantly inhibited the proliferation of PAAD cells and markedly reduced migration and invasion compared with the negative controls. miR­32­5p was shown to target TLDC1, with miR­32­5p expression in PAAD being negatively correlated with TLDC1 expression. High TLDC1 expression levels were associated with a poorer prognosis compared with low TLDC1 expression levels. Co­transfection of miR­32­5p mimic and pcDNA/TLDC1 demonstrated that TLDC1 significantly reversed miR­32­5p­mediated inhibition of the proliferation, migration and invasion of PAAD cells. Overall, the present study demonstrated that miR­32­5p may serve as a tumor­suppressor gene by inhibiting the proliferation and migration and invasion of PAAD cells via the downregulation of TLDC1. Therefore, miR­32­5p may serve as a potential diagnostic or prognostic marker for PAAD.

Treatment of severe acute pancreatitis via endoscopic pancreatic stenting and nasopancreatic drainage: Case reports.

Severe acute pancreatitis (SAP) is associated with high mortality. SAP is generally treated by conservative management at the early phase, and removal of the pancreatic and peripancreatic necrotic tissue at the late phase. However, studies have suggested that the surgical treatment of SAP should focus on pressure reduction and drainage. In this case report, 3 SAP patients of 44, 30 and 60 years of age were treated at the General Hospital of Ningxia Medical University. They underwent emergency endoscopic pancreatic stenting at the early phase and nasopancreatic drainage at the late phase when peripancreatic encapsulated effusion was observed. All patients were successfully treated and discharged from the hospital. The disease duration of the patients was 71, 58, and 88 days, respectively. Our cases suggested that the surgical strategy of endoscopic pancreatic stenting at the early phase and nasopancreatic drainage at the late phase is promising for the treatment of SAP.

Alisertib induces cell cycle arrest and autophagy and suppresses epithelial-to-mesenchymal transition involving PI3K/Akt/mTOR and sirtuin 1-mediated signaling pathways in human pancreatic cancer cells.

Pancreatic cancer is the most aggressive cancer worldwide with poor response to current therapeutics. Alisertib (ALS), a potent and selective Aurora kinase A inhibitor, exhibits potent anticancer effects in preclinical and clinical studies; however, the effect and underlying mechanism of ALS in the pancreatic cancer treatment remain elusive. This study aimed to examine the effects of ALS on cell growth, autophagy, and epithelial-to-mesenchymal transition (EMT) and to delineate the possible molecular mechanisms in human pancreatic cancer PANC-1 and BxPC-3 cells. The results showed that ALS exerted potent cell growth inhibitory, pro-autophagic, and EMT-suppressing effects in PANC-1 and BxPC-3 cells. ALS remarkably arrested PANC-1 and BxPC-3 cells in G2/M phase via regulating the expression of cyclin-dependent kinases 1 and 2, cyclin B1, cyclin D1, p21 Waf1/Cip1, p27 Kip1, and p53. ALS concentration-dependently induced autophagy in PANC-1 and BxPC-3 cells, which may be attributed to the inhibition of phosphatidylinositol 3-kinase (PI3K)/protein kinase B (Akt)/mammalian target of rapamycin (mTOR), p38 mitogen-activated protein kinase (p38 MAPK), and extracellular signal-regulated kinases 1 and 2 (Erk1/2) but activation of 5'-AMP-dependent kinase signaling pathways. ALS significantly inhibited EMT in PANC-1 and BxPC-3 cells with an increase in the expression of E-cadherin and a decrease in N-cadherin. In addition, ALS suppressed the expression of sirtuin 1 (Sirt1) and pre-B cell colony-enhancing factor/visfatin in both cell lines with a rise in the level of acetylated p53. These findings show that ALS induces cell cycle arrest and promotes autophagic cell death but inhibits EMT in pancreatic cancer cells with the involvement of PI3K/Akt/mTOR, p38 MAPK, Erk1/2, and Sirt1-mediated signaling pathways. Taken together, ALS may represent a promising anticancer drug for pancreatic cancer treatment. More studies are warranted to investigate other molecular targets and mechanisms and verify the efficacy and safety of ALS in the treatment of pancreatic cancer.

miR-34c-3p inhibits cell proliferation, migration and invasion of hepatocellular carcinoma by targeting MARCKS.

Recent studies have shown that microRNA-34c-3p (miR-34c-3p) is down-regulated in various types of cancers and involved in tumor growth, invasion and metastasis. However, the roles of miR-34c-3p in hepatocellular carcinoma (HCC) are poorly understood. In this study, the expression profile of miR-34c-3pin HCC tissues and cell lines were examined by quantitative real-time polymerase chain reaction (qRT-PCR). The correlations of miR-34c-3p expression and clinicopathological characteristics were analyzed. The biological role of MiR-34c-3pin cell proliferation, migration and invasion was examined. In addition, the targets of miR-34c-3p were identified. The results showed that miR-34c-3p expression was significantly down-regulated in HCC tissues and cell lines; low expression level of miR-34c-3p was correlated with vascular invasion and advanced TNM stage. In vitro functional assays showed that overexpression of miR-34c-3pin HepG2 and Huh7 cells significantly reduced cell proliferation, migration and invasion. Furthermore, target analysis and luciferase assay identified myristoylated alanine-rich protein kinase c substrate (MARCKS) as a specific target of miR-34c-3p. Knockdown of MARCKS in HepG2 cells reduced cell migration and invasion, but not cell proliferation. Taken together, our findings implicate the potential application of miR-34c-3p as a tumor suppressor in cancer therapy.

Effect of the GLP-1 analog exendin-4 and oxaliplatin on intrahepatic cholangiocarcinoma cell line and mouse model.

The influence of Glucagon-like peptide-1 (GLP-1) and Exendin-4 on development of intrahepatic cholangiocarcinoma (ICC) is evaluated in the study. In vitro tests, including acute toxicity test, cell colony formation assays, cells proliferation and apoptosis, transwell assay, were performed. An ICC in situ tumor animal model was established. Then, animals were randomly divided into four groups (n = 6): control, Exendin-4 treatment, oxaliplatin treatment and Exendin-4-oxaliplatin treatment. Animals in the Exendin-4 treatment and Exendin-4-oxaliplatin treatment groups received a subcutaneous injection of Exendin-4 (100 µg/kg/day) for 1 week, and then received oxaliplatin (10 mg/kg/week) by tail vein injection. Animals in the control group received PBS. Immunohistochemistry tests were used for PCNA, Ki67, Caspase 3 expression in tumor tissue. Results show that that, after incubation of human cholangiocarcinoma cell lines, HuCCTI and GLP-1, or HuCCTI and Exendin-4, colony formation number was sharply decreased. However, GLP-1, HuCCTI or Exendin-4 did not affect the colony of normal cells. Combination treatment with oxaliplatin and Exendin-4 can significantly inhibit tumor cells' proliferation and promote apoptosis. The combined effect is stronger than that of oxaliplatin or Exendin-4. Combination treatment with oxaliplatin and Exendin4 can significantly decrease Ki67 and PCNA proteins' expression in subcutaneous tumors of nude mice. The inhibitory effect of Combination treatment with oxaliplatin and Exendin4 is clearly stronger than that of oxaliplatin. In addition, Combination treatment with oxaliplatin and Exendin4 can significantly increase Caspase3 protein positive expression. In short, these results show that combination treatment with oxaliplatin and Exendin4 can inhibit tumor cells' proliferation, and promote apoptosis.

Evaluation of antitumour activity of tea carbohydrate polymers in hepatocellular carcinoma animals.

Box-Behnken design criterion was applied to identify the significant effects of various extraction parameters such as temperature, time, and solvent-solid ratio on extraction of tea carbohydrate. Among the three variables tested extraction temperature, and solvent-solid ratio were found to have significant effect on tea carbohydrate extraction. The most suitable condition for extraction of tea carbohydrate was found to be a single step extraction at extraction temperature 90°C, extraction time 30 min, and solvent-solid ratio 5:1. At these optimum extraction parameters, the maximum yield of tea carbohydrate obtained experimentally was found to be very close to its predicted value of 3.47% dry weight of root. Then, we have studied the influence of tea carbohydrate on biochemical parameters in hepatocellular carcinoma (HCC) animals. Hepatocellular carcinoma was induced by the injection of 1×10(5) H22 hepatocarcinoma cells into right hind thigh muscle in experimental animals. Tea carbohydrate could inhibit tumour growth and decrease microvessel density in tumour tissue. The altered amount of serum white blood cells (WBC), Interferon-gamma (IFN-γ) and tumor necrosis factor-alpha (TNF-α) in HCC animals were dose-dependently increased, whereas activities of serum alanine transaminase (ALT), aspartate transaminase (AST) and alkaline phosphatase (ALP) were dose-dependently decreased in the drug treated animals. In addition, tea carbohydrate administration could decrease expression of vascular endothelial growth factor (VEGF) and proliferating cell nuclear antigen (PCNA) in H22 tumor tissue. It can be concluded that tea carbohydrate displayed strong antitumour activity in animals.

Benford's law and number selection in fixed-odds numbers game.

In fixed-odds numbers games, the prizes and the odds of winning are known at the time of placement of the wager. Both players and operators are subject to the vagaries of luck in such games. Most game operators limit their liability exposure by imposing a sales limit on the bets received for each bet type, at the risk of losing the rejected bets to the underground operators. This raises a question--how should the game operator set the appropriate sales limit? We argue that the choice of the sales limit is intimately related to the ways players select numbers to bet on in the games. There are ample empirical evidences suggesting that players do not choose all numbers with equal probability, but have a tendency to bet on (small) numbers that are closely related to events around them (e.g., birth dates, addresses, etc.). To the best of our knowledge, this is the first paper to quantify this phenomenon and examine its relation to the classical Benford's law. We use this connection to develop a choice model, and propose a method to set the appropriate sales limit in these games.