Effect of hyperbaric oxygen and ulinastatin on plasma endotoxin, soluble CD14, endotoxin-neutralizing capacity and cytokines in acute necrotizing pancreatitis.

BACKGROUND: We sought to study the effect of a combination therapy comprised of hyperbaric oxygen (HBO) and ulinastatin on the plasma levels of endotoxin, soluble CD14 (sCD14), endotoxin neutralizing capacity (ENC) and cytokines in acute necrotizing pancreatitis (ANP) in rats. METHODS: We randomly allocated 90 Sprague-Dawley rats into 6 groups: group 1 (ordinary control), group 2 (sham operation), group 3 (ANP), group 4 (ANP with HBO), group 5 (ANP with ulinastatin) and group 6 (ANP with HBO and ulinastatin). We induced ANP by retrograde injection of 3.5% sodium taurocholate (2.5 mL/kg) via the pancreatic duct. Five minutes after induction, animals in groups 5 and 6 were infused with ulinastatin (20 000 U/kg) via the portal vein. Thirty minutes after induction, animals in groups 4 and 6 received HBO therapy. We collected samples 3, 6 and 10 hours after induction of ANP. RESULTS: We found that the plasma level of endotoxin in group 3 was significantly higher than in group 4 (3, 6 h, both p < 0.001), group 5 (3 h, p < 0.001; 6 h, p = 0.014) and group 6 (both p < 0.001). The level of plasma sCD14 in group 3 was significantly higher than in group 4 (3, 6 h, both p < 0.001), group 5 (3, 6 h, both p = 0.001) and group 6 (3 h, p < 0.001; 6 h, p = 0.001). The plasma endotoxin and sCD14 levels in group 6 were significantly lower than in groups 4 and 5. The plasma ENC level in group 6 was significantly higher than in groups 3, 4 and 5 (p < 0.001). The ENC level in groups 4 and 5 were higher than in group 3, but there was no significant difference. The plasma level of tumour necrosis factor-alpha (TNF-alpha) and IL-6 in group 6 were significantly lower than in groups 3, 4 and 5 (p < 0.001). The TNF-alpha and IL-6 levels in groups 4 and 5 were lower than in group 3, but there was no significant difference. CONCLUSION: The use of an early combination therapy of HBO and ulinastatin was more effective than either therapy alone in the treatment of ANP.

[Effect of mycophenolic acid on maturation, endocytotic capacity and allostimulatory properties of human myeloid dendritic cell].

OBJECTIVE: To study the effect of mycophenolic acid (MPA) on maturation, endocytotic capacity and allostimulatory properties of human myeloid dendritic cell (MDC). METHODS: The peripheral blood mononuclear cells were isolated freshly from healthy volunteers (n = 15). The study group was treated by MPA. Co-stimulatory and adhesion molecular on MDC were analyzed by flow cytometry. After isolation of blood dendritic cell antigen-1(+) (BDCA-1(+)), the cellular uptake of FITC-dextran was measured by flow cytometry. After mixed lymphocyte reaction, the percentage of allogenic CD(4)(+) T cell in G(0) phase was analyzed by flow cytometry. RESULTS: (1) Maturation: Compared to control group, MPA down-regulated the level of CD40, CD62L, HLA-DR, CD54, CD80, CD83 and CD86 on MDC significantly in study group (t = -3.713, P > 0.010). (2) Endocytotic capacity: MPA enhanced significantly the endocytotic capacity of MDC in study group (t = 10.171, P = 0.000). (3) Allostimulatory capacity: Compared to control group, MPA reduced the ability of MDC to stimulate markedly the T cell proliferation in study group. CONCLUSION: MPA may enhance the phagocytic capacity of human peripheral MDC, inhibit the maturity of MDC and impair its allostimulatory capacity of CD(4)(+) T lymphocyte.

Potential of utilization of albumin as a delivery module in cancer model.

PURPOSE: This study compared the potential of 2 nanoparticles, namely albumin and gold nanoparticles, for the specific delivery of 99mTC- resveratrol in colon cancer. The aim of adding radioactive tag (99mTC) to resveratrol was to utilize it for imaging purposes. METHODS: We compared the potential of albumin loaded 99mTC- resveratrol with gold nanoparticles loaded 99mTc-resveratrol for tumor specific delivery. Twenty male Wistar rats were used for the formation of colon cancer model. Both delivery molecules were tested for tumor specific delivery. RESULTS: The results showed efficient delivery of 99mTC-resveratrol with albumin in comparison to gold nanoparticles, as confirmed by single-photon emission computed tomography (SPECT). CONCLUSIONS: Albumin is a superior molecule for the efficient delivery of 99mTC- resveratrol at tumor sites in comparison to gold nanoparticles.

DNA Methylation Biomarkers Predict Objective Responses to PD-1/PD-L1 Inhibition Blockade.

Immune checkpoint inhibitor (ICI) treatment could bring long-lasting clinical benefits to patients with metastatic cancer. However, only a small proportion of patients respond to PD-1/PD-L1 blockade, so predictive biomarkers are needed. Here, based on DNA methylation profiles and the objective response rates (ORRs) of PD-1/PD-L1 inhibition therapy, we identified 269 CpG sites and developed an initial CpG-based model by Lasso to predict ORRs. Notably, as measured by the area under the receiver operating characteristic curve (AUC), our model can produce better performance (AUC = 0.92) than both a model based on tumor mutational burden (TMB) (AUC = 0.77) and a previously reported TMB model (AUC = 0.71). In addition, most CpGs also have additional synergies with TMB, which can achieve a higher prediction accuracy when joined with TMB. Furthermore, we identified CpGs that are associated with TMB at the individual level. DNA methylation modules defined by protein networks, Kyoto Encylopedia of Genes and Genomes (KEGG) pathways, and ligand-receptor gene pairs are also associated with ORRs. This method suggested novel immuno-oncology targets that might be beneficial when combined with PD-1/PD-L1 blockade. Thus, DNA methylation studies might hold great potential for individualized PD1/PD-L1 blockade or combinatory therapy.

[Differential gene expression of liver carcinoma cell after transfection of by ARL-1 with microarray].

OBJECTIVE: To study the difference of the gene expression profile and to identify the different expression after transfection of the ARL-1 gene. METHODS: The cDNA probes were synthesized from total RNA of study group and control group, which was differentially hybridized to cDNA chips and confirmed by a gene specific semiquantitative reverse transcription polymerase chain reaction (RT-PCR). RESULTS: Six kinds of gene expression were increased and 9 kinds of gene expression were decreased. The findings were correlated with protein metabolism, signal pathway, metastasis, and drug resistance. CONCLUSIONS: cDNA chips showed that gene expression profile of liver carcinoma cell was changed after transfection of the ARL-1 gene. It is a useful method in understanding the mechanism of drug resistance.

Impact of fish oil enriched total parenteral nutrition on elderly patients after colorectal cancer surgery.

BACKGROUND: Polyunsaturated omega-3 fatty acids may beneficially influence healing processes and patient outcomes. The aim of this research was to study the clinical efficacy of fish oil enriched total parenteral nutrition in elderly patients after colorectal cancer surgery. METHODS: Fifty-seven elderly patients with colorectal cancer were enrolled in this prospective, randomized, double-blind, controlled clinical trial. All patients received isocaloric and isonitrogenous total parenteral nutrition by continuous infusion (20 - 24 hours per day) for seven days after surgery. The control group (n = 28) received 1.2 g/kg soybean oil per day, whereas the treatment group (n = 29) received 0.2 g/kg fish oil and 1.0 g/kg soybean oil per day. Blood samples were taken pre-operatively, and at days one and eight after the operation. The plasma levels of CD4, CD8, CD4/CD8, interleukin 6 (IL-6) and tumor necrosis factor α (TNF-α) were measured. Clinical outcomes were then analysed. RESULTS: Patient characteristics were comparable between the two groups. At day eight post-surgery, IL-6, TNF-α and CD8 titres were lower in the treatment group when compared to the control group; these results reached statistical significance. In the treatment group, there were fewer infectious complications and incidences of systemic inflammatory response syndrome (SIRS), and shorter lengths of hospital stay were observed. The total cost of medical care was comparable for the two groups. No serious adverse events occurred in either group. CONCLUSIONS: Fish oil 0.2 g/kg per day administrated to elderly patients after colorectal surgery was safe and may shorten the length of hospital stay and improve clinical outcomes.

[Screening of the drug resistance-associated gene in HepG2 cell line transfected with aldose reductase like gene-1 (ARL-1)].

BACKGROUND & OBJECTIVE: Aldose reductase like gene-1 overexpresses in hepatocellular carcinomas; it might be the drug resistance- associated gene to anti-tumor drugs containing carbonyl groups on hepatocellular carcinomas. This study was designed to establish human hepatocellular carcinoma cell line (HepG2) transfected with aldose reductase like gene-1 (ARL-1), then to observe the changes of drug resistance to anti-tumor drugs with carbonyl group and screen the drug resistance-associated genes through cDNA chip. METHODS: PBK/ARL-1 plasmid was transfected to HepG2 cells by liposome to establish HepG2 monoclonal cells with high expression of ARL-1. The HepG2 cells with high expression of ARL-1 were determined by reverse transcription-polymerase chain reaction (RT-PCR),flow cytometric analysis (FCA), and immunohistochemical staining. MTT assay and cell apoptosis analysis were performed to determine the drug resistance ability of the cells to Adriamycin (ADM) and mytomycin (MMC), which contain carbonyl group. 5-fluorouracil (5-FU) that has no carbonyl group was used as control. Then two kinds of cDNA probes were made from HepG2 cells and ARL-HepG2 cells and labeled with Cy3 and Cy5 dyes. The probes were hybridized to the human liver cDNA chip and differentially expressed genes from the two cells were screened. The genes could be involved in drug resistance were confirmed by RT-PCR and Western blot. RESULTS: Compared with HepG2 cells, HepG2 cells transfected with ARL-1 gene have an increase in expression of ARL-1. Drug resistance ability of HepG2 cells transfected with ARL-1 gene to ADM and MMC increased 2.6 and 3.47 folds, respectively (t=6.39, P< 0.05 in ADM group; t=30.06, P< 0.01 in MMC group). Drug resistance to 5-FU had no statistical difference between them (t=0.684,P >0.05 in 5-FU group). 15 genes down-regulated and 9 genes up-regulated after transfected with ARL-1 gene were found by cDNA chip scanned. Some differentially displayed genes such as ubiquitin and MDR confirmed by RT-PCR and Western blot were consistent to the results of cDNA chip. CONCLUSION: The HepG2 cells with high expression of ARL-1 have more drug resistance to anti-tumor drugs with carbonyl group. Up-regulated MDR expression and down-regulated ubiquitin expression may contribute to the drug resistance of HepG2 cells.