Correlation between liver cancer pain and the HIF-1 and VEGF expression levels.

A possible correlation between liver cancer pain and the hypoxia-inducible factor (HIF)-1 and vascular endothelial growth factor (VEGF) expression levels was examined. From January, 2015 to January, 2016, 30 patients suffering from liver cancer with pain, 30 patients with liver cancer without pain and 30 hepatitis patients with pain were enrolled in the study. Pain level was evaluated by visual analogue scale (VAS), the expression levels of HIF-1 and VEGF mRNA were determined by RT-PCR and the expression levels of HIF-1 and VEGF proteins were examined by ELISA. Before intervention, the VAS in the hepatitis group was significantly higher than that of the liver cancer pain group. However, after intervention the VAS in the two groups was reduced. HIF-1 and VEGF mRNA expression levels in the liver cancer pain group were significantly higher than those in the liver cancer group before and after intervention. The expression levels of HIF-1 and VEGF mRNA in the hepatitis group were the lowest. The expression levels of HIF-1 and VEGF mRNA in the liver cancer pain group considerably increased after intervention. The expression levels of HIF-1 and VEGF mRNA in the other two groups showed no changes before or after intervention. Before and after the intervention, VAS in the liver cancer pain group was positively correlated to the expression levels of HIF-1 and VEGF. Thus, pain occurrence and the pain level in liver cancer patients were correlated with the expression levels of HIF-1 and VEGF. As the regular three-step medicine analgesic ladder is ineffective in these cases, verification of HIF-1 and VEGF expression levels may be considered the new target for pain release.

Amelioration of dementia induced by Aß 22-35 through rectal delivery of undecapeptide-hEGF to mouse brain.

A group of growth factors have been shown to play important roles in amelioration of the malfunction of the neurodegenerative diseases. However, the proteins or polypeptides passing across the blood-brain barrier (BBB) to access the brain parenchyma are relatively few so that it hinders the therapies in clinic. Here a genetically reconstructed fusion peptide of human epidermal growth factor (hEGF) with an undecapeptide YGRKKRRQRRR (P11) was used to investigate the permeability between the cell membrane and the BBB via rectal administration. The efficiency to rescue the Aß 22-35-induced dementia in mice was assessed after administration of P11-hEGF per rectal. Our results showed that P11-hEGF permeates across not only the 3T3 cell membrane in vitro, but also the endothelia of vessels after intravenous injection (IV), and the mucosa of the rectum after per rectal administration. Further results showed that the circulating P11-hEGF allowed penetrating through the blood-brain barrier and then getting into the brain manifesting biological responses. In the animal experiments, treatment with P11-hEGF not only ameliorated the dementia induced by Aß 22-35 but also rescued the dementia of the aged mice, no matter how it was administrated (IV or per rectal). These results suggest that the rectal non-invasive delivery of the P11 polypeptide-conjugated growth factor is an efficient way for BBB transduction, thus raises the hope of real therapeutic progress against neurodegenerative diseases.

[Expression of procalcitonin and characterization of antibodies against PCT].

AIM: To construct the expression vectors of procalcitonin (PCT), prepare polyclonal antibodies (pAbs) and monoclonal antibodies (mAbs) against PCT and identify their specific biological activity. METHODS: The recombinant expression plasmids of pGEX-4T-1-PCT and PET-32a-PCT were constructed using thyroid carcinoma cell line (TT cell) cDNA as template. The fusion protein of His-PCT was expressed in E.coli and used as immunogen. The specificity of antiserum against human PCT was characterized by ELISA, Western blot and indirect immunofluorescence. The mAbs against human PCT were identified by Western blot and indirect immunofluorescence. RESULTS: The recombinant expression plasmids of pGEX-4T-1-PCT and PET-32a-PCT were constructed and the fusion protein of His-PCT was expressed and purified. The antiserum against human PCT was prepared and the titer detected by ELISA was 1:256 000. The pAb specifically recognized the recombinant human PCT. Eight hybridoma cell lines secreting specific mAbs against PCT were established. The mAbs recognized the recombinant human PCT and four of them recognized the native PCT of TT cytoplasm in immunofluorescent assay. CONCLUSION: The successful preparation of polyclonal and monoclonal antibodies against human PCT is beneficial to further research into the pathological and physiological functions of PCT in severe bacterial infection and sepsis.