Cloning the prophenoloxidase cDNA and monitoring the expression of proPO mRNA in prawns (Macrobrachium rosenbergii) stimulated in vivo by CpG oligodeoxynucleotides.

The prophenoloxidase (proPO) system forms the basis of the non-specific defence system in crustaceans. The aim of this study was to develop an RT-PCR procedure to determine proPO gene expression. We used several degenerate primers designed from the conserved regions in amino acid sequences of proPOs from other species to clone the possible cDNA(s) from the haemocytes of the prawn Macrobrachium rosenbergii. One DNA fragment, 2016 bp long, was cloned by a combination of reverse transcriptase-polymerase chain reaction (RT-PCR) and rapid amplification of cDNA ends (3'/5'-RACE). This fragment could encode a putative polypeptide with 671 amino acids. Further analysis showed that this putative polypeptide contained six histidine residues and a thiol ester-like motif (GCGWPRHM) just like the structural features of proPOs from the shrimp Penaeus monodon. A partial fragment of the sequence with 934 bp containing three histidine residues and the thiol ester-like motif was used as a target to monitor the activation of the proPO gene by the semi-quantitative RT-PCR analysis. The result showed that the highest level of proPO mRNA was detected at 1h after in vivo injection with 5 microg of CpG oligodeoxynucleotide 2006 per prawn; in the same experiment, the highest PO activity was detected at 6 h after injection. In the control, a continuous and slow elevation of PO activity was observed during the experimental period, but such elevation of proPO mRNA was not observed. From our previous study and the time course of gene expression of proPO and enzymatic activity of PO in this study, it can be concluded that the enhancement effect was through its transcriptional level, its translational level and then its post-translational level sequentially. These results suggest that, both for reliability, sensitivity and for the timing of sampling, the change in proPO mRNA is more useful than the PO activity in monitoring the activation of the proPO non-specific defence system of prawns after treatment with stimulants.

[An experience in nursing an acute lymphocytic leukemia patient with Peripherally Inserted Central Catheter].

The Peripherally Inserted Central Catheter (PICC) is still in its infancy in Taiwan. It has many advantages, including safety, simplicity, and few complications during the insertion process. It is suitable for patients who need long-term parenteral administration of medication or chemotherapy. However, the duration of PICC implantation is around six months, or even up to one year. As a result, it is very important to educate patients to take care of themselves throughout the implantation period in order to prevent complications. This report describes the experience of nursing a forty-four-year old male patient who suffered from acute lymphocytic leukemia and received a PICC implantation while undergoing chemotherapy treatment. As a PICC case manager, the author utilized self-efficacy theory and devoted herself to nursing care, interviews and phone calls to collect valuable information. During the process of PICC implantation, the author analyzed the characteristics of this case and assisted the patient and his primary care giver in choosing a suitable catheter and self-care techniques to achieve minimal complications during implantation. It is hoped that, by sharing in her experience, nursing staff may enhance their ability to assist patients with PICC and to help them to maintain their quality of life.