Postobstructive pulmonary edema that developed immediately after the removal of an endobronchial foreign body.

The patient was a 5-year-old boy who was transported to our hospital for a paroxysmal cough, disturbance of consciousness, tonic-clonic convulsions and labored breathing. The patient's respiratory failure persisted after the convulsions remitted, and the presence of an endobronchial foreign body was suspected based on the findings of chest CT performed the following day. A peanut was subsequently removed from the right main bronchus using a bronchoscope with tracheal intubation and bag valve mask ventilation. Immediately after removal, the patient rapidly developed exacerbated hypoxemia, and a reduction in right lung lucency was noted on chest radiography. He was therefore diagnosed with type II postobstructive pulmonary edema, and his condition improved within a short period of time.

Specific Echocardiographic Findings Useful for the Diagnosis of Common Pulmonary Vein Atresia.

In this paper, we report a case of common pulmonary vein atresia, which is a very rare disease characterized by cyanosis, heart failure and pulmonary hypertension. Reverse flow in the pulmonary artery at end-diastole as well as in the isthmus of the aorta from early systole to end-diastole detected by echocardiography were found to be specific features useful in diagnosing the disease.

Traumatic dislocation of the hip in a child caused by trivial force for age.

Traumatic hip dislocation in children has a relatively rare occurrence. There are some residual complications, such as avascular necrosis of the femoral head, growth disturbance caused by premature fusion, neurological injury, recurrent dislocation, and posttraumatic arthritis. There is no consensus in the literature about the period of non-weight bearing after reduction. A rare case of a 13-year-old boy of hip dislocation caused by trivial force for age is reported followed by review of the pediatric literatures with treatment recommendation.

[Prognostic study of preoperative serum levels of CEA and CA 19-9 in colorectal cancer].

PURPOSE: Carcinoembrionic Antigen (CEA) and carbohydrate antigen 19-9 (CA 19-9) are the most frequently used tumor markers in the clinical setting of colorectal cancer. The aim of this study is to evaluate the prognostic value of preoperative serum levels of CEA and CA 19-9 in colorectal cancer patients. METHODS: Serum levels of CEA and CA 19-9 were examined in 586 patients with colorectal cancer. Cut-off levels were calculated at reference value:<2.5 ng/mL (group A) versus >2.5 ng/mL (group B) for CEA and, <37 U/mL (group A) versus >37 U/mL (group B) for CA 19-9. RESULTS: According to tumor progression, each marker tended to show a higher level. Group A showed a significantly better prognosis than group B in both CEA and CA 19-9. In Dukes classification A, B and C, only CEA showed a better prognosis in group A than group B. At the time of recurrence compared to the pre-operative point, the CEA and CA 19-9 levels were significantly higher in both group A and B, however. In relation to the necessity of adjuvant chemotherapy (5-FU containing regimen) in Dukes A, the cases without adjuvant chemotherapy in group B of CEA showed a poor prognosis. CONCLUSION: The measurement of preoperative serum CEA and CA 19-9 is useful for prognostic prediction in colorectal cancer. Cut-off levels calculated at the reference value reflect the prognosis in this study. Especially, preoperative CEA reveals a potential high risk group in Dukes A which should be carefully treated by adjuvant chemotherapy to avoid recurrence.

Design of PCR-amplified DNA fragments for in vivo gene delivery: size-dependency on stability and transgene expression.

PCR-amplified DNA fragments can be more efficient and safer vectors than conventional plasmid DNA because of their smaller size and fewer numbers of immunostimulatory cytosine-phosphate-guanine (CpG) motifs. In the present study, the expression unit of plasmid DNA encoding farnesylated enhanced green fluorescent protein (EGFPF; pEGFP-F) or firefly luciferase (pLuc) was amplified by polymerase chain reaction (PCR) to obtain DNA fragments (EGFPF-mini, Luc-mini). EGFPF-mini was as effective as pEGFP-F on the basis of the number of EGFPF-expressing cells after intravenous injection into mice by the hydrodynamics-based procedure. Then, the effects of the length of DNA fragments on transgene expression were examined using luciferase-expressing DNA preparations. Luc-mini preparations showed high levels of luciferase activity in cultured cells as well as in mouse liver, even although the levels did not exceed that of pLuc. An elongation of the DNA fragment on either side of the minimal expression unit was effective in increasing the transgene expression and the stability against nucleases. PCR-amplified DNA fragments showed a sustained luciferase activity in mouse liver compared with pLuc, indicating that they are effective in achieving a prolonged expression. Their stabilization against nucleases will further increase the potential of such short, structure-controlled and synthetic DNA fragments for in vivo gene delivery.

Hydrodynamics-based procedure involves transient hyperpermeability in the hepatic cellular membrane: implication of a nonspecific process in efficient intracellular gene delivery.

BACKGROUND: The mechanisms underlying the efficient gene transfer by a large-volume and high-speed intravenous injection of naked plasmid DNA (pDNA), a so-called hydrodynamics-based procedure, remain unclear and require further investigation. In this report, we have investigated possible mechanisms for the intracellular transport of naked pDNA by this procedure. METHODS: Propidium iodide (PI), a fluorescent indicator for cell membrane integrity, and luciferase- or green fluorescent protein (GFP)-expressing pDNA were injected into mice by the hydrodynamics-based procedure. RESULTS: PI was efficiently taken up by hepatocytes which appeared to be viable following the hydrodynamics-based procedure. Pre-expressed GFP in the cytosol was rapidly eliminated from the hepatocytes by a large-volume injection of saline. The profiles of plasma ALT and AST showed a steady decline with the highest values observed immediately after the hydrodynamics-based procedure. These results suggest that the hydrodynamics-based procedure produces a transient increase in the permeability of the cell membrane. The cellular uptake process appeared nonspecific, since simultaneous injection of an excess of empty vector did not affect the transgene expression. Sequential injections of a large volume of pDNA-free saline followed by naked pDNA in a normal volume revealed that the increase in membrane permeability was transient, with a return to normal conditions within 30 min. Transgene expression was observed in hepatocyte cultures isolated 10 min after pDNA delivery and in the liver as early as 10 min after luciferase-expressing RNA delivery, indicating that pDNA delivered immediately by the hydrodynamics-based procedure has the potential to produce successful transgene expression. CONCLUSIONS: These findings suggest that the mechanism for the hydrodynamics-based gene transfer would involve in part the direct cytosolic delivery of pDNA through the cell membrane due to transiently increased permeability.

Hepatic delivery of particulates in the submicron range by a hydrodynamics-based procedure: implications for particulate gene delivery systems.

BACKGROUND: A large-volume intravenous (i.v.) injection of DNA, i.e. a hydrodynamics-based transfection procedure, is known to be an efficient and liver-specific method of in vivo gene delivery. However, little is available on an applicable particle size in the procedure. METHODS: We examined the effect of particle size on the hepatic delivery by the hydrodynamics-based procedure, using fluorescein isothiocyanate labeled polystyrene microspheres (MS) of 50, 200 or 500 nm in diameter. MS were injected intravenously to mice by a conventional (normal) or the hydrodynamics-based procedure and their degree of hepatic uptake was determined fluorometrically. RESULTS: For all sizes tested, the two procedures were similar in terms of the apparent degree of hepatic uptake, whereas the intrahepatic localization of MS was apparently different between the procedures as shown by an examination of frozen tissue sections. In mice with gadolinium chloride induced Kupffer cell blockade, the hepatic uptake of MS following the normal procedure was decreased while that of the hydrodynamics-based procedure was less affected. This phenomenon of enhanced hepatic delivery seemed to be more effective for larger particles. Confocal microscopic observation of hepatocyte suspensions indicated that part of the injected MS-50 was delivered intracellularly following the hydrodynamics-based procedure, whereas almost all the observed MS-200 and MS-500 were detected in the extracellular compartment or on the surface of the cells. This was supported by the fact that most of the injected MS existed pericellularly around the transgene-expressing cells. CONCLUSIONS: The hydrodynamics-based procedure facilitated extravasation and hepatic delivery of MS. Larger MS were more efficiently extravasated and trapped by the liver, whereas intracellular delivery hardly occurred with them.

Vector-based in vivo RNA interference: dose- and time-dependent suppression of transgene expression.

RNA interference (RNAi) induced by delivery of a small-interfering RNA (siRNA)-expressing vector was characterized in mice. siRNA-expressing plasmid DNA (pDNA) was injected by a hydrodynamics-based procedure along with pDNA encoding an exogenous target luciferase gene. A comparative study showed that stem-loop-type siRNA-expressing pDNA was superior, in terms of the transgene suppressive efficacy, to the tandem-type in the liver following systemic delivery of these pDNAs. Transgene suppression occurred in the liver, kidney, and lung as well as muscle. The degree of suppression was dependent on the dose of siRNA-expressing pDNA and the time at which transgene expression was determined following simultaneous injection of siRNA-expressing and target pDNAs. A reduction in transgene expression became apparent at 1 day after injection, whereas a lower degree of inhibition was obtained before this, as early as 6 h even in mice treated with an excess of siRNA-expressing pDNA. These results suggest that delivery of siRNA-expressing pDNA requires a period of time for induction of RNAi. A study of sequential injections revealed that prior injection of siRNA-expressing pDNA produced a significant suppression for at least 1 day, which disappeared within 4 days. Confocal microscopic studies indicated that the localization of the cells with successful delivery of transgene was different between primary and secondary hydrodynamics-based injections, accounting for the less effective inhibition following the sequential injections. Taken together, these results demonstrate that vector-based in vivo RNAi is a dose- and time-dependent process and offers the possibility of suppressing endogenous targets in a variety of somatic cells.