In vivo electroporation has emerged as a leading technology for developing nonviral gene therapies, and the various technical parameters governing electroporation efficiency have been optimized by both theoretical and experimental analysis. However, most electroporation parameters focused on the electric conditions and the preferred vehicle for plasmid DNA injections has been normal saline. We hypothesized that salts in vehicle for plasmid DNA must affect the efficiency of DNA transfer because cations would alter ionic atmosphere, ionic strength, and conductivity of their medium. Here, we show that half saline (71 mM) is an optimal vehicle for in vivo electroporation of naked DNA in skeletal muscle. With various salt concentrations, two reporter genes, luciferase and beta-galactosidase were injected intramuscularly under our optimal electric condition (125 V/cm, 4 pulses x 2 times, 50 ms, 1 Hz). Exact salt concentrations of DNA vehicle were measured by the inductively coupled plasma-atomic emission spectrometer (ICP-AES) and the conductivity change in the tissue induced by the salt in the medium was measured by Low-Frequency (LF) Impedance Analyzer. Luciferase expression in-creased as cation concentration of vehicle dec-reased and this result can be visualized by X-Gal staining. However, at lower salt concentration, transfection efficiency was diminished because the hypoosmotic stress and electrical injury by low conductivity induced myofiber damage. At optimal salt concentration (71 mM), we observed a 3-fold average increase in luciferase expression in comparison with the normal saline condition (p < 0.01). These results provide a valuable experimental parameter for in vivo gene therapy mediated by electroporation.
Animals ; Comparative Study ; DNA/*administration & dosage/metabolism ; Drug Delivery Systems ; Electric Conductivity ; Electroporation/methods ; Escherichia coli/genetics ; Female ; Gene Therapy/*methods ; *Gene Transfer Techniques ; Genes, Reporter ; Injections, Intramuscular ; Luciferase/metabolism ; Mice ; Mice, Inbred BALB C ; Muscle, Skeletal/drug effects/*metabolism/pathology ; Osmolar Concentration ; Plasmids/genetics/*metabolism ; Sodium Chloride/*pharmacology ; Transfection ; Vehicles/*administration & dosage ; beta-Galactosidase/metabolism

In vivo electroporation has emerged as a leading technology for developing nonviral gene therapies, and the various technical parameters governing electroporation efficiency have been optimized by both theoretical and experimental analysis. However, most electroporation parameters focused on the electric conditions and the preferred vehicle for plasmid DNA injections has been normal saline. We hypothesized that salts in vehicle for plasmid DNA must affect the efficiency of DNA transfer because cations would alter ionic atmosphere, ionic strength, and conductivity of their medium. Here, we show that half saline (71 mM) is an optimal vehicle for in vivo electroporation of naked DNA in skeletal muscle. With various salt concentrations, two reporter genes, luciferase and beta-galactosidase were injected intramuscularly under our optimal electric condition (125 V/cm, 4 pulses x 2 times, 50 ms, 1 Hz). Exact salt concentrations of DNA vehicle were measured by the inductively coupled plasma-atomic emission spectrometer (ICP-AES) and the conductivity change in the tissue induced by the salt in the medium was measured by Low-Frequency (LF) Impedance Analyzer. Luciferase expression in-creased as cation concentration of vehicle dec-reased and this result can be visualized by X-Gal staining. However, at lower salt concentration, transfection efficiency was diminished because the hypoosmotic stress and electrical injury by low conductivity induced myofiber damage. At optimal salt concentration (71 mM), we observed a 3-fold average increase in luciferase expression in comparison with the normal saline condition (p < 0.01). These results provide a valuable experimental parameter for in vivo gene therapy mediated by electroporation.
Animals ; Comparative Study ; DNA/*administration & dosage/metabolism ; Drug Delivery Systems ; Electric Conductivity ; Electroporation/methods ; Escherichia coli/genetics ; Female ; Gene Therapy/*methods ; *Gene Transfer Techniques ; Genes, Reporter ; Injections, Intramuscular ; Luciferase/metabolism ; Mice ; Mice, Inbred BALB C ; Muscle, Skeletal/drug effects/*metabolism/pathology ; Osmolar Concentration ; Plasmids/genetics/*metabolism ; Sodium Chloride/*pharmacology ; Transfection ; Vehicles/*administration & dosage ; beta-Galactosidase/metabolism

Monoamniotic twin pregnancies are relatively rare, but perinatal mortality is high about 30-70%. The major cause of fetal death is cord accident, congenital anomalies, twin-twin transfusion syndrome, and intrauterine growth restriction. Especially cord entanglement occurs; perinatal mortality is about 50%. So the active management of monoamniotic twins to reduce the complication of cord entanglement is important. This is a case of monoamniotic twin pregnancy with cord entanglement and both fetuses survival by elective cesarean section at 32+4 weeks after medical amnioreduction and intensive fetal surveillance.
Cesarean Section ; Female ; Fetal Death ; Fetus ; Humans ; Perinatal Mortality ; Pregnancy ; Pregnancy, Twin

OBJECTIVE: To evaluate efficacy and safety of local injection of potassium chloride (KCl) and methotrexate (MTX) in the treatment of cornual pregnancy. METHODS: We retrospectively reviewed all cases of cornual pregnancy treated conservatively from 2005 through August 2007. Eight cases were identified and two cases were heterotopic interstitial pregnancy. All cases were managed with local injection of KCl or methotrexate under ultrasound guidance and 2 cases were treated with intra-muscular injection of MTX in combination. After the procedure all cases with cardiac activity was confirmed to be aborted by ultrasound. Also serial follow-up sonographic examination and serum beta-hCG measurement were performed. RESULTS: The mean initial beta-hCG level was 53,331.6 mIU/mL and ranged from 14,332 mIU/mL to 125,721 mIU/mL. Mean gestational age was 48.3 days from 40 to 65 days. All cases were aborted successfully and follow up beta-hCG were declined abruptly. Two cases of heterotopic pregnancy resulted in successful deliveries at full term. CONCLUSION: Cornual resection or hysterectomy should no longer be the first line of treatment for hemodynamically stable patients with cornual pregnancy. They can be successfully treated through local injection of MTX or KCl by ultrasound guidance.
Follow-Up Studies ; Gestational Age ; Humans ; Hysterectomy ; Methotrexate ; Potassium ; Potassium Chloride ; Pregnancy ; Pregnancy, Heterotopic ; Retrospective Studies

PURPOSE: Liver transplantation is the therapy of choice for patients with acute and acute-on-chronic severe liver failure or hepatocellular carcinoma. But a suitable liver is not always available for transplantation due to limited donor numbers. To increase the number of available liver for transplantation, a non-heart-beating donor (NHBD) liver transplant program is started. In NHBD liver transplantation, warm ischemic injury of liver occurs. The duration of warm ischemia is thought to be the most important risk factor for postoperative complications such as primary nonfunction or severe hepatic dysfunction. Recent evidence indicates that hepatocyte growth factor (HGF) plays an important role as a cytoprotector against hepatic injury by anti-apoptotic effect and mitogen in liver regeneration. Therefore studies also were performed to examine whether HGF influenced the viability and regeneration of hepatocytes from rats, subjected to prolonged warm ischemic injury. METHODS: Male Sprague- Dawley rats were subjected to non-heart-beating death by cervical spine fracture. Rats left in room temperature directly after, 30-minutes, 1-hours before surgery and perfusion was performed for isolating hepatocyte. Among three groups, hepatocyte viability was compared by trypan blue stain. And isolated hepatocytes from 30-minutes warm ischemic group were cultured for 24-hours, which were treated with no HGF and addition of various doses (5 ng/mL, 10 ng/mL, 20 ng/ mL, 40 ng/mL, 100 ng/mL) of HGF. Anti-apoptosis and regeneration of hepatocyte were compared by LDH assay, MTS assay, western blot, and immunocyto-chemistry after a 24-hours culture. RESULTS: The results of hepatocyte viability along the prolonged warm ischemic groups in isolated hepatocytes decreased sequentially 74.8+/-12.6%, 45.0+/-5.4%, 37.8+/-10.4% along directly after, 30-minutes, 1-hours in trypan blue stain (P<0.01). And 24-hour-cultured hepatocytes from 30-minutes warm ischemic group were treated with HGF. The results of LDH assay, MTS assay did not have relation with HGF addition. But the results of western blot and immunocytochemistry shown that HGF doses dependent anti-apoptosis and regeneration of hepatocyte increased. That indicates HGF presumably inhibites apoptotic pathway by phosphorylation. And HGF also makes hepatocyte hypertrophy and albumin synthesis. CONCLUSION: HGF was a potent cytoprotector against hepatic injury by anti- apoptotic effect and mitogen of liver regeneration in NHBD liver animal model. HGF facilitates recovery of the liver from prolong warm ischemic injury. If the more clinical studies and large animal studies are performed, NHBD using liver transplantation will be available with more chances by HGF.
Animals ; Blotting, Western ; Carcinoma, Hepatocellular ; Diminazene ; Hepatocyte Growth Factor ; Hepatocytes ; Humans ; Hypertrophy ; Immunohistochemistry ; Liver ; Liver Failure ; Liver Regeneration ; Liver Transplantation ; Male ; Models, Animal ; Perfusion ; Phosphorylation ; Postoperative Complications ; Rats ; Regeneration ; Risk Factors ; Spine ; Tissue Donors ; Transplants ; Trypan Blue ; Warm Ischemia

The vast majority of ampullary tumors are epithelial in nature and are either adenoma or adenocarcinoma. Rarely lymphoma or mucinous and squamous elements may be found. Primary lymphoma of pancreaticobiliary region are exceedingly rare, and mostly nodal. Recently, we have experienced a 29-year-old woman with primary malignant lymphoma of the ampulla of Vater with recurrent acute pancreatitis and jaundice. On duodenoscopy, bulging ampulla with normal overlying duodenal mucosa was observed. Endoscopic sphincterotomy was done and tumor inside the ampulla was exposed. ERCP showed high grade biliary and pancreatic ductal strictures extending from the papillary orifice, with upstream ductal dilatation, respectively. With endoscopic biopsy alone, the tumor was confirmed as B-cell lymphoma histologically.
Adenocarcinoma ; Adenoma ; Adult ; Ampulla of Vater* ; Biopsy ; Cholangiopancreatography, Endoscopic Retrograde ; Constriction, Pathologic ; Dilatation ; Duodenoscopy ; Female ; Humans ; Jaundice ; Lymphoma* ; Lymphoma, B-Cell ; Mucins ; Mucous Membrane ; Pancreatic Ducts ; Pancreatitis ; Sphincterotomy, Endoscopic

PURPOSE: Croup is known to have epidemics in seasonal and biennial trends, and to be strongly associated with epidemics of parainfluenza virus. However, seasonal and annual epidemics of croup have not been clearly reported in Korea. This study aimed to examine the seasonal/annual patterns and etiologies of childhood croup in Korea during a consecutive 6-year period. METHODS: Pediatric croup data were collected from 23 centers in Korea from 1 January 2010 to 31 December 2015. Electronic medical records, including multiplex reverse transcription polymerase chain reaction (RT-PCR) results, demographics and clinical information were cross-sectionally reviewed and analyzed. RESULTS: Overall, 2,598 childhood croup patients requiring hospitalization were identified during the study period. Among them, a total of 927 who underwent RT-PCR were included in the analysis. Males (61.5%) predominated, and most (63.0%) of them were younger than 2 years of age (median, 19 months; interquartile range, 11–31 months). Peak hospitalization occurred in 2010 and 2012 in even-numbered years, and parainfluenza virus (PIV, 39.7%) was the most common cause of childhood croup requiring hospitalization, followed by respiratory syncytial virus (14.9%), human rhinovirus (12.5%), Mycoplasma pneumonaie (10.6%), and human coronavirus (7.3%). CONCLUSION: It is concluded that croup hospitalization has a biennial pattern in even-numbered years. PIV may be the most common cause of childhood croup; however, croup epidemics could be attributed to other viruses.
Child ; Coronavirus ; Croup ; Demography ; Electronic Health Records ; Hospitalization ; Humans ; Korea ; Male ; Mycoplasma ; Paramyxoviridae Infections ; Polymerase Chain Reaction ; Respiratory Syncytial Viruses ; Retrospective Studies ; Reverse Transcription ; Rhinovirus ; Seasons

Purpose: Mycoplasma pneumoniae pneumonia (MP) is a major cause of community-acquired pneumonia (CAP) in children and is associated with extrapulmonary manifestations (EPM). The incidence and risk factors for EPM in children are unknown. Methods: This was a retrospective study involving 65,243 pediatric patients with CAP between 2010 and 2015 at 23 nationwide hospitals in South Korea. Medical records were reviewed to collect information regarding the clinical characteristics, radiological results, and laboratory findings. Logistic regression with multivariate analysis was performed to evaluate the risk factors associated with EPM in MP. Results: The incidence of EPM was 23.9%, including elevation of liver enzymes (18.1%), mucocutaneous manifestations (4.4%), proteinuria (4.1%), cardiovascular and neurological manifestations (0.4%), hematologic manifestations (0.2%), and arthritis (0.2%). Statistical analysis showed that mucocutaneous manifestations significantly increased with elevated alanine aminotransferase (adjusted odds ratio [aOR], 3.623; 95% confidence interval [CI], 1.933-6.790) and atopic sensitization (aOR, 2.973; 95% CI, 1.615–5.475) and decreased with respiratory virus coinfection (aOR, 0.273; 95% CI, 0.084–0.887). Elevated liver enzymes were significantly associated with elevated lactate dehydrogenase (aOR, 3.055; 95% CI, 2.257–4.137), presence of pleural effusion (aOR, 2.635; 95% CI, 1.767–3.930), and proteinuria with respiratory virus coinfection (aOR, 2.245; 95% CI, 1.113–4.527). Conclusion Approximately 24% of pediatric patients with MP had various EPM. As the risk factors associated with each EPM were different, it is necessary to evaluate the various clinical aspects and findings of MP to predict and prepare for the occurrence of EPM.