The outcomes of intensified 5-fluorouracil plus leucovorin chemotherapy for preoperative chemoradiation in rectal cancers

PURPOSE: The clinical benefit of intensified neoadjuvant chemoradiotherapy (CRT) in rectal cancer has not been proved. We investigated clinical outcomes of intensified 5-fluorouracil plus leucovorin (5-FU/LV) chemotherapy.METHODS: We retrospectively analyzed 45 patients with locally advanced rectal adenocarcinoma who underwent neoadjuvant CRT between 2010 and 2015. Intensified group took additional 1 cycle of 5-FU/LV chemotherapy after radiation completion (resting period) before surgery, compared to conventional group.RESULTS: Eighteen patients were in conventional group and 27 were in intensified group. Median follow-up duration was 33.7 months (range, 7.8–75.6 months). Complete response rate was 11.4% (5/45). Twelve patients in conventional group and 16 patients in intensified group achieved downstaging (P=0.435). In aspect of toxicity, anemia and thrombocytopenia tended to be more frequent in intensified group without statistical difference. There was also no difference in survival between two groups.CONCLUSION: The intensified CRT with additional 1 cycle of 5-FU/LV in rectal cancer revealed no clinical benefit compared to conventional regimen. Considering that the adverse event was minimal and generally acceptable, further research with additional cycles of 5-FU/LV is needed to prove a real benefit of intensified CRT.

Increase in Anti-Gal IgM Level is Associated With Early Graft Failure in Intraportal Porcine Islet Xenotransplantation

BACKGROUND: Anti-Gal is a major antibody induced in non-human primates (NHPs) after xenotransplantation. To understand the mechanism of graft rejection, we investigated the association between anti-Gal responses and graft failure in NHP recipients of porcine islet transplantation (PITx). METHODS: Intraportal PITx was performed in 35 diabetic NHPs, and graft function was monitored. Early graft failure (EGF) was defined as loss of graft function within a month after PITx. Seven, 19, nine NHPs received immunosuppression (IS) without CD40 pathway blockade (Group I), with anti-CD154 (Group II), and with anti-CD40 (Group III), respectively. The anti-Gal levels on day 0 and day 7 of PITx were measured by ELISA. RESULTS: The frequency of EGF was significantly lower in Group II (26.3%) than in Group I (100%, P=0.0012) and Group III (77.8%, P=0.0166). While levels of anti-Gal IgG in Group I and anti-Gal IgM in Group III increased on day 7 compared with day 0 (P=0.0156 and 0.0273), there was no increase in either on day 7 in Group II. The ratio of anti-Gal IgM or IgG level on day 7 to that on day 0 (Ratio7/0) was significantly higher in recipients with EGF than without EGF (P=0.0009 and 0.0027). ROC curve analysis of anti-Gal IgM Ratio7/0 revealed an area under the curve of 0.789 (P=0.0003). CONCLUSIONS: IS with anti-CD154 suppressed anti-Gal responses and prevented EGF in PITx. Anti-Gal IgM Ratio7/0, being associated with EGF, is a predictive marker for EGF.

The Feasibility of Fundal Retraction of the Gallbladder in the Single Port Laparoscopic Cholecystectomy: Comparison between a 4-instrument Fundal Retraction Group and a 3-instrument Group

PURPOSE: Owing to the accumulation of surgical experience, the indications of single port laparoscopic cholecystectomy (SLC) have increased. To overcome the difficulties and limitations of SLC, we included an additional instrument for use in retracting the gallbladder fundus. The aim of this study was to investigate the feasibility of 4-instrument fundal retraction SLC. METHODS: We retrospectively analyzed 134 patients who had undergone SLC for benign gallbladder disease. We compared the clinical outcome between patients who had undergone SLC without fundal retraction (3-instrument SLC, n=102) and those who had undergone SLC with fundal retraction (4-instrument fundal retraction SLC, n=32). RESULTS: Of 134 patients, 47 were male and 87 were female. A significantly higher proportion of patients in the 4-instrument fundal retraction group had gallbladder distention and wall thickening than patients in the 3-instrument SLC group. No statistically significant difference in the incidence of pericholecystic inflammation, adhesion, and gallbladder perforation; duration of operation, the incidence of complications, and duration of postoperative hospital stay was observed between the two groups. In univariate analysis to perform 4-instrument fundal retraction SLC, higher BMI, the presence of gallbladder distension, and wall thickening were significant factors. In multivariate analysis, gallbladder distention and the presence of concurrent operation during SLC were independently significant factors for performing 4-instrument fundal retraction SLC. CONCLUSION: Four-instrument fundal retraction SLC is a feasible and safe surgical procedure, particularly in patients with a high BMI, gallbladder distention, wall thickening, inflammation, or adhesions. If difficulties are encountered during 3-instrument SLC, simple fundal retraction using an additional instrument may be the preferred option prior to converting the operation to conventional laparoscopic cholecystectomy.

Erratum: Correction of the Grant Number

We found an error in our published article.

Application of the Multiplex Cytokine Analysis to Monitor Xenogeneic Immune Responses to the Porcine Islet Graft in Non-Human Primate

Non-human primate studies must be conducted prior to the clinical trial of xenotransplantation. In order to develop clinically applicable immune-modulatory regimen through non-human primate studies, close monitoring of xenogeneic immune responses is required. We adopted multiplex cytokine analysis in assessment of the immune status during the course of pig-to-non-human primate islet transplantation. This study aimed to assess the feasibility of this multiplex cytokine assay in the development of immune-modulatory regimen. Using this assay, we were able to detect different cytokines with a minimal usage of blood samples, and this allowed us to detect various immunological situations in the recipients. Detection of TNF-alpha surge (347.8 pg/mL) guided us to block TNF-alpha in the early phase of transplantation. Supportive information for in vivo efficacy of cytokine neutralizing antibody could be speculated by in vitro neutralization assay (1,250 pg/mL --> 0 pg/mL). In addition, periodic monitoring of cytokines in peripheral blood allowed the detection of the infection episode prior to other routine assays. These benefits of multiplex cytokine assay may be generally applied to other pre-clinical research, which is a prerequisite for clinical trials.

T Cell Immunoglobulin Mucin Domain (TIM)-3 Promoter Activity in a Human Mast Cell Line

T cell immunoglobulin mucin domain (TIM)-3 is an immunomodulatory molecule and upregulated in T cells by several cytokines. TIM-3 also influences mast cell function but its transcriptional regulation in mast cells has not been clarified. Therefore, we examined the transcript level and the promoter activity of TIM-3 in mast cells. The TIM-3 transcript level was assessed by real-time RT-PCR and promoter activity by luciferase reporter assay. TIM-3 mRNA levels were increased in HMC-1, a human mast cell line by TGF-beta1 stimulation but not by stimulation with interferon (IFN)-alpha, IFN-lambda, TNF-alpha, or IL-10. TIM-3 promoter -349~+144 bp region relative to the transcription start site was crucial for the basal and TGF-beta1-induced TIM-3 promoter activities in HMC-1 cells. TIM-3 promoter activity was increased by overexpression of Smad2 and Smad4, downstream molecules of TGF-beta1 signaling. Our results localize TIM-3 promoter activity to the region spanning -349 to +144 bp in resting and TGF-beta1 stimulated mast cells.

Application of Regulatory T Cells in Transplantation Field / 대한이식학회지

The development of immunosuppressant treatments has enabled remarkable progress in the tissue and organ transplantation field by helping to prevent acute graft rejection. However, complications related to transplantation, such as infection by bacteria and viruses, and the occurrence of cancers resulting from prolonged immune suppression are major obstacles to overcome. Therefore, transplantation immunology research efforts should focus on the induction of donor-specific immune tolerance which preserves patient immune competence which promotes infection and cancer surveillance. Additionally, lifelong administration of immunosuppressants should be forgone in preference to short term therapies. In the 1990s, Dr. Shimon Sakaguchi identified the CD4+CD25+ regulatory T cells which develop in the thymus, and demonstrated that these cells play crucial roles in the maintenance of immune self tolerance. Studies which followed proved that these regulatory T cells are important to the control of autoimmune disease and prevention of graft rejection. Regulatory T cells have also been found to induce immune tolerance in rodent models. In this review, we discuss several considerations for the use of regulatory T cell therapy in the clinical transplantation field.

Current Status and Future Perspectives of Xenotransplantation / 대한이식학회지

Xenotransplantation using pigs as the transplant source holds great promise to resolve the severe shortage of human organ donors. Although stem-cell-derived organ and tissue regeneration have a potential to solve this as well for the future, it still remains as very early experimental phase. Likewise, artificial organs and mechanical devices have been simply used for bridge therapy to transplant. Therefore, xenotransplantation might provide the most imminent solution to the scarcity of human organ donors. In the last two decades, major progress has been made in understanding the mechanisms of xenografts rejection, zoonotic infections including porcine endogenous retrovirus (PERV) and production of genetically engineered pigs including alpha1,3-galactosyltransferase-deficient pigs. With these elaborations, it is now on the threshold of first clinical application. Particularly promising first target is porcine pancreatic islet xenotransplantation. Graft survival has been prolonged to almost one year in the non-human primate study and is waiting for the development of relatively non-toxic or clinically applicable immunosuppressive or tolerance-inducing regimens. This review highlights the currently known obstacles to translate xenotransplantation into clinical therapies and the possible strategies to overcome these hurdles, as well as current status and future perspective for clinical xenotransplantaion.

Depletion of Cytoplasmic Tail of UL18 Enhances and Stabilizes the Surface Expression of UL18

BACKGROUND: Human cytomegalovirus UL18, a MHC class I homologue, has been considered a natural killer (NK) cell decoy. It ligates LIR-1/ILT2 (CD85j), an NK inhibitory receptor, to prevent lysis of infected target cells. However, precise role of UL18 to NK cell cytotoxicity is yet elusive. Difficulty in clarifying the function of UL18 lies in complication in detecting UL18 mainly due to low level expression of UL18 on the surface and gradual loss of its expression. METHODS: To overcome this hurdle, cDNA of cytoplasmic tail-less UL18 was constructed and expressed in swine endothelial cell (SEC). The expression level and its stability in the cell surface were monitored with FACS analysis. RESULTS: Surface expression of UL18 is up-regulated by removing cytoplasmic tail portion from UL18F (a full sequence of UL18). SECs transfected with a cDNA of UL18CY (a cytoplasmic tail-less UL18) stably expressed UL18 molecule on the surface without gradual loss of its expression during 6 week continuous cultures. In the NK cytotoxicity assay, UL18 functions either inhibiting or activating NK cell cytotoxicity according to the source of NK cells. We found that there is individual susceptibility in determining whether the engagement of NK cell and UL18 results in overall inhibiting or activating NK cell cytotoxicity. CONCLUSION: In this study, we found that cytoplasmic tail is closely related to the regulatory function for controlling surface expression of UL18. Furthermore, by constructing stable cell line in which UL18 expression is up-regulated and stable, we provided a useful tool to clarify exact functions of UL18 on various immune cells having ILT2 receptor.

Characterization of the CD11c Promoter Which Is Expressed in the Mouse Dendritic Cells

BACKGROUND: CD11c, also known as integrin alpha x, is one of the optimum markers of dendritic cells. However, the regulation of the CD11c expression in mouse has not been identified yet. In this study, in order to analyze the regulation of CD11c expression, the promoter of CD11c was cloned and characterized. METHODS: To identify the promoter portion, various sizes of what are considered to be CD11c promoter fragments was amplified by polymerase chain reaction (PCR), using mouse genomic DNA as a template. After sequence was obtained, these fragments were transfected into various cell lines including mouse dendritic cell lines such as JAWSII and DC2.4 and L929 as control cell line.. The promoter activity of three promoter fragments was measured and compared by luciferase activity in the transfected cells. RESULTS: Three clones with size of 1kb, 3kb and 6kb were obtained from mouse genomic DNA. Flow cytometry analysis of JAWSII cells revealed that 52% of the cells expressed CD11c, which was confirmed by RT-PCR analysis. On the contrary, L929 and DC 2.4 cells did not express CD11c. The CD11c+ JAWSII cells were enriched from 52% to 90% with cell sorter. The comparative luciferase activity analyisis demonstrated that the region responsible for tissue specific expression was contained within ?3 kb and the clone with size of 3 kb particularly showed higher luciferase activity than 6 kb and 1 kb clones. CONCLUSION: The CD11c promoter region containing the region responsible for tissue specificity was successfully cloned and ?3 kb region showed the highest activity.

Generation and Characterization of Monoclonal Antibodies against Human Interferon-lambda1

BACKGROUND: Members belonging to the interferon-lambda (IFN-lambda) family exert protective action against viral infection; however, the mechanisms of their action have remained elusive. To study IFN-lambda biology, such as endocytosis of IFN-lambda, we produced monoclonal antibodies (Abs) against human IFN-lambda and examined their usefulness. METHODS: We purified recombinant human IFN-lambda1 expressed in Escherichia coli by using affinity columns. Then, we generated hybridoma cells by fusing myeloma cells with splenocytes from IFN-lambda1- immunized mice. For evaluating the neutralizing activity of the monoclonal Abs against IFN-lambda1, we performed RT-PCR for the MxA transcript. In order to study the binding activity of IFN-lambda and the monoclonal Ab complex on HepG2 cells, we labeled the monoclonal Ab with rhodamine and determined the fluorescence intensity. RESULTS: Four hybridoma clones secreting Abs specific to IFN-lambda1 were generated and designated as HL1, HL2, HL3, and HL4. All the Abs reacted with IFN-lambda1 in the denatured form as well as in the native form. Abs produced by HL1, HL3, and HL4 did not neutralize the induction of the MxA gene by IFN-lambda1. We also demonstrated the binding of the HL1 monoclonal anbitody and IFN-lambda complex on HepG2 cells. CONCLUSION: Monoclonal Abs against IFN-lambda1 were produced. These Abs can be used to study the cellular binding and internalization of IFN-lambda.

Generation and Characterization of Monoclonal Antibodies against Human Interferon-lambda1

BACKGROUND: Members belonging to the interferon-lambda (IFN-lambda) family exert protective action against viral infection; however, the mechanisms of their action have remained elusive. To study IFN-lambda biology, such as endocytosis of IFN-lambda, we produced monoclonal antibodies (Abs) against human IFN-lambda and examined their usefulness. METHODS: We purified recombinant human IFN-lambda1 expressed in Escherichia coli by using affinity columns. Then, we generated hybridoma cells by fusing myeloma cells with splenocytes from IFN-lambda1- immunized mice. For evaluating the neutralizing activity of the monoclonal Abs against IFN-lambda1, we performed RT-PCR for the MxA transcript. In order to study the binding activity of IFN-lambda and the monoclonal Ab complex on HepG2 cells, we labeled the monoclonal Ab with rhodamine and determined the fluorescence intensity. RESULTS: Four hybridoma clones secreting Abs specific to IFN-lambda1 were generated and designated as HL1, HL2, HL3, and HL4. All the Abs reacted with IFN-lambda1 in the denatured form as well as in the native form. Abs produced by HL1, HL3, and HL4 did not neutralize the induction of the MxA gene by IFN-lambda1. We also demonstrated the binding of the HL1 monoclonal anbitody and IFN-lambda complex on HepG2 cells. CONCLUSION: Monoclonal Abs against IFN-lambda1 were produced. These Abs can be used to study the cellular binding and internalization of IFN-lambda.

Current Status and Future Perspectives of Xenotransplantation and Stem Cell Research in Transplantation Field

The severe shortage of human organ donors is one of the biggest problems with organ transplantation. The solution for this problem would be development of artificial organs or mechanical devices, stem cell derived organs, and xenogeneic organs. Artificial organs may provide a short term life or functional support, but they cannot be considered as a life-long curative therapeutic modality in the near future. Although considerable efforts have been invested in the production of lab-grown organs using stem cells, clinical application of these organs will demand many years of research and investment. Currently, stem cells are clinically applied in cell replacement therapy. Therefore, xenotransplantation would be the most imminent solution for the organ shortage. Recent advances in our understanding of the mechanisms of xenograft rejection, zoonotic infections including PERV (porcine endogenous retrovirus), and production of alpha-1,3-galactosyltransferase-deficient pigs, put xenotransplantation closer to the clinical reality. At this stage, pancreatic islet xenotransplantation would be the first target for clinical application, the efficacy of which has been proven in non-human primate study and is waiting for the development of relatively non-toxic or clinically applicable immunosuppressive or tolerance-inducing regimens.

A Case of Preeclampsia Complicating a Pregnancy with Malignant Neurofibromatosis / 대한주산의학회잡지

Neurofibromatosis type 1 (NF-1) is one of the most common genetic disorders, occurring once in every 4000 births. The prevalence of neurofibromatosis associated with pregnancy was one in 5000~18500 deliveries. Higher rates of maternal and neonatal complications have been reported in NF-1 patients including as hypertension, intrauterine growth restriction (IUGR), preterm labor, stillbirth and cesarean section rates. Thus, pregnant patients with NF-1 are required close antenatal observation at high risk tertiary centers in order to detect complication early. We report a case of malignant neurofibromatosis associated with pregnancy with a review of literature.

A new scoring system for differentiation between benign and malignant ovarian masses / 대한산부인과학회지

OBJECTIVE: The purpose of this study was to design a new scoring system for differentiation between benign and malignant ovarian masses and to assess effectiveness of new scoring system comparing other scoring systems. METHODS: This study was based on 199 women who visited Soonchunhyang Hospital for surgery of ovarian mass. Ultrasonography and scoring system based on De Priest, Sassone, Ferrazi and Alcazar was performed the day before operation. Pathologic diagnosis after operation was directly compared with diagnosis of scoring system. The cut-off level of the new index is 11 points. This study was evaluated by sensitivity, specificity, positive predictive value (PPV) and negative predictive value (NPV). RESULTS: Parameters of new scoring system were wall thickness, number of septum, volume of mass, irregularity, vascularity and echogenicity. The new scoring system had sensitivity 73.9%, specificity 97.7% and negative and positive predictive values of 96.6% and 80.9% respectively. CONCLUSION: Sensitivity of new scoring system was similar to previous ones but specificity of that is higher. Our new scoring system shows better to differentiate benign from malignant ovarian mass than four other scoring systems.

A case of congenital retroperitoneal immature teratoma: Prenatally diagnosed as Neuroblastoma / 대한산부인과학회지

Through a routine antenatal ultrasound examination of 37-year-old woman at 38 weeks' gestation, a 6 cm-diameter mixed solid and cystic mass was found in the left suprarenal area of her fetus. The following antenatal magnetic resonance image showed a larger cystic mass with a central solid component. The surgical exploration of the mass was performed in 50 days after delivery and by the pathologic examination a grade III immature teratoma was found. The retroperitoneal immature teratomas are extremely rare in childhood. To our Knowledge this is the first reported case about congenital retroperitoneal immature teratoma. The immature teratoma should be included in the differential diagnosis of fetal suprarenal mass and the serum alpha-fetoprotein level should be included in the initial laboratory examination at birth.

Effect of titanium surface roughness on adhesion and differentiation of osteoblasts / 대한치주과학회지

The success of an implant is determined by its integration into the tissue surrounding the biomaterial. Surface roughness is considered to influence the behavior of adherent cells. The aim of this in vitro study was to determine the effect of surface roughness on Saos-2 osteoblast-like cells. Titanium disks blasted with 75 micrometer aluminum oxide particles and machined titanium disks were prepared. Saos-2 were plated on the disks at a density of 50,000 cells per well in 48-well dishes. After 1 hour, 1 day, 6 days cell numbers were counted. One day, 6 days after plating, alkaline phosphatase(ALPase) activity was determined. Compared to experimental group, the number of cells was significantly higher on control group. The stimulatory effect of surface roughness on ALPase was more pronounced on the experimental group than on control group. These results demonstrate that surface roughness alters proliferation and differentiation of osteoblasts. The results also suggest that implant surface roughness may play a role in determining phenotypic expression of cells.

Emergency Cervical Cerclage: A Retrospective Study of 5 Years' Practice in 71 Cases / 대한산부인과학회잡지

OBJECTIVE: We investigated the outcome of emergency cervical cerclage in women with cervical incompetence. Cervical incompetence was diagnosed when cervical dilatation exceeded 2 cm with intact but bulging membranes. METHODS: Retrospective chart review of 71 cases of patients who underwent emergency cervical cerclage using Mcdonald suture after amnioreduction performed for cervical incompetence with cervical dilatation and membrane bulging from March 1998 through August 2003 at Kang-Nam Sacred Heart Hospital, Hallym University. Clinical variables evaluated included gestational age at cerclage, cervical dilatationa at cerclage, prolongation of pregnancy, and neonatal outcome. RESULTS: Emergency cerclage was performed successfully in 67 cases (94%). Gestational age at cerclage ranged from 16 weeks to 29 weeks, with the mean being 23.6 +/- 3.3 weeks. Cervical dilatation at cerclage was between 2 cm and 9 cm (mean 3.8 +/- 1.6 cm). Gestational age at delivery ranged from 16 to 40 weeks (mean 28.2 +/- 6.0 weeks). Prolongation of pregnancy following cerclage varied from between 1 to 134 days (mean 31.5 +/- 33.1 days). The median birth weight was 1370.29 +/- 72.6 g (range 140-3640 g). Thirty-seven babies were born live, and 30 of them survived (survival rate 49%). CONCLUSION: The possibility of a 49% survival rate is considered a good result for emergency cerclage. Emergency cervical cerclage can prolong pregnancy and influence the outcome of pregnancy favorably, and may be considered one potential method of treatment in such cases.

A case of typhlitis developed after anticancer chemotherapy in a patient with solid tumor / 대한내과학회지

Typhlitis is a necrotizing enterocolitis of the cecum, ascending colon and terminal ileum. Typhlits has been reported in the severely neutropenic patients and likely results from a combination of neutropenia and defects in the bowel mucosa related to cytotoxic chemotherapy. This disease is most common in patients with leukemia who have undergone intensive myeloablative chemotherapy. Presumptive diagnostic criteria for typhlitis include fever, abdominal pain and tenderness, and radiologic evidence of right-sided colonic inflammation in patients with neutropenia. Recently, this disease is also reported in patients with solid tumor due to increasing challenges of high dose chemotherapy. We report a case of typhlitis developed in the circumstance of neutropenia induced by chemotherapy in a patient with malignant testicular tumor.

A Study on the Effects of CO2 Pneumoperitoneum and Trendelenburg position on Arterial Acid-Base Balance and Middle Cerebral Artery Hemodynamics in Gynecologic Laparoscopic Surgery / 대한산부인과학회잡지

OBJECTIVE: The purpose of this study was to identify arterial acid-balance and cerebral hemodynamics in patients undergoing gynecologic laparoscopic operations according to induction of CO2 pneumoperitoneum and Trendelenburg position. METHODS: Twenty patients without cardiopulmonary disease undergoing various laparoscopic pelvic surgeries were resulted in data of arterial blood and hemodynamic parameters such as systolic blood pressure (SBP), diastolic blood pressure (DBP), heart rate (HR), pCO2, pH, pO2, middle cerebral blood flow velocity (MCABFV), pulsatile index (PI), resistance index (RI). Under the standardized setting of general anesthesia, arterial blood and hemodynamic parameters were determined in supine position (base line) before CO2 insufflation, and 15 minutes, 30 minutes, 45 minutes in Trendelenburg position after CO2 insufflation, and 10 minutes in supine position after CO2 defflation. RESULTS: The arterial BP, pCO2 and MCABFV increased significantly in Trendelenburg position after CO2 insufflation and persisted even after 10 minutes in supine position after CO2 defflation. PI decreased significantly compared with control in Trendelenburg position after CO2 insufflation and persisted even after 10 minutes in supine position after CO2 defflation. RI decreased significantly as compared with control in Trendelenburg position after CO2 insufflation and persisted even after 10 minutes in supine position after CO2 defflation. CONCLUSION: Intraoperative arterial blood gas analysis is required in patients with high risk of hypercarbia during gynecologic laparoscopic surgery, and clinicians should anticipate an increase in cerebral blood flow and decrease in cerebral vascular resistance after CO2 pneumoperitoneum and Trendelenburg position, and gynecologic laparoscopy may be safe in view point of cerebral hemodynamics.