Ex vivo Expansion of Human Natural Killer Cells from Blood Retained in a Disposable Platelet Apheresis Set / 대한수혈학회지

BACKGROUND: Natural killer cells expanded from human peripheral blood (PB) have been used in cancer immunotherapy research. Although most research teams have access to human PB, it is necessary to find a source of blood that can be easily obtained. We have tested the possibility of using blood retained in a disposable platelet apheresis set as an alternative source, with special interest in expansion of NK cells for use in cancer immunotherapy research. METHODS: For expansion of NK cells, peripheral blood mononuclear cells (PBMCs) were isolated from an MCS+ platelet apheresis kit (Haemonetics, Braintree, USA) and PB from the same donor (n=7) and co-cultured with 100-Gy gamma ray-irradiated K562 cells expressing the 4-1BB ligand and membrane-bound IL-15 for three weeks in RPMI1640 medium in the presence of IL-2 and IL-15. Cytotoxicity was measured using WST-1 at 1:1, 2:1, and 4:1 effector-to-target (E:T) ratios for a period of four hours. RESULTS: Mean rate of expansion of NK cells was 1,097-fold and their purity was 94.4% from blood retained in a disposable platelet apheresis set; mean rate of expansion of NK cells was 953-fold and their purity was 92.0% from PB after a period of three weeks. No differences in cytotoxicity against K562, 697, Raji, and RPMI8226 were observed between NK cells expanded from two blood sources. CONCLUSION: Blood retained in a disposable platelet apheresis set is a useful and convenient source for expansion of NK cells for use in cancer immunotherapy research.

Application of Real-Time PCR for ABO Genotyping for Large-scale Population Screening / 대한수혈학회지

BACKGROUND: For large-scale population screening, the method of ABO genotyping needs to be simple, accurate and cost-effective. The real-time PCR method has been introduced and it is suitable for dealing with large numbers of specimens. In this study, we examined the ABO genotyping of 1,700 residents of Jeollanam-do for an epidemiologic study by applying the real-time PCR method. METHODS: Genomic DNA was extracted from the peripheral blood samples of 1,700 residents of Jeollanam-do between July 2004 and January 2006 and these samples were stored at -70degrees C. The ABO genotype in all the samples was determined by four-color real-time PCR using displacing probes and three cases that had an atypical real time PCR pattern were confirmed by direct sequencing and PCR-based cloning of exons 6&7 of the ABO gene. RESULTS: The genotyping results of 1,700 samples included O/O (25.6%), A/A (9.1%), A/O (29.1%), B/B (4.5%), B/O (19.8%) and A/B (11.9%), and the allele frequencies of O, A and B were 50.1%, 29.5% and 20.4%, respectively. The frequency of the O allele was lower in the residents of Jeollanam-do than that previously reported for the residents of Kangwon-do (P=0.014), while the frequency of the A allele was higher in the residents of Jeollanam-do than that previously reported for the residents of Kangwon-do (P=0.003). The three cases with atypical results were revealed to be B101/O24, Bvar(296C>T)/O01 and B101/Ovar(801G>T). It takes 6 days to perform ABO genotyping on 1,700 samples by a calculation per test. CONCLUSION: ABO genotyping by real-time PCR using displacing probes can be useful for mass screening for ABO genotyping. In Korea, the frequency of the ABO allele was significantly different among different regions.

New Variant of O02 Allele Found in the Family of Cis-AB: Report of Three Cases in a Family / 대한수혈학회지

In preoperative ABO typing, we observed an ABO discrepancy in a 61 year-old patient with A2B3 phenotype. Standard serologic tests for the ABO blood group phenotypes and ABO gene direct sequencing for the exons 6 and 7 were done as part of the patient's family study. We found that the patient's genotype was identical to cis-AB01/O02, except for a 703 G>A polymorphism at exon 7. An allele-separation by cloning and subsequent sequencing of the heterozygote was carried out. We identified a novel O02 variant allele characterized by a 703 G>A polymorphism in the patient and in her 2 daughters.

The M142T Mutation Causes B3 Phenotype: Three Cases and an in vitro Expression Study / 대한진단검사의학회지

The B3 phenotype is the most common B subtype in Korea. The B305 allele (425 T>C, M142T) was first reported in 2 Chinese individuals; however, it has not yet been reported in the Koreans, and the impact of the M142T mutation on the expression of the B3 phenotype has also not been studied. To resolve an ABO discrepancy between a group O neonate and her group O father and A(1)B(3) mother, blood samples from these individuals and other family members were referred to our laboratory for ABO gene analysis. The B305 allele was discovered in the neonate (B305/O01), her mother (A102/ B305), and her maternal aunt (B305/O02), while her father was typed as O01/O02. Transient transfection experiments were performed in HeLa cells using the B305 allele synthesized by site-directed mutagenesis; flow cytometric analysis revealed that this transfect expressed 35.5% of the total B antigen produced by the B101 allele transfect. For comparison, Bx01 allele transfects were also created, and they expressed 11.4% of the total B antigen expressed on the surface of B101 transfects. These experiments demonstrate that the M142T (425 T>C) mutation is responsible for the B subtype phenotype produced by the B305 allele.

The Bacterial Surface Expression of SARS Viral Epitope using Salmonella typhi Cytolysin A

The cytolysin A (ClyA) is a 34 kDa pore-forming cytotoxic protein and expressed by some enteric bacteria including Salmonella typhi. This toxin is transported on the bacterial surface and secreted without posttranslational modification. Using the surface display of ClyA, the expression vectors for 193-aa immunogenic antigen of spike protein (termed S1E) from severe acute respiratory syndrome coronavirus (SARS-CoV) were constructed. The vectors carried a gene encoding S. typhi ClyA conjugated to S1E at the C terminus (termed ClyA-S1E) and asd gene in pGEM-T and pBR322, named pGApLCS1E and pBApLCS1E, respectively. An asd-mutated E. coli transformed with these vectors could grow without diaminopimelic acid (DAP), indicating that they were stably maintained in such mutants. ClyA-S1E recombinant proteins from these vectors were expressed on the surface of the attenuated S. typhimurium deficient of global virulence gene regulator, ppGpp. However, they did not show the hemolytic activity on the blood agar plate and cytotoxicity against HeLa cells. To examine whether bacteria expressing ClyA-S1E induced the immune response against S1E, S. typhimurium deficient of ppGpp and Asd was transformed with these vectors and orally immunized in mice. In the western blotting against GST-conjugated S1E using the immunized mouse sera, it was shown that the significant band was detected in the mouse serum by the bacteria transformed with pGApLCS1E but not with pBApLCS1E. It indicates that the immune response producing antibody was dependent on the expression level of ClyA-S1E. Therefore, ClyA delivery system can be used for SARS vaccine development.

Estimation of Individualized Probabilities of Developing Breast Cancer for Korean Women

PURPOSE: Based on the results of the relative risk model of Korean breast cancer, the aim of this study was to develop a Korean breast cancer risk assessment tool which would display the absolute breast cancer risks of Korean women. METHODS: The tool was developed in the three steps: selection of risk factors and relative risks, calculation of baseline breast cancer incidences, and estimation of absolute breast cancer risks. The risk factors used in this tool were age, family history of first- and second-degree relatives, body mass index, age at first delivery, history of breast-feeding, and a special test on the breasts. A program was developed in an HTML file, which was used for input of the risk factors, and a CGI file, which was used to calculate the risk and display the results. RESULTS: The program was stored in the Internet web page, http://home.dankook.ac.kr/breast/brca/brca.htm. After receiving an input of risk factors, the program was able to calculate the relative risk compared to all the age groups, the estimated absolute risks following 5 and 10 years, and the estimated absolute risks up to ages 64 and 74 years. The estimated risks of Korean women using this tool were less than those reported by the NCI risk assessment tool. The risk of breast cancer was highest in the fifth decade. CONCLUSION: In this study, we developed a web page containing a Korean breast cancer risk assessment tool. This program may be useful for the assessment of individual breast cancer risks, the selection of screening tools, and the evaluation of preventive options for risk reduction.

Estimation of Individualized Probabilities of Developing Breast Cancer for Korean Women

PURPOSE: Based on the results of the relative risk model of Korean breast cancer, the aim of this study was to develop a Korean breast cancer risk assessment tool which would display the absolute breast cancer risks of Korean women. METHODS: The tool was developed in the three steps: selection of risk factors and relative risks, calculation of baseline breast cancer incidences, and estimation of absolute breast cancer risks. The risk factors used in this tool were age, family history of first- and second-degree relatives, body mass index, age at first delivery, history of breast-feeding, and a special test on the breasts. A program was developed in an HTML file, which was used for input of the risk factors, and a CGI file, which was used to calculate the risk and display the results. RESULTS: The program was stored in the Internet web page, http://home.dankook.ac.kr/breast/brca/brca.htm. After receiving an input of risk factors, the program was able to calculate the relative risk compared to all the age groups, the estimated absolute risks following 5 and 10 years, and the estimated absolute risks up to ages 64 and 74 years. The estimated risks of Korean women using this tool were less than those reported by the NCI risk assessment tool. The risk of breast cancer was highest in the fifth decade. CONCLUSION: In this study, we developed a web page containing a Korean breast cancer risk assessment tool. This program may be useful for the assessment of individual breast cancer risks, the selection of screening tools, and the evaluation of preventive options for risk reduction.

A Comparative Study of Outcomes between Emergency and Elective Surgeries for Colon Cancer

PURPOSE: The purpose of this study was to compare the efficacy of curative emergency surgery for complicated colon cancer in terms of tumor recurrence and survival compared with that of elective surgery. METHODS: A total of 238 primary surgeries for colon cancer were performed. All patients were deemed to have undergone a curative resection. Patients were classified into an emergency surgery group for complicated colon cancers (n=40) and an elective surgery group for uncomplicated colon cancers (n=198). RESULTS: Emergency colonic cancers present at a more advanced stage (P=0.002). The postoperative mortality rate in the emergency group was significantly higher than it was in the elective group (15.0% vs. 2.5%, P= 0.004). There were differences between the two groups in tumor recurrence (32.5% vs. 13.1%, P=0.003), overall survival (52.5% vs. 71.7%, P=0.017), and disease-free survival (50.0% vs. 69.7%, P=0.016). However, after the patients were stratified according to tumor stage, no statistical differences were observed. CONCLUSIONS: When compared with uncomplicated colon cancers, complicated colon cancers present at a more advanced stage with a higher postoperative mortality and an overall worse prognosis. However, the difference decreases when patients are stratified according to the tumor stage. The negative prognostic efficacy of emergency surgery for complicated colon cancers appears to be confined to the perioperative period. Despite the more advanced stage of tumors in patients undergoing emergency surgery, the aim of the surgeon should be to offer a curative resection for better survival, if possible.

Results and Survivorship of High Tibial Osteotomy / 대한정형외과학회잡지

PURPOSE: To determine survivorship and establish the ideal correction angle in high tibial osteotomy for primary osteoarthritis through retrospective study. MATERIALS AND METHODS: One hundred and eleven valgus osteotomies of the tibia were performed in seventy-nine patients from 1985 to 1997. The average follow-up period was 9 year 6 months (range, 2.4 to 14.1 years). Failure I was defined as the need for conversion of a high tibial osteotomy to a total knee arthroplasty, and Failure II as the need for conversion or in a patient with less than 60 HSS knee score points. The probability of survival was estimated using the Kaplan-Meier survivorship method. RESULTS: The HSS knee score averaged 60 points preoperatively, 94 points at the 1 year follow-up and 87 points at the last followup. The 4 year and 14 year survival rates were 99% and 85.0% using the first definition of failure, and 96.4% and 75.1% using the second. CONCLUSION: High tibial osteotomy is a reliable method for treating unicompartmental osteoarthrtis, provided that the postoperative femorotibial angle is corrected by more than 7degrees of valgus.

Expression of Vascular Endothelial Growth Factor Correlated with Recurrence in Gastric Carcinomas

PURPOSE: Angiogenesis is essential for tumor growth and metastasis and depends on the production of angiogenic factors that are secreted by tumor cells. Vascular endothelial growth factor (VEGF) is the most significant angiogenic factor and a selective mitogen for endothelial cells. VEGF, also known as the vascular permeability factor, acts on endothelial cells to increase microvascular permeability and directly stimulate the growth of new blood vessels. Several studies have reported that the expression of VEGF is correlated with hematogenous recurrence via angiogenesis in gastric carcinomas. This research evaluated the relationship between the expression of VEGF and hepatic and peritoneal recurrence in gastric carcinomas. MATENRIALS AND METHODS: Thirty specimens resected from patients with primary gastric carcinomas who had undergone curative resections were divided into three group: Group I, early gastric carcinomas without recurrence; Group II, advanced gastric carcinomas with hepatic recurrence; and Group III, advanced gastric carcinomas with peritoneal recurrence. The expression of VEGF and the density of the microvessel count were examined using immunohistochemistry. RESULTS: 1) The expression of VEGF in Group II and Group III (63.2+/-24.3%) was stronger than that in Group I(7+/-4.2%). The expression of VEGF in Group II (76.5+/-13.2%) was stronger than that of the Group III (50+/-14.2%) (P<0.05). 2) The microvessel count in Group II (49.9+/-14.5) was more than that in Group I (8.6+/-2.6) and Group III (29.1+/-18.1) (P<0.05). 3) The microvessel count was increased significantly with increasing the expression of VEGF. CONCLUSION: The expression of VEGF is associated with advanced stomach cancer and hepatic recurrence has a higher expression of VEGF than peritoneal recurrence with neovascularization. Thus the expression of VEGF can be considered to be a useful indicator of recurrence in gastric carcinoma and especially in hepatic recurrence.

Revision Total Knee Arthroplasty of Failed Unicompartmental Knee Arthroplasty

PURPOSE: To analyze the failure mechanism, complexity of surgery, complication and outcome of revision of failed unicompartmental knee arthroplasty(UKA). MATERIALS AND METHODS: 14 patients(16 knees) were followed up a mean 70 months. The patients were a mean age of 58.2 years at time of UKA and predominantly female(85%). Revision surgery was done 14 to 104 months with an average time of 4 years 10 months after UKA. The UKA component that were revised include 10 Microloc, 4 Allegretto, 1 Oxford and 1 Marmor modular II. Clinical and radiographic evaluation were completed preoperatively and at last follow-up. The scoring system used in this study is Hospital for Special Surgery(HSS) knee score. Radiographic assessment was performed using Bauer's method(tibio-femoral angle) and American knee society roentgenographic evaluation system. RESULTS: The predominant failure mechanism was polyethylene wear and femoral component loosening. At the last follow-up, the average HSS knee score significantly improved from 60 to 86. Average tibio-femoral angle was corrected from varus 2o to valgus 5o. The implant type used for the revision was PFC-PS. 11 knees required local autograft but no allograft were used. Femoral block augment and tibial metal wedges with stem were also used to reconstruct the defect. There were 2 cases of complications, subluxation of patella and deep infection after intraarticular injection at local clinic. CONCLUSION: Polyethylene wear and femoral component loosening was common failure mechanism in revision of UKA. Bone defects needed to reconstruct were detected in 11 cases but autograft was enough to reconstruct the defect. Result of failed UKA compares favorably with those of total knee revision.

Results of the Posterolateral Fusion Using a Pedicle Screw for the Spondylolisthesis / 대한정형외과학회잡지

PURPOSE: This is a retrospective study to analyze the clinical and radiological results of posterolateral fusion with a pedicle screw fixation for lumbar spondylolisthesis. MATERIAL AND METHOD: From January 1994 to June 1997, forty-seven patients with spondylolisthesis were treated by posterolateral fusion with pedicle screw fixation. The clinical results were analyzed according to Kirkaldy-Willis & Kim's criteria and radiological union, reduction of slippage, change of disc space, change of lumbar and segmental angle were assessed by simple X-ray. RESULTS: The good bony union rate of formation and satisfactory clinical results were obtained, but the reduction of slippage, the restoration of disc height and the change of sagittal angle were not sustained. There was a significant relationship between the clinical results and the loss of anterior disc height. CONCLUSIONS: The satisfactory clinical results were seen to be in follow-up observation, and the maintenance of anterior disc height significantly influenced clinical results. We think that the additional operation method is necessary in maintaining the disc height.

Localized Pigmented Villonodular Synovitis Causing Anterior Knee Pain: A Case Report

No Abstract Available.

Use of Structureal Allograft in Revision Total Knee Arthroplasty

PURPOSE: To analyze the clinical and radiological results after revision total knee arthroplasty(TKA) using structural allograft for severe bone defect, MATERIALS AND METHODS: Between l992 and 1997, we used structural allograft for revision TKA in 20 knees of 18 patients with severe bone defect, The average age at surgery was of 61.6 years. The follow-up period averaged 43 months(range, 1 3-80 months). All patients had severe bone defect of the femur and/or tibia. Ten distal femur and thirteen proximal tibia required allografting. Fresh frozen femoral heads were used in all patients. We used posterior stabilized stemmed PFC knee component in all cases except one case with constrained condylar type. Patients were evaluated with physical examination, radiographs and the Hospital for Special Surgery knee rating scale. RESULTS: Average range of motion before surgery was 87degrees and after surgery 107degrees. Average knee score was 65 before and 90.4 after surgery. Preoperative alignment averaged 1.8degrees valgus, ranging from 17degrees varus to 13degrees valgus, and postoperative alignment averaged 7,2 degrees velgus, ranging from 3degrees valgus to 10 val-gus, All patient improved pain and stability. Postoperatively average HSS score improved from 65(rang- ing from 46 to 92) to 90.4(ranging from 75 to 100). All of the allografts united with host bone. There was no radiolucent line, lysis, fracture and infection. CONCLUSION: Structural allograft can be a satisfactory method of managing large bone defects in the failed total knee arthroplasty.

Chondrolipoma of the Knee: A Case Report

Chondrolipoma is a benign tumor composed of mature adipose tissue and mature chondrocytes, and very rarely found. The patient, a 36-year-old woman, presented with a firm, walnut sized, mild tender mass at her left knee. There had been dull pain after long walking. There were no limitation of motion in physical examination. On plane lateral radiograph, radioopaque calcific densities were seen in elliptical soft tissue shadow below inferior pole of the patella. Magnetic resonance image demonstrated well defined mass of mixed signal intensity consistent with fat tissue and cartilaginous tissue. Gross specimen showed that the tumor appeared to be a round, encapsulated and firm nodule with milky whitish yellow cut surface. Microscopically the tumor consisted of nodular cartilage and mature fat tissue. The cartilage cells did not show mitotic figure and were often arranged in small clusters. But there were no cytologic features of malignancy. Thus the authors diagnosed this tumor as chondrolipoma, and report it with the literature review.